When a drop is subjected to an external flow, the balance between the interfacial tension and the flow forcing determines the drop shape while the imbalance between them leads to drop breakup. We numerically investigate deformation of a three-dimensional viscous drop forced by a potential vortex and other time-dependent extensional flows. Such flows represent oscillating forces present in multiphase flows such as due to turbulent eddies. The Simulation is performed at non-zero Reynolds numbers to explore the effects of inertia using a front-tracking finite difference method. We investigate the effects of interfacial tension, viscosity, density ratio, periodicity, and inertia on the drop deformation. Introduction of inertia and time dependence to the imposed flow lead to some unusual dynamics in the drop deformation such as increased deformation with increased surface tension–resonance. Such phenomena are analyzed and explained with the help of a simple ordinary differential equation model. A new mechanism for frequency-specific drop breakup in turbulence flow is suggested.

The viscoelastic flow computation has historically been riddled with severe numerical problems at high Weissenberg numbers, with single phase two-dimensional benchmark flows being actively pursued as late as in the 90s and 2000s. Our group is one of the first few to simulate multiphase viscoelastic flow computation. We have developed a front-tracking finite difference tool with a robust scheme for viscoelastic constitutive relations (Oldroyd B, FENE etc) that mitigates many of the numerical problems. There have been contraditory observations regarding effects of viscoelasticity on drop deformation–whether it increases or decreases. We have numerically simulated and with perturbative analysis explained the nonmonotonic nature of the deformation and breakup resolving this controversy. Currently, the focus is the effects of viscoelasticity on drop migration. The physics is simultaneously being used to applications such as biological problems (blood, cells, vescicles etc) involving viscoelasticity.


Related Publication

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Singh R, Sarkar K 2015 “Hydrodynamic interactions between pairs of capsules and drops in a simple shear: effects of viscosity ratio and heterogeneous collision,” Physical Review E, 92, 063029.

    Hydrodynamic interactions between a pair of capsules in simple shear are numerically investigated using afront-tracking finite difference method. The membrane of the capsule is modeled using different hyperelasticconstitutive relations. We also compare the pair interactions between drops to those between capsules. Anincreased viscosity ratio leads to a reduced net cross-stream separation between capsules as well as drops aftercollision. At low viscosity ratios, for the same capillary number drop-pairs show higher cross-stream separationthan those for capsule-pairs, while substantially large viscosity ratios result in almost the same value for bothcases. We investigate pair-collisions between two heterogeneous capsules C1and C2with two different capillarynumbers. The maximum deformation of C1was seen to increase with increasing stiffness (decreasing capillarynumber) of C2, even though the stiffness of C1was kept fixed. The findings are similar for a drop-pair, however,with a smaller maximum deformation for the same combinations of capillary numbers. The final cross-streamdrift of the trajectory of C1decreases with the increasing stiffness of C2, but the relative trajectory betweenthe capsules remains unchanged. The maximum deformation and the cross-stream drift of the trajectory of C1are shown to approximately vary with power-law functions of the ratio of the capillary numbers of C1andC2. An analytical explanation of the dependence on the two capillary numbers is offered. Different membraneconstitutive laws result in similar deformation and drift in trajectory.

  • Singh R, Sarkar K 2015 “Hydrodynamic interactions between pairs of capsules and drops in a simple shear: effects of viscosity ratio and heterogeneous collision,” Physical Review E, 92, 063029.

    Hydrodynamic interactions between a pair of capsules in simple shear are numerically investigated using afront-tracking finite difference method. The membrane of the capsule is modeled using different hyperelasticconstitutive relations. We also compare the pair interactions between drops to those between capsules. Anincreased viscosity ratio leads to a reduced net cross-stream separation between capsules as well as drops aftercollision. At low viscosity ratios, for the same capillary number drop-pairs show higher cross-stream separationthan those for capsule-pairs, while substantially large viscosity ratios result in almost the same value for bothcases. We investigate pair-collisions between two heterogeneous capsules C1and C2with two different capillarynumbers. The maximum deformation of C1was seen to increase with increasing stiffness (decreasing capillarynumber) of C2, even though the stiffness of C1was kept fixed. The findings are similar for a drop-pair, however,with a smaller maximum deformation for the same combinations of capillary numbers. The final cross-streamdrift of the trajectory of C1decreases with the increasing stiffness of C2, but the relative trajectory betweenthe capsules remains unchanged. The maximum deformation and the cross-stream drift of the trajectory of C1are shown to approximately vary with power-law functions of the ratio of the capillary numbers of C1andC2. An analytical explanation of the dependence on the two capillary numbers is offered. Different membraneconstitutive laws result in similar deformation and drift in trajectory.

  • Singh R, Sarkar K 2015 “Hydrodynamic interactions between pairs of capsules and drops in a simple shear: effects of viscosity ratio and heterogeneous collision,” Physical Review E, 92, 063029.

    Hydrodynamic interactions between a pair of capsules in simple shear are numerically investigated using afront-tracking finite difference method. The membrane of the capsule is modeled using different hyperelasticconstitutive relations. We also compare the pair interactions between drops to those between capsules. Anincreased viscosity ratio leads to a reduced net cross-stream separation between capsules as well as drops aftercollision. At low viscosity ratios, for the same capillary number drop-pairs show higher cross-stream separationthan those for capsule-pairs, while substantially large viscosity ratios result in almost the same value for bothcases. We investigate pair-collisions between two heterogeneous capsules C1and C2with two different capillarynumbers. The maximum deformation of C1was seen to increase with increasing stiffness (decreasing capillarynumber) of C2, even though the stiffness of C1was kept fixed. The findings are similar for a drop-pair, however,with a smaller maximum deformation for the same combinations of capillary numbers. The final cross-streamdrift of the trajectory of C1decreases with the increasing stiffness of C2, but the relative trajectory betweenthe capsules remains unchanged. The maximum deformation and the cross-stream drift of the trajectory of C1are shown to approximately vary with power-law functions of the ratio of the capillary numbers of C1andC2. An analytical explanation of the dependence on the two capillary numbers is offered. Different membraneconstitutive laws result in similar deformation and drift in trajectory.

  • Singh R, Li X, Sarkar K 2014 “Lateral migration of an elastic capsule in a wall-bounded shear,” Journal of Fluid Mechanics, 739, 421-443.

    The migration of a capsule enclosed by an elastic membrane in a wall-bounded linearshear is investigated using a front-tracking method. A detailed comparison with themigration of a viscous drop is presented varying the capillary number (in the caseof a capsule, the elastic capillary number) and the viscosity ratio. In both cases,the deformation breaks the flow reversal symmetry and makes them migrate awayfrom the wall. They quickly go through a transient evolution to eventually reach aquasi-steady state where the dynamics becomes independent of the initial positionand only depends on the wall distance. Previous analytical theories predicted thatfor a viscous drop, in the quasi-steady state, the migration and slip velocities scaleapproximately with the square of the inverse of the drop–wall separation, whereasthe drop deformation scales as the inverse cube of the separation. These power lawrelations are shown to hold for a capsule as well. The deformation and inclinationangle of the capsule and the drop at the same wall separation show a crossoverin their variation with the capillary number: the capsule shows a steeper variationthan that of the drop for smaller capillary numbers and slower variation than thedrop for larger capillary numbers. Using the Green’s function of Stokes flow, asemi-analytic theory is presented to show that the far-field stresslet that causes themigration has two distinct contributions from the interfacial stresses and the viscosityratio, with competing effects between the two defining the dynamics. It predicts thescaling of the migration velocity with the capsule–wall separation, however, matchingwith the simulated result very well only away from the wall. A phenomenologicalcorrelation for the migration velocity as a function of elastic capillary number, walldistance and viscosity ratio is developed using the simulation results. The effects ofdifferent membrane hyperelastic constitutive equations – neo-Hookean, Evans–Skalak,and Skalak – are briefly investigated to show that the behaviour remains similar fordifferent equations.

  • Mukherjee S, Sarkar K 2014 “Lateral migration of a viscoelastic drop in a Newtonian fluid in a shear flow near a wall,” Physics of Fluids, 26, 103102.

    Wall induced lateral migration of a viscoelastic (FENE-MCR) drop in a Newtonianfluid is investigated. Just like a Newtonian drop, a viscoelastic drop reaches a quasi-steady state where the lateral velocity only depends on the instantaneous distancefrom the wall. The drop migration velocity and the deformation scale inversely withthe square and the cube of the distance from the wall, respectively. The migration ve-locity varies non-monotonically with increasing viscoelasticity (increasing Deborahnumber); initially increasing and then decreasing. An analytical explanation has beengiven of the effects by computing the migration velocity as arising from an imagestresslet field due to the drop. The semi-analytical expression matches well with thesimulated migration velocity away from the wall. It contains a viscoelastic stressletcomponent apart from those arising from interfacial tension and viscosity ratio. Themigration dynamics is a result of the competition between the viscous (interfacialtension and viscosity ratio) and the viscoelastic effects. The viscoelastic stressletcontribution towards the migration velocity steadily increases. But the interfacialstresslet—arising purely from the drop shape—first increases and then decreases withrising Deborah number causing the migration velocity to be non-monotonic. The ge-ometric effect of the interfacial stresslet is caused by a corresponding nonmonotonicvariation of the drop inclination. High viscosity ratio is briefly considered to showthat the drop viscoelasticity could stabilize a drop against breakup, and the increase inmigration velocity due to viscoelasticity is larger compared to the viscosity-matchedcase.

  • Srivastava P, Malipeddi Reddy A, Sarkar K 2016 “Steady shear rheology of a viscous emulsion in the presence of finite inertia at moderate volume fractions: sign reversal of normal stress differences,” Journal of Fluid Mechanics, 85, 494-522.

    The shear rheology of an emulsion of viscous drops in the presence of finite inertiais investigated using direct numerical simulation. In the absence of inertia, emulsionsdisplay a non-Newtonian rheology with positive first and negative second normalstress differences. However, recently it was discovered that a small amount ofdrop-level inertia alters their signs – the first normal stress difference becomesnegative and the second one becomes positive, each in a small range of capillarynumbers (Li & Sarkar,J. Rheol., vol. 49, 2005, pp. 1377–1394). Sign reversal wasshown numerically and analytically, but only in the limit of a dilute emulsion wheredrop–drop interactions were neglected. Here, we compute the rheology of a density-and viscosity-matched emulsion, accounting for the interactions in the volume fractionrange of 5 %–27 % and Reynolds number range of 0.1–10. The computed rheologicalproperties (effective shear viscosity and first and second normal stress differences) inthe Stokes limit match well with previous theoretical (Choi–Schowalter in the dilutelimit) and simulated results (for concentrated systems) using the boundary elementmethod. The two distinct components of the rheology arising from the interfacialstresses at the drop surface and the perturbative Reynolds stresses are investigated asfunctions of the drop Reynolds number, capillary number and volume fraction. Thesign change is caused by the increasing drop inclination in the presence of inertia,which in turn directly affects the interfacial stresses. Increase of the volume fractionor capillary number increases the critical Reynolds number for sign reversals due toenhanced alignment of the drops with the flow directions. The effect of increasingthe volume fraction on the rheology is explained by relating it to interactions andspecifically to the contact pair-distribution function computed from the simulation.The excess stresses are seen to show an approximately linear behaviour with theReynolds number in the range of 0.1–5, while with the capillary number and volumefraction, the variation is weakly quadratic.

  • Aliabouzar M, Zhang LG, Sarkar K, 2016 “Lipid-coated microbubbles and low intensity pulsed ultrasound enhance chondrogenesis of human mesenchymal stem cells in 3D printed scaffolds,” Nature Scientific Report, 6, 37728.

    Lipid-coated microbubbles are used to enhance ultrasound imaging and drug delivery. Here we apply these microbubbles along with low intensity pulsed ultrasound (LIPUS) for the first time to enhance proliferation and chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in a 3D printed poly-(ethylene glycol)-diacrylate (PEG-DA) hydrogel scaffold. The hMSC proliferation increased up to 40% after 5 days of culture in the presence of 0.5% (v/v) microbubbles and LIPUS in contrast to 18% with LIPUS alone. We systematically varied the acoustic excitation parameters—excitation intensity, frequency and duty cycle—to find 30 mW/cm2, 1.5 MHz and 20% duty cycle to be optimal for hMSC proliferation. A 3-week chondrogenic differentiation results demonstrated that combining LIPUS with microbubbles enhanced glycosaminoglycan (GAG) production by 17% (5% with LIPUS alone), and type II collagen production by 78% (44% by LIPUS alone). Therefore, integrating LIPUS and microbubbles appears to be a promising strategy for enhanced hMSC growth and chondrogenic differentiation, which are critical components for cartilage regeneration. The results offer possibilities of novel applications of microbubbles, already clinically approved for contrast enhanced ultrasound imaging, in tissue engineering.

  • Aliabouzar M, Kumar KN, Sarkar K, 2018Acoustic vaporization threshold of lipid coated perfluoropentane droplets,Journal of the Acoustical Society of America, 143, 2001-2012.

    Phase shift droplets vaporizable by acoustic stimulation offer the advantages of producing micro-bubbles as contrast agentsin situas well as higher stability and the possibility of achieving smallersizes. Here, the acoustic droplet vaporization (ADV) threshold of a suspension of droplets with aperfluoropentane (PFP) core (diameter 400–3000 nm) is acoustically measured as a function of theexcitation frequency in a tubeless setup at room temperature. The changes in scattered responses—fundamental, sub-, and second harmonic—are investigated, a quantitative criterion is used to deter-mine the ADV phenomenon, and findings are discussed. The average threshold obtained using threedifferent scattered components increases with frequency—1.0560.28 MPa at 2.25 MHz,1.8960.57 MPa at 5 MHz, and 2.3460.014 MPa at 10 MHz. The scattered response from vapor-ized droplets was also found to qualitatively match with that from an independently prepared lipid-coated microbubble suspension in magnitude as well as trends above the determined ADV thresh-old value.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Karandish F, Haldar MK, Xia L, Gange KN, Feng L, You S, Choi Y, Sarkar K, Mallik S  2018Nucleus-targeted, echogenic polymersomes for delivering a cancer stemness inhibitor to pancreatic cancer cells,Biomacromolecules, 19,4122-4132.

    Chemotherapeutic agents for treating cancers show considerable sideeffects, toxicity, and drug resistance. To mitigate the problems, we designed nucleus-targeted, echogenic, stimuli-responsive polymeric vesicles (polymersomes) to transport andsubsequently release the encapsulated anticancer drugs within the nuclei of pancreaticcancer cells. We synthesized an alkyne-dexamethasone derivative and conjugated it to N3−polyethylene glycol (PEG)−polylactic acid (PLA) copolymer employing the Cu2+catalyzed“Click”reaction. We prepared polymersomes from the dexamethasone−PEG−PLA conjugate along with a synthesized stimuli-responsive polymer PEG−S−S−PLA. Thedexamethasone group dilates the nuclear pore complexes and transports the vesicles to thenuclei. We designed the polymersomes to release the encapsulated drugs in the presence ofa high concentration of reducing agents in the nuclei of pancreatic cancer cells. Weobserved that the nucleus-targeted, stimuli-responsive polymersomes released 70% ofencapsulated contents in the nucleus-mimicking environment in 80 min. We encapsulatedthe cancer stemness inhibitor BBI608 in the vesicles and observed that the BBI608encapsulated polymersomes reduced the viability of the BxPC3 cells to 43% in three-dimensional spheroid cultures. Thepolymersomes were prepared following a special protocol so that they scatter ultrasound, allowing imaging by a medicalultrasound scanner. Therefore, these echogenic, targeted, stimuli-responsive, and drug-encapsulated polymersomes have thepotential for trackable, targeted carrier of chemotherapeutic drugs to cancer cell nuclei.

  • Osborn J, Aliabouzar A, Zhou X, Rao R, Zhang LG, Sarkar K 2018 “Ultrasound and microbubbles enhance osteogenic differentiation of human mesenchymal stem cells on 3D printed scaffolds,” Advanced Biosystems, 2, 1800257.

    Lipid-coated microbubbles, clinically approved as contrast enhancing agents for ultrasound imaging, are investigated for the first time for their possible applications in bone tissue engineering. Effects of microbubbles (average diameter 1.1 μm) coated by a mixture of lipids (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000], and 1,2-dipalmitoyl-3-trimethylmmonium-propane) in the presence of low intensity pulsed ultrasound (LIPUS) on human mesenchymal stem cells seeded on 3D printed poly(lactic acid) porous scaffolds are investigated. LIPUS stimulation (30 mW cm−2, 1.5 MHz, 20% duty cycle) for 3 min a day with 0.5% v/v microbubbles results in a significant increase in proliferation (up to 19.3%) when compared to control after 1, 3, and 5 d. A 3-week osteogenic differentiation study shows a significant increase in total protein content (up to 27.5%), calcium deposition (up to 4.3%), and alkaline phosphatase activity (up to 43.1%) initiated by LIPUS with and without the presence of microbubbles. The microbubbles are found to remain stable during exposure, and their sustained oscillations demonstrably help focus the LIPUS energy toward enhanced cellular response. Integrating LIPUS and microbubbles promises to be a novel and effective strategy for bone tissue engineering and regeneration therapies.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Singh R, Li X, Sarkar K 2014 “Lateral migration of an elastic capsule in a wall-bounded shear,” Journal of Fluid Mechanics, 739, 421-443.

    The migration of a capsule enclosed by an elastic membrane in a wall-bounded linearshear is investigated using a front-tracking method. A detailed comparison with themigration of a viscous drop is presented varying the capillary number (in the caseof a capsule, the elastic capillary number) and the viscosity ratio. In both cases,the deformation breaks the flow reversal symmetry and makes them migrate awayfrom the wall. They quickly go through a transient evolution to eventually reach aquasi-steady state where the dynamics becomes independent of the initial positionand only depends on the wall distance. Previous analytical theories predicted thatfor a viscous drop, in the quasi-steady state, the migration and slip velocities scaleapproximately with the square of the inverse of the drop–wall separation, whereasthe drop deformation scales as the inverse cube of the separation. These power lawrelations are shown to hold for a capsule as well. The deformation and inclinationangle of the capsule and the drop at the same wall separation show a crossoverin their variation with the capillary number: the capsule shows a steeper variationthan that of the drop for smaller capillary numbers and slower variation than thedrop for larger capillary numbers. Using the Green’s function of Stokes flow, asemi-analytic theory is presented to show that the far-field stresslet that causes themigration has two distinct contributions from the interfacial stresses and the viscosityratio, with competing effects between the two defining the dynamics. It predicts thescaling of the migration velocity with the capsule–wall separation, however, matchingwith the simulated result very well only away from the wall. A phenomenologicalcorrelation for the migration velocity as a function of elastic capillary number, walldistance and viscosity ratio is developed using the simulation results. The effects ofdifferent membrane hyperelastic constitutive equations – neo-Hookean, Evans–Skalak,and Skalak – are briefly investigated to show that the behaviour remains similar fordifferent equations.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Singh R, Li X, Sarkar K 2014 “Lateral migration of an elastic capsule in a wall-bounded shear,” Journal of Fluid Mechanics, 739, 421-443.

    The migration of a capsule enclosed by an elastic membrane in a wall-bounded linearshear is investigated using a front-tracking method. A detailed comparison with themigration of a viscous drop is presented varying the capillary number (in the caseof a capsule, the elastic capillary number) and the viscosity ratio. In both cases,the deformation breaks the flow reversal symmetry and makes them migrate awayfrom the wall. They quickly go through a transient evolution to eventually reach aquasi-steady state where the dynamics becomes independent of the initial positionand only depends on the wall distance. Previous analytical theories predicted thatfor a viscous drop, in the quasi-steady state, the migration and slip velocities scaleapproximately with the square of the inverse of the drop–wall separation, whereasthe drop deformation scales as the inverse cube of the separation. These power lawrelations are shown to hold for a capsule as well. The deformation and inclinationangle of the capsule and the drop at the same wall separation show a crossoverin their variation with the capillary number: the capsule shows a steeper variationthan that of the drop for smaller capillary numbers and slower variation than thedrop for larger capillary numbers. Using the Green’s function of Stokes flow, asemi-analytic theory is presented to show that the far-field stresslet that causes themigration has two distinct contributions from the interfacial stresses and the viscosityratio, with competing effects between the two defining the dynamics. It predicts thescaling of the migration velocity with the capsule–wall separation, however, matchingwith the simulated result very well only away from the wall. A phenomenologicalcorrelation for the migration velocity as a function of elastic capillary number, walldistance and viscosity ratio is developed using the simulation results. The effects ofdifferent membrane hyperelastic constitutive equations – neo-Hookean, Evans–Skalak,and Skalak – are briefly investigated to show that the behaviour remains similar fordifferent equations.

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Singh R, Li X, Sarkar K 2014 “Lateral migration of an elastic capsule in a wall-bounded shear,” Journal of Fluid Mechanics, 739, 421-443.

    The migration of a capsule enclosed by an elastic membrane in a wall-bounded linearshear is investigated using a front-tracking method. A detailed comparison with themigration of a viscous drop is presented varying the capillary number (in the caseof a capsule, the elastic capillary number) and the viscosity ratio. In both cases,the deformation breaks the flow reversal symmetry and makes them migrate awayfrom the wall. They quickly go through a transient evolution to eventually reach aquasi-steady state where the dynamics becomes independent of the initial positionand only depends on the wall distance. Previous analytical theories predicted thatfor a viscous drop, in the quasi-steady state, the migration and slip velocities scaleapproximately with the square of the inverse of the drop–wall separation, whereasthe drop deformation scales as the inverse cube of the separation. These power lawrelations are shown to hold for a capsule as well. The deformation and inclinationangle of the capsule and the drop at the same wall separation show a crossoverin their variation with the capillary number: the capsule shows a steeper variationthan that of the drop for smaller capillary numbers and slower variation than thedrop for larger capillary numbers. Using the Green’s function of Stokes flow, asemi-analytic theory is presented to show that the far-field stresslet that causes themigration has two distinct contributions from the interfacial stresses and the viscosityratio, with competing effects between the two defining the dynamics. It predicts thescaling of the migration velocity with the capsule–wall separation, however, matchingwith the simulated result very well only away from the wall. A phenomenologicalcorrelation for the migration velocity as a function of elastic capillary number, walldistance and viscosity ratio is developed using the simulation results. The effects ofdifferent membrane hyperelastic constitutive equations – neo-Hookean, Evans–Skalak,and Skalak – are briefly investigated to show that the behaviour remains similar fordifferent equations.

  • Singh R, Sarkar K 2015 “Hydrodynamic interactions between pairs of capsules and drops in a simple shear: effects of viscosity ratio and heterogeneous collision,” Physical Review E, 92, 063029.

    Hydrodynamic interactions between a pair of capsules in simple shear are numerically investigated using afront-tracking finite difference method. The membrane of the capsule is modeled using different hyperelasticconstitutive relations. We also compare the pair interactions between drops to those between capsules. Anincreased viscosity ratio leads to a reduced net cross-stream separation between capsules as well as drops aftercollision. At low viscosity ratios, for the same capillary number drop-pairs show higher cross-stream separationthan those for capsule-pairs, while substantially large viscosity ratios result in almost the same value for bothcases. We investigate pair-collisions between two heterogeneous capsules C1and C2with two different capillarynumbers. The maximum deformation of C1was seen to increase with increasing stiffness (decreasing capillarynumber) of C2, even though the stiffness of C1was kept fixed. The findings are similar for a drop-pair, however,with a smaller maximum deformation for the same combinations of capillary numbers. The final cross-streamdrift of the trajectory of C1decreases with the increasing stiffness of C2, but the relative trajectory betweenthe capsules remains unchanged. The maximum deformation and the cross-stream drift of the trajectory of C1are shown to approximately vary with power-law functions of the ratio of the capillary numbers of C1andC2. An analytical explanation of the dependence on the two capillary numbers is offered. Different membraneconstitutive laws result in similar deformation and drift in trajectory.

  • Mukherjee S, Sarkar K 2014 “Lateral migration of a viscoelastic drop in a Newtonian fluid in a shear flow near a wall,” Physics of Fluids, 26, 103102.

    Wall induced lateral migration of a viscoelastic (FENE-MCR) drop in a Newtonianfluid is investigated. Just like a Newtonian drop, a viscoelastic drop reaches a quasi-steady state where the lateral velocity only depends on the instantaneous distancefrom the wall. The drop migration velocity and the deformation scale inversely withthe square and the cube of the distance from the wall, respectively. The migration ve-locity varies non-monotonically with increasing viscoelasticity (increasing Deborahnumber); initially increasing and then decreasing. An analytical explanation has beengiven of the effects by computing the migration velocity as arising from an imagestresslet field due to the drop. The semi-analytical expression matches well with thesimulated migration velocity away from the wall. It contains a viscoelastic stressletcomponent apart from those arising from interfacial tension and viscosity ratio. Themigration dynamics is a result of the competition between the viscous (interfacialtension and viscosity ratio) and the viscoelastic effects. The viscoelastic stressletcontribution towards the migration velocity steadily increases. But the interfacialstresslet—arising purely from the drop shape—first increases and then decreases withrising Deborah number causing the migration velocity to be non-monotonic. The ge-ometric effect of the interfacial stresslet is caused by a corresponding nonmonotonicvariation of the drop inclination. High viscosity ratio is briefly considered to showthat the drop viscoelasticity could stabilize a drop against breakup, and the increase inmigration velocity due to viscoelasticity is larger compared to the viscosity-matchedcase.

  • Srivastava P, Malipeddi Reddy A, Sarkar K 2016 “Steady shear rheology of a viscous emulsion in the presence of finite inertia at moderate volume fractions: sign reversal of normal stress differences,” Journal of Fluid Mechanics, 85, 494-522.

    The shear rheology of an emulsion of viscous drops in the presence of finite inertiais investigated using direct numerical simulation. In the absence of inertia, emulsionsdisplay a non-Newtonian rheology with positive first and negative second normalstress differences. However, recently it was discovered that a small amount ofdrop-level inertia alters their signs – the first normal stress difference becomesnegative and the second one becomes positive, each in a small range of capillarynumbers (Li & Sarkar,J. Rheol., vol. 49, 2005, pp. 1377–1394). Sign reversal wasshown numerically and analytically, but only in the limit of a dilute emulsion wheredrop–drop interactions were neglected. Here, we compute the rheology of a density-and viscosity-matched emulsion, accounting for the interactions in the volume fractionrange of 5 %–27 % and Reynolds number range of 0.1–10. The computed rheologicalproperties (effective shear viscosity and first and second normal stress differences) inthe Stokes limit match well with previous theoretical (Choi–Schowalter in the dilutelimit) and simulated results (for concentrated systems) using the boundary elementmethod. The two distinct components of the rheology arising from the interfacialstresses at the drop surface and the perturbative Reynolds stresses are investigated asfunctions of the drop Reynolds number, capillary number and volume fraction. Thesign change is caused by the increasing drop inclination in the presence of inertia,which in turn directly affects the interfacial stresses. Increase of the volume fractionor capillary number increases the critical Reynolds number for sign reversals due toenhanced alignment of the drops with the flow directions. The effect of increasingthe volume fraction on the rheology is explained by relating it to interactions andspecifically to the contact pair-distribution function computed from the simulation.The excess stresses are seen to show an approximately linear behaviour with theReynolds number in the range of 0.1–5, while with the capillary number and volumefraction, the variation is weakly quadratic.

  • Aliabouzar M, Zhang LG, Sarkar K, 2016 “Lipid-coated microbubbles and low intensity pulsed ultrasound enhance chondrogenesis of human mesenchymal stem cells in 3D printed scaffolds,” Nature Scientific Report, 6, 37728.

    Lipid-coated microbubbles are used to enhance ultrasound imaging and drug delivery. Here we apply these microbubbles along with low intensity pulsed ultrasound (LIPUS) for the first time to enhance proliferation and chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in a 3D printed poly-(ethylene glycol)-diacrylate (PEG-DA) hydrogel scaffold. The hMSC proliferation increased up to 40% after 5 days of culture in the presence of 0.5% (v/v) microbubbles and LIPUS in contrast to 18% with LIPUS alone. We systematically varied the acoustic excitation parameters—excitation intensity, frequency and duty cycle—to find 30 mW/cm2, 1.5 MHz and 20% duty cycle to be optimal for hMSC proliferation. A 3-week chondrogenic differentiation results demonstrated that combining LIPUS with microbubbles enhanced glycosaminoglycan (GAG) production by 17% (5% with LIPUS alone), and type II collagen production by 78% (44% by LIPUS alone). Therefore, integrating LIPUS and microbubbles appears to be a promising strategy for enhanced hMSC growth and chondrogenic differentiation, which are critical components for cartilage regeneration. The results offer possibilities of novel applications of microbubbles, already clinically approved for contrast enhanced ultrasound imaging, in tissue engineering.

  • Aliabouzar M, Kumar KN, Sarkar K, 2018Acoustic vaporization threshold of lipid coated perfluoropentane droplets,Journal of the Acoustical Society of America, 143, 2001-2012.

    Phase shift droplets vaporizable by acoustic stimulation offer the advantages of producing micro-bubbles as contrast agentsin situas well as higher stability and the possibility of achieving smallersizes. Here, the acoustic droplet vaporization (ADV) threshold of a suspension of droplets with aperfluoropentane (PFP) core (diameter 400–3000 nm) is acoustically measured as a function of theexcitation frequency in a tubeless setup at room temperature. The changes in scattered responses—fundamental, sub-, and second harmonic—are investigated, a quantitative criterion is used to deter-mine the ADV phenomenon, and findings are discussed. The average threshold obtained using threedifferent scattered components increases with frequency—1.0560.28 MPa at 2.25 MHz,1.8960.57 MPa at 5 MHz, and 2.3460.014 MPa at 10 MHz. The scattered response from vapor-ized droplets was also found to qualitatively match with that from an independently prepared lipid-coated microbubble suspension in magnitude as well as trends above the determined ADV thresh-old value.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Karandish F, Haldar MK, Xia L, Gange KN, Feng L, You S, Choi Y, Sarkar K, Mallik S  2018Nucleus-targeted, echogenic polymersomes for delivering a cancer stemness inhibitor to pancreatic cancer cells,Biomacromolecules, 19,4122-4132.

    Chemotherapeutic agents for treating cancers show considerable sideeffects, toxicity, and drug resistance. To mitigate the problems, we designed nucleus-targeted, echogenic, stimuli-responsive polymeric vesicles (polymersomes) to transport andsubsequently release the encapsulated anticancer drugs within the nuclei of pancreaticcancer cells. We synthesized an alkyne-dexamethasone derivative and conjugated it to N3−polyethylene glycol (PEG)−polylactic acid (PLA) copolymer employing the Cu2+catalyzed“Click”reaction. We prepared polymersomes from the dexamethasone−PEG−PLA conjugate along with a synthesized stimuli-responsive polymer PEG−S−S−PLA. Thedexamethasone group dilates the nuclear pore complexes and transports the vesicles to thenuclei. We designed the polymersomes to release the encapsulated drugs in the presence ofa high concentration of reducing agents in the nuclei of pancreatic cancer cells. Weobserved that the nucleus-targeted, stimuli-responsive polymersomes released 70% ofencapsulated contents in the nucleus-mimicking environment in 80 min. We encapsulatedthe cancer stemness inhibitor BBI608 in the vesicles and observed that the BBI608encapsulated polymersomes reduced the viability of the BxPC3 cells to 43% in three-dimensional spheroid cultures. Thepolymersomes were prepared following a special protocol so that they scatter ultrasound, allowing imaging by a medicalultrasound scanner. Therefore, these echogenic, targeted, stimuli-responsive, and drug-encapsulated polymersomes have thepotential for trackable, targeted carrier of chemotherapeutic drugs to cancer cell nuclei.

  • Osborn J, Aliabouzar A, Zhou X, Rao R, Zhang LG, Sarkar K 2018 “Ultrasound and microbubbles enhance osteogenic differentiation of human mesenchymal stem cells on 3D printed scaffolds,” Advanced Biosystems, 2, 1800257.

    Lipid-coated microbubbles, clinically approved as contrast enhancing agents for ultrasound imaging, are investigated for the first time for their possible applications in bone tissue engineering. Effects of microbubbles (average diameter 1.1 μm) coated by a mixture of lipids (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000], and 1,2-dipalmitoyl-3-trimethylmmonium-propane) in the presence of low intensity pulsed ultrasound (LIPUS) on human mesenchymal stem cells seeded on 3D printed poly(lactic acid) porous scaffolds are investigated. LIPUS stimulation (30 mW cm−2, 1.5 MHz, 20% duty cycle) for 3 min a day with 0.5% v/v microbubbles results in a significant increase in proliferation (up to 19.3%) when compared to control after 1, 3, and 5 d. A 3-week osteogenic differentiation study shows a significant increase in total protein content (up to 27.5%), calcium deposition (up to 4.3%), and alkaline phosphatase activity (up to 43.1%) initiated by LIPUS with and without the presence of microbubbles. The microbubbles are found to remain stable during exposure, and their sustained oscillations demonstrably help focus the LIPUS energy toward enhanced cellular response. Integrating LIPUS and microbubbles promises to be a novel and effective strategy for bone tissue engineering and regeneration therapies.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Aliabouzar M, Kumar KN, Sarkar K, 2018Acoustic vaporization threshold of lipid coated perfluoropentane droplets,Journal of the Acoustical Society of America, 143, 2001-2012.

    Phase shift droplets vaporizable by acoustic stimulation offer the advantages of producing micro-bubbles as contrast agentsin situas well as higher stability and the possibility of achieving smallersizes. Here, the acoustic droplet vaporization (ADV) threshold of a suspension of droplets with aperfluoropentane (PFP) core (diameter 400–3000 nm) is acoustically measured as a function of theexcitation frequency in a tubeless setup at room temperature. The changes in scattered responses—fundamental, sub-, and second harmonic—are investigated, a quantitative criterion is used to deter-mine the ADV phenomenon, and findings are discussed. The average threshold obtained using threedifferent scattered components increases with frequency—1.0560.28 MPa at 2.25 MHz,1.8960.57 MPa at 5 MHz, and 2.3460.014 MPa at 10 MHz. The scattered response from vapor-ized droplets was also found to qualitatively match with that from an independently prepared lipid-coated microbubble suspension in magnitude as well as trends above the determined ADV thresh-old value.

  • Karandish F, Haldar MK, Xia L, Gange KN, Feng L, You S, Choi Y, Sarkar K, Mallik S  2018Nucleus-targeted, echogenic polymersomes for delivering a cancer stemness inhibitor to pancreatic cancer cells,Biomacromolecules, 19,4122-4132.

    Chemotherapeutic agents for treating cancers show considerable sideeffects, toxicity, and drug resistance. To mitigate the problems, we designed nucleus-targeted, echogenic, stimuli-responsive polymeric vesicles (polymersomes) to transport andsubsequently release the encapsulated anticancer drugs within the nuclei of pancreaticcancer cells. We synthesized an alkyne-dexamethasone derivative and conjugated it to N3−polyethylene glycol (PEG)−polylactic acid (PLA) copolymer employing the Cu2+catalyzed“Click”reaction. We prepared polymersomes from the dexamethasone−PEG−PLA conjugate along with a synthesized stimuli-responsive polymer PEG−S−S−PLA. Thedexamethasone group dilates the nuclear pore complexes and transports the vesicles to thenuclei. We designed the polymersomes to release the encapsulated drugs in the presence ofa high concentration of reducing agents in the nuclei of pancreatic cancer cells. Weobserved that the nucleus-targeted, stimuli-responsive polymersomes released 70% ofencapsulated contents in the nucleus-mimicking environment in 80 min. We encapsulatedthe cancer stemness inhibitor BBI608 in the vesicles and observed that the BBI608encapsulated polymersomes reduced the viability of the BxPC3 cells to 43% in three-dimensional spheroid cultures. Thepolymersomes were prepared following a special protocol so that they scatter ultrasound, allowing imaging by a medicalultrasound scanner. Therefore, these echogenic, targeted, stimuli-responsive, and drug-encapsulated polymersomes have thepotential for trackable, targeted carrier of chemotherapeutic drugs to cancer cell nuclei.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Osborn J, Aliabouzar A, Zhou X, Rao R, Zhang LG, Sarkar K 2018 “Ultrasound and microbubbles enhance osteogenic differentiation of human mesenchymal stem cells on 3D printed scaffolds,” Advanced Biosystems, 2, 1800257.

    Lipid-coated microbubbles, clinically approved as contrast enhancing agents for ultrasound imaging, are investigated for the first time for their possible applications in bone tissue engineering. Effects of microbubbles (average diameter 1.1 μm) coated by a mixture of lipids (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000], and 1,2-dipalmitoyl-3-trimethylmmonium-propane) in the presence of low intensity pulsed ultrasound (LIPUS) on human mesenchymal stem cells seeded on 3D printed poly(lactic acid) porous scaffolds are investigated. LIPUS stimulation (30 mW cm−2, 1.5 MHz, 20% duty cycle) for 3 min a day with 0.5% v/v microbubbles results in a significant increase in proliferation (up to 19.3%) when compared to control after 1, 3, and 5 d. A 3-week osteogenic differentiation study shows a significant increase in total protein content (up to 27.5%), calcium deposition (up to 4.3%), and alkaline phosphatase activity (up to 43.1%) initiated by LIPUS with and without the presence of microbubbles. The microbubbles are found to remain stable during exposure, and their sustained oscillations demonstrably help focus the LIPUS energy toward enhanced cellular response. Integrating LIPUS and microbubbles promises to be a novel and effective strategy for bone tissue engineering and regeneration therapies.

  • Aliabouzar M, Zhang LG, Sarkar K, 2016 “Lipid-coated microbubbles and low intensity pulsed ultrasound enhance chondrogenesis of human mesenchymal stem cells in 3D printed scaffolds,” Nature Scientific Report, 6, 37728.

    Lipid-coated microbubbles are used to enhance ultrasound imaging and drug delivery. Here we apply these microbubbles along with low intensity pulsed ultrasound (LIPUS) for the first time to enhance proliferation and chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in a 3D printed poly-(ethylene glycol)-diacrylate (PEG-DA) hydrogel scaffold. The hMSC proliferation increased up to 40% after 5 days of culture in the presence of 0.5% (v/v) microbubbles and LIPUS in contrast to 18% with LIPUS alone. We systematically varied the acoustic excitation parameters—excitation intensity, frequency and duty cycle—to find 30 mW/cm2, 1.5 MHz and 20% duty cycle to be optimal for hMSC proliferation. A 3-week chondrogenic differentiation results demonstrated that combining LIPUS with microbubbles enhanced glycosaminoglycan (GAG) production by 17% (5% with LIPUS alone), and type II collagen production by 78% (44% by LIPUS alone). Therefore, integrating LIPUS and microbubbles appears to be a promising strategy for enhanced hMSC growth and chondrogenic differentiation, which are critical components for cartilage regeneration. The results offer possibilities of novel applications of microbubbles, already clinically approved for contrast enhanced ultrasound imaging, in tissue engineering.

  • Srivastava P, Malipeddi Reddy A, Sarkar K 2016 “Steady shear rheology of a viscous emulsion in the presence of finite inertia at moderate volume fractions: sign reversal of normal stress differences,” Journal of Fluid Mechanics, 85, 494-522.

    The shear rheology of an emulsion of viscous drops in the presence of finite inertiais investigated using direct numerical simulation. In the absence of inertia, emulsionsdisplay a non-Newtonian rheology with positive first and negative second normalstress differences. However, recently it was discovered that a small amount ofdrop-level inertia alters their signs – the first normal stress difference becomesnegative and the second one becomes positive, each in a small range of capillarynumbers (Li & Sarkar,J. Rheol., vol. 49, 2005, pp. 1377–1394). Sign reversal wasshown numerically and analytically, but only in the limit of a dilute emulsion wheredrop–drop interactions were neglected. Here, we compute the rheology of a density-and viscosity-matched emulsion, accounting for the interactions in the volume fractionrange of 5 %–27 % and Reynolds number range of 0.1–10. The computed rheologicalproperties (effective shear viscosity and first and second normal stress differences) inthe Stokes limit match well with previous theoretical (Choi–Schowalter in the dilutelimit) and simulated results (for concentrated systems) using the boundary elementmethod. The two distinct components of the rheology arising from the interfacialstresses at the drop surface and the perturbative Reynolds stresses are investigated asfunctions of the drop Reynolds number, capillary number and volume fraction. Thesign change is caused by the increasing drop inclination in the presence of inertia,which in turn directly affects the interfacial stresses. Increase of the volume fractionor capillary number increases the critical Reynolds number for sign reversals due toenhanced alignment of the drops with the flow directions. The effect of increasingthe volume fraction on the rheology is explained by relating it to interactions andspecifically to the contact pair-distribution function computed from the simulation.The excess stresses are seen to show an approximately linear behaviour with theReynolds number in the range of 0.1–5, while with the capillary number and volumefraction, the variation is weakly quadratic.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Singh R, Sarkar K 2015 “Hydrodynamic interactions between pairs of capsules and drops in a simple shear: effects of viscosity ratio and heterogeneous collision,” Physical Review E, 92, 063029.

    Hydrodynamic interactions between a pair of capsules in simple shear are numerically investigated using afront-tracking finite difference method. The membrane of the capsule is modeled using different hyperelasticconstitutive relations. We also compare the pair interactions between drops to those between capsules. Anincreased viscosity ratio leads to a reduced net cross-stream separation between capsules as well as drops aftercollision. At low viscosity ratios, for the same capillary number drop-pairs show higher cross-stream separationthan those for capsule-pairs, while substantially large viscosity ratios result in almost the same value for bothcases. We investigate pair-collisions between two heterogeneous capsules C1and C2with two different capillarynumbers. The maximum deformation of C1was seen to increase with increasing stiffness (decreasing capillarynumber) of C2, even though the stiffness of C1was kept fixed. The findings are similar for a drop-pair, however,with a smaller maximum deformation for the same combinations of capillary numbers. The final cross-streamdrift of the trajectory of C1decreases with the increasing stiffness of C2, but the relative trajectory betweenthe capsules remains unchanged. The maximum deformation and the cross-stream drift of the trajectory of C1are shown to approximately vary with power-law functions of the ratio of the capillary numbers of C1andC2. An analytical explanation of the dependence on the two capillary numbers is offered. Different membraneconstitutive laws result in similar deformation and drift in trajectory.

  • Mukherjee S, Sarkar K 2014 “Lateral migration of a viscoelastic drop in a Newtonian fluid in a shear flow near a wall,” Physics of Fluids, 26, 103102.

    Wall induced lateral migration of a viscoelastic (FENE-MCR) drop in a Newtonianfluid is investigated. Just like a Newtonian drop, a viscoelastic drop reaches a quasi-steady state where the lateral velocity only depends on the instantaneous distancefrom the wall. The drop migration velocity and the deformation scale inversely withthe square and the cube of the distance from the wall, respectively. The migration ve-locity varies non-monotonically with increasing viscoelasticity (increasing Deborahnumber); initially increasing and then decreasing. An analytical explanation has beengiven of the effects by computing the migration velocity as arising from an imagestresslet field due to the drop. The semi-analytical expression matches well with thesimulated migration velocity away from the wall. It contains a viscoelastic stressletcomponent apart from those arising from interfacial tension and viscosity ratio. Themigration dynamics is a result of the competition between the viscous (interfacialtension and viscosity ratio) and the viscoelastic effects. The viscoelastic stressletcontribution towards the migration velocity steadily increases. But the interfacialstresslet—arising purely from the drop shape—first increases and then decreases withrising Deborah number causing the migration velocity to be non-monotonic. The ge-ometric effect of the interfacial stresslet is caused by a corresponding nonmonotonicvariation of the drop inclination. High viscosity ratio is briefly considered to showthat the drop viscoelasticity could stabilize a drop against breakup, and the increase inmigration velocity due to viscoelasticity is larger compared to the viscosity-matchedcase.

  • Singh R, Li X, Sarkar K 2014 “Lateral migration of an elastic capsule in a wall-bounded shear,” Journal of Fluid Mechanics, 739, 421-443.

    The migration of a capsule enclosed by an elastic membrane in a wall-bounded linearshear is investigated using a front-tracking method. A detailed comparison with themigration of a viscous drop is presented varying the capillary number (in the caseof a capsule, the elastic capillary number) and the viscosity ratio. In both cases,the deformation breaks the flow reversal symmetry and makes them migrate awayfrom the wall. They quickly go through a transient evolution to eventually reach aquasi-steady state where the dynamics becomes independent of the initial positionand only depends on the wall distance. Previous analytical theories predicted thatfor a viscous drop, in the quasi-steady state, the migration and slip velocities scaleapproximately with the square of the inverse of the drop–wall separation, whereasthe drop deformation scales as the inverse cube of the separation. These power lawrelations are shown to hold for a capsule as well. The deformation and inclinationangle of the capsule and the drop at the same wall separation show a crossoverin their variation with the capillary number: the capsule shows a steeper variationthan that of the drop for smaller capillary numbers and slower variation than thedrop for larger capillary numbers. Using the Green’s function of Stokes flow, asemi-analytic theory is presented to show that the far-field stresslet that causes themigration has two distinct contributions from the interfacial stresses and the viscosityratio, with competing effects between the two defining the dynamics. It predicts thescaling of the migration velocity with the capsule–wall separation, however, matchingwith the simulated result very well only away from the wall. A phenomenologicalcorrelation for the migration velocity as a function of elastic capillary number, walldistance and viscosity ratio is developed using the simulation results. The effects ofdifferent membrane hyperelastic constitutive equations – neo-Hookean, Evans–Skalak,and Skalak – are briefly investigated to show that the behaviour remains similar fordifferent equations.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Aliabouzar M, Kumar KN, Sarkar K, 2018Acoustic vaporization threshold of lipid coated perfluoropentane droplets,Journal of the Acoustical Society of America, 143, 2001-2012.

    Phase shift droplets vaporizable by acoustic stimulation offer the advantages of producing micro-bubbles as contrast agentsin situas well as higher stability and the possibility of achieving smallersizes. Here, the acoustic droplet vaporization (ADV) threshold of a suspension of droplets with aperfluoropentane (PFP) core (diameter 400–3000 nm) is acoustically measured as a function of theexcitation frequency in a tubeless setup at room temperature. The changes in scattered responses—fundamental, sub-, and second harmonic—are investigated, a quantitative criterion is used to deter-mine the ADV phenomenon, and findings are discussed. The average threshold obtained using threedifferent scattered components increases with frequency—1.0560.28 MPa at 2.25 MHz,1.8960.57 MPa at 5 MHz, and 2.3460.014 MPa at 10 MHz. The scattered response from vapor-ized droplets was also found to qualitatively match with that from an independently prepared lipid-coated microbubble suspension in magnitude as well as trends above the determined ADV thresh-old value.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Karandish F, Haldar MK, Xia L, Gange KN, Feng L, You S, Choi Y, Sarkar K, Mallik S  2018Nucleus-targeted, echogenic polymersomes for delivering a cancer stemness inhibitor to pancreatic cancer cells,Biomacromolecules, 19,4122-4132.

    Chemotherapeutic agents for treating cancers show considerable sideeffects, toxicity, and drug resistance. To mitigate the problems, we designed nucleus-targeted, echogenic, stimuli-responsive polymeric vesicles (polymersomes) to transport andsubsequently release the encapsulated anticancer drugs within the nuclei of pancreaticcancer cells. We synthesized an alkyne-dexamethasone derivative and conjugated it to N3−polyethylene glycol (PEG)−polylactic acid (PLA) copolymer employing the Cu2+catalyzed“Click”reaction. We prepared polymersomes from the dexamethasone−PEG−PLA conjugate along with a synthesized stimuli-responsive polymer PEG−S−S−PLA. Thedexamethasone group dilates the nuclear pore complexes and transports the vesicles to thenuclei. We designed the polymersomes to release the encapsulated drugs in the presence ofa high concentration of reducing agents in the nuclei of pancreatic cancer cells. Weobserved that the nucleus-targeted, stimuli-responsive polymersomes released 70% ofencapsulated contents in the nucleus-mimicking environment in 80 min. We encapsulatedthe cancer stemness inhibitor BBI608 in the vesicles and observed that the BBI608encapsulated polymersomes reduced the viability of the BxPC3 cells to 43% in three-dimensional spheroid cultures. Thepolymersomes were prepared following a special protocol so that they scatter ultrasound, allowing imaging by a medicalultrasound scanner. Therefore, these echogenic, targeted, stimuli-responsive, and drug-encapsulated polymersomes have thepotential for trackable, targeted carrier of chemotherapeutic drugs to cancer cell nuclei.

  • Osborn J, Aliabouzar A, Zhou X, Rao R, Zhang LG, Sarkar K 2018 “Ultrasound and microbubbles enhance osteogenic differentiation of human mesenchymal stem cells on 3D printed scaffolds,” Advanced Biosystems, 2, 1800257.

    Lipid-coated microbubbles, clinically approved as contrast enhancing agents for ultrasound imaging, are investigated for the first time for their possible applications in bone tissue engineering. Effects of microbubbles (average diameter 1.1 μm) coated by a mixture of lipids (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000], and 1,2-dipalmitoyl-3-trimethylmmonium-propane) in the presence of low intensity pulsed ultrasound (LIPUS) on human mesenchymal stem cells seeded on 3D printed poly(lactic acid) porous scaffolds are investigated. LIPUS stimulation (30 mW cm−2, 1.5 MHz, 20% duty cycle) for 3 min a day with 0.5% v/v microbubbles results in a significant increase in proliferation (up to 19.3%) when compared to control after 1, 3, and 5 d. A 3-week osteogenic differentiation study shows a significant increase in total protein content (up to 27.5%), calcium deposition (up to 4.3%), and alkaline phosphatase activity (up to 43.1%) initiated by LIPUS with and without the presence of microbubbles. The microbubbles are found to remain stable during exposure, and their sustained oscillations demonstrably help focus the LIPUS energy toward enhanced cellular response. Integrating LIPUS and microbubbles promises to be a novel and effective strategy for bone tissue engineering and regeneration therapies.

  • Aliabouzar M, Kumar KN, Sarkar K, 2019 “Effects of size and boiling point of perfluorocarbon droplets on the frequency dependence of vaporization threshold, Journal of the Acoustical Society of America, 145, 1105-1106.

    Phase shift liquid perfluorocarbon (PFC) droplets vaporizable by ultrasound into echogenicmicrobubble above a threshold pressure, termed acoustic droplet vaporization (ADV), are usedfor therapeutic and diagnostic applications. This study systematically investigated the effect ofexcitation frequency (2.25, 10, and 15 MHz) on the ADV and inertial cavitation (IC) thresholds oflipid-coated PFC droplets of three different liquid cores—perfluoropentane (PFP), perfluorohexane(PFH), and perfluorooctyl bromide (PFOB)—and of two different sizes—average diameters smallerthan 3lm and larger than 10lm—in a tubeless setup. This study found that the ADV thresholdincreases with frequency for the lowest boiling point liquid, PFP, for both large and small sizedroplets. For higher boiling point liquids, PFH and PFOB, this study did not detect vaporization forsmall size droplets at the excitation levels (maximum 4 MPa peak negative) studied here. The largePFOB droplets experienced ADV only at the highest excitation frequency 15 MHz. For large PFHdroplets, ADV threshold decreases with frequency that could possibly be due to the superharmonicfocusing being a significant effect at larger sizes and the higher excitation pressures. ADV thresh-olds at all the frequencies studied here occurred at lower rarefactional pressures than IC thresholdsindicating that phase transition precedes inertial cavitation.

  • Aliabouzar M, Kumar KN, Sarkar K, 2018Acoustic vaporization threshold of lipid coated perfluoropentane droplets,Journal of the Acoustical Society of America, 143, 2001-2012.

    Phase shift droplets vaporizable by acoustic stimulation offer the advantages of producing micro-bubbles as contrast agentsin situas well as higher stability and the possibility of achieving smallersizes. Here, the acoustic droplet vaporization (ADV) threshold of a suspension of droplets with aperfluoropentane (PFP) core (diameter 400–3000 nm) is acoustically measured as a function of theexcitation frequency in a tubeless setup at room temperature. The changes in scattered responses—fundamental, sub-, and second harmonic—are investigated, a quantitative criterion is used to deter-mine the ADV phenomenon, and findings are discussed. The average threshold obtained using threedifferent scattered components increases with frequency—1.0560.28 MPa at 2.25 MHz,1.8960.57 MPa at 5 MHz, and 2.3460.014 MPa at 10 MHz. The scattered response from vapor-ized droplets was also found to qualitatively match with that from an independently prepared lipid-coated microbubble suspension in magnitude as well as trends above the determined ADV thresh-old value.

  • Kulkarni P, Haldar MK, Karandish F, Confeld M, Hossain R, Borowicz P, Gange KN, Xia L, Sarkar K, Mallik S 2018Tissue-penetrating, hypoxia-responsive echogenic polymersomes for drug delivery to solid tumors,Chemistry A European Journal, 24, 12490-12494.

    Hypoxia in solid tumors facilitates the progres-sion of the disease, develops resistance to chemo and radiotherapy, and contributes to relapse. Due to the lack of tumor penetration, most of the reported drug carriers are unable to reach the hypoxic niches of the solid tumors. We have developed tissue-penetrating, hypoxia-responsive echogenic polymersomes to deliver anti cancer drugs to solid tumors. The polymersomes are composed of a hy-poxia-responsive azobenzene conjugated and a tissue penetrating peptide functionalized polylactic acid-polyethylene glycol polymer. The drug-encapsulated, hypoxia-responsive polymersomes substantially decreased the viability of pancreatic cancer cells in spheroidal cultures. Under normoxic conditions, polymersomes were echogenic at diagnostic ultrasound frequencies but lose the echogenicity under hypoxia. In vivo imaging studies with xenograft mouse model further confirmed the ability of the polymersomes to target, penetrate, and deliver the encapsulated contents in hypoxic pancreatic tumor tissues.

  • Karandish F, Haldar MK, Xia L, Gange KN, Feng L, You S, Choi Y, Sarkar K, Mallik S  2018Nucleus-targeted, echogenic polymersomes for delivering a cancer stemness inhibitor to pancreatic cancer cells,Biomacromolecules, 19,4122-4132.

    Chemotherapeutic agents for treating cancers show considerable sideeffects, toxicity, and drug resistance. To mitigate the problems, we designed nucleus-targeted, echogenic, stimuli-responsive polymeric vesicles (polymersomes) to transport andsubsequently release the encapsulated anticancer drugs within the nuclei of pancreaticcancer cells. We synthesized an alkyne-dexamethasone derivative and conjugated it to N3−polyethylene glycol (PEG)−polylactic acid (PLA) copolymer employing the Cu2+catalyzed“Click”reaction. We prepared polymersomes from the dexamethasone−PEG−PLA conjugate along with a synthesized stimuli-responsive polymer PEG−S−S−PLA. Thedexamethasone group dilates the nuclear pore complexes and transports the vesicles to thenuclei. We designed the polymersomes to release the encapsulated drugs in the presence ofa high concentration of reducing agents in the nuclei of pancreatic cancer cells. Weobserved that the nucleus-targeted, stimuli-responsive polymersomes released 70% ofencapsulated contents in the nucleus-mimicking environment in 80 min. We encapsulatedthe cancer stemness inhibitor BBI608 in the vesicles and observed that the BBI608encapsulated polymersomes reduced the viability of the BxPC3 cells to 43% in three-dimensional spheroid cultures. Thepolymersomes were prepared following a special protocol so that they scatter ultrasound, allowing imaging by a medicalultrasound scanner. Therefore, these echogenic, targeted, stimuli-responsive, and drug-encapsulated polymersomes have thepotential for trackable, targeted carrier of chemotherapeutic drugs to cancer cell nuclei.

  • Malipeddy Reddy A, Sarkar K, 2019 “Shear-induced collective diffusivity down a concentration gradient in a viscous emulsion,” Journal of Fluid Mechanics, 868, 5-25.

    The shear-induced collective diffusivity down a concentration gradient in a viscous
    emulsion is computed using direct numerical simulation. A layer of randomly packed
    drops subjected to a shear flow, shows the layer width to increase with the 1=3
    power of time, consistent with a semi-dilute theory that assumes a diffusivity linear
    with concentration. This characteristic scaling and the underlying theory are used
    to compute the collective diffusivity coefficient. This is the first ever computation
    of this quantity for a system of deformable particles using fully resolved numerical
    simulation. The results match very well with previous experimental observations.
    The coefficient of collective diffusivity varies non-monotonically with the capillary
    number, due to the competing effects of increasing deformation and drop orientation.
    A phenomenological correlation for the collective diffusivity coefficient as a function
    of capillary number is presented. We also apply an alternative approach to compute
    collective diffusivity, developed originally for a statistically homogeneous rigid sphere
    suspension – computing the dynamic structure factor from the simulated droplet
    positions and examining its time variation at small wavenumber. We show that
    the results from this alternative approach qualitatively agree with our computation
    of collective diffusivity including the prediction of the non-monotonic variation of
    diffusivity with the capillary number.

  • Malipeddy Reddy A, Sarkar K, 2019 “Shear-induced collective diffusivity down a concentration gradient in a viscous emulsion,” Journal of Fluid Mechanics, 868, 5-25.

    The shear-induced collective diffusivity down a concentration gradient in a viscous
    emulsion is computed using direct numerical simulation. A layer of randomly packed
    drops subjected to a shear flow, shows the layer width to increase with the 1=3
    power of time, consistent with a semi-dilute theory that assumes a diffusivity linear
    with concentration. This characteristic scaling and the underlying theory are used
    to compute the collective diffusivity coefficient. This is the first ever computation
    of this quantity for a system of deformable particles using fully resolved numerical
    simulation. The results match very well with previous experimental observations.
    The coefficient of collective diffusivity varies non-monotonically with the capillary
    number, due to the competing effects of increasing deformation and drop orientation.
    A phenomenological correlation for the collective diffusivity coefficient as a function
    of capillary number is presented. We also apply an alternative approach to compute
    collective diffusivity, developed originally for a statistically homogeneous rigid sphere
    suspension – computing the dynamic structure factor from the simulated droplet
    positions and examining its time variation at small wavenumber. We show that
    the results from this alternative approach qualitatively agree with our computation
    of collective diffusivity including the prediction of the non-monotonic variation of
    diffusivity with the capillary number.

  • Malipeddy Reddy A, Sarkar K, 2019 “Shear-induced collective diffusivity down a concentration gradient in a viscous emulsion,” Journal of Fluid Mechanics, 868, 5-25.

    The shear-induced collective diffusivity down a concentration gradient in a viscous
    emulsion is computed using direct numerical simulation. A layer of randomly packed
    drops subjected to a shear flow, shows the layer width to increase with the 1=3
    power of time, consistent with a semi-dilute theory that assumes a diffusivity linear
    with concentration. This characteristic scaling and the underlying theory are used
    to compute the collective diffusivity coefficient. This is the first ever computation
    of this quantity for a system of deformable particles using fully resolved numerical
    simulation. The results match very well with previous experimental observations.
    The coefficient of collective diffusivity varies non-monotonically with the capillary
    number, due to the competing effects of increasing deformation and drop orientation.
    A phenomenological correlation for the collective diffusivity coefficient as a function
    of capillary number is presented. We also apply an alternative approach to compute
    collective diffusivity, developed originally for a statistically homogeneous rigid sphere
    suspension – computing the dynamic structure factor from the simulated droplet
    positions and examining its time variation at small wavenumber. We show that
    the results from this alternative approach qualitatively agree with our computation
    of collective diffusivity including the prediction of the non-monotonic variation of
    diffusivity with the capillary number.

  • Srivastava P, Malipeddi Reddy A, Sarkar K 2016 “Steady shear rheology of a viscous emulsion in the presence of finite inertia at moderate volume fractions: sign reversal of normal stress differences,” Journal of Fluid Mechanics, 85, 494-522.

    The shear rheology of an emulsion of viscous drops in the presence of finite inertiais investigated using direct numerical simulation. In the absence of inertia, emulsionsdisplay a non-Newtonian rheology with positive first and negative second normalstress differences. However, recently it was discovered that a small amount ofdrop-level inertia alters their signs – the first normal stress difference becomesnegative and the second one becomes positive, each in a small range of capillarynumbers (Li & Sarkar,J. Rheol., vol. 49, 2005, pp. 1377–1394). Sign reversal wasshown numerically and analytically, but only in the limit of a dilute emulsion wheredrop–drop interactions were neglected. Here, we compute the rheology of a density-and viscosity-matched emulsion, accounting for the interactions in the volume fractionrange of 5 %–27 % and Reynolds number range of 0.1–10. The computed rheologicalproperties (effective shear viscosity and first and second normal stress differences) inthe Stokes limit match well with previous theoretical (Choi–Schowalter in the dilutelimit) and simulated results (for concentrated systems) using the boundary elementmethod. The two distinct components of the rheology arising from the interfacialstresses at the drop surface and the perturbative Reynolds stresses are investigated asfunctions of the drop Reynolds number, capillary number and volume fraction. Thesign change is caused by the increasing drop inclination in the presence of inertia,which in turn directly affects the interfacial stresses. Increase of the volume fractionor capillary number increases the critical Reynolds number for sign reversals due toenhanced alignment of the drops with the flow directions. The effect of increasingthe volume fraction on the rheology is explained by relating it to interactions andspecifically to the contact pair-distribution function computed from the simulation.The excess stresses are seen to show an approximately linear behaviour with theReynolds number in the range of 0.1–5, while with the capillary number and volumefraction, the variation is weakly quadratic.

  • Aliabouzar M, Zhang LG, Sarkar K, 2016 “Lipid-coated microbubbles and low intensity pulsed ultrasound enhance chondrogenesis of human mesenchymal stem cells in 3D printed scaffolds,” Nature Scientific Report, 6, 37728.

    Lipid-coated microbubbles are used to enhance ultrasound imaging and drug delivery. Here we apply these microbubbles along with low intensity pulsed ultrasound (LIPUS) for the first time to enhance proliferation and chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in a 3D printed poly-(ethylene glycol)-diacrylate (PEG-DA) hydrogel scaffold. The hMSC proliferation increased up to 40% after 5 days of culture in the presence of 0.5% (v/v) microbubbles and LIPUS in contrast to 18% with LIPUS alone. We systematically varied the acoustic excitation parameters—excitation intensity, frequency and duty cycle—to find 30 mW/cm2, 1.5 MHz and 20% duty cycle to be optimal for hMSC proliferation. A 3-week chondrogenic differentiation results demonstrated that combining LIPUS with microbubbles enhanced glycosaminoglycan (GAG) production by 17% (5% with LIPUS alone), and type II collagen production by 78% (44% by LIPUS alone). Therefore, integrating LIPUS and microbubbles appears to be a promising strategy for enhanced hMSC growth and chondrogenic differentiation, which are critical components for cartilage regeneration. The results offer possibilities of novel applications of microbubbles, already clinically approved for contrast enhanced ultrasound imaging, in tissue engineering.

  • Singh R, Sarkar K 2015 “Hydrodynamic interactions between pairs of capsules and drops in a simple shear: effects of viscosity ratio and heterogeneous collision,” Physical Review E, 92, 063029.

    Hydrodynamic interactions between a pair of capsules in simple shear are numerically investigated using afront-tracking finite difference method. The membrane of the capsule is modeled using different hyperelasticconstitutive relations. We also compare the pair interactions between drops to those between capsules. Anincreased viscosity ratio leads to a reduced net cross-stream separation between capsules as well as drops aftercollision. At low viscosity ratios, for the same capillary number drop-pairs show higher cross-stream separationthan those for capsule-pairs, while substantially large viscosity ratios result in almost the same value for bothcases. We investigate pair-collisions between two heterogeneous capsules C1and C2with two different capillarynumbers. The maximum deformation of C1was seen to increase with increasing stiffness (decreasing capillarynumber) of C2, even though the stiffness of C1was kept fixed. The findings are similar for a drop-pair, however,with a smaller maximum deformation for the same combinations of capillary numbers. The final cross-streamdrift of the trajectory of C1decreases with the increasing stiffness of C2, but the relative trajectory betweenthe capsules remains unchanged. The maximum deformation and the cross-stream drift of the trajectory of C1are shown to approximately vary with power-law functions of the ratio of the capillary numbers of C1andC2. An analytical explanation of the dependence on the two capillary numbers is offered. Different membraneconstitutive laws result in similar deformation and drift in trajectory.

  • Singh R, Li X, Sarkar K 2014 “Lateral migration of an elastic capsule in a wall-bounded shear,” Journal of Fluid Mechanics, 739, 421-443.

    The migration of a capsule enclosed by an elastic membrane in a wall-bounded linearshear is investigated using a front-tracking method. A detailed comparison with themigration of a viscous drop is presented varying the capillary number (in the caseof a capsule, the elastic capillary number) and the viscosity ratio. In both cases,the deformation breaks the flow reversal symmetry and makes them migrate awayfrom the wall. They quickly go through a transient evolution to eventually reach aquasi-steady state where the dynamics becomes independent of the initial positionand only depends on the wall distance. Previous analytical theories predicted thatfor a viscous drop, in the quasi-steady state, the migration and slip velocities scaleapproximately with the square of the inverse of the drop–wall separation, whereasthe drop deformation scales as the inverse cube of the separation. These power lawrelations are shown to hold for a capsule as well. The deformation and inclinationangle of the capsule and the drop at the same wall separation show a crossoverin their variation with the capillary number: the capsule shows a steeper variationthan that of the drop for smaller capillary numbers and slower variation than thedrop for larger capillary numbers. Using the Green’s function of Stokes flow, asemi-analytic theory is presented to show that the far-field stresslet that causes themigration has two distinct contributions from the interfacial stresses and the viscosityratio, with competing effects between the two defining the dynamics. It predicts thescaling of the migration velocity with the capsule–wall separation, however, matchingwith the simulated result very well only away from the wall. A phenomenologicalcorrelation for the migration velocity as a function of elastic capillary number, walldistance and viscosity ratio is developed using the simulation results. The effects ofdifferent membrane hyperelastic constitutive equations – neo-Hookean, Evans–Skalak,and Skalak – are briefly investigated to show that the behaviour remains similar fordifferent equations.

  • Mukherjee S, Sarkar K 2014 “Lateral migration of a viscoelastic drop in a Newtonian fluid in a shear flow near a wall,” Physics of Fluids, 26, 103102.

    Wall induced lateral migration of a viscoelastic (FENE-MCR) drop in a Newtonianfluid is investigated. Just like a Newtonian drop, a viscoelastic drop reaches a quasi-steady state where the lateral velocity only depends on the instantaneous distancefrom the wall. The drop migration velocity and the deformation scale inversely withthe square and the cube of the distance from the wall, respectively. The migration ve-locity varies non-monotonically with increasing viscoelasticity (increasing Deborahnumber); initially increasing and then decreasing. An analytical explanation has beengiven of the effects by computing the migration velocity as arising from an imagestresslet field due to the drop. The semi-analytical expression matches well with thesimulated migration velocity away from the wall. It contains a viscoelastic stressletcomponent apart from those arising from interfacial tension and viscosity ratio. Themigration dynamics is a result of the competition between the viscous (interfacialtension and viscosity ratio) and the viscoelastic effects. The viscoelastic stressletcontribution towards the migration velocity steadily increases. But the interfacialstresslet—arising purely from the drop shape—first increases and then decreases withrising Deborah number causing the migration velocity to be non-monotonic. The ge-ometric effect of the interfacial stresslet is caused by a corresponding nonmonotonicvariation of the drop inclination. High viscosity ratio is briefly considered to showthat the drop viscoelasticity could stabilize a drop against breakup, and the increase inmigration velocity due to viscoelasticity is larger compared to the viscosity-matchedcase.

  • Sarkar K, Singh R 2013Spatial ordering due to hydrodynamic interactions between a pair of colliding drops in a confined shear,Physics of Fluids, 25, 051702.

    Pair-collision between viscous drops in a confined shear is simulated to show that the confinement alters the trajectories of the drops spatially ordering them at a finite separation in the center of the domain. In contrast to free shear where drops eventually adopt free streamlines with a finite cross-stream separation, here they move towards the centerline achieving zero cross-stream separation but a net stream-wise separation. The latter varies as inverse of capillary number and cube of the confinement (distance between the walls). The final stream-wise separation does not depend on the initial positions of the drops when the drops are in the same shear plane. The separation decreases approximately linearly with the initial separation in the vorticity direction. An analytical theory explaining the phenomenon is presented. Effects of the ratio of drop to matrix viscosity are briefly investigated

     

  • Karandish F, Haldar MK, Xia L, Gange KN, Feng L, You S, Choi Y, Sarkar K, Mallik S  2018Nucleus-targeted, echogenic polymersomes for delivering a cancer stemness inhibitor to pancreatic cancer cells,Biomacromolecules, 19,4122-4132.

    Chemotherapeutic agents for treating cancers show considerable sideeffects, toxicity, and drug resistance. To mitigate the problems, we designed nucleus-targeted, echogenic, stimuli-responsive polymeric vesicles (polymersomes) to transport andsubsequently release the encapsulated anticancer drugs within the nuclei of pancreaticcancer cells. We synthesized an alkyne-dexamethasone derivative and conjugated it to N3−polyethylene glycol (PEG)−polylactic acid (PLA) copolymer employing the Cu2+catalyzed“Click”reaction. We prepared polymersomes from the dexamethasone−PEG−PLA conjugate along with a synthesized stimuli-responsive polymer PEG−S−S−PLA. Thedexamethasone group dilates the nuclear pore complexes and transports the vesicles to thenuclei. We designed the polymersomes to release the encapsulated drugs in the presence ofa high concentration of reducing agents in the nuclei of pancreatic cancer cells. Weobserved that the nucleus-targeted, stimuli-responsive polymersomes released 70% ofencapsulated contents in the nucleus-mimicking environment in 80 min. We encapsulatedthe cancer stemness inhibitor BBI608 in the vesicles and observed that the BBI608encapsulated polymersomes reduced the viability of the BxPC3 cells to 43% in three-dimensional spheroid cultures. Thepolymersomes were prepared following a special protocol so that they scatter ultrasound, allowing imaging by a medicalultrasound scanner. Therefore, these echogenic, targeted, stimuli-responsive, and drug-encapsulated polymersomes have thepotential for trackable, targeted carrier of chemotherapeutic drugs to cancer cell nuclei.

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