Sachin S. Velankar

Preparation and rheological characterization of a gel form of the porcine urinary bladder matrix.

Biologic scaffolds composed of extracellular matrix (ECM) have been used to facilitate the repair and reconstruction of a variety of tissues in clinical and pre-clinical studies. The clinical utility of such scaffolds can be limited by the geometric and mechanical properties of the tissue or organ from which the ECM is harvested. An injectable gel form of ECM could potentially conform to any three-dimensional shape and could be delivered to sites of interest by minimally invasive techniques. The objectives of the present study were to prepare a gel form of ECM harvested from the urinary bladder (urinary bladder matrix or UBM), to characterize the rheological properties of the gel, and finally to evaluate the ability of the gel to support in vitro growth of smooth muscle cells. Following enzymatic solubilization with pepsin, UBM was induced to self-assemble into a gel when brought to physiological conditions. The UBM gel supported the adhesion and growth of rat aortic smooth muscle cells when cultured under static in vitro conditions. The present study showed that an intact form of UBM can be successfully solubilized without purification steps and induced to repolymerize into a gel form of the UBM biologic scaffold material.

A hydrogel derived from decellularized dermal extracellular matrix.

The ECM of mammalian tissues has been used as a scaffold to facilitate the repair and reconstruction of numerous tissues. Such scaffolds are prepared in many forms including sheets, powders, and hydrogels. ECM hydrogels provide advantages such as injectability, the ability to fill an irregularly shaped space, and the inherent bioactivity of native matrix. However, material properties of ECM hydrogels and the effect of these properties upon cell behavior are neither well understood nor controlled. The objective of this study was to prepare and determine the structure, mechanics, and the cell response in vitro and in vivo of ECM hydrogels prepared from decellularized porcine dermis and urinary bladder tissues. Dermal ECM hydrogels were characterized by a more dense fiber architecture and greater mechanical integrity than urinary bladder ECM hydrogels, and showed a dose dependent increase in mechanical properties with ECM concentration. In vitro, dermal ECM hydrogels supported greater C2C12 myoblast fusion, and less fibroblast infiltration and less fibroblast mediated hydrogel contraction than urinary bladder ECM hydrogels. Both hydrogels were rapidly infiltrated by host cells, primarily macrophages, when implanted in a rat abdominal wall defect. Both ECM hydrogels degraded by 35 days in vivo, but UBM hydrogels degraded more quickly, and with greater amounts of myogenesis than dermal ECM. These results show that ECM hydrogel properties can be varied and partially controlled by the scaffold tissue source, and that these properties can markedly affect cell behavior.

Rheology and morphology of compatibilized polymer blends

Research into the compatibilization of immiscible polymer blends has, during the past few years, begun to focus on the role of block co-polymers (bcp) on various morphological processes. Considerable advances have been made, from both a theoretical and an experimental point of view, in relating the presence of compatibilizers to structure development during flow.

Hydrogels derived from central nervous system extracellular matrix

Biologic scaffolds composed of extracellular matrix (ECM) are commonly used repair devices in preclinical and clinical settings; however the use of these scaffolds for peripheral and central nervous system (CNS) repair has been limited. Biologic scaffolds developed from brain and spinal cord tissue have recently been described, yet the conformation of the harvested ECM limits therapeutic utility. An injectable CNS-ECM derived hydrogel capable of in vivo polymerization and conformation to irregular lesion geometries may aid in tissue reconstruction efforts following complex neurologic trauma. The objectives of the present study were to develop hydrogel forms of brain and spinal cord ECM and compare the resulting biochemical composition, mechanical properties, and neurotrophic potential of a brain derived cell line to a non-CNS-ECM hydrogel, urinary bladder matrix. Results showed distinct differences between compositions of brain ECM, spinal cord ECM, and urinary bladder matrix. The rheologic modulus of spinal cord ECM hydrogel was greater than that of brain ECM and urinary bladder matrix. All ECMs increased the number of cells expressing neurites, but only brain ECM increased neurite length, suggesting a possible tissue-specific effect. All hydrogels promoted three-dimensional uni- or bi-polar neurite outgrowth following 7 days in culture. These results suggest that CNS-ECM hydrogels may provide supportive scaffolding to promote in vivo axonal repair.

Interfacial elasticity and coalescence suppression in compatibilized polymer blends

Shear-induced coalescence was studied in immiscible blends of polydimethylsiloxane (PDMS) and polyisoprene (PI) with a droplet-matrix morphology, using both rheology and scanning electron microscopy. Dynamic moduli of the blends compatibilized with different amounts of a PDMS–PI diblock were measured. The experimental results indicate that the blend response is characterized by two relaxation mechanisms. The general Palierne model with an interfacial shear modulus was used to analyze the data, since this model can describe the dynamic response of polymer blends in which interfacial tension gradients induce an extra relaxation mechanism besides droplet relaxation. Scanning electron microscopy was used to investigate the droplet size evolution in the blends during coalescence. For systems with a high amount of compatibilizer, it is shown that coalescence is completely suppressed under the conditions studied here.

Fabrication of Large‐Area Two‐Dimensional Colloidal Crystals

Nanoparticle coating: A suspension of colloidal particles in a water/propanol solution was layered onto a water surface, where the particles self-assembled into ordered two-dimensional hexagonal crystal arrays (>280 cm 2) within two minutes. These arrays were transferred from the water surface to other substrates (see picture) and embedded in a chitosan hydrogel for visual detection of the pH value. Copyright

Pressure drops for droplet flows in microfluidic channels

Designing on-chip microfluidic pumps requires that the dependence of pressure drop across the device on the fluid flow rate be known. This relationship can be complicated for the case of two-phase droplet type flows. We characterized the flow-pressure relationship for the single-file flow of water drops in oil in microfluidic channels of rectangular cross sections. The pressure for such droplet flows was always larger—sometimes over 50% larger—than that for corresponding single-phase flows of the continuous phase. This is in spite of the fact that the water drops had a substantially lower viscosity than the continuous phase oil. The excess pressure was found to correlate reasonably well with the size of the drops relative to the size of the channels. This correlation for the excess pressure, as well as a correlation presented here for the size of drops in microchannels, should provide convenient guidelines in designing microfluidic devices for two-phase flows.

Effect of compatibilization on the breakup of polymeric drops in shear flow

A block copolymer may be added as a compatibilizer during polymer processing in order to promote intimate mixing of thermodynamically immiscible homopolymers. The action of this compatibilizer can only partially be attributed to its effect on the interfacial tension between the immiscible homopolymers. Here the additional contributions of the compatibilizer are directly probed by measuring the capillary number during coalescence experiments. Model blends consisting of polyisobutylene (PIB) and polydimethylsiloxane (PDMS), compatibilized with various amounts of a PIB–PDMS diblock copolymer, are used for this purpose. The mean capillary number of the droplets is determined from the mechanical frequency response of the blends. With increasing amounts of compatibilizer, a systematic increase in steady shear capillary number is seen, to values well above the critical capillary number for droplet breakup of uncompatibilized systems. This indicates that a simple decrease in interfacial tension is not the only effect of adding the compatibilizer to these immiscible blends. Past simulations suggest that these results are associated with gradients in interfacial tension (Marangoni stresses) induced by the gradients of compatibilizer concentration due to flow. Direct evidence of the presence of such interfacial tension gradients along the surface of compatibilized drops was obtained by optical microscopy.A block copolymer may be added as a compatibilizer during polymer processing in order to promote intimate mixing of thermodynamically immiscible homopolymers. The action of this compatibilizer can only partially be attributed to its effect on the interfacial tension between the immiscible homopolymers. Here the additional contributions of the compatibilizer are directly probed by measuring the capillary number during coalescence experiments. Model blends consisting of polyisobutylene (PIB) and polydimethylsiloxane (PDMS), compatibilized with various amounts of a PIB–PDMS diblock copolymer, are used for this purpose. The mean capillary number of the droplets is determined from the mechanical frequency response of the blends. With increasing amounts of compatibilizer, a systematic increase in steady shear capillary number is seen, to values well above the critical capillary number for droplet breakup of uncompatibilized systems. This indicates that a simple decrease in interfacial tension is not the only ef...

A Photonic Crystal Protein Hydrogel Sensor for Candida albicans

We report two-dimensional (2D) photonic crystal (PC) sensing materials that selectively detect Candida albicans (C. albicans). These sensors utilize Concanavalin A (Con A) protein hydrogels with a 2D PC embedded on the Con A protein hydrogel surface, that multivalently and selectively bind to mannan on the C. albicans cell surface to form crosslinks. The resulting crosslinks shrink the Con A protein hydrogel, reduce the 2D PC particle spacing, and blue-shift the light diffracted from the PC. The diffraction shifts can be visually monitored, measured with a spectrometer, or determined from the Debye diffraction ring diameter. Our unoptimized hydrogel sensor has a detection limit of around 32 CFU/mL for C. albicans. This sensor distinguishes between C. albicans and those microbes devoid of cell-surface mannan such as the gram-negative bacterium E. coli. This sensor provides a proof-of-concept for utilizing recognition between lectins and microbial cell surface carbohydrates to detect microorganisms in aqueous environments.

Extracellular Matrix Hydrogels from Decellularized Tissues: Structure and Function.

Extracellular matrix (ECM) bioscaffolds prepared from decellularized tissues have been used to facilitate constructive and functional tissue remodeling in a variety of clinical applications. The discovery that these ECM materials could be solubilized and subsequently manipulated to form hydrogels expanded their potential in vitro and in vivo utility; i.e. as culture substrates comparable to collagen or Matrigel, and as injectable materials that fill irregularly-shaped defects. The mechanisms by which ECM hydrogels direct cell behavior and influence remodeling outcomes are only partially understood, but likely include structural and biological signals retained from the native source tissue. The present review describes the utility, formation, and physical and biological characterization of ECM hydrogels. Two examples of clinical application are presented to demonstrate in vivo utility of ECM hydrogels in different organ systems. Finally, new research directions and clinical translation of ECM hydrogels are discussed. STATEMENT OF SIGNIFICANCE More than 70 papers have been published on extracellular matrix (ECM) hydrogels created from source tissue in almost every organ system. The present manuscript represents a review of ECM hydrogels and attempts to identify structure-function relationships that influence the tissue remodeling outcomes and gaps in the understanding thereof. There is a Phase 1 clinical trial now in progress for an ECM hydrogel.