Development and characterization of a collagen-based matrix for vascularization and cell delivery

Abstract

Since the development of the Edmonton protocol, islet transplantation is increasingly encouraging as a treatment for type 1 diabetes. Strategies to ameliorate problems with the intraportal site include macroencapsulating the islets in diverse biomaterials. Characterization of these biomaterials is important to optimally tune the properties to support islets and promote vascularization. In this study, we characterize the cross-linker-dependent properties of collagen-based matrices containing chondroitin-6-sulfate, chitosan, and laminin, cross-linked with 7.5, 30, or 120 mM of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide. The swelling ratio was found to be significantly negatively correlated with increasing cross-linker concentrations (p<0.0001; R2=0.718). The matrix released insulin in a reproducible logarithmic manner (R2 of 0.99 for all concentrations), demonstrating cross-linker-dependent control of drug release. The matrices with the highest cross-linker concentrations resisted degradation by collagenase for longer than the lowest concentrations (58.13%±2.22% vs. 13.69%±7.67%; p<0.05). Scanning electron microscopy images of the matrices revealed that the matrices had uniform topography and porosity, indicating efficient cross-linking and incorporation of the polymer components. Matrices were transplanted subcutaneously in naive BALB/c mice, and the number and size of vessels were quantified using von Willebrand factor staining; matrices with higher cross-linking concentrations had significantly larger capillaries at every time point up to 4 weeks after transplantation compared to the lowest cross-linker concentration group. CD31 staining visualized the capillaries at each time point. Taken together, these data show that this collagen-based matrix is reproducible with cross-linking-dependent properties that can be optimized to support vascularization and islet function.