Customized Heparinized Alginate and Collagen Hydrogels for Tunable, Local Delivery of Angiogenic Proteins

Therapeutic protein delivery has ushered in a promising new generation of disease treatment, garnering more recognition for its clinical potential than ever. However, proteins' limited stability, extremely short average half-lives, and evidenced toxicity following systemic delivery continue to undercut their efficacy. Biomaterial-based protein delivery, however, demonstrates the potential to overcome these obstacles. To this end, we have developed a heparinized alginate and collagen hydrogel for the local, sustained delivery of therapeutic proteins. In an effort to match this ubiquitous application of protein delivery to various disease states and target tissues with sufficient versatility, we identified three distinct delivery modes as design targets. A shear-thinning, low-viscosity injectable for minimal tissue damage, a higher-viscosity gel plug for subcutaneous injection, and a submillimeter-thickness film for solid-form implantation were optimized and characterized in this work. In vitro assessments confirmed feasible injection control, mechanical stability for up to 6 h of unsubmerged storage, and isotropic early collagen fibril assembly. Release kinetics were assessed both in vitro and in vivo, demonstrating up to 14 days of functional vascular endothelial growth factor delivery. Rodent models of pulmonary hypertension, subcutaneous injection, and myocardial infarction, three promising applications of protein therapeutics, were used to assess the feasible delivery and biocompatibility of the injectable gel, gel plug, and film, respectively. Histological evaluation of the delivered materials and surrounding tissue showed high biocompatibility with cell and blood vessel infiltration, remodeling, and integration with the host tissue. Our successful customization of the biomaterial to heterogeneous delivery modes demonstrates its versatile capacity for the local, sustained delivery of therapeutic proteins for a diverse array of regenerative medicine applications.