A novel porcine fibrinogen formulation for fabricating biodegradable hybrid scaffolds by electrospinning with poly(L-lactic acid-co-ε-caprolactone)

摘要

Introduction: Fibrinogen has been used as a surgical sealant in the clinical setting for decades. The primary ingredient in commercial fibrin sealants is fibrinogen isolated from pooled human plasma and subjected to heat inactivation and/or solvent/detergent extraction to reduce the risk of viral contamination. However, sterilization can also reduce the ability of fibrinogen to assemble aggregate structures. The high risk of priori/virus transmission in human plasma and the thermo/radio labile properties of fibrinogen during sterilization and storage have limited the application of fibrin sealant. Methods: We previously developed a novel arginine-formulated fibrinogen from cryoprecipitates of porcine plasma. To further characterize this material, we also used this formulation of fibrinogen to construct a biodegradable hybrid scaffold by electrospinning with poly(L-lactic acid-co-ε-caprolactone) (P(LLA-CL)). Results: Circular dichroism and SDS-PAGE were used to confirm that this formulated fibrinogen was stable even after radiation sterilization, which can denature unformulated fibrinogen. The hybrid scaffolds possessed the benefits associated with the individual components, including a moderate mechanical strength and excellent biocompatibility. Endothelial cells seeded onto the hybrid scaffolds achieved a relatively high level of cell attachment after 3 days of in vitro culture, and their proliferation rate was significantly increased after 7 days of culture compared with cells on pure fibrinogen or P(LLA-CL) scaffolds. The in vivo effects of hybrid and pure P(LLA-CL) patches were determined by implantation in a rat model of abdominal wall defects. The implants were degraded by 6 months in both groups, but muscle reconstruction was only observed in the hybrid patch group. Discussion: This formulated fibrinogen overcame the instability problems of labile fibrinogen that occur during storage and transportation. It can also be reconstituted within 10 min, greatly facilitating presurgical preparation. Furthermore, the performance of the hybrid scaffold was significantly improved by the addition of fibrinogen, including better biocompatibility, surface hydrophilicity, and tissue regeneration. Conclusion: These data indicate that this fibrinogen product may be more economical, readily available, and convenient to use, while having a decreased risk of human blood-borne pathogen transmission. The material exhibited excellent stability during sterilization and better hemostalic efficacy than a leading commercial hemostatic product in both animal and clinical studies. The P(LLA-CL)/fibrinogen hybrid scaffold was evaluated after transplantation in vivo as an abdominal wall replacement, with a pure P(LLA-CL) scaffold as the control group. The implants from both groups were completely degraded by 6 months. Reconstructed muscle tissue was observed in the hybrid scaffold group, but only large bundles of collagen fibers were found in the pure P(LLA-CL) scaffold group.