Development of a Silk Cable-Reinforced Gelatin/Silk Fibroin Hybrid Scaffold for Ligament Tissue Engineering

Abstract

The objective of this study was to develop a silk cable-reinforced gelatin/silk fibroin hybrid scaffold for ligament tissue engineering. The scaffold was fabricated by lyophilizing the cross-linked gelatin and silk fibroin mixture with braided silk cables. Scanning electronic microscopy (SEM) observation showed that microporous gelatin/silk fibroin sponges formed around silk cables mimicked the microstructures of ligament extracellular matrix (ECM). The silk cables significantly increased the tensile strength of the scaffold to meet the mechanical requirements for ligament tissue engineering. The scaffold possessed good cell adhesion property, and when mesenchymal stem cells (MSCs) were seeded on it, cells proliferated profusely. After 2 weeks of culture, seeded MSCs were distributed uniformly throughout the scaffold and were highly viable. Occurrence of cell death during culture was not significant. Deposition of collagen on the scaffold was found to increase with time. Differentiation of MSCs into ligament fibroblasts was verified by expressions of ligament ECM specific genes including collagen type I, collagen type III, and tenascin-C in mRNA and protein level. Immunohistochemistry stains also confirmed the production of key ligament ECM components on the scaffold. The results demonstrate that silk cable-reinforced gelatin/silk fibroin scaffold possesses the appropriate mechanical properties and has enlarged surface area. It is also capable of supporting cell proliferation and differentiation for ligament tissue engineering.