Biomolecule incorporated PCL nanofibrous scaffolds supporting meniscal cell attachment and proliferation.

Tissue engineered constructs with rapid restoration of mechanical and biological properties remain a challenge, emphasizing the need to develop novel scaffolds. Here, we present a multicomponent composite three-dimensional scaffold structure with biomimetic reinforcement and biomolecule functionalization for meniscus tissue engineering. The scaffold structure was developed using 3:1 silk fibroin (SF) and polyvinyl alcohol (PVA). Autoclaved eggshell membrane (AESM) powder (1-3%w/v) was used as reinforcement to enhance biomechanical properties. Further to improve cell attachment and proliferation, these scaffolds were functionalized using an optimized unique combination of biomolecules. Comprehensive analysis of scaffolds was carried out on morphological, structural, mechanical and biological functionalities. Their mechanical properties were compared with different native human menisci. The results indicated that, functionalized SF-PVA with 3%AESM has shown similar order of magnitude of compressive and dynamic mechanical properties as in human meniscus. Moreover, 3% AESM based scaffolds were found to support better primary human meniscal cellular proliferation and extracellular matrix secretion. Immunohistochemical analysis revealed angiogenesis and biocompatibility with minimal inflammatory response for subcutaneously implanted scaffolds in New Zealand white rabbits. The developed reinforced and functionalized SF-PVA scaffolds can uniquely combine the potential for load-bearing properties with improved in vitro and in vivo support for meniscus tissue regeneration. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1722-1731, 2018.

In this study, silk fibroin nanofibrous scaffolds were developed to investigate the attachment and proliferation of primary human meniscal cells. Silk fibroin (SF)-polyvinyl alcohol (PVA) blended electrospun nanofibrous scaffolds with different blend ratios (2:1, 3:1, and 4:1) were prepared. Morphology of the scaffolds was characterized using atomic force microscopy (AFM). The hybrid nanofibrous mats were crosslinked using 25 % (v/v) glutaraldehyde vapor. In degradation study, the crosslinked nanofiber showed slow degradation of 20 % on weight after 35 days of incubation in simulated body fluid (SBF). The scaffolds were characterized with suitable techniques for its functional groups, porosity, and swelling ratio. Among the nanofibers, 3:1 SF:PVA blend showed uniform morphology and fiber diameter. The blended scaffolds had fluid uptake and swelling ratio of 80 % and 458 ± 21 %, respectively. Primary meniscal cells isolated from surgical debris after meniscectomy were subcultured and seeded onto these hybrid nanofibrous scaffolds. Meniscal cell attachment studies confirmed that 3:1 SF:PVA nanofibrous scaffolds supported better cell attachment and growth. The DNA and collagen content increased significantly with 3:1 SF:PVA. These results clearly indicate that a blend of SF:PVA at 3:1 ratio is suitable for meniscus cell proliferation when compared to pure SF-PVA nanofibers.

Enhanced human primary meniscal cell proliferation in autoclaved egg shell membrane.

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