| Objective: To fabricate decellularized annulus fibrosus matrix (DAFM), and thenexplore the role of DAFM on the proliferation and differentiation potential of bonemarrow-derived mesenchymal stem cells (BMSCs).Methods: The annulus fibrosus of pig was obtained and ground into powder, whichwas consequently treated with trypsin, nuclease, and Triton X-100to remove cells from thematrix. The matrix was then dissolved in0.5M acetic acid to obtain a solution, which wasfurther used to make DAFM films. DAPI staining was used to confirm the completeremoval of cells from the DAFM. Scanning electron microscopy was used to examine themicrostructures of DAFM film. Two-month old New Zealand white rabbits were used toobtain BMSCs through whole bone marrow culture. BMSCs were cultured on DAFMfilms and plastic surfaces, respectively. Cell proliferation was tested by MTT assay.Multi-differentiation potential of BMSCs on DAFM films and plastic surfaces waschecked, including adipogenesis, osteogenesis, and chondrogenesis.Results: DAFM which was cell-free and well retained collagen microfibrils has beensuccessfully prepared. BMSCs that expressed expression of stem cell markers Oct-4andnucleostemin and were capable of induced adipogenesis, osteogenesis, and chondrogenesiswere isolated. BMSCs proliferated well on both DAFM films and plastic surfaces. BMSCsgrown on DAFM films and plastic surfaces with various induction media were stainedusing Oil Red O for adipogenesis, Alizarin Red for osteogenesis, and Safranin O forchondrogenesis. The number of lipid drops, calcium nodules, glycosaminoglycansproduced by BMSCs on DAFM films was more than that on plastic surfaces. Conclusion: Decellularized annulus fibrosus matrix were fabricated, which preservedcollagen fibrils. During culture, DAFM films well maintained the morphology of BMSCs.In addition, BMSCs grown on DAFM films exhibited enhanced proliferation andmulti-differentiation potential. |
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