The Preparation Of Decellularized Meniscal Extracellular Matrix And The Effects On Meniscal Fibrochondrocytes

Objective:Functional restoration of injuried meniscus remains a substantial challenge, the aim of this study was to produce a novel decellularized meniscal extracellular matrix (MECM), observe the cell dedifferentiation during rabbit meniscus fibrochondrocytes passaging monolayer culture, and further investigate the effect of coated MECM surface on passaged meniscal fibrochondrocytes in vitro.Method:The porcine meniscus was performed decellularization process to prepare MECM. Rabbit passaged fibrochondrocytes were investigated in terms of cell appearance and gene expression changes during monolayer culture. CCK-8 was used to evaluate the effect of MECM on cell proliferation capacity. The various biomimetic surfaces were coated, including chondroitin sulfate (CS) surface, MECM surface, and MECM/CS surface (ratio of 5:1). Passaged meniscal fibrochondrocytes were cultured on these coated surfaces in vitro for 7,14 days and assayed the cell adherence, viability, cellularity, matrix production and gene expression in comparison with the control group (TCP).Results:MECM can well preserve meniscal extracellular matrix (ECM), including collagens and sulfated glycosaminoglycans (GAG), and remove DNA. Passaged cells rapidly occur to dedifferentiate on monolayer culture, followed by changing from round cells to elongated cells, collagen I mRNA up-regulation and collagen II mRNA down-regulation. MECM can enhance the proliferation capacity of passaged meniscal cells. MECM surface positively and significantly affected meniscal fibrochondrocytes viability, adherence, cellularity, increased the expression levels of collagen type II (12-fold than TCP), proteoglycans and collagen production. The passaged meniscal fibrochondrocytes on MECM coated surface could redifferentiate as convinced by safranin O and toluidine blue staining.Conclusion:MECM can preserve well native meniscal ECM, and remove DNA. Passaged cells rapidly occur to dedifferentiate on monolayer culture. MECM can enhance both the proliferation and redifferentiation capacity of passaged meniscal cells, making it a potential candidate biomaterial for future meniscal tissue engineering applications.