Decellularized matrix (DCM) derived from native tissues may be a promising supporting material to induce cellular differentiation by sequestered bioactive factors. However, no previous study has investigated the use of human meniscus-derived DCM to re-differentiate human meniscus fibrochondrocytes (MFCs) to form meniscus-like extracellular matrix (ECM). We expanded human MFCs and seeded them upon a cadaveric meniscus-derived DCM prepared by physical homogenization under hypoxia. To assess the bioactivity of the DCM, we used conditions with and without chondrogenic factor TGF-β3 and set up a cell pellet culture model as a biomaterial-free control. We found that the DCM supported chondrogenic re-differentiation and ECM formation of MFCs only in the presence of exogenous TGF-β3. Chondrogenic re-differentiation was more robust at the protein level in the pellet model as MFCs on the DCM appeared to favour a more proliferative phenotype. Interestingly, without growth factors, the DCM tended to promote expression of hypertrophic differentiation markers relative to the pellet model. Therefore, the human meniscus-derived DCM prepared by physical homogenization contained insufficient bioactive factors to induce appreciable ECM formation by human MFCs.

中文翻译：

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人半月板纤维软骨细胞的再分化在三维细胞聚集体和脱细胞的人半月板基质支架上有所不同。

源自天然组织的脱细胞基质（DCM）可能是一种有前途的支持材料，可以通过螯合的生物活性因子诱导细胞分化。但是，以前没有研究调查使用人类半月板来源的DCM重新分化人类半月板纤维软骨细胞（MFCs）以形成半月板样细胞外基质（ECM）。我们扩增了人类MFC，并将其接种在尸体半月板衍生的DCM上，该DCM是在缺氧条件下通过物理均质化制备的。为了评估DCM的生物活性，我们使用了有或没有软骨生成因子TGF-β3的条件，并建立了细胞沉淀培养模型作为无生物材料的对照。我们发现，仅在存在外源性TGF-β3的情况下，DCM支持MFC的成软骨再分化和ECM形成。在颗粒模型中，软骨生成的再分化在蛋白质水平上更为稳健，因为DCM上的MFCs似乎更倾向于增殖表型。有趣的是，没有生长因子，相对于颗粒模型，DCM倾向于促进肥大分化标记物的表达。因此，通过物理均质化制备的人半月板来源的DCM包含的生物活性因子不足，无法诱导人MFC形成明显的ECM。