In vitro engineered cartilage constructs produced by press-coating biodegradable polymer with human mesenchymal stem cells
In vitro engineered cartilage constructs produced by press-coating biodegradable polymer with human mesenchymal stem cells
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DOI：
10.1089/107632702753503126
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发表时间：
2002-02-01
影响因子：
--
通讯作者：
Tuan, RS
中科院分区：
文献类型：
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作者：
Nöth, U;Tuli, R;Tuan, RS
通过将 1 x 0.5 x 0.5 cm 的 D,D-L,L-聚乳酸聚合物块压涂到从接受全髋关节置换术的患者股骨头中分离的 1.5 x 10(6) 人类间充质干细胞 (mhMSC) 的高密度细胞颗粒上来制造软骨结构。将细胞沉淀附着到聚合物表面后，通过将构建体在补充有转化生长因子β-1（TGF-β1）的无血清、化学成分确定的软骨形成分化培养基中培养3周来诱导软骨形成。组织化学分析表明，压制包被的颗粒形成了由形态独特的软骨细胞样细胞组成的细胞层，周围环绕着纤维状、富含硫酸化蛋白聚糖的细胞外基质。免疫组织化学分析检测到细胞外基质内有 II 型胶原蛋白和软骨蛋白聚糖连接蛋白。通过 RT-PCR 检测软骨特异性标记基因 II、IX、X 和 XI 型胶原以及聚集蛋白聚糖的表达。扫描电子显微镜显示细胞聚合物结构内有组织且空间上不同的细胞区域，表面层类似于致密的透明软骨。用 mhMSC 压制涂覆可生物降解聚合物的制造方法允许在体外生产软骨结构，而无需从完整的关节软骨表面收获软骨细胞。这些结构可用作重建人类关节软骨缺陷的原型。
Cartilage constructs were fabricated by press-coating D,D-L,L-polylactic acid polymer blocks of 1 x 0.5 x 0.5 cm onto high-density cell pellets of 1.5 x 10(6) human mesenchymal stem cells (mhMSCs) isolated from the femoral head of patients undergoing total hip arthroplasty. Following attachment of the cell pellets to the polymer surfaces, chondrogenesis was induced by culturing the constructs for 3 weeks in a serum-free, chemically defined, chondrogenic differentiation medium supplemented with transforming growth factor beta-1 (TGF-beta1). Histochemical analysis showed that the press-coated pellets formed cell layers composed of morphologically distinct, chondrocyte-like cells, surrounded by a fibrous, sulfated proteoglycan-rich extracellular matrix. Immunohistochemical analysis detected collagen type II and cartilage proteoglycan link protein within the extracellular matrix. Expression of the cartilage-specific marker genes collagen types II, IX, X, and XI, and aggrecan was detected by RT-PCR. Scanning electron microscopy revealed organized and spatially distinct zones of cells within the cell-polymer constructs, with the superficial layer resembling compact hyaline cartilage. The fabrication method of press-coating biodegradable polymers with mhMSCs allows the in vitro production of cartilage constructs without harvesting chondrocytes from intact articular cartilage surfaces. These constructs may be applicable as prototypes for the reconstruction of articular cartilage defects in humans.