Three-dimensional cartilage tissue engineering using adult stem cells from osteoarthritis patients
Three-dimensional cartilage tissue engineering using adult stem cells from osteoarthritis patients
复制标题
DOI：
10.1002/art.22285
复制
发表时间：
2007-01-01
影响因子：
--
通讯作者：
Hollander, Anthony P.
中科院分区：
文献类型：
--
作者：
Kafienah, Wael;Mistry, Sanjay;Hollander, Anthony P.
Objective.目的：探讨骨关节炎（OA）患者骨髓间充质干细胞（BMSCs）能否构建三维透明软骨。将来自髋关节OA患者的扩增的BMSC接种到聚乙醇酸支架上，并在存在或不存在甲状旁腺相关蛋白（PTHrP）的情况下使用转化生长因子β 3进行分化以调节肥大。使用相同的细胞建立微团沉淀培养物用于比较。在培养结束时，通过定量实时逆转录-聚合酶链反应处理构建体或沉淀物用于信使RNA（mRNA）分析。基质蛋白用特异性分析法进行分析。从BMSC工程化的软骨素构建体的重量至少是等效沉淀培养物的5倍。组织学，mRNA，和生化分析的结构显示广泛的蛋白多糖和II型胶原蛋白的合成，但只有低水平的I型胶原蛋白。蛋白质含量与牛鼻软骨细胞工程软骨几乎相同。X型胶原mRNA的分析显示，与未分化的BMSCs相比，软骨形成结构中的mRNA水平较高，表明组织工程细胞肥大的风险增加。然而，在培养期间以1 μ M或10 μ M的剂量包含PTHrP导致X型胶原mRNA表达的显著抑制和碱性磷酸酶活性的显著降低，而软骨特异性基质蛋白没有任何损失。利用骨关节炎患者的骨髓间充质干细胞可以构建三维透明软骨。因此，该方法可用于修复软骨损伤。
Objective. To determine whether it is possible to engineer 3-dimensional hyaline cartilage using mesenchymal stem cells derived from the bone marrow (BMSCs) of patients with osteoarthritis (OA).Methods. Expanded BMSCs derived from patients with hip OA were seeded onto polyglycolic acid scaffolds and differentiated using transforming growth factor beta 3 in the presence or absence of parathyroid hormone-related protein (PTHrP) to regulate hypertrophy. Micromass pellet cultures were established using the same cells for comparison. At the end of culture, the constructs or pellets were processed for messenger RNA (mRNA) analysis by quantitative real-time reverse transcription-polymerase chain reaction. Matrix proteins were analyzed using specific assays.Results. Cartilage constructs engineered from BMSCs were at least 5 times the weight of equivalent pellet cultures. Histologic, mRNA, and biochemical analyses of the constructs showed extensive synthesis of proteoglycan and type II collagen but only low levels of type I collagen. The protein content was almost identical to that of cartilage engineered from bovine nasal chondrocytes. Analysis of type X collagen mRNA revealed a high level of mRNA in chondrogenic constructs compared with that in undifferentiated BMSCs, indicating an increased risk of hypertrophy in the tissue-engineered cells. However, the inclusion of PTHrP at a dose of 1 mu M or 10 mu M during the culture period resulted in significant suppression of type X collagen mRNA expression and a significant decrease in alkaline phosphatase activity, without any loss of the cartilage-specific matrix proteins.Conclusion. Three-dimensional hyaline cartilage can be engineered using BMSCs from patients with OA. This method could thus be used for the repair of cartilage lesions.