Berner A, Siebenlist S, Reichert J C, Hendrich C, Nöth U
Abteilung für Unfallchirurgie, Klinikum der Universität Regensburg, Regensburg.
Z Orthop Unfall. 2010 Jan;148(1):31-8. doi: 10.1055/s-0029-1240753. Epub 2010 Feb 11.
Mesenchymal stem cells have a high therapeutic potential for the reconstruction of articular cartilage defects. In this study, a cartilage-polymer construct using mesenchymal stem cells from trabecular bone and a polylactic acid polymer was fabricated with a press-coating technique. We investigated whether cells from human trabecular bone fragments have the same chondrogenic differentiation potential as mesenchymal stem cells derived from bone marrow and whether it is possible to reconstruct an osteochondral lesion in the nude rat with the fabricated construct.
Cells were obtained from the femoral head of patients undergoing total hip arthroplasty. The fabrication of the constructs was performed by centrifugation of 1.5x10(6) cells to a cell pellet which was then placed in a polymer block. The fabricated cell constructs were cultivated for 3 weeks in a serum-free medium, supplemented with transforming growth factor beta1. Every third day, the chondrogenic differentiation was analysed using chondrogenic and osteogenic marker genes. After three weeks the constructs were implanted into 5 mm osteochondral defects of the knee joint of nude rats. After 4 and 12 weeks histochemical and immunohistochemical analyses were performed.
At the end of the culture period the constructs showed a proteoglycan-rich extracellular matrix with the expression of collagen types II, IX and X as well as aggrecan und COMP (cartilage oligomeric matrix protein). No osteogenic markers except collagen type I could be detected. The analysis of the in vivo experiment showed a good defect filling with a reconstructed cartilage surface along with increasing resorption of the polymer.
We have shown that it is possible to fabricate cartilage-polymer constructs from trabecular bone-derived cells, and that the cells have the same chondrogenic differentiation potential as mesenchymal stem cells derived from bone marrow. With the fabricated cartilage-polymer construct it is possible to reconstruct an osteochondral defect in the knee joint of the nude rat.
间充质干细胞在关节软骨缺损修复方面具有很高的治疗潜力。在本研究中,采用压涂技术制备了一种使用来自松质骨的间充质干细胞和聚乳酸聚合物的软骨-聚合物构建体。我们研究了来自人松质骨碎片的细胞是否具有与源自骨髓的间充质干细胞相同的软骨形成分化潜能,以及用制备的构建体在裸鼠中重建骨软骨损伤是否可行。
从接受全髋关节置换术患者的股骨头获取细胞。通过将1.5×10⁶个细胞离心形成细胞沉淀,然后将其置于聚合物块中来制备构建体。将制备的细胞构建体在补充有转化生长因子β1的无血清培养基中培养3周。每隔一天,使用软骨形成和成骨标记基因分析软骨形成分化情况。3周后,将构建体植入裸鼠膝关节5mm的骨软骨缺损处。4周和12周后进行组织化学和免疫组织化学分析。
在培养期结束时,构建体显示出富含蛋白聚糖的细胞外基质,伴有II型、IX型和X型胶原蛋白以及聚集蛋白聚糖和软骨寡聚基质蛋白(COMP)的表达。除I型胶原蛋白外,未检测到成骨标记物。体内实验分析显示缺损填充良好,有重建的软骨表面,同时聚合物的吸收增加。
我们已经表明,从松质骨来源的细胞制备软骨-聚合物构建体是可行的,并且这些细胞具有与源自骨髓的间充质干细胞相同的软骨形成分化潜能。使用制备的软骨-聚合物构建体可以在裸鼠膝关节中重建骨软骨缺损。