Human amniotic membrane as a delivery matrix for articular cartilage repair
Human amniotic membrane as a delivery matrix for articular cartilage repair
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DOI：
10.1089/ten.2006.0184
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发表时间：
2007-04-01
影响因子：
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通讯作者：
Min, Byoung-Hyun
中科院分区：
文献类型：
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作者：
Jin, Cheng Zhe;Park, So Ra;Min, Byoung-Hyun
本研究的目的是通过评估人羊膜（HAM）作为软骨细胞载体的体外增殖和表型维持以及体内软骨再生的可行性。用0.1%胰蛋白酶-乙二胺四乙酸（EDTA）处理完整的火腿15分钟，去除上皮细胞，制成去皮火腿。然后将兔关节软骨细胞接种在三种不同的火腿基质上：完整火腿的上皮侧（IHE），剥去的火腿的基底侧（DHB）和剥去的火腿的基质侧（DHS）。这些细胞基质标本培养4周，在第1周和第4周检测细胞增殖率和表型稳定性。当软骨细胞在IHE和DHB底物表面以单层方式生长时，DHS中植入的细胞渗透并扩散到整个间质层厚度。DHB软骨细胞增殖活性持续上调。DHS在第一周也有类似的增殖活性，并在4周内保持稳定。DHS组II型胶原的表达随时间逐渐增加，而DHB组II型胶原的表达逐渐减少或IHE组II型胶原的表达完全不存在。这些结果表明，剥去的HAM能够在体外支持软骨细胞增殖和维持表型，当使用DHS时似乎更有利。在此基础上，采用带软骨细胞的DHS覆盖基质侧朝内的兔骨软骨缺损。植入8周后，红素- o染色和国际软骨修复学会（ICRS）评分显示缺损区再生成功，形成透明软骨。总之，我们的研究结果表明，剥去的HAM可能是软骨再生的理想细胞载体基质之一。
The purpose of this study is to evaluate the feasibility of human amniotic membrane (HAM) as a chondrocyte carrier by assessing cell proliferation and maintenance of phenotype in vitro and cartilage regeneration in vivo. Intact HAM was treated with 0.1% trypsin-ethylenediaminetetraacetic acid (EDTA) for 15 min and the epithelial cells removed to make a denuded HAM. Rabbit articular chondrocytes were then seeded on three different HAM substrates: the epithelial side of intact HAM (IHE), basement side of denuded HAM (DHB), and stromal side of denuded HAM (DHS). These cell-substrate specimens were cultured for up to 4 weeks, and cell proliferation rate and phenotypic stability were examined at weeks 1 and 4. While chondrocytes grew in monolayer fashion on the surface of IHE and DHB substrates, the cells seeded in DHS penetrated and spread into the whole thickness of the stromal layer. The proliferating activity of chondrocytes in DHB was continuously up-regulated. A similar proliferating activity was observed in DHS in the first week, which remained stable for up to 4 weeks. The expression of type II collagen gradually increased with time in the DHS group, while it gradually decreased in the DHB group or was not detected at all in the IHE group. These results suggested that denuded HAM was able to support chondrocyte proliferation and maintenance of phenotype in vitro, seemingly more favorable when DHS was used. Based on this data, the DHS with chondrocytes was used to cover rabbit osteochondral defect with the stromal side facing in. The defect area was successfully regenerated with hyaline cartilage in the Safranin-O stain and International Cartilage Repair Society (ICRS) scoring after 8 weeks of implantation. In conclusion, our findings suggest that denuded HAM could be one of the ideal cell carrier matrices for cartilage regeneration.