A Human Amnion-Derived Extracellular Matrix-Coated Cell-Free Scaffold for Cartilage Repair: In Vitro and In Vivo Studies.
A Human Amnion-Derived Extracellular Matrix-Coated Cell-Free Scaffold for Cartilage Repair: In Vitro and In Vivo Studies.
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DOI：
10.1089/ten.tea.2015.0285
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发表时间：
2016-03
影响因子：
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通讯作者：
Makiko Nogami;Tomoatsu Kimura;Shoji Seki;Y. Matsui;Toshiko Yoshida;Chika Koike-Soko;M. Okabe;H. Motomura;R. Gejo;T. Nikaido
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文献类型：
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作者：
Makiko Nogami;Tomoatsu Kimura;Shoji Seki;Y. Matsui;Toshiko Yoshida;Chika Koike-Soko;M. Okabe;H. Motomura;R. Gejo;T. Nikaido
目的人羊膜间充质细胞来源的细胞外基质（ECM）具有多种生物活性。在这项研究中，我们开发了一种新的HAM衍生的ECM涂层聚乳酸-羟基乙酸共聚物（ECM-PLGA）支架，检查其对间充质细胞的性能，并研究其作为软骨修复的无细胞支架的潜力。材料和方法ECM-PLGA支架是由EQUIPHAM在PLGA上开发的。在通过照射脱细胞后，检查累积的ECM。采用细胞贴壁增殖实验和逆转录-聚合酶链反应（RT-PCR）检测大鼠骨髓间充质干细胞（MSCs）在ECM-PLGA上的体外生长和分化情况。将无细胞的ECM-PLGA支架植入大鼠膝关节滑车沟骨软骨缺损处。在4、12或24周后，处死动物并对收获的组织进行组织学检查。结果ECM-PLGA含有模拟天然羊膜基质的ECM，其含有I型胶原、纤连蛋白、透明质酸和硫酸软骨素。该材料具有良好的细胞粘附和增殖能力。接种在ECM-PLGA支架上的MSC在培养3周后显示II型胶原mRNA表达加速。将ECM-PLGA植入大鼠膝关节骨软骨缺损处，诱导组织逐渐再生，导致透明软骨修复，这优于空白对照组。结论体外和体内实验表明，由HAM来源的ECM和PLGA组成的无细胞支架为MSCs提供了良好的生长环境，促进了软骨修复过程。ECM-PLGA有望成为软骨修复治疗的“现成”生物材料。
OBJECTIVE Extracellular matrix (ECM) derived from human amniotic mesenchymal cells (HAMs) has various biological activities. In this study, we developed a novel HAM-derived ECM-coated polylactic-co-glycolic acid (ECM-PLGA) scaffold, examined its property on mesenchymal cells, and investigated its potential as a cell-free scaffold for cartilage repair. MATERIALS AND METHODS ECM-PLGA scaffolds were developed by inoculating HAM on a PLGA. After decellularization by irradiation, accumulated ECM was examined. Exogenous cell growth and differentiation of rat mesenchymal stem cells (MSCs) on the ECM-PLGA were analyzed in vitro by cell attachment/proliferation assay and reverse transcription-polymerase chain reaction. The cell-free ECM-PLGA scaffolds were implanted into osteochondral defects in the trochlear groove of rat knees. After 4, 12, or 24 weeks, the animals were sacrificed and the harvested tissues were examined histologically. RESULTS The ECM-PLGA contained ECM that mimicked natural amniotic stroma that contains type I collagen, fibronectin, hyaluronic acid, and chondroitin sulfates. The ECM-PLGA showed excellent properties of cell attachment and proliferation. MSCs inoculated on the ECM-PLGA scaffold showed accelerated type II collagen mRNA expression after 3 weeks in culture. The ECM-PLGA implanted into an osteochondral defect in rat knees induced gradual tissue regeneration and resulted in hyaline cartilage repair, which was better than that in the empty control group. CONCLUSION These in vitro and in vivo experiments show that the cell-free scaffold composed of HAM-derived ECM and PLGA provides a favorable growth environment for MSCs and facilitates the cartilage repair process. The ECM-PLGA may become a "ready-made" biomaterial for cartilage repair therapy.