Biomaterials Research Center, Korea Institute of Science and Technology, P.O. Box 131, Cheongryang, Seoul 130-650, Korea.
J Control Release. 2010 Oct 1;147(1):84-91. doi: 10.1016/j.jconrel.2010.06.020. Epub 2010 Jul 3.
In this study, a composite hydrogel using Pluronic F127 derivatives and crosslinked hyaluronic acid (X-HA) was investigated, exploring the benefits in the induction of chondrogenic differentiation of human adipose-derived stem cells (ASCs). F127 was chemically modified through a series of reactions that produced multiple F127 derivatives. A chondrogenic growth factor, transforming growth factor-beta 1 (TGF-β1) was then coupled to the heparin-conjugated F127. X-HA was used as a physical stabilizer of the composite hydrogel (X-HA/F127). The chemical structures of F127 derivatives were analyzed using (1)H-NMR and ATR-FTIR. Sol-gel transition of the composite hydrogel was identified at body temperature. The conjugated TGF-β1 was moderately released in vitro from the composite hydrogel. Cell viability of human ASCs in the hydrogels was about 50% after in vitro culture for 3 days. As the ASCs/hydrogel were injected into nude mice subcutaneously, DAPI staining of the retrieved constructs showed that ASCs were dispersed through the hydrogel matrix. From the immunofluorescent staining of type II collagen, the TGF-conjugated group exhibited more active green signals than the others. In addition, when those constructs were loaded into the full-thickness defect of rabbit knee articular cartilage, Alcian blue staining identified the formation of cartilaginous matrix from the TGF-conjugated hydrogel. The present work indicated that X-HA/F127 composite hydrogel was thermoreversible and biodegradable, and that the TGF-conjugated hydrogel could be effective in inducing chondrogenesis of human ASCs.
在这项研究中，我们研究了一种使用 Pluronic F127 衍生物和交联透明质酸（X-HA）的复合水凝胶，探讨了其对人脂肪来源干细胞（ASCs）诱导软骨分化的益处。F127 通过一系列反应进行化学修饰，生成了多种 F127 衍生物。然后将转化生长因子-β1（TGF-β1）这种软骨生长因子偶联到肝素化的 F127 上。X-HA 用作复合水凝胶（X-HA/F127）的物理稳定剂。使用 1H-NMR 和 ATR-FTIR 分析了 F127 衍生物的化学结构。复合水凝胶在体温下可发生溶胶-凝胶转变。TGF-β1 从复合水凝胶中体外适度释放。人 ASC 在水凝胶中的体外培养 3 天后，细胞活力约为 50%。当将 ASC/水凝胶注入裸鼠皮下时，回收的构建体的 DAPI 染色显示 ASC 分散在水凝胶基质中。从 II 型胶原的免疫荧光染色可以看出，TGF 缀合组的绿色信号比其他组更活跃。此外，当将这些构建体加载到兔膝关节关节软骨的全层缺损中时，阿利新蓝染色鉴定出 TGF 缀合水凝胶形成了软骨基质。本研究表明，X-HA/F127 复合水凝胶具有温度可逆性和生物可降解性，TGF 缀合水凝胶可有效诱导人 ASC 软骨形成。
