Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Biomaterials. 2010 Mar;31(8):2193-200. doi: 10.1016/j.biomaterials.2009.11.092. Epub 2010 Jan 19.
Three-dimensionally woven poly(epsilon-caprolactone) (PCL) scaffolds were combined with adult human mesenchymal stem cells (hMSC) to engineer mechanically functional cartilage constructs in vitro. The specific objectives were to: (i) produce PCL scaffolds with cartilage-like mechanical properties, (ii) demonstrate that hMSCs formed cartilage after 21 days of culture on PCL scaffolds, and (iii) study effects of scaffold structure (loosely vs. tightly woven), culture vessel (static dish vs. oscillating bioreactor), and medium composition (chondrogenic additives with or without serum). Aggregate moduli of 21-day constructs approached normal articular cartilage for tightly woven PCL cultured in bioreactors, were lower for tightly woven PCL cultured statically, and lowest for loosely woven PCL cultured statically (p<0.05). Construct DNA, total collagen, and glycosaminoglycans (GAG) increased in a manner dependent on time, culture vessel, and medium composition. Chondrogenesis was verified histologically by rounded cells within a hyaline-like matrix that immunostained for collagen type II but not type I. Bioreactors yielded constructs with higher collagen content (p<0.05) and more homogenous matrix than static controls. Chondrogenic additives yielded constructs with higher GAG (p<0.05) and earlier expression of collagen II mRNA if serum was not present in medium. These results show feasibility of functional cartilage tissue engineering from hMSC and 3D-woven PCL scaffolds.
三维编织的聚己内酯（PCL）支架与成人间充质干细胞（hMSC）结合，在体外构建具有机械功能的软骨组织工程。具体目标是：（i）制备具有类似软骨机械性能的 PCL 支架，（ii）证明 hMSC 在 PCL 支架上培养 21 天后可形成软骨，以及（iii）研究支架结构（疏松与紧密编织）、培养容器（静态培养皿与振荡生物反应器）和培养基组成（含或不含血清的软骨形成添加剂）对细胞的影响。在生物反应器中培养的紧密编织 PCL 构建体的 21 天聚集模量接近正常关节软骨，在静态培养的紧密编织 PCL 中较低，在静态培养的疏松编织 PCL 中最低（p<0.05）。构建体的 DNA、总胶原和糖胺聚糖（GAG）随时间、培养容器和培养基组成而增加。组织学上通过免疫染色为 II 型胶原但不是 I 型胶原的透明样基质内的圆形细胞证实了软骨形成。生物反应器产生的构建体具有更高的胶原含量（p<0.05）和更均匀的基质，优于静态对照。如果培养基中不含血清，软骨形成添加剂可产生具有更高 GAG（p<0.05）和更早表达胶原 II mRNA 的构建体。这些结果表明从 hMSC 和 3D 编织 PCL 支架构建功能性软骨组织工程的可行性。
