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壳聚糖改性聚(L-丙交酯-共-ε-己内酯)支架上提高间充质干细胞黏附及体外软骨组织形成。

Improved mesenchymal stem cells attachment and in vitro cartilage tissue formation on chitosan-modified poly(L-lactide-co-epsilon-caprolactone) scaffold.

机构信息

Tissue Engineering Program, Life Sciences Institute, National University of Singapore, Singapore.

出版信息

Tissue Eng Part A. 2012 Feb;18(3-4):242-51. doi: 10.1089/ten.TEA.2011.0315. Epub 2011 Dec 22.

Abstract

Considering the load-bearing physiological requirement of articular cartilage, scaffold for cartilage tissue engineering should exhibit appropriate mechanical responses as natural cartilage undergoing temporary deformation on loading with little structural collapse, and recovering to the original geometry on unloading. A porous elastomeric poly l-lactide-co-ɛ-caprolactone (PLCL) was generated and crosslinked at the surface to chitosan to improve its wettability. Human bone marrow derived mesenchymal stem cells (MSC) attachment, morphological change, proliferation and in vitro cartilage tissue formation on the chitosan-modified PLCL scaffold were compared with the unmodified PLCL scaffold. Chitosan surface promoted more consistent and even distribution of the seeded MSC within the scaffold. MSC rapidly adopted a distinct spread-up morphology on attachment on the chitosan-modified PLCL scaffold with the formation of F-actin stress fiber which proceeded to cell aggregation; an event much delayed in the unmodified PLCL. Enhanced cartilage formation on the chitosan-modified PLCL was shown by real-time PCR analysis, histological and immunochemistry staining and biochemical assays of the cartilage extracellular matrix components. The Young's modulus of the derived cartilage tissues on the chitosan-modified PLCL scaffold was significantly increased and doubled that of the unmodified PLCL. Our results show that chitosan modification of the PLCL scaffold improved the cell compatibility of the PLCL scaffold without significant alteration of the physical elastomeric properties of PLCL and resulted in the formation of cartilage tissue of better quality.

摘要

考虑到关节软骨的承载生理要求,软骨组织工程支架应表现出适当的机械响应,类似于天然软骨在加载时会发生暂时变形而不会发生结构崩溃,并在卸载时恢复到原始几何形状。我们生成了一种多孔弹性聚 L-丙交酯-共-ε-己内酯(PLCL),并在表面交联壳聚糖以提高其润湿性。比较了壳聚糖修饰的 PLCL 支架和未修饰的 PLCL 支架对人骨髓间充质干细胞(MSC)黏附、形态变化、增殖和体外软骨组织形成的影响。壳聚糖表面促进了更多一致且均匀的种子 MSC 在支架内的分布。MSC 在壳聚糖修饰的 PLCL 支架上黏附后迅速呈现出明显的伸展形态,并形成 F-肌动蛋白应力纤维,导致细胞聚集;而在未修饰的 PLCL 中,这一事件延迟了很多。实时 PCR 分析、组织学和免疫化学染色以及软骨细胞外基质成分的生化分析表明,壳聚糖修饰的 PLCL 支架上形成了增强的软骨组织。壳聚糖修饰的 PLCL 支架上衍生的软骨组织的杨氏模量显著增加,是未修饰 PLCL 的两倍。我们的结果表明,壳聚糖修饰 PLCL 支架提高了 PLCL 支架的细胞相容性,而对 PLCL 的物理弹性特性没有显著改变,从而形成了质量更好的软骨组织。

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