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Chitosan-cartilage extracellular matrix hybrid scaffold induces chondrogenic differentiation to adipose-derived stem cells.
Regenerative Therapy ( IF 4.3 ) Pub Date : 2020-05-15 , DOI: 10.1016/j.reth.2020.03.014
I-Chan Lin,Tsung-Jen Wang,Chien-Liang Wu,Dai-Hua Lu,Yi-Ru Chen,Kai-Chiang Yang

Introduction

Adipose-derived stem cells (ASCs) are potential cell sources for cartilage tissue engineering. Chitosan has been shown to enhance the stemness and differentiation capability of ASCs, and the native extracellular matrix (ECM) derived from articular cartilage has been also reported to induce chondrogenic differentiation of ASCs. Here we tested the hypothesis that a porous three-dimensional (3D) hybrid scaffold composed of chitosan and cartilage ECM can provide a better environment to induce ASC chondrogenesis.

Methods

Mixed solution composed of chitosan and cartilage ECM was frozen and lyophilized to form a composite construct. The porous 3D scaffolds were further crosslinked by genipin and used for ASC culture.

Results

Cultivation of ASCs in the chitosan/cartilage ECM composite 3D scaffolds induced the formation of cell spheroids with profound glycosaminoglycan production after 14 and 28 days culture. Chondrogenesis of ASCs seeded in the 3D scaffolds was also evident by mRNA expressions of cartilage-specific gene COL2A1 and ACAN on day 14. Histology and immunohistochemistry on day 28 also showed abundant cartilage-specific macromolecules, namely collagen type II and proteoglycan, deposited in a surface layer of the composite scaffold with tangential layer, transitional layer, and lacunae-like structures. Otherwise, hypertrophic markers collagen type I and X were concentrated in the area beneath the surface.

Conclusion

Our findings demonstrated spatial chondrogenic differentiation of ASCs in the chitosan-cartilage ECM composite scaffolds. This 3D hybrid scaffold exhibits great potentials for ASC-based cartilage tissue engineering.



中文翻译:

壳聚糖-软骨细胞外基质混合支架诱导软骨向脂肪干细胞分化。

介绍

脂肪干细胞 (ASC) 是软骨组织工程的潜在细胞来源。壳聚糖已被证明可增强 ASCs 的干性和分化能力,并且据报道,源自关节软骨的天然细胞外基质 (ECM) 可诱导 ASCs 的软骨分化。在这里,我们测试了由壳聚糖和软骨 ECM 组成的多孔三维 (3D) 混合支架可以提供更好的环境来诱导 ASC 软骨形成的假设。

方法

将壳聚糖和软骨ECM组成的混合溶液冷冻并冻干以形成复合结构。多孔 3D 支架通过京尼平进一步交联并用于 ASC 培养。

结果

在壳聚糖/软骨 ECM 复合 3D 支架中培养 ASC,在培养 14 天和 28 天后诱导形成具有大量糖胺聚糖产生的细胞球体。在第 14 天,软骨特异性基因COL2A1ACAN的 mRNA 表达也证明了接种在 3D 支架中的 ASC 的软骨形成。第 28 天的组织学和免疫组织化学也显示出丰富的软骨特异性大分子,即 II 型胶原蛋白和蛋白聚糖,沉积在复合支架的表层具有切向层、过渡层和腔隙状结构。否则,I 型和 X 型胶原蛋白的肥大标志物集中在表面下方的区域。

结论

我们的研究结果证明了壳聚糖-软骨 ECM 复合支架中 ASC 的空间软骨形成分化。这种 3D 混合支架在基于 ASC 的软骨组织工程中表现出巨大的潜力。

更新日期:2020-05-15
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