Collagen Type II enhances chondrogenic differentiation in agarose-based modular microtissues.
Collagen Type II enhances chondrogenic differentiation in agarose-based modular microtissues.
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DOI：
10.1016/j.jcyt.2015.10.015
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发表时间：
2016-02
期刊：
影响因子：
4.5
通讯作者：
Stegemann JP
中科院分区：
文献类型：
--
作者：
Tiruvannamalai Annamalai R;Mertz DR;Daley EL;Stegemann JP
基于细胞的疗法对骨关节炎的治疗产生了影响，然而厚软骨缺损的修复和再生是一个重要且日益增长的临床问题。将联合收割机细胞与生物材料相结合的下一代疗法可能会提供更好的结果。我们已经开发了模块化的微环境，模拟关节软骨的组成作为一个交付系统，一贯分化的细胞。使用油包水乳液技术，将人骨髓来源的间充质干细胞（MSC）包埋在由琼脂糖（AG）组成的模块化微珠中，所述琼脂糖（AG）补充有0%、10%和20%的II型胶原（COL-II）。AG和AG/COL-II微珠的特征在于其结构完整性，尺寸分布和蛋白质含量。还评估了包埋的MSC的活力及其在培养物中三周内分化成成骨、成脂和成软骨谱系的能力。用<20% COL-II制备的微珠是坚固的，通常为球形，直径为80 ± 10 µm。MSC在微珠中的活力在培养物中持续一周以上高，而在相应的散装水凝胶中的活力随着COL-II含量的增加而降低。MSC的成骨分化在AG和AG/COL-II微珠中得到适度支持，而成脂分化在含有COL-II的微珠中受到强烈抑制。与纯AG基质相比，含有COL-II的微珠明显促进了MSC的软骨分化。微珠形式的琼脂糖基质中包含II型胶原蛋白可以增强人MSC的软骨形成分化。这样的组合物定制的微组织可能会发现在下一代软骨修复疗法中的细胞递送的效用。
Cell-based therapies have made an impact on the treatment of osteoarthritis, however the repair and regeneration of thick cartilage defects is an important and growing clinical problem. Next-generation therapies that combine cells with biomaterials may provide improved outcomes. We have developed modular microenvironments that mimic the composition of articular cartilage as a delivery system for consistently differentiated cells. Human bone marrow-derived mesenchymal stem cells (MSC) were embedded in modular microbeads consisting of agarose (AG) supplemented with 0%, 10%, and 20% collagen Type II (COL-II) using a water-in-oil emulsion technique. AG and AG/COL-II microbeads were characterized in terms of their structural integrity, size distribution, and protein content. The viability of embedded MSC and their ability to differentiate into osteogenic, adipogenic and chondrogenic lineages over three weeks in culture were also assessed. Microbeads made with <20% COL-II were robust, generally spheroidal in shape and 80 ± 10 µm in diameter. MSC viability in microbeads was consistently high over a week in culture, while viability in corresponding bulk hydrogels decreased with increasing COL-II content. Osteogenic differentiation of MSC was modestly supported in both AG and AG/COL-II microbeads, while adipogenic differentiation was strongly inhibited in COL-II containing microbeads. Chondrogenic differentiation of MSC was clearly promoted in microbeads containing COL-II, compared to pure AG matrices. Inclusion of collagen Type II in agarose matrices in microbead format can potentiate chondrogenic differentiation of human MSC. Such compositionally tailored microtissues may find utility for cell delivery in next-generation cartilage repair therapies.