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旋转微重力生物反应器内封装的大鼠间充质干细胞的成骨分化:体内外评估

Osteogenic differentiation of encapsulated rat mesenchymal stem cells inside a rotating microgravity bioreactor: in vitro and in vivo evaluation.

作者信息

Koç Demir Aysel, Elçin Ayşe Eser, Elçin Yaşar Murat

机构信息

Tissue Engineering, Biomaterials and Nanobiotechnology Laboratory, Faculty of Science and Stem Cell Institute, Ankara University, Degol Caddesi, Tandogan, 06100, Ankara, Turkey.

Biovalda Health Technologies, Inc., Ankara, Turkey.

出版信息

Cytotechnology. 2018 Oct;70(5):1375-1388. doi: 10.1007/s10616-018-0230-8. Epub 2018 Jun 25.

Abstract

The objective of this study is to evaluate the in vitro and in vivo osteogenic potential of rat bone marrow mesenchymal stem cells (BM-MSCs) using chitosan/hydroxyapatite (C/HAp) microbeads as encapsulation matrix under osteoinductive medium and dynamic culture conditions. The degradation characteristics of C/HAp microbeads were evaluated under in vitro and in vivo conditions for 180 days. BM-MSCs were encapsulated in C/HAp microbeads with > 85% viability, and were cultured in a slow turning lateral vessel-type rotating bioreactor simulating microgravity conditions for 28 days, under the effect of osteogenic inducers. MTT assay showed that the metabolic activity of encapsulated cells was preserved > 80% after a week. In vitro experiments confirmed that the encapsulated BM-MSCs differentiated into osteoblastic cells, formed bone-like tissue under osteogenic microgravity bioreactor conditions. Preliminary in vivo study indicated C/HAp microbeads containing BM-MSCs were able to repair the surgically-created small bone defects in the rat femur. BM-MSCs-C/HAp composite microbeads may have potential for modular bone regeneration.

摘要

本研究的目的是在骨诱导培养基和动态培养条件下,使用壳聚糖/羟基磷灰石(C/HAp)微珠作为包封基质,评估大鼠骨髓间充质干细胞(BM-MSCs)的体外和体内成骨潜力。在体外和体内条件下评估C/HAp微珠180天的降解特性。将BM-MSCs封装在C/HAp微珠中,存活率>85%,并在模拟微重力条件的慢速旋转侧血管型旋转生物反应器中在成骨诱导剂的作用下培养28天。MTT试验表明,封装细胞的代谢活性在一周后保持>80%。体外实验证实,封装的BM-MSCs分化为成骨细胞,在成骨微重力生物反应器条件下形成类骨组织。初步体内研究表明,含有BM-MSCs的C/HAp微珠能够修复大鼠股骨手术造成的小骨缺损。BM-MSCs-C/HAp复合微珠可能具有模块化骨再生的潜力。

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