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聚乳酸-纳米羟基磷灰石纤维支架中共培养细胞的成骨与破骨细胞分化

Osteogenic and osteoclastogenic differentiation of co-cultured cells in polylactic acid-nanohydroxyapatite fiber scaffolds.

作者信息

Morelli Sabrina, Salerno Simona, Holopainen Jani, Ritala Mikko, De Bartolo Loredana

机构信息

Institute on Membrane Technology, National Research Council of Italy, ITM-CNR, via P. Bucci cubo 17/C, I-87030 Rende, CS Italy.

Laboratory of Inorganic Chemistry, Department of Chemistry, University of Helsinki, PO Box 55, FI-00014 Helsinki, Finland.

出版信息

J Biotechnol. 2015 Jun 20;204:53-62. doi: 10.1016/j.jbiotec.2015.03.023. Epub 2015 Apr 6.

Abstract

The design of bone substitutes involves the creation of a microenvironment supporting molecular cross-talk between cells and scaffolds during tissue formation and remodelling. Bone remodelling process includes the cooperation of bone-building cells and bone-resorbing cells. In this paper we developed polylactic acid (PLA) and composite PLA-nanohydroxyapatite (nHA) scaffolds with 20 and 50wt.% of nHA by electrospinning technique to be used in bone tissue engineering. The developed scaffolds have different fiber diameter, porosity with interconnected pores and mechanical properties. Taking cues from the bone environment features we investigated the differentiation of human mesenchymal stem cells (hMSCs) from bone marrow in osteoblasts and the osteoclastogenesis in the developed scaffolds in homotypic and in co-culture up to 46 days. PLA and composite PLA-nHA scaffolds induced osteogenic and osteoclastogenic differentiation. Both osteoblasts and osteoclasts displayed high expression of specific markers (osteopontin, osteocalcin, RANK, RANKL) and functions such as secretion of ALP, cathepsin K and TRAP activity on composite scaffolds especially on PLA-nHA containing 20wt.% of nHA. The heterotypic interactions between osteoblasts and osteoclasts co-cultured in the developed scaffolds triggered their functional differentiation and activation.

摘要

骨替代物的设计涉及在组织形成和重塑过程中创建一个支持细胞与支架之间分子相互作用的微环境。骨重塑过程包括成骨细胞和破骨细胞的协同作用。在本文中,我们通过静电纺丝技术制备了聚乳酸(PLA)以及含有20wt.%和50wt.%纳米羟基磷灰石(nHA)的PLA-nHA复合支架,用于骨组织工程。所制备的支架具有不同的纤维直径、孔隙率以及相互连通的孔隙和机械性能。借鉴骨环境特征,我们研究了来自骨髓的人间充质干细胞(hMSCs)在成骨细胞中的分化以及在同型和共培养条件下,所制备支架中破骨细胞生成长达46天的情况。PLA和PLA-nHA复合支架诱导了成骨和成破骨细胞分化。在复合支架上,尤其是在含有20wt.% nHA的PLA-nHA支架上,成骨细胞和破骨细胞均表现出特定标志物(骨桥蛋白、骨钙素、RANK、RANKL)的高表达以及诸如碱性磷酸酶分泌、组织蛋白酶K分泌和抗酒石酸酸性磷酸酶活性等功能。在制备的支架中共培养的成骨细胞和破骨细胞之间的异型相互作用触发了它们的功能分化和激活。

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