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用于骨组织工程的丙烯酸接枝罗望子核多糖基水凝胶,无需任何骨诱导因子

Acrylic acid grafted tamarind kernel polysaccharide-based hydrogel for bone tissue engineering in absence of any osteo-inducing factors.

作者信息

Kumar Satish, Majhi Rakesh Kumar, Sanyasi Sridhar, Goswami Chandan, Goswami Luna

机构信息

a School of Biotechnology , KIIT University , Bhubaneswar , India.

b School of Biological Sciences , National Institute of Science Education and Research , Bhubaneswar , Orissa , India.

出版信息

Connect Tissue Res. 2018 Dec;59(sup1):111-121. doi: 10.1080/03008207.2018.1442444.

Abstract

PURPOSE

With increased life expectancy, disorders in lifestyle and other clinical conditions, and the changes in the connective tissues such as in bone, impose diverse biomedical problems. Cells belong to osteogenic lineages are extremely specific for their surface requirements. Therefore, suitable surfaces are the critical bottle neck for successful bone tissue engineering. This study involves assessment of polysaccharide-based hydrogel which effectively allows growth, differentiation and mineralisation of osteogenic cells even in the absence of osteogenic inducing factors.

MATERIALS AND METHODS

Tamarind Kernel Polysaccharide was grafted with acrylic acid at different mole ratio. The critical parameter, surface morphology for bio application was assessed by SEM. MTT assay has been performed with hydrogels on Saos-2 cells. The biocompatibility and adhesion of different cell lines (F-11, Saos-2, Raw 264.7 and MSCs) on hydrogel surface was performed by Phalloidin and DAPI staining. Further the differentiation, mineralization and expression of different osteogenic markers, ALP assay, Alizarin Red staining and q-PCR was performed.

RESULTS

The hydrogels show highly porous and interconnected pores. MTT assay demonstrates the hydrogel have no cytotoxicity towards Saos-2 cells and are suitable for proliferation of different lineage of cell lines. ALP, Alizarin red staining and q-PCR assay shows that the hydrogel surface enhances the differentiation, mineralization and expression of different osteogenic genes in Saos-2 cells in the absence of any osteogenic inducing factors. Conclusion Synthesized hydrogel surface triggers signalling events towards osteogenesis even in the absence of added growth factors. We proposed that this material can be used for effective bone tissue engineering in vitro at low cost.

摘要

目的

随着预期寿命的增加、生活方式紊乱及其他临床状况,以及诸如骨骼等结缔组织的变化,带来了各种生物医学问题。属于成骨谱系的细胞对其表面需求极为特殊。因此,合适的表面是成功进行骨组织工程的关键瓶颈。本研究涉及评估基于多糖的水凝胶,即使在没有成骨诱导因子的情况下,该水凝胶也能有效促进成骨细胞的生长、分化和矿化。

材料与方法

将罗望子核多糖与丙烯酸以不同摩尔比接枝。通过扫描电子显微镜评估生物应用的关键参数——表面形态。用该水凝胶对Saos-2细胞进行MTT检测。通过鬼笔环肽和DAPI染色检测不同细胞系(F-11、Saos-2、Raw 264.7和间充质干细胞)在水凝胶表面的生物相容性和黏附情况。进一步进行不同成骨标志物的分化、矿化和表达检测、碱性磷酸酶检测、茜素红染色和定量聚合酶链反应。

结果

水凝胶呈现出高度多孔且相互连通的孔隙。MTT检测表明该水凝胶对Saos-2细胞无细胞毒性,适合不同细胞系的增殖。碱性磷酸酶、茜素红染色和定量聚合酶链反应检测表明,在没有任何成骨诱导因子的情况下,水凝胶表面可增强Saos-2细胞中不同成骨基因的分化、矿化和表达。结论:即使在没有添加生长因子的情况下,合成的水凝胶表面也能触发成骨信号事件。我们认为这种材料可用于低成本的体外有效骨组织工程。

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