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兔骨组织工程中明胶基生物材料的成骨细胞研究。

Osteoblast studied on gelatin based biomaterials in rabbit Bone Bioengineering.

机构信息

School of Biochemical Engineering, IIT (BHU), Varanasi 221005, India.

出版信息

Mater Sci Eng C Mater Biol Appl. 2019 Nov;104:109892. doi: 10.1016/j.msec.2019.109892. Epub 2019 Jun 16.

Abstract

The bone-derived-osteoblast seeded biomaterials scaffold in tissue engineering, have displayed prominence in the treatment of the osseous medical condition. In vitro osteogenesis of rabbit osteoblast cell (rOb) from bone tissue (rT) and MSC-derived rOb from bone marrow (rM) on Gelatin-Hydroxyapatite (HG) based biomaterials was investigated. In this work, lyophilized biomaterial was prepared by the addition of amorphous chitosan ('C') to 'H' dispersed in 'G' matrix, to find its role in biomaterials biocompatibility. Isolated rOb seeded biomaterials were studied using CLSM and flow cytometry for proliferation potential. The biomaterial's core and surface morphology was studied from SEM-EDX and AFM respectively. Upon co-culture with HCG, rT over rM showed rabbit bone extracellular matrix (ECM) mimicking properties both in in vitro studies and biomaterials micro architecture. The in vitro metabolic behaviour was studied by Alamar Blue (AB) assay, DNA content using Hoechst 33258, potency via the activity of Alkaline Phosphatase (ALP), Calcium's relative content by Alizarin Red S (ARS) assay. A novel combination of biomaterials-cell interaction was observed when rT was co-cultured with HCG and proved effective in osteogenesis with regard to Bone Bioengineering.

摘要

在组织工程中,源于骨的成骨细胞接种生物材料支架在治疗骨医学病症方面表现出了突出的效果。本研究旨在研究兔骨组织来源的成骨细胞(rOb)和骨髓间充质干细胞来源的成骨细胞(rM)在基于明胶-羟基磷灰石(HG)的生物材料上的体外成骨情况。在这项工作中,通过将无定形壳聚糖(“C”)添加到分散在“G”基质中的“H”中,制备了冻干生物材料,以研究其在生物材料生物相容性中的作用。使用 CLSM 和流式细胞术研究了接种生物材料的分离 rOb 的增殖潜力。分别通过 SEM-EDX 和 AFM 研究了生物材料的核心和表面形态。在与 HCG 共培养时,rT 比 rM 表现出了在体外研究和生物材料微观结构中均具有兔骨细胞外基质(ECM)模拟特性。通过 Alamar Blue(AB)测定法、Hoechst 33258 测定法中的 DNA 含量、碱性磷酸酶(ALP)活性测定法中的活力以及茜素红 S(ARS)测定法中的钙相对含量来研究体外代谢行为。当 rT 与 HCG 共培养时,观察到了一种新型生物材料-细胞相互作用的组合,并且在骨生物工程方面,这种组合在成骨方面被证明是有效的。

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