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体外构建仿生成骨细胞龛及其生物相容性评价

In Vitro Fabrication and Biocompatibility Assay of a Biomimetic Osteoblastic Niche.

机构信息

State Key Laboratory of Fine Chemicals, Dalian R&D Center for Stem Cell and Tissue Engineering, Dalian University of Technology, Dalian, 116024, China.

Burns Research Group, ANZAC Research Institute, University of Sydney, Concord, New South Wales, 2139, Australia.

出版信息

Appl Biochem Biotechnol. 2019 Oct;189(2):471-484. doi: 10.1007/s12010-019-03015-z. Epub 2019 May 2.

Abstract

A novel HAp-CS/Gel biomimetic osteoblastic niche was fabricated by freeze-drying, and its mechanical strength and biocompatibility were characterized. HAp-CS/Gel scaffolds in various ratios of 100:5, 100:10, and 100:20 (CS/Gel to HAp) were prepared by freeze-drying prior to chemical cross-linking followed by infrared spectrum analysis, EDS, FITR, SEM, fluorescence microscopy, MTT, and ALP experiments. Results from the infrared spectrum analysis showed that HAp doping remained the surface morphology and the architecture of scaffold with interconnected pores in the size range of 135 to 150 μm. The HAp doping ratio of 100:20 was found to be optimal based on its high porosity of 90%, better water uptake folds of 19.1. In addition, EDS and FITR analyses demonstrated that HAps were uniformly distributed on the surface of a scaffold with aggregates and particles, which has sufficient roughness for cell attachment and proliferation of osteoblasts. Under SEM and fluorescent microscopy, osteoblasts seeded onto the scaffold showed evenly distributed viable cells, which is believed to form a biomimetic niche. In the present study, we further demonstrate that osteoblasts can maintain their function and grow well on the scaffold through MTT and ALP tests. Thus, the scaffold has favorable physical properties and biocompatibility to support the proliferation and differentiation of osteoblasts and further to support the constructs of biomimetic osteoblastic niche.

摘要

通过冷冻干燥制备了一种新型 HAp-CS/Gel 仿生成骨细胞龛,并对其力学性能和生物相容性进行了表征。通过冷冻干燥制备了不同比例的 HAp-CS/Gel 支架(CS/Gel 与 HAp 的比例分别为 100:5、100:10 和 100:20),然后进行化学交联、红外光谱分析、EDS、FITR、SEM、荧光显微镜、MTT 和 ALP 实验。红外光谱分析结果表明,HAp 掺杂后仍保持支架的表面形貌和结构,具有 135 至 150μm 大小的连通孔。根据高孔隙率(90%)和更好的吸水率(19.1 倍),发现 HAp 掺杂比为 100:20 时最佳。此外,EDS 和 FITR 分析表明,HAps 均匀分布在支架表面,有团聚物和颗粒,具有足够的粗糙度,有利于细胞附着和成骨细胞增殖。在 SEM 和荧光显微镜下,接种在支架上的成骨细胞显示出均匀分布的存活细胞,这被认为形成了仿生龛。在本研究中,我们进一步通过 MTT 和 ALP 试验证明,成骨细胞可以在支架上保持其功能并良好生长。因此,该支架具有良好的物理性能和生物相容性,可支持成骨细胞的增殖和分化,并进一步支持仿生成骨细胞龛的构建。

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