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首页> 中文期刊> 《中国医药生物技术》 >半月板细胞外基质-海藻酸水凝胶的制备及其对半月板细胞的影响

半月板细胞外基质-海藻酸水凝胶的制备及其对半月板细胞的影响

         
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摘要

目的 制备脱细胞半月板细胞外基质-海藻酸(AMECM-Alg)水凝胶,并研究AMECM-Alg水凝胶对半月板细胞增殖、再分化的影响.方法 联合物理化学方法制备AMECM,以AMECM和海藻酸为原料制备5种不同浓度的AMECM-Alg水凝胶(0%、0.5%、1%、2%和4%);复合P3代半月板细胞,通过细胞死活染色、CCK8实验、DNA定量评估水凝胶促进细胞增殖的能力,通过胶原、糖胺多糖定量,RT-PCR评估水凝胶促进细胞再分化的能力.结果 死活细胞染色、CCK8实验和DNA定量实验表明AMECM-Alg水凝胶均能促进细胞的增殖,其中以2%浓度组作用最强;胶原和糖胺多糖定量结果表明AMECM-Alg水凝胶均能促进半月板细胞分泌细胞外基质,其中2%浓度组明显优于其他组.RT-PCR结果显示半月板细胞在2%浓度组水凝胶中collagen Ia2、collagen IIa1、SOX9和aggrecan基因表达量明显高于其他组.结论 AMECM-Alg水凝胶具有良好的细胞相容性,既能促进细胞增殖,又能促进细胞再分化,其中以2%浓度组的作用最强,在未来组织工程半月板生物材料研究方面具有很好的应用前景.%Objective To manufacture accellular meniscus extracellular matrix-alginate (AMECM-Alg) hydrogel scaffolds and investigate their biological effect on the cell viability, proliferation, and re-differentiation of the meniscal fibrochondrocytes. Methods Combined physical and chemical methods were used to prepare accellular meniscus extracellular matrix. Five concentrations of AMECM-Alg hydrogel were manufactured by AMECM and alginate with different ratios (0%, 0.5%, 1%, 2%, 4%). The microstructrue of AMECM-Alg hydrogel was investigated by scanning electron microscopy (SEM). Passaged meniscus fibrochondrocytes were seeded in the five different hydrogel scaffolds, and then cultured for 1, 4, 7 and 14 d. Live/dead staining was used to evaluate the cell viability. The cell proliferation on different hydrogel scaffolds was studied by CCK8 test and DNA quantification. Re-differentiation of the passaged meniscal fibrochondrocytes was determined by biochemical assays for GAG and collagen as well as RT-PCR. Results SEM results showed that the porosity of the hydrogel scaffolds became smaller as the AMECM concentration increased. Live/dead staining results indicated that meniscal fibrochondrocytes could maintain good viability in the different hydrogel scaffolds. The results of CCK8 test and DNA quantification revealed that AMECM-Alg hydrogels could promote cell proliferation, with 2% group having the strongest effect. GAG and collagen content assessment results demonstrated that AMECM-Alg hydrogels enhanced collagen and GAGs production, with 2% group inducing the greatest effect. RT-PCR results revealed that AMECM-Alg hydrogels up-regulated the expression of collagen Ia2, collagen IIa1, SOX9, aggrecan, with 2% group exhibiting the robustest effect. Conclusion AMECM-Alg hydrogel provides a suitable microenvironment for seed cells which may maintain cell viability and promote the cell proliferation and re-differentiation, with 2% group having the strongest effect. In conclusion, 2% AMECM-Alg hydrogel scaffold is a promising candidate biomaterial for meniscus tissue engineering.

著录项

  • 来源
    《中国医药生物技术》 |2017年第6期|505-512|共8页
  • 作者

    陈明学; 刘舒云; 郝立波; 韩纲; 郭全义; 郭维民; 沈师; 王振勇; 王泽昊; 张增增; 李旭; 张学亮; 荆晓光;

  • 作者单位

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

    100853 北京,中国人民解放军总医院骨科研究所;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类
  • 关键词

    半月板,胫骨; 组织工程; 水凝胶; 细胞外基质; 海藻酸;

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