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基于细胞的椎间盘组织工程:人类椎间盘细胞在选定细胞载体中的基因表达和基质产生的体外研究

Cell-based tissue engineering for the intervertebral disc: in vitro studies of human disc cell gene expression and matrix production within selected cell carriers.

作者信息

Gruber Helen E, Leslie Kelly, Ingram Jane, Norton H James, Hanley Edward N

机构信息

Department of Orthopaedic Surgery, Carolinas Medical Center, 1542 Garden Terrace, Charlotte, NC 28203, USA.

出版信息

Spine J. 2004 Jan-Feb;4(1):44-55. doi: 10.1016/s1529-9430(03)00425-x.

Abstract

BACKGROUND CONTEXT

Little is known about how disc cells attach, proliferate and form extracellular matrix (ECM) within carrier materials. Such information is needed to help formulate criteria for successful cell-carrier interactions in tissue engineering.

PURPOSE

To compare proliferation, ECM production and gene expression in annulus cells cultured in a variety of cell carrier materials with potential application in tissue engineering of the disc.

STUDY DESIGN

Human intervertebral disc cells from the annulus were used in a prospective study of proliferation, ECM production and gene expression within selected cell carriers.

METHODS

Annulus cells from discs of 29 individuals were tested in collagen sponge, collagen gel, agarose, alginate or fibrin gel formulations. In situ hybridization assessed ECM gene expression of Types I and II collagen, aggrecan and chondroitin-6 sulfotransferase. Cell proliferation, cell shape, attachment and ECM production were evaluated.

RESULTS

Collagen sponges provided the best microenvironment for disc cell ECM production and gene expression. Although collagen gels often could support good cell growth, such constructs did not result in either abundant ECM production or ECM gene expression, as shown by in situ hybridization. Growth and ECM production and gene expression in alginate, agarose and fibrin microenvironments were inferior.

CONCLUSIONS

Tissue engineering techniques open new therapeutic possibilities for use of autologous disc cells, but fundamental questions on how these cells interact with cell carriers are unexplored. Results provide novel data on disc cell gene expression within diverse microenvironments. The collagen sponge proved to be a superior microenvironment.

摘要

背景

关于椎间盘细胞如何在载体材料内附着、增殖并形成细胞外基质(ECM),我们了解甚少。而此类信息对于制定组织工程中细胞与载体成功相互作用的标准至关重要。

目的

比较在多种具有椎间盘组织工程潜在应用价值的细胞载体材料中培养的纤维环细胞的增殖、ECM产生及基因表达情况。

研究设计

采用来自纤维环的人类椎间盘细胞,对选定细胞载体中的细胞增殖、ECM产生及基因表达进行前瞻性研究。

方法

对来自29名个体椎间盘的纤维环细胞在胶原海绵、胶原凝胶、琼脂糖、藻酸盐或纤维蛋白凝胶制剂中进行测试。原位杂交评估I型和II型胶原、聚集蛋白聚糖及软骨素-6-硫酸转移酶的ECM基因表达。评估细胞增殖、细胞形态、附着及ECM产生情况。

结果

胶原海绵为椎间盘细胞ECM产生及基因表达提供了最佳微环境。尽管胶原凝胶通常能支持良好的细胞生长,但原位杂交显示,此类构建物并未导致大量的ECM产生或ECM基因表达。藻酸盐、琼脂糖及纤维蛋白微环境中的细胞生长、ECM产生及基因表达较差。

结论

组织工程技术为自体椎间盘细胞的应用开辟了新的治疗可能性,但关于这些细胞如何与细胞载体相互作用的基本问题尚未得到探索。研究结果提供了关于不同微环境中椎间盘细胞基因表达的新数据。胶原海绵被证明是一种优越的微环境。

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