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改善颞下颌关节盘组织工程的培养条件。

Improving culture conditions for temporomandibular joint disc tissue engineering.

作者信息

Johns D E, Athanasiou K A

机构信息

Department of Bioengineering, Rice University, Houston, Texas 77251, USA.

出版信息

Cells Tissues Organs. 2007;185(4):246-57. doi: 10.1159/000102173.

Abstract

BACKGROUND

The temporomandibular joint (TMJ) is extremely important for activities like eating and talking, which can become painful and difficult for patients with TMJ dysfunction. Tissue engineering is a potential alternative to current surgical interventions through replacement of diseased or injured tissue with a functional construct. Since research with TMJ disc cells began relatively recently, optimal culturing conditions must be determined.

METHODS

Metabolic additives, L-glutamine, L-alanyl-L-glutamine, sodium pyruvate, and insulin, were examined for their effects on TMJ disc cells in monolayer. Effects of L-proline were examined in three-dimensional (3-D) culture at concentrations of 0, 25 and 100 mg/l.

RESULTS

The combination of L-glutamine, sodium pyruvate, and insulin improved cell proliferation rates without affecting collagen production or gene expression. No differences were observed in mechanical properties of the engineered constructs; however, collagen and glycosaminoglycan quantities normalized to cell number decreased at the highest concentration of L-proline.

CONCLUSION

This work identified supplements for 2-D monolayer expansion. Other supplements or culture conditions still need to be investigated for 3-D tissue production. This work improves upon porcine TMJ disc cell culturing conditions, taking us closer to being able to engineer the TMJ disc.

摘要

背景

颞下颌关节(TMJ)对于进食和说话等活动极为重要,而颞下颌关节功能障碍患者会感到疼痛且活动困难。组织工程是一种潜在的替代当前外科干预的方法,通过用功能性构建体替换患病或受损组织来实现。由于对颞下颌关节盘细胞的研究相对较新,因此必须确定最佳培养条件。

方法

研究了代谢添加剂、L-谷氨酰胺、L-丙氨酰-L-谷氨酰胺、丙酮酸钠和胰岛素对单层培养的颞下颌关节盘细胞的影响。在三维(3-D)培养中,研究了浓度为0、25和100mg/l的L-脯氨酸的作用。

结果

L-谷氨酰胺、丙酮酸钠和胰岛素的组合提高了细胞增殖率,且不影响胶原蛋白的产生或基因表达。在工程构建体的力学性能方面未观察到差异;然而,在L-脯氨酸的最高浓度下,归一化到细胞数量的胶原蛋白和糖胺聚糖量减少。

结论

这项工作确定了二维单层扩增的补充剂。对于三维组织生产,仍需研究其他补充剂或培养条件。这项工作改进了猪颞下颌关节盘细胞的培养条件,使我们更接近能够构建颞下颌关节盘。

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