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在椎间盘退变的细胞移植治疗中使用人髓核细胞系作为细胞来源的可行性。

Feasibility of using a human nucleus pulposus cell line as a cell source in cell transplantation therapy for intervertebral disc degeneration.

作者信息

Iwashina Toru, Mochida Joji, Sakai Daisuke, Yamamoto Yukihiro, Miyazaki Takeshi, Ando Kiyoshi, Hotta Tomomitsu

机构信息

Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa, Japan.

出版信息

Spine (Phila Pa 1976). 2006 May 15;31(11):1177-86. doi: 10.1097/01.brs.0000217687.36874.c4.

Abstract

STUDY DESIGN

Assessment of the potential use of an immortalized human nucleus pulposus cell line as an alternative cell source in cell transplantation therapy for intervertebral disc degeneration.

OBJECTIVES

To evaluate the effect of transplanting the human nucleus pulposus cell line into a disc degeneration model in rabbits and to define whether it is capable of becoming an alternative cell source for cell transplantation therapy for disc degeneration.

SUMMARY OF BACKGROUND DATA

Interest in cell transplantation therapy for disc degeneration has been growing for several years, and a range of different cell types have been examined as possible donor cells. In addition, the establishment of a novel cell line that possesses some of the major characteristics of a normal human nucleus pulposus cells has been reported.

METHODS

Human nucleus pulposus cell line was established, and cells were transplanted into a rabbit disc degeneration model. At 4, 8, and 24 weeks after transplantation, inhibition of intervertebral disc degeneration was assessed by examining the disc height, macroscopic appearance, histologic findings, and immunohistochemistry. In addition, aggrecan, versican, and Type II collagen gene expression in the nucleus pulposus were measured semiquantitatively at the mRNA level. Furthermore, the survival of transplanted cells was examined using immunohistochemistry for Simian Virus 40 T antigen, and the presence of graft-versus-host reaction was assessed by immunohistochemistry for CD4 and CD58.

RESULTS

The disc height was significantly greater in the transplanted group than in the degenerative group's disc from 4 weeks' posttransplantation. Macroscopically, the nucleus pulposus was absent and there was loss of disc height in the degenerative group at 24 weeks after transplantation, whereas the nucleus pulposus was preserved in the transplanted group. Histologic examination showed that the structure of the inner anulus fibrosus was significantly preserved in the transplanted group, and the boundary between the nucleus and anulus could be clearly visualized. Expression of mRNAs of the nucleus pulposus matrix, aggrecan, and Type II collagen was significantly greater in the transplanted group than in the degenerative group. This indicates that transplantation of human nucleus pulposus cell line helped to preserve the matrix of the nucleus pulposus. Thus, transplantation of a human nucleus pulposus cell line was shown to delay disc degeneration in this rabbit model.

CONCLUSION

The human nucleus pulposus cell line may become an alternative cell source for cell transplantation therapy of intervertebral disc degeneration.

摘要

研究设计

评估永生化人髓核细胞系作为椎间盘退变细胞移植治疗中替代细胞来源的潜在用途。

目的

评估将人髓核细胞系移植到兔椎间盘退变模型中的效果,并确定其是否能够成为椎间盘退变细胞移植治疗的替代细胞来源。

背景资料总结

数年来,对椎间盘退变细胞移植治疗的兴趣不断增长,一系列不同类型的细胞已被作为可能的供体细胞进行研究。此外,已有报道建立了一种具有正常人髓核细胞一些主要特征的新型细胞系。

方法

建立人髓核细胞系,并将细胞移植到兔椎间盘退变模型中。在移植后4周、8周和24周,通过检查椎间盘高度、宏观外观、组织学发现和免疫组织化学来评估椎间盘退变的抑制情况。此外,在mRNA水平上半定量测量髓核中聚集蛋白聚糖、多功能蛋白聚糖和II型胶原基因的表达。此外,使用针对猿猴病毒40 T抗原的免疫组织化学检查移植细胞的存活情况,并通过针对CD4和CD58的免疫组织化学评估移植物抗宿主反应的存在。

结果

移植组在移植后4周时的椎间盘高度显著高于退变组的椎间盘。宏观上,退变组在移植后24周时髓核缺失且椎间盘高度降低,而移植组的髓核得以保留。组织学检查显示,移植组纤维环内层结构得到显著保留,髓核与纤维环之间的边界清晰可见。移植组髓核基质、聚集蛋白聚糖和II型胶原的mRNA表达显著高于退变组。这表明人髓核细胞系的移植有助于保留髓核基质。因此,在该兔模型中,人髓核细胞系的移植显示出可延缓椎间盘退变。

结论

人髓核细胞系可能成为椎间盘退变细胞移植治疗的替代细胞来源。

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