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上皮细胞条件培养基对狭窄成纤维细胞增殖和迁移的抑制作用

Inhibition of proliferation and migration of stricture fibroblasts by epithelial cell-conditioned media.

作者信息

Nath Nilima, Saraswat Sumit K, Jain Shashank, Koteshwar Sridhar

机构信息

Department of Cell Therapy, Sri Research for Tissue Engineering Pvt. Ltd., Sri Shankara Research Center, Rangadore Memorial Hospital, 1 Cross, Shankarapuram, Bangalore, India.

出版信息

Indian J Urol. 2015 Apr-Jun;31(2):111-5. doi: 10.4103/0970-1591.152809.

Abstract

INTRODUCTION

Urethral stricture is characterized by urethral lumen narrowing due to fibrosis. Urethroplasty of the urethral stricture involves excision of scar, and may be followed by reconstruction of the urethra using split-thickness skin, buccal mucosa, urethral mucosa or, more recently, tissue-engineered grafts. The stricture wound healing process after urethroplasty is known to be mediated by an interaction between keratinocyte and fibroblasts; however, the underlying mechanisms are not studied in detail yet. We investigated the influence of epithelial cell-conditioned medium (ECCM) (obtained from confluent penile skin, buccal mucosa and urethral cell cultures) on the proliferation and migration of stricture fibroblasts using an in vitro scratch assay.

MATERIALS AND METHODS

ECCM was collected from confluent primary epithelial cell cultures of three different human biopsies (penile skin, buccal mucosa and urethral mucosa), whereas stricture fibroblasts were isolated from human urethral stricture biopsies. The effect of ECCM on stricture fibroblasts' proliferation and migration into the scratch was observed using a standard in vitro scratch assay over a period of 3 days. Four experiments were performed independently using four stricture fibroblasts from four patients and ECCM was collected from 12 different patients' primary cell cultures.

RESULTS

ECCM from primary epithelial cells cultures obtained from penile skin, buccal mucosa and urethra inhibited stricture fibroblasts' proliferation and migration in the in vitro scratch assay.

CONCLUSION

These results demonstrate the ability of ECCM to inhibit the proliferation and migration of stricture fibroblasts and present it as an effective adjunct in urethroplasty, which may influence stricture wound healing and inhibit the recurrence of stricture.

摘要

引言

尿道狭窄的特征是由于纤维化导致尿道腔狭窄。尿道狭窄的尿道成形术包括切除瘢痕,随后可能使用断层皮片、颊黏膜、尿道黏膜或最近的组织工程移植物重建尿道。已知尿道成形术后狭窄伤口愈合过程是由角质形成细胞和成纤维细胞之间的相互作用介导的;然而,其潜在机制尚未得到详细研究。我们使用体外划痕试验研究了上皮细胞条件培养基(ECCM)(从汇合的阴茎皮肤、颊黏膜和尿道细胞培养物中获得)对狭窄成纤维细胞增殖和迁移的影响。

材料与方法

ECCM取自三种不同人类活检组织(阴茎皮肤、颊黏膜和尿道黏膜)的汇合原代上皮细胞培养物,而狭窄成纤维细胞则从人类尿道狭窄活检组织中分离得到。使用标准的体外划痕试验在3天内观察ECCM对狭窄成纤维细胞增殖和向划痕处迁移的影响。使用来自四名患者的四种狭窄成纤维细胞独立进行了四项实验,ECCM取自12名不同患者的原代细胞培养物。

结果

在体外划痕试验中,来自阴茎皮肤、颊黏膜和尿道的原代上皮细胞培养物的ECCM抑制了狭窄成纤维细胞的增殖和迁移。

结论

这些结果证明了ECCM抑制狭窄成纤维细胞增殖和迁移的能力,并表明其作为尿道成形术中的一种有效辅助手段,可能影响狭窄伤口愈合并抑制狭窄复发。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f6ca/4397546/9987347c47aa/IJU-31-111-g001.jpg

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