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[脱细胞角膜基质的生物相容性与组织工程角膜上皮移植]

[Biocompatibility of acellular corneal stroma and transplantation of tissue-engineered corneal epithelium].

作者信息

Fang Xing-feng, Zhao Jing, Shi Wei-yun, Xie Li-xin

机构信息

Shandong Eye Institute, Shandong Provincial Key Lab of Ophthalmology, Qingdao 266071, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2008 Oct;44(10):934-42.

Abstract

OBJECTIVE

To evaluate the biocompatibility of xenogeneic acellular corneal stroma, the feasibility of tissue engineered corneal epithelial transplantation, and verify the long term survival of epithelial allograft.

METHODS

It was a experimental study. Porcine corneal stroma was treated by dispase followed by Triton-X-100 detergent. Treated porcine corneal stroma (group A) or fresh corneal stroma (group B) were put into the sac of rabbit cornea. Rabbit cornea without implantation of porcine corneal stroma was used as the control group (group C). Immunological rejection was evaluated in morphology, histopathology and immunohistochemistry in month 1, 3, 6. Female rabbit underwent lamellar keratoplasty (LK) using male tissue engineered corneal epithelium as donors, and immunological rejection after LK was analyzed. The corneas were collected at day 1, 3, 7, 15 and 30 after LK and evaluated by histopathology, immunohistochemistry and sex-determining region of Y-chromosome (SRY)-PCR analysis.

RESULTS

All corneas became transparent gradually after the transplantation of treated porcine corneal stroma and were not rejected. The epithelium, stroma, endothelium, Bowman's and Descemet's membrane were preserved in all corneas of group A and B in histological observations, collagen fibers were parallel, a few keratocytes presented in the acellular and fresh corneal stromas. The corneas of group C were normal in histological sections. No significant immune rejection was noted in any of the corneas. The corneas in the study of transplantation of tissue engineered cornea epithelium recovered smoothly in 3 or 4 days, turned transparent in 15 or 20 days after surgery and were not stained by fluorescein. Well-differentiated corneal epithelium were recognized at 15 and 30 days after LK. Many keratocytes infiltrated into the scaffold. SRY-PCR analysis showed that allogenic donor corneal epithelium cells could survive in recipients after a long-term observation.

CONCLUSIONS

Acellular porcine corneal stroma shows a satisfying biocompatibility. Tissue engineered corneal epithelium using acellular porcine corneal stroma as carrier could be used as donors for LK with satisfactory results. Donor cells have the potential to survive in recipients after long-term observation.

摘要

目的

评估异种脱细胞角膜基质的生物相容性、组织工程角膜上皮移植的可行性,并验证同种异体上皮移植的长期存活情况。

方法

这是一项实验研究。用 dispase 处理猪角膜基质,随后用 Triton-X-100 去污剂处理。将处理后的猪角膜基质(A 组)或新鲜角膜基质(B 组)植入兔角膜囊内。未植入猪角膜基质的兔角膜作为对照组(C 组)。在第 1、3、6 个月通过形态学、组织病理学和免疫组织化学评估免疫排斥反应。雌性兔接受使用雄性组织工程角膜上皮作为供体的板层角膜移植术(LK),并分析 LK 后的免疫排斥反应。在 LK 后第 1、3、7、15 和 30 天收集角膜,通过组织病理学、免疫组织化学和 Y 染色体性别决定区(SRY)-PCR 分析进行评估。

结果

处理后的猪角膜基质移植后所有角膜逐渐变透明,未发生排斥反应。组织学观察显示,A 组和 B 组所有角膜的上皮、基质、内皮、Bowman 膜和 Descemet 膜均得以保留,胶原纤维平行排列,脱细胞和新鲜角膜基质中有少量角膜细胞。C 组角膜组织学切片正常。所有角膜均未观察到明显的免疫排斥反应。组织工程角膜上皮移植研究中的角膜在 3 或 4 天顺利恢复,术后 15 或 20 天变透明,荧光素染色阴性。LK 后 15 和 30 天可见分化良好 的角膜上皮。许多角膜细胞浸润到支架中。SRY-PCR 分析显示,长期观察后同种异体供体角膜上皮细胞可在受体中存活。

结论

脱细胞猪角膜基质具有良好的生物相容性。以脱细胞猪角膜基质为载体的组织工程角膜上皮可作为 LK 的供体,效果满意。长期观察后供体细胞有在受体中存活的潜力。

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