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[脱细胞异种真皮基质用于眼睑重建的实验研究]

[Experimental study on eyelid reconstruction with acellular xenogenic dermal matrix].

作者信息

Ren Bai-Chao, Zhao Jia-Qi, Zhang Juan

机构信息

Department of Ophthalmology, The Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an, China.

出版信息

Zhonghua Yan Ke Za Zhi. 2007 Oct;43(10):906-11.

Abstract

OBJECTIVE

To investigate the histocompatibility of acellular xenogenic pig dermal matrix (Xeno-ADM) after transplanted into the rabbit eyelids.

METHODS

Thirty-eight New Zealand rabbits were divided into two groups randomly. After excising a 5 mm x 4 mm tarso-conjunctiva flap from one lower eyelid of each rabbit, Xeno-ADM and allogenic sclera were implanted in these two groups separately. Samples of implanted material were collected for histological examination in 1, 2, 4, 6, 8, 12 and 16 weeks after the operation. RIA method was employed to determine the contents of IL-2 and IL-6 in the homogenate fluid from local tissues and in peripheral blood.

RESULTS

Compared to the control group, after the operation, the conjunctival congestion, effusion, inflammatory reaction and red swelling of the eyelid subsided faster in the treated group. No rejection or necrosis occurred in transplanted rabbits. At 16 weeks after operation, SNK-q test showed there was no obvious difference in the average breadth and height of the eyelids (P > 0.05) between these two groups and normal rabbits. Histological examination of sections of eyelid tissues stained by HE staining and observed under light microscope showed that at 1, 2, 4, 6, 8, 12 and 16 weeks postoperatively, eyelids implanted with Xeno-ADM had less inflammatory reaction, fewer lymphocytes infiltrating and higher vascularization with faster ingrowths of new collagen fibers, as compared with the sclera-implanted group. This also indicated that Xeno-ADM had a good compatibility. Statistical analysis of immunogenic indexes indicated that the level of IL-2 and IL-6 in the homogenate fluid at 6-8 weeks postoperatively was greater than those in 1, 2, 4 weeks after implantation. The level of IL-2 and IL-6 rose significantly after the operation and reached a peak value at the 6 th week (experimental group, IL-2, 0.292 81 ng/ml) and 8 th week (experimental group, IL-6, 118.258 pg/ml; control group, IL-2, 0.277 99 ng/ml and IL-6, 255.871 pg/ml). The level of IL-2 and IL-6 dropped significantly at the 12th week and raised again at the 16th week, therefore, the concentration achieved another peak value in the control group (IL-2, 1.363 41 ng/ml; IL-6, 622.863 pg/ml), which was much higher than those in the first peak. The level of IL-2 and IL-6 in peripheral blood 1 week after the transplantation was higher than the preoperative level. These two indexes increased gradually with marked fluctuation, particularly in 4 - 8 weeks after implantation, both of them reached the peak values at that period, which was similar to the inflammation reaction changes in the implanted groups. With replicated testing and analysis of variance (ANOVA), neither the type of treatment nor the time factor had any effect on the levels of IL-2 and IL-6 in the serum, and there was no significant difference between experimental group and control group (P > 0.05).

CONCLUSIONS

Xeno-ADM shows good histocompatibility to New Zealand rabbits, which can induce neovascular and collagen fibers grow into implanted tissues. Therefore, this can be used to support the eyelids as a substitution for the tarsus. High levels of IL-2 and IL-6 can be detected in local tissues and in the serum after Xeno-ADM transplantation, but no statistical differences present between the dynamic changes of the level of both ILs in Xeno-ADM group and allogenic sclera group. Xeno-ADM could be an ideal substitute for allogenic sclera in the reconstruction of eye lids and could be used broadly in ophthalmology. However, as a hetero-transplantation material used clinically, further ethical and immunological studies are required.

摘要

目的

研究脱细胞异种猪真皮基质(Xeno-ADM)植入兔眼睑后的组织相容性。

方法

将38只新西兰兔随机分为两组。从每只兔的一侧下眼睑切除一块5mm×4mm的睑结膜瓣后,分别在两组植入Xeno-ADM和同种异体巩膜。术后1、2、4、6、8、12和16周采集植入材料样本进行组织学检查。采用放射免疫分析法(RIA法)测定局部组织匀浆和外周血中白细胞介素-2(IL-2)和白细胞介素-6(IL-6)的含量。

结果

与对照组相比,术后治疗组结膜充血、渗出、炎症反应及眼睑红肿消退更快。移植兔未发生排斥或坏死。术后16周,SNK-q检验显示两组眼睑的平均宽度和高度与正常兔相比无明显差异(P>0.05)。苏木精-伊红(HE)染色的眼睑组织切片在光学显微镜下观察的组织学检查显示,术后1、2、4、6、8、12和16周,植入Xeno-ADM的眼睑炎症反应较轻,淋巴细胞浸润较少,血管化程度较高,新生胶原纤维向内生长较快,与植入巩膜组相比。这也表明Xeno-ADM具有良好的相容性。免疫原性指标的统计分析表明,术后6 - 8周局部组织匀浆中IL-2和IL-6水平高于植入后1、2、4周。术后IL-2和IL-6水平显著升高,在第6周(实验组,IL-2,0.292 81 ng/ml)和第8周(实验组,IL-6,118.258 pg/ml;对照组,IL-2,0.277 99 ng/ml和IL-6,255.871 pg/ml)达到峰值。第12周IL-2和IL-6水平显著下降,第16周再次升高,因此,对照组浓度达到另一个峰值(IL-2,1.363 41 ng/ml;IL-6,622.863 pg/ml),远高于第一个峰值。移植后1周外周血中IL-2和IL-6水平高于术前水平。这两个指标逐渐升高且波动明显,尤其是在植入后4 - 8周,两者在此期间均达到峰值,这与植入组的炎症反应变化相似。经重复检验和方差分析(ANOVA),治疗类型和时间因素对血清中IL-2和IL-6水平均无影响,实验组与对照组之间无显著差异(P>0.05)。

结论

Xeno-ADM对新西兰兔显示出良好的组织相容性,可诱导新生血管和胶原纤维长入植入组织。因此,可作为睑板替代物用于支撑眼睑。Xeno-ADM移植后局部组织和血清中可检测到高水平的IL-2和IL-6,但Xeno-ADM组和同种异体巩膜组两者水平的动态变化无统计学差异。Xeno-ADM可能是眼睑重建中同种异体巩膜的理想替代物,可广泛应用于眼科。然而,作为临床使用的异种移植材料,还需要进一步的伦理学和免疫学研究。

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