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丝素膜移植兔眼睑原位重建的研究

Experimental study on the eyelid reconstruction in situ with the silk fibroin membrane

摘要背景 利用自体或异体组织进行活体组织重建存在着供体材料少、易发生排斥反应等问题.研究证实,作为组织工程细胞支架,丝素膜有良好的生物相容性,但其是否可作为组织替代物尚未见报道.目的 探讨应用丝素膜行眼睑原位重建术的可行性.方法健康新西兰大白兔18只,双眼上眼睑制作眼睑板缺损4 mmx3 mm模型,右眼采用再生丝素膜材料进行上睑板重建术为丝素膜组,左眼采用同种异体巩膜材料行睑板重建为异体巩膜组,活体观察4周.采用随机数字表法将实验兔分为3组,每组6只,分别于术后第1、2、4周用空气栓塞法处死实验兔,收集带有移植片的兔眼睑制作石蜡切片,苏木精—伊红染色观察术区炎性细胞的浸润情况;用Masson染色法观察植片区胶原组织的形成和分布情况;应用免疫组织化学法检测植入区碱性成纤维细胞生长因子(bFGF)的表达;采用ImagePro Plus分析软件对免疫组织化学结果进行半定量分析.结果所有兔眼眼睑缺损均Ⅰ期愈合,睑结膜面光滑,眼表炎症反应不明显,但异体巩膜组睑缘切迹较丝素膜组明显.组织学检查结果显示,术后随着时间的延长,移植区炎性细胞浸润逐渐减轻,术后4周丝素膜组术区胶原纤维排列整齐,结缔组织增生不明显;异体巩膜组术区胶原纤维排列相对紊乱,瘢痕组织增生较明显.术后第1、2、4周,丝素膜组术区组织中bFGF的表达量(A值)分别为0.027 67±0.004 69、0.051 73±0.008 72、0.058 72±0.006 88,巩膜组分别为0.056 48±0.009 14、0.072 83±0.009 17和0.078 73±0.010 84,两组各时间点bFGF表达量的差异均有统计学意义(t=-6.38、t=-4.99、t=-2.87,P<0.05).结论丝素膜作为眼睑原位重建术睑板的替代物组织相容性好,能有效恢复睑板形态,可替代异体巩膜重建眼睑.

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abstractsBackground Autologous and allograft renal transplantation exist some disadvantages of less donor source and rejection.As a scaffold of cell in tissue engineering,fibroin was determined to have a good biocompatibility.But whether the fibroin membrane can become a substitution for tissue defect is seldom reported.Objective This experiment aimed to investigate the feasibility of silk fibroin membrane in the rabbit eyelid reconstruction in situ.Methods A 4 mmx3 mm tarsi defect model was created on the upper eyelids of 18 healthy New Zealand white rabbits.The eyelid reconstruction in situ was performed with regenerated silk fibroin membrane material in the right upper eyelids (silk fibroin group ) and allogenic sclera material (sclera group ) on the upper eyelids of fellow eyes.The grafts were clinically examined for the evaluation of inflammation and implant exposure at the first,second and forth week after operation.The inflammation response and collagen distribution were examined by hematoxylin & eosin staining and Masson staining.Expression of basic fibroblast growth factor(bFGF) in the grafts was detected by immunohistochemistry,and ImagePro Plus software was used for statistical analysis.Results All eyelid defects showed a primary healing.The surface of palpebral conjunctival was smooth and the inflammation of ocular surface was mild.The eyelid margin in the sclera group was more notch than that in the silk fibroin group.Results of pathological examination revealed that the arrangement of collagen fibers in the sclera group was more disordered,but that in the silk fibroin group was regular.The expression level(A value) of b-FGF in the operative area in silk fibroin group were 0.027 67±0.004 69,0.051 73±0.008 72,0.058 72±0.006 88,and those in the sclera group were 0.056 48±0.009 14,0.072 83 ± 0.009 17 and 0.078 73 ±0.010 84 in 1,2,4 weeks after operation,showing statistically significant differences between two groups in various time points ( t =- 6.38,t =- 4.99,t =- 2.87,P <0.05 ).Conclusions Silk fibroin membrane can reconstruct the eyelid shape in situ with the less inflammation response and good biocompatibility.Silk fibroin membrane could be used to support the eyelid as a new tarsal repairing materials.

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中华实验眼科杂志

中华实验眼科杂志

2011年29卷11期

968-972页

ISTICPKUCSCDCA

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