中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2011年
12期
1065-1070
,共6页
戎慧丰%赵青%颜华%刘贵培%刘源%刘文广
戎慧豐%趙青%顏華%劉貴培%劉源%劉文廣
융혜봉%조청%안화%류귀배%류원%류문엄
角膜移植%重组人Ⅲ型胶原/聚[3-(甲基丙烯酰胺)丙基—二甲基(3-磺丙)胺]互穿聚合物网络水凝胶%组织工程角膜%神经生长因子%生物相容性
角膜移植%重組人Ⅲ型膠原/聚[3-(甲基丙烯酰胺)丙基—二甲基(3-磺丙)胺]互穿聚閤物網絡水凝膠%組織工程角膜%神經生長因子%生物相容性
각막이식%중조인Ⅲ형효원/취[3-(갑기병희선알)병기—이갑기(3-광병)알]호천취합물망락수응효%조직공정각막%신경생장인자%생물상용성
Keratoplasty%Recombinant human type-Ⅲ collagen/poly ( (3- (methacryloylamino) propyl)dimethyl (3-sulfopropyl) ammonium hydroxide) interpenetrating polymer network hydrogel%Tissue-engineered cornea%Nerve growth factor%Biocompatibility
背景 组织工程角膜由于更接近活体角膜形态,植入体内易于成活而成为当今眼科学关注的热点.目的 探讨组织工程角膜重组人Ⅲ型胶原(RHC-Ⅲ)/聚[3-(甲基丙烯酰胺)丙基—二甲基(3-磺丙)胺] (PMPDSAH)互穿聚合物网络(IPN)(RHC-Ⅲ/PMPDSAH IPN)水凝胶植入兔眼的生物相容性及其作为角膜替代物的可行性.方法 中国大耳白兔108只按照随机数字表法分为2个组,实验组90只,正常对照组3只,其他15只兔(30只眼)为同种异体移植组提供供体角膜.实验组根据角膜材料的不同分为RHC-Ⅲ/PMPDSAH IPN组、负载神经生长因子(NGF)的NGF RHC-Ⅲ/PMPDSAH IPN组和同种异体移植组,每组各30只兔,均取右眼为手术眼.将实验组3种材料移植入兔角膜前板层,术后裂隙灯下观察有无排斥反应,对角膜透明度、角膜新生血管(CNV)的变化进行评分,记录各组兔术眼角膜上皮化时间,观察期为6个月.分别于术后3d、1周、2周及术后1、3、6个月每组各取5只兔角膜行苏木精—伊红染色,术后6个月采用免疫组织化学法检测角膜上皮细胞特异性标志蛋白角蛋白K3的表达.对实验组3个亚组各时间点角膜透明度和CNV评分的差异比较进行多组独立样本的Kraskal-WallisH检验,3个亚组角膜上皮化时间比较采用单因素方差分析及LSD-t检验.结果 实验的6个月中,RHC-Ⅲ/PMPDSAH IPN组、NGF RHC-Ⅲ/PMPDSAH IPN组和同种异体移植组兔术眼的植片均未脱出,未见免疫排斥反应.术后6个月,RHC-Ⅲ/PMPDSAH IPN组、NGFRHC-Ⅲ/PMPDSAH IPN组和同种异体移植组角膜透明度、CNV与正常对照组比较差异均无统计学意义(H=4.34,P=0.23;H=2.60,P=0.46).NGF RHC-Ⅲ/PMPDSAH IPN组角膜上皮化时间最短,平均为(4.97±0.63)d,与RHC-Ⅲ/PMPDSAH IPN组和同种异体移植组比较差异均有统计学意义(t=11.97,P=0.00；t=5.80,P=0.00)；RHC-Ⅲ/PMPDSAH IPN组和同种异体移植组比较差异有统计学意义(t=6.32,P=0.00).RHC-Ⅲ/PMPDSAH IPN组和NGF RHC-Ⅲ/PMPDSAH IPN组移植材料与兔角膜融合较好,术后2周材料部分降解,术后1个月材料均完全降解,术后6个月新生胶原纤维排列整齐,角膜上皮细胞中K3表达阳性.结论 RHC-Ⅲ/PMPDSAH IPN与兔眼的生物相容性较好,NGF可以有效地促进角膜创口愈合和上皮再生,提高其力学强度后,有望作为一种安全有效的角膜替代物.
揹景 組織工程角膜由于更接近活體角膜形態,植入體內易于成活而成為噹今眼科學關註的熱點.目的 探討組織工程角膜重組人Ⅲ型膠原(RHC-Ⅲ)/聚[3-(甲基丙烯酰胺)丙基—二甲基(3-磺丙)胺] (PMPDSAH)互穿聚閤物網絡(IPN)(RHC-Ⅲ/PMPDSAH IPN)水凝膠植入兔眼的生物相容性及其作為角膜替代物的可行性.方法 中國大耳白兔108隻按照隨機數字錶法分為2箇組,實驗組90隻,正常對照組3隻,其他15隻兔(30隻眼)為同種異體移植組提供供體角膜.實驗組根據角膜材料的不同分為RHC-Ⅲ/PMPDSAH IPN組、負載神經生長因子(NGF)的NGF RHC-Ⅲ/PMPDSAH IPN組和同種異體移植組,每組各30隻兔,均取右眼為手術眼.將實驗組3種材料移植入兔角膜前闆層,術後裂隙燈下觀察有無排斥反應,對角膜透明度、角膜新生血管(CNV)的變化進行評分,記錄各組兔術眼角膜上皮化時間,觀察期為6箇月.分彆于術後3d、1週、2週及術後1、3、6箇月每組各取5隻兔角膜行囌木精—伊紅染色,術後6箇月採用免疫組織化學法檢測角膜上皮細胞特異性標誌蛋白角蛋白K3的錶達.對實驗組3箇亞組各時間點角膜透明度和CNV評分的差異比較進行多組獨立樣本的Kraskal-WallisH檢驗,3箇亞組角膜上皮化時間比較採用單因素方差分析及LSD-t檢驗.結果 實驗的6箇月中,RHC-Ⅲ/PMPDSAH IPN組、NGF RHC-Ⅲ/PMPDSAH IPN組和同種異體移植組兔術眼的植片均未脫齣,未見免疫排斥反應.術後6箇月,RHC-Ⅲ/PMPDSAH IPN組、NGFRHC-Ⅲ/PMPDSAH IPN組和同種異體移植組角膜透明度、CNV與正常對照組比較差異均無統計學意義(H=4.34,P=0.23;H=2.60,P=0.46).NGF RHC-Ⅲ/PMPDSAH IPN組角膜上皮化時間最短,平均為(4.97±0.63)d,與RHC-Ⅲ/PMPDSAH IPN組和同種異體移植組比較差異均有統計學意義(t=11.97,P=0.00；t=5.80,P=0.00)；RHC-Ⅲ/PMPDSAH IPN組和同種異體移植組比較差異有統計學意義(t=6.32,P=0.00).RHC-Ⅲ/PMPDSAH IPN組和NGF RHC-Ⅲ/PMPDSAH IPN組移植材料與兔角膜融閤較好,術後2週材料部分降解,術後1箇月材料均完全降解,術後6箇月新生膠原纖維排列整齊,角膜上皮細胞中K3錶達暘性.結論 RHC-Ⅲ/PMPDSAH IPN與兔眼的生物相容性較好,NGF可以有效地促進角膜創口愈閤和上皮再生,提高其力學彊度後,有望作為一種安全有效的角膜替代物.
배경 조직공정각막유우경접근활체각막형태,식입체내역우성활이성위당금안과학관주적열점.목적 탐토조직공정각막중조인Ⅲ형효원(RHC-Ⅲ)/취[3-(갑기병희선알)병기—이갑기(3-광병)알] (PMPDSAH)호천취합물망락(IPN)(RHC-Ⅲ/PMPDSAH IPN)수응효식입토안적생물상용성급기작위각막체대물적가행성.방법 중국대이백토108지안조수궤수자표법분위2개조,실험조90지,정상대조조3지,기타15지토(30지안)위동충이체이식조제공공체각막.실험조근거각막재료적불동분위RHC-Ⅲ/PMPDSAH IPN조、부재신경생장인자(NGF)적NGF RHC-Ⅲ/PMPDSAH IPN조화동충이체이식조,매조각30지토,균취우안위수술안.장실험조3충재료이식입토각막전판층,술후렬극등하관찰유무배척반응,대각막투명도、각막신생혈관(CNV)적변화진행평분,기록각조토술안각막상피화시간,관찰기위6개월.분별우술후3d、1주、2주급술후1、3、6개월매조각취5지토각막행소목정—이홍염색,술후6개월채용면역조직화학법검측각막상피세포특이성표지단백각단백K3적표체.대실험조3개아조각시간점각막투명도화CNV평분적차이비교진행다조독립양본적Kraskal-WallisH검험,3개아조각막상피화시간비교채용단인소방차분석급LSD-t검험.결과 실험적6개월중,RHC-Ⅲ/PMPDSAH IPN조、NGF RHC-Ⅲ/PMPDSAH IPN조화동충이체이식조토술안적식편균미탈출,미견면역배척반응.술후6개월,RHC-Ⅲ/PMPDSAH IPN조、NGFRHC-Ⅲ/PMPDSAH IPN조화동충이체이식조각막투명도、CNV여정상대조조비교차이균무통계학의의(H=4.34,P=0.23;H=2.60,P=0.46).NGF RHC-Ⅲ/PMPDSAH IPN조각막상피화시간최단,평균위(4.97±0.63)d,여RHC-Ⅲ/PMPDSAH IPN조화동충이체이식조비교차이균유통계학의의(t=11.97,P=0.00；t=5.80,P=0.00)；RHC-Ⅲ/PMPDSAH IPN조화동충이체이식조비교차이유통계학의의(t=6.32,P=0.00).RHC-Ⅲ/PMPDSAH IPN조화NGF RHC-Ⅲ/PMPDSAH IPN조이식재료여토각막융합교호,술후2주재료부분강해,술후1개월재료균완전강해,술후6개월신생효원섬유배렬정제,각막상피세포중K3표체양성.결론 RHC-Ⅲ/PMPDSAH IPN여토안적생물상용성교호,NGF가이유효지촉진각막창구유합화상피재생,제고기역학강도후,유망작위일충안전유효적각막체대물.
Background The tissue-engineered cornea is becoming the hot spot in the ophthalmologic field,while the research of corneal substitute is in the ascendant,because it is more similar to the corneal morpha and easy to survive in vivo.Objective This study was to investigate the biocompatibility of recombinant human type-Ⅲ collagen/poly9 ( 3-( methacryloylamino ) propyl dimethyl ( 3-sulfopropyl ) ammonium hydroxide ) ( PMPDSAH ) interpenetrating polymer network (IPN) (RHC-Ⅲ/PMPDSAH IPN) hydrogel as a tissue-engineered cornea in rabbit eye and its feasibility as the corneal substitute.Methods One hundred and eight rabbits were randomly divided into experimental group( 90 rabbits) and normal control group ( 3 rabbits),and 15 rabbits ( 30 eyes ) used as the donor corneas.RHC-Ⅲ/PMPDSAH IPN,NGF PMPDSAH IPN and corneal grafts were lamellarly transplanted into the right eyes in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group respectively.The corneal transparency and neovascularization were examined and scored under the slim lamp and compared among three groups using Kraskal-Wallis H test.The corneal epithelization time was observed and compared among these three groups using one way analysis of variance and LSD-t test.The histological examination of corneas was performed at the 3rd day,1st and 2nd week,1 st,3rd and 6th month after the surgery.The immunohistochemistry was used to detect the expression of K3 in cornea at the 6th month.Results The grafts were well attached in RHC-Ⅲ/PMPDSAH IPN group,NGF PMPDSAH IPN group and allograft group,and no rejection reaction was found throughout 6-month following up.Compared with normal control group,no significant differences were found in the scores of corneal opacification and neovescularization in these three groups (x2 =4.34,P =0.23 ;x2 =2.60,P =0.46 ) at the 6th month.NGF PMPDSAH IPN group achieved reepithelialization in (4.97±0.63) days and was obviously shorted than that in RHC-Ⅲ/PMPDSAH IPN group and allograft group ( t =11.97,P =0.00; t =5.80,P =0.00).The re-epithelialization time in RHC-Ⅲ/PMPDSAH IPN was (6.86±0.71) days,and that of allograft group was (5.87±0.43 ) days,showing a significant difference ( t =6.32,P =0.00).Hematoxylin-eosin staining results demonstrated that implanted materials integrated into the host corneal tissue well and support corneal epithelialization.Part of the material degraded at the 2nd week and degraded completely 1 month later.Regular alignment and distribution of collagen fibers were seen in the regenerated cornea and were similar to those of the normal stroma in 6 months.Immunohistochemistry showed the positive expression of keratin-3 in corneal epithelial cells.Conclusions RHC-Ⅲ/PMPDSAH IPN has a good biocompatibility without toxicity to corneal tissue.Furthermore,NGF can promote the corneal wound-healing and re-epithelialization.The material can be used as safe and reliable corneal substitute after improving the mechanical strength.
