羊膜与SK GEL生物胶在兔非穿透性小梁切除术中的应用

目的探讨保存人羊膜组织、sKGEL生物胶（交键透明质酸钠凝胶）在兔非穿透性小梁切除术中抗瘢痕化的安全性及有效性。方法新西兰白兔18只，随机分为A、B、c3组，每组各6只，每组兔的单眼行非穿透性小梁切除术。A组术中植入羊膜，B组术中植入SKGEL生物胶，C组术中无植入物。观察各组滤过泡的形态、内部结构及维持时间。结果A、B组滤过泡的形态及维持时间均优于c组。病理切片显示滤过空腔的保持与植入物有关，术后12周时A、B组眼空腔仍维持较好．c组眼瘢痕形成明显。结论保存人羊膜、SKGEL生物胶在兔非穿透性小梁切除术中应用均可减轻手术区滤过腔瘢痕的形成，安全有效。[眼科新进展2007；27（2）：106—109]     

蛇毒制剂对兔眼滤过术的影响

目的评价蛇毒制剂在兔眼滤过术后抗瘢痕形成的效果和眼毒性.方法对10只兔双眼行巩膜全层咬切术,术后实验组球结膜下注射5×10-2U/ml的蛇毒制剂0.5ml,对照组注射等量生理盐水,裂隙灯观察眼前段.术后第21天,用鼠尾胶原作为房水外流示踪剂行前房恒压灌注及原位固定,对手术区组织行功能及组织形态学检查.结果术后14,21天,实验组有5眼、3眼可见滤过泡,对照组无1眼有滤过泡.滤过泡消失眼的滤过口被瘢痕组织封闭,手术区结膜下纤维组织明显增生.有滤过泡眼的滤过口仍显开放,房水示踪剂经此开口流入滤过泡.结论蛇毒制剂能抑制兔眼滤过术后结膜下过分瘢痕化,延长滤过泡的存在时间和功能,其眼毒性不明显.     

羊膜植片在穿透性滤过手术中抑制巩膜瓣下纤维增殖的研究

目的探讨羊膜植片在兔眼穿透性滤过手术中抗纤维增殖情况。方法将羊膜上皮面朝外折叠成双层植入穿透性滤过手术的兔眼浅层巩膜瓣下。于术后不同时间点,行眼球组织学切片及运用显微图像分析系统对滤过道内成纤维细胞进行记数、分析。结果术后2周滤过道内开始出现成纤维细胞,随时间延长成纤维细胞数目逐渐增多,但无统计学差异(P>0.05)。对照组由术后1周开始出现纤维增殖,且逐渐加重,术后2周滤过道大部分封闭,术后3周已有新生血管长入。实验组和对照组的成纤维细胞数在术后1、2、3、4周分别进行比较,有显著性差异(P<0.05)。结论羊膜植片应用于穿透性滤过手术,可明显抑制滤过道使成纤维细胞增殖,滤过道保持通畅。     

羊膜对兔眼小梁切除术后滤过泡的影响

目的探讨羊膜对小梁切除术后滤过泡的影响。 方法对12只新西兰白兔双眼行小梁切除术,术中l眼于巩膜和巩膜瓣间放置6mm×4mm的羊 膜植片,另一眼作为对照。术后1周、2周、3周检查滤过泡形态和功能,光镜下观察滤过泡下巩膜 表面瘢痕形成情况。 结果实验组术后1、2、3周滤过泡轻微隆起弥散,有较好的滤过功能,对照组术后2、3周滤过泡 区瘢痕形成,滤过功能差。组织切片观察显示实验组滤过泡未形成瘢痕,成纤维细胞少,但有炎性 细胞浸润;对照组滤过泡区被增生的纤维组织所代替,成纤维细胞丰富。 结论羊膜能改善滤过泡功能,减少瘢痕形成。眼科学报2000;1673-76。     

非穿透性小梁切除术联合低浓度丝裂霉素C的实验研究

目的探讨应用低浓度丝裂霉素C的非穿透性小梁切除术后手术区的形态学变化及房水引流途径.方法新西兰白兔12只(24眼),随机分为实验组9只(18眼)和正常对照组3只(6眼).实验组兔双眼行非穿透性小梁切除术,分剐于术后1d、14d、1个月时随机选择3只兔,一眼前房注入辣根过氧化物酶(horseradish pemxidase,HRP),另一眼前房注入相对分子质量为70 000四甲基罗丹明标记的葡聚糖(tetramethyl rhodamine-dextran,TMR-D).60 min后经颈动脉灌注固定,摘取眼球,冰冻切片,观察示踪剂的分布及手术区的形态学变化.正常对照组3只兔,双眼未行手术,示踪剂前房注入方法同前.结果实验组兔眼滤过泡及浅层巩膜瓣下的滤过腔于术后1d及14d时可见HRP强染色.术后14d时,滤过腔内未见明显纤维组织形成,滤过腔周围观膜内有HRP散在染色.术后1个月时,滤过腔局限,少量HRP颗粒分布,滤过腔见纤维组织增生.正常对照组结膜下仅微弱HRP显色,巩膜内无HRP颗粒分布.实验组及正常对照组脉络膜均可见TMR-D红色荧光.结论非穿透性小梁切除术后房水引流途径涉及结膜下外滤过、巩膜内吸收及葡萄膜巩膜引流.     

异体角膜缘上皮细胞培养后移植的实验研究

目的观察含培养的角膜缘上皮细胞的羊膜片移植到角膜缘干细胞损伤的异体兔眼角膜上的效果。方法把羊膜为载体组织块培养的角膜缘上皮细胞移植到角膜缘干细胞损伤的异体兔眼角膜上,术后观察角膜混浊度、角膜新生血管、角膜上皮等情况,定期兔进行病理组织学检查。结果角膜缘上皮细胞在羊膜上培养16d形成3～4层,用含有培养的异体角膜上皮细胞的羊膜片移植的12只兔眼,术后第5天,损伤的角膜表面全部上皮化,第16天开始,12只兔眼逐渐出现排斥反应。结论含培养的角膜缘上皮细胞羊膜片异体移植术后早期角膜全部上皮化,晚期则出现排斥反应。     

羊膜植片应用于兔眼穿透性滤过术局部变化特点

目的:探讨羊膜移植应用于兔眼穿透性滤过手术手术区细胞病理学变化特点。方法:选取科研用白色家兔20只,双眼行穿透性滤过手术,术中将羊膜上皮面朝外折叠成双层植入于浅层巩膜瓣下。于术后不同时间点观察巩膜滤过道内细胞病理学改变。结果:实验组术后1wk手术区大量炎细胞,随后炎细胞数量减少,并有少量杆状核细胞出现,术后3wk炎症反应达最高峰,随后炎细胞数量再次减少,术后8wk羊膜完全溶解,滤过道内仅存少量异物巨细胞,并于术后12wk完全消失,滤过道保持开放。在整个实验观察过程中,实验组手术区成纤维细胞数量无明显变化(P >0.05),但与对照组相比较有明显统计学差异(P <0.05)。结论:羊膜植片在术后8wk完全溶解。在实验动物中将羊膜应用于穿透性滤过手术可明显抑制成纤维细胞增殖,保持滤过道开放。     

非穿透性小梁手术联合羊膜移植治疗开角型青光眼

目的 :探讨非穿透性小梁手术 (NPTS)联合羊膜移植治疗开角型青光眼的机制及疗效评价。寻找透明质酸钠凝胶植入材料替代物。方法 :对 18例 2 5只开角型青光眼患者行非穿透性小梁手术联合羊膜移植 ,术后观察视力、眼压、滤过泡及眼内反应 ,随访时间最短 30天 ,最长 4 2 0天。结果 :术前 2 5只眼平均眼压为 (38 2± 14 4 )mmHg ,2 5只眼术后 3只眼眼压高于 2 1mmHg ,经降眼压药物治疗后 2只眼仍高达 2 8mmHg以上 ,余眼压均控制在 7～ 2 0mmHg之内 ,术后 2个月手术成功率为 88% ,条件成功率为 92 % ,术后 3个月手术成功率为 82 % ,条件成功率为 88%。术后视力均有显著提高 ,滤过泡扁平弥漫 ,轻微充血。术中、术后未出现浅前房、前房出血、玻璃体脱出、脉络膜脱离等并发症。结论 :非穿透性小梁手术联合羊膜移植能安全、有效地降低眼压 ,为非穿透性小梁手术提供了有效安全植入材料。可广泛用于开角型青光眼的治疗     

羊膜移植治疗大泡性角膜病变的临床应用

目的：观察羊膜移植治疗大泡性角膜病变的疗效。方法：对22例22只眼大泡性角膜病变，其中白内障术后无晶状体眼2例，人工晶状体12例，穿透性角膜移植术后3例，绝对期青光眼3例，青光眼白内障术后2例，进行羊膜移植手术并观察治疗效果。结果：18只眼术后第1天症状缓解，所有病例术后5天症状消失，19只眼角膜缺损区在3周内愈合，经6个月－2年的随访，无一例复发，亦未见新生血管和翼状胬肉的发生。结论：羊膜移植是治疗大泡性角膜病变可选择的有效方法。     

羊膜移植联合鸡角膜缘上皮移植治疗兔全角膜表层损伤的组织学改变

目的：研究羊膜移植术和联合鸡角膜缘上皮移植（联合移植）术治疗兔全角膜表层损伤的组织学改变。方法：兔39只分为3组，每组13只。对照组，将角膜及角膜缘外3mm的表层组织去除；羊膜移植组，在去除表层组织的创面上移植羊膜；联合移植组，在羊膜移植片表面角膜缘区移植鸡角膜缘上皮。手术当天各组处死1只兔，7、28、90、105天各组处死3只兔，摘除眼球分别进行光镜、免疫组化，透射电镜及扫描电镜检查。结果：对照组：角膜紊乱，表面不平，上皮表面微绒毛稀少，基质纤维血管化。羊膜移植组及联合移植组：羊膜结构逐渐被基质胶元纤维取代和改建，基质纤维平行排列，角膜表面光滑，上皮表面微绒毛丰富，细胞之间可见桥粒连接。结论：羊膜移植术和联合鸡角膜缘移植术治疗兔角膜广泛损伤，能抑制新生血管和纤维组织向角膜生长，较快促进眼表重建。     

青光眼滤过术中羊膜、丝裂霉素和透明质酸钠抗增殖作用的比较

目的:探讨青光眼滤过术中羊膜、丝裂霉素(mitomycin C,MMC)和透明质酸(Healon)抗增殖作用。方法:健康同种系新西兰大白兔15只30眼随机选5只为1组,共3组:羊膜组、MMC组和Healon组。均行小梁切除术,在作巩膜瓣后,羊膜组在巩膜瓣下放置保存羊膜;MMC组在巩膜瓣下放置0.25g/L的MMC海绵片4min;Healon组在巩膜瓣下放置透明质酸钠。术后观察滤过泡、角膜和眼底情况。结果:术后28d取滤过泡周围组织作病理检查,羊膜组、MMC组多数形成功能性滤过泡,但羊膜组无任何并发症,病理未见结缔组织胶原化。结论:羊膜在青光眼滤过术中抗增殖作用较MMC,Healon效果好,无副作用,是一种安全有效的抗增殖方法,值得推广。     

兔眼滤过手术模型制作探析

目的:探讨建立长期有效的兔眼滤过手术模型.方法:对18只36眼家兔行青光眼滤过手术,其中右眼植入单层生物羊膜,左眼植入双层生物羊膜,术后观察眼压、滤过泡形态.结果:家兔术后巩膜瓣修复快,滤过通道难以长期有效维持,易导致造模失败.术中植入双层羊膜,可明显延长有效滤过通道的维持时间,部分模型有效滤过可维持14~30d.双层羊膜组眼压控制也较理想.结论:兔眼小梁切除术中植入双层羊膜,可以提高滤过模型的成功率.     

Inhibitory effect of biopolymer materials on scar formation following trabeculectomy

AIM: To investigate the inhibitory effects of amniotic membrane, polylactic acid membrane and chitosan membrane on scar formation following trabeculectomy. METHODS: A total of 24 New Zealand white rabbits (48 eyes) were randomly divided into 4 groups: amniotic membrane group, polylactic acid membrane group, chitosan membrane group, and control group, with 6 rabbits (12 eyes) in each group. The left eyes underwent routine trabeculectomy, and the right eyes were considered as controls. Amniotic membrane, polylactic acid membrane and chitosan membrane were respectively installed under sclera flap in three groups, but any treatment was not applied in control group. Intraocular pressure, conjunctival filtering bleb, and anterior chamber inflammation responses were monitored at day 1, 3, 7, 14, 28 and 56 post-operatively. Eyeball tissue underwent histopathological examination at day 56 post-operatively. RESULTS: Fibrocytes and inflammatory cells were reduced in amniotic membrane, polylactic acid membrane and chitosan membrane groups compared to that in control group. At day 1 post-operatively, intraocular pressure was decreased in three membrane groups compared to that in control group. At day 14 post-operatively, the intraocular pressure was decreased significantly, while it of three membrane groups was significantly lower than that of preoperative (P<0.01). There were no significant differences among three membrane groups (P>0.05). Filtering bleb of four groups was clearly observed at day 7 post-operatively, but there was no significant difference in pair-wise comparison. At day 28 and 56 post-operatively, filtering bleb in control group was significantly narrowed compared to that in three membrane groups (P<0.05), but there was no significant difference in pair-wise comparison of three membrane groups. CONCLUSION: All amniotic membrane, polylactic acid membrane and chitosan membrane can effectively inhibit scar formation following trabeculectomy, the effect of amniotic membrane is the best.     

兔眼小梁切除术中巩膜瓣下植入陈旧生物羊膜的病理变化

目的:评价青光眼滤过手术中应用陈旧生物羊膜抗纤维增殖作用。方法:采取随机对照动物实验研究方法,选日本大耳兔30只,共60眼,采用随机数字表法将其分为两组。两组中右眼(30眼)均行小梁切除术,术中巩膜瓣层间植入陈旧生物羊膜,组A左眼(15眼)行小梁切除术联合应用干扰素α-2b,组B左眼(15眼)仅行单纯小梁切除术。并于术后3,7,14,30,60d分别处死两组中各3只动物,摘除眼球作病理切片,观察羊膜降解时间、成纤维细胞和炎症细胞计数。结果:羊膜组早期即可见大量成纤维细胞增殖,成纤维细胞处于功能活跃状态。羊膜组炎性细胞数目、成纤维细胞数目,术后3,7,14d均高于其它两组。羊膜大致在14～21d内降解。结论:兔眼小梁切除术中应用陈旧的生物羊膜诱发了兔眼的免疫排斥反应。     

Inhibitory effect of three biopolymer materials on scar formation following trabeculectomy

AIM: To investigate the inhibitory effects of amniotic membrane, polylactic acid membrane and chitosan membrane on scar formation following trabeculectomy. METHODS: A total of 24 New Zealand white rabbits (48 eyes) were randomly divided into 4 groups: amniotic membrane group, polylactic acid membrane group, chitosan membrane group, and control group, with 6 rabbits (12 eyes) in each group. The left eyes underwent routine trabeculectomy, and the right eyes were considered as controls. Amniotic membrane, polylactic acid membrane and chitosan membrane were respectively installed under sclera flap in three groups, but any treatment was not applied in control group. Intraocular pressure, conjunctival filtering bleb, and anterior chamber inflammation responses were monitored at day 1, 3, 7, 14, 28 and 56 post-operatively. Eyeball tissue underwent histopathological examination at day 56 post-operatively. RESULTS: Fibrocytes and inflammatory cells were reduced in amniotic membrane, polylactic acid membrane and chitosan membrane groups compared to that in control group. At day 1 post-operatively, intraocular pressure was decreased in three membrane groups compared to that in control group. At day 14 post-operatively, the intraocular pressure was decreased significantly, while it of three membrane groups was significantly lower than that of preoperative (P <0.01). There were no significant differences among three membrane groups (P >0.05). Filtering bleb of four groups was clearly observed at day 7 post-operatively, but there was no significant difference in pair-wise comparison. At day 28 and 56 post-operatively, filtering bleb in control group was significantly narrowed compared to that in three membrane groups (P <0.05), but there was no significant difference in pair-wise comparison of three membrane groups. CONCLUSION: All amniotic membrane, polylactic acid membrane and chitosan membrane can effectively inhibit scar formation following trabeculectomy, the effect of amniotic membrane is the best.     

Lack of Corneal Toxicity of Interferon Alpha—2b Administered Subconjunctivally after Sclerectomy

Purpose: To evaluate the corneal toxicity of subconjunctival injection interferon α-2bat filtering bleb after sclerectomy in white rabbits.Methods: Eight rabbits which had been performed sclerectomy were randomlydivided into two groups. Each group consisted of four rabbits. Eight eyes in group 1were subconjunctivally received interferon α-2b 5 × 105IU/0. 2ml into filtering blebfrom the edge of the filtering site immediately after operation and every postoperativeday. The other eight eyes in group 2 were injected with 0. 2ml normal saline. All ofthe eyes underwent daily examination by slip-lamp microscopy and directophthalmoloscopy. Sodium fluorescein was used to assess corneal epithelial integrity.On day 3,4,7 and 14, every two rabbits (group 1 and 2 each, respectively) werekilled and removed cornea immediately to take examination of the viability of cornealendothelium by dual staining with typan blue and alizanin red S.Results: No sign of toxicity in corneal epithelium and endothelium were foundfoll     

兔角膜碱烧伤新鲜羊膜移植后的角膜病理学及超微结构观察

目的观察兔角膜碱烧伤立即行新鲜羊膜移植后的角膜病理学和超微结构变化。方法40只新西兰白兔中随机取4只作为正常对照组,剩余36只制作双眼碱烧伤模型,右眼立即行新鲜羊膜移植术,左眼不做羊膜移植。分别在术后7、14、28d取材分别行HE染色和透射电镜观察。结果光镜、电镜结果显示:羊膜移植组与未移植组相比,上皮愈合好,基质纤维排列整齐,炎细胞浸润轻,新生血管少。结论兔角膜碱烧伤急性期行新鲜羊膜移植可以减少炎症反应、促进角膜上皮愈合、减轻角膜瘢痕形成、减少角膜新生血管形成。     

难治性青光眼小梁切除术中应用生物羊膜与丝裂霉素C的对比研究

目的:探讨生物羊膜与MMC应用于青光眼小梁切除术中治疗难治性青光眼的临床疗效。方法:采用随机对照的方法,将临床收治的40例(57眼)难治性青光眼患者分为羊膜组32眼和丝裂霉素组25眼,前者施行小梁切除术联合巩膜瓣下生物羊膜植入术,后者在术中一次性应用MMC,浓度为0.2～0.4g/L,时间2～4min,随访1a,比较两种手术方式的临床疗效。结果:术后与术前相比:两组眼压均明显降低(P值均<0.01);术后1～2wk内MMC组视力下降者占45%,而羊膜组视力下降者仅占10%,两组间差异有显著性(P<0.05)。术后并发症:羊膜组眼部的副作用小,引起的并发症主要有术后浅前房;MMC引起的眼部并发症主要有薄壁滤过泡、滤过泡渗漏、低眼压性脉络膜脱离。结论:生物羊膜应用于难治性青光眼小梁切除术可有效地防止滤过泡的瘢痕组织形成,且并发症较应用MMC少,是治疗难治性青光眼安全、有效的手术方法。     

Adenoviral-mediated gene transfer to the filtering bleb in rabbits.

PURPOSE: To determine if genes can be transferred to fibroblasts in the filtering bleb using adenoviral vectors. MATERIALS AND METHODS: Twelve New Zealand albino rabbits underwent bilateral full-thickness sclerostomies. On postoperative day 1 an adenoviral vector carrying a reporter gene (lacZ) was injected into the right-eye bleb and saline was injected into the left eye bleb of each rabbit. Three rabbits were euthanized on each of the after days (days 3, 7, 14, and 21). The eyes were enucleated and tissue samples from the filtering bleb were processed for beta-galactosidase activity (a marker for lacZ gene expression) and expression of vimentin (a fibroblast marker). The median number of cells per high-power field with both beta-galactosidase activity and vimentin expression on days 3, 7, 14, and 21 in the right and left eyes were counted to determine adenoviral-mediated gene expression in fibroblasts. RESULTS: In the adenoviral vector-treated eyes, the median number of cells per high-power field with both beta-galactosidase and vimentin expression on days 3, 7, 14, and 21 was 83,100, 1, and 0, respectively. No beta-galactosidase activity was noted in the saline-treated eyes. CONCLUSIONS: Adenoviral vectors can transfer genes to fibroblasts in filtering blebs after glaucoma surgery. The peak efficiency of gene transfer to fibroblasts occurred 7 days after glaucoma surgery. These studies show a potential for genetic manipulation of fibroblast activity in filtering blebs after glaucoma surgery.     

曲安奈德/聚乳酸己内酯载药静电纺丝膜的抗兔眼滤过泡瘢痕化作用

背景 青光眼滤过术后滤过道的瘢痕形成是手术失败的主要原因,近年来多采用小梁切除术中局部使用抗瘢痕形成药物的方法来保持滤过道的开放,但药物可引起并发症.研究证实,聚乳酸己内酯(PLCA)材料制作的静电纺丝膜可作为负药载体以达到药物缓释的目的,但其在青光眼滤过术中的作用尚未证实. 目的 探讨小梁切除术联合巩膜层间填充负载曲安奈德(TA)静电纺丝膜在兔眼穿透性小梁切除术中抗瘢痕化的作用及可行性.方法 利用静电纺丝技术制备TA/PLCA载药静电纺丝膜,扫描电子显微镜下检测其表面超微结构,采用高效液相色谱仪检测体外药物缓释性能.采用前房注射卡波姆法制备单眼青光眼动物模型,将造模成功的40只新西兰白兔均行穿透性小梁切除术并采用随机数字表法分为5个组,每组8只眼.TA/PLCA载药静电纺丝膜组、PLCA静电纺丝膜组和羊膜组在术中于巩膜瓣下分别植入TA/PLCA载药静电纺丝膜、PLCA静电纺丝膜或羊膜.TA组术眼术毕在结膜下注射40 mg/ml TA溶液而术中不植入任何植片,单纯小梁切除术组仅行小梁切除术.分别于术后1、2、4、8和12周测量术眼眼压,并采用裂隙灯显微镜观察滤过泡的形态变化;于术后12周制备术眼滤过泡组织切片,采用苏木精-伊红染色法检查各组术眼滤过泡的组织病理学变化.结果 制备的PLCA静电纺丝纤维直径为0.5 ～1.5 μm,呈空间立体网状结构,而TA/PLCA静电纺丝膜结构与PLCA静电纺丝膜相近,可体外缓释药物14d.单纯小梁切除组术眼滤过泡于术后8周前全部消失,术后12周TA/PLCA载药静电纺丝膜组8只眼均存在功能性滤过泡,PLCA静电纺丝膜组中维持功能性滤过泡者5只眼,羊膜组和TA组中分别为4只眼.术后12周各组术眼的眼压明显不同,TA/PLCA载药静电纺丝膜组术眼眼压最低,而单纯小梁切除术组术眼眼压最高,各组间比较差异均有统计学意义(均P=0.000).组织病理学检查显示,术后12周TA/PLCA载药静电纺丝膜组术眼滤过道存在,滤过腔表面上皮化;PLCA静电纺丝膜组、羊膜组、TA组术眼滤过腔存在微小空隙,可见周围组织增生,而单纯小梁切除术组滤过腔消失,组织增生呈瘢痕化. 结论 TA/PLCA载药静电纺丝膜具有纳米级微观结构,体外药物缓释性能较好.小梁切除术中联合巩膜瓣下TA/PLCA载药静电纺丝膜植入可明显抑制手术区滤过腔瘢痕化,可能是TA抑制瘢痕作用与静电纺丝膜支架作用共同作用的结果.