生物羊膜联合自体角膜缘移植术治疗眼表疾病

目的观察生物羊膜联合自体角膜缘移植术治疗眼表疾病的疗效。方法对33例(39只眼)不同类型眼表疾病患者行生物羊膜联合自体角膜缘移植,随访3～18个月。结果术后移植片全部存活,角膜上皮愈合。术后视力均有不同程度提高。随访期间,眼表稳定,无新生血管形成及复发。结论生物羊膜移植联合自体角膜缘移植术能有效治疗眼表疾病。     

羊膜联合角膜缘干细胞移植对眼表疾病的治疗作用研究

目的:分析羊膜联合角膜缘干细胞移植对眼表疾病的治疗作用,为临床提供参考。方法:选取我院2013年4月至2014年4月收治的80例眼表疾病的患者作为研究对象,随机将其分成实验组和对照组,实验组40例,对照组40例,对照组患者采取自体角膜缘干细胞移植术治疗眼表疾病;实验组患者采取羊膜联合角膜缘干细胞移植治疗眼表疾病。结果:对照组患者总有效率为80.00%,实验组患者总有效率为97.50%,与对照组相比,差异具有显著性意义(P0.05)。结论:与自体角膜缘干细胞移植术相比,羊膜联合角膜缘干细胞移植治疗眼表疾病的临床效果更好,利于患者早日恢复健康,值得临床推广使用。     

培养兔角膜缘干细胞自体移植治疗眼表碱烧伤研究

目的：探讨培养兔角膜缘干细胞，自体移值治疗眼表碱烧伤的可行性。方法：体外培养兔角膜缘干细胞，传代于去上皮羊膜，自体移植治疗眼表碱烧伤；术后应用裂隙灯观察、免疫组化、透视电镜等方法，与单纯羊膜移植组疗效进行对比。结果：体外培养细胞既有角膜上皮特性，又具有强的增殖潜力。培养细胞移植组术后角膜上皮完整，炎症化、血管化减轻，与单纯羊膜移植组各项指标差异有显著性意义。结论：培养角膜缘干细胞移植术可有效重建眼表。     

新鲜羊膜联合自体角膜缘移植术治疗眼表疾病的疗效观察

【目的】观察新鲜羊膜联合自体角膜缘移植术治疗眼表疾病的疗效。【方法】对33例(39只眼)不同类型眼表疾病患行新鲜羊膜联合自体角膜缘移植，随访3～18m。【结果】术后移植片全部存活，角膜上皮愈合。术后视力均有不同程度提高。随访期问，眼表稳定，无新生血管形成及复发等。【结论】新鲜羊膜移植联合自体角膜缘移植术能有效治疗眼表疾病。     

角膜缘干细胞移植及其联合新鲜羊膜移植治疗复发性翼状胬肉

翼状胬肉是一种常见的眼表疾病,手术是其最常用的治疗方法,但术后常可复发[1],而随着复发次数及手术次数的增加,常导致睑球粘连的发生.近年来采用角膜缘干细胞移植及角膜缘干细胞联合新鲜羊膜移植治疗复发性翼状胬肉治疗取得了良好效果,我院分别采用角膜缘干细胞移植及角膜缘干细胞联合新鲜羊膜移植两种术式对复发性翼状胬肉进行治疗,现将不同术式的疗效报告如下.     

人羊膜移植联合自体角膜缘干细胞移植治疗中重度眼部碱烧伤

目的 观察人羊膜移植联合自角膜缘干细胞移植治疗中重度眼部碱烧伤的临床效果。方法 对8眼中重度眼部碱烧伤早期进行人羊膜移植联合自体角膜缘干细胞移植。结果 8眼中,7眼眼表面重建成功,4眼角膜透明,3眼角膜半透明,仅1眼发生轻度睑球粘连。结论 人羊膜移植联合自体角膜缘干细胞移植治疗中重度眼部碱烧伤。对于重建眼表面,提高视功能,早期应用疗效较好。     

培养的异体人角膜缘干细胞移植治疗眼表疾病

目的探讨培养的异体人角膜缘干细胞移植治疗眼表疾病的方法和疗效。方法采用消化培养法,对角膜移植术后剩余角膜缘组织进行体外培养,载体为冰冻保存人羊膜,培养的细胞形成单层后,移植到已去除病变组织的眼球表面,观察眼表结构的愈合情况。结果31例(34眼)眼表疾病患者进行了培养的异体人角膜缘干细胞移植,所有病例术后早期(2月内)均形成良好眼表,但随访3~41(平均15.4±11.6)个月后,有3例复发性翼状胬肉患者再次复发,1例急性眼碱烧伤于术后2月时发现角膜轻度血管化和结膜化,其他患者在随访期内均维持稳定的眼表情况,视力也有不同程度的提高。结论培养的异体角膜缘干细胞移植术是治疗因角膜缘干细胞缺乏或功能障碍引起的眼表疾病的有效方法。     

培养角膜上皮和自体角膜缘移植治疗眼表损伤的比较

目的比较培养角膜上皮移植和自体角膜缘移植治疗严重眼表损伤的效果。方法对去除全角膜缘的新西兰大白兔行培养自体角膜上皮和自体角膜缘联合羊膜移植,观察术后角膜形态改变,并进行组织学和免疫细胞化学检测。结果培养角膜上皮移植后兔眼表面平整稳定,角膜中央无新生血管生长,自体角膜缘联合羊膜移植后兔眼表面不同程度结膜上皮化,两者治疗效果有显著性差异。结论培养角膜上皮移植能有效治疗以角膜缘干细胞受损为特征的严重眼表损伤。     

同种异体保存羊膜在复杂眼表疾病中的应用

目的：探讨同种异体保存关膜移植在复杂眼表疾病中的治疗作用。方法：采用同种异体保存羊膜或联合角膜缘上皮移植治疗复杂性眼表疾病，并进行随访。结果：完成手术39例（40眼），均治愈，视力明显提高，减少睑球粘连，角膜新和血管增生等并发症症。结论：同种异体保存羊膜能有效地重建角膜表面，改善角缘基质，减轻炎症反应，从而阻止新生血管，利于眼表重建。     

培养角膜上皮和自体角膜缘移植治疗眼表损伤的比较

目的比较培养角膜上皮移植和自体角膜缘移植治疗严重眼表损伤的效果.方法对去除全角膜缘的新西兰大白兔行培养自体角膜上皮和自体角膜缘联合羊膜移植,观察术后角膜形态改变,并进行组织学和免疫细胞化学检测.结果培养角膜上皮移植后兔眼表面平整稳定,角膜中央无新生血管生长,自体角膜缘联合羊膜移植后兔眼表面不同程度结膜上皮化,两者治疗效果有显著性差异.结论培养角膜上皮移植能有效治疗以角膜缘干细胞受损为特征的严重眼表损伤.     

保存人羊膜联合自体角膜缘移植治疗严重眼表疾病

目的：观察和评价羊膜移植在重建眼表面手术中的应用价值。方法：采用保存人羊膜联合自体或亲属健康眼的角膜缘移植治疗眼表面疾病４８例５２只眼。结果：术后随访６０～９２周,５２只眼中,４７只眼获得稳定的眼表面（除周边部有少量纤维血管翳外,中央部无新生血管及复发性上皮缺损发生）,３７只眼角膜透明或半透明,３１只眼脱盲。结论：羊膜移植不是上皮移植,而是改良基底的手术,以保存人羊膜为基底膜,自体或亲属健康眼的角膜缘为上皮细胞来源,重建眼表面治疗眼表面疾病,初步结果显示是一种有价值的方法,其作用机制尚不清楚。     

角膜缘干细胞移植联合丝裂霉素和血管抑素治疗严重眼表碱烧伤

目的探讨角膜缘干细胞移植联合丝裂霉素和血管抑素治疗严重眼表碱烧伤的临床疗效。方法8例(8眼)严重眼表碱烧伤患者,行角膜缘干细胞移植术,术中应用丝裂霉素、术后应用血管抑素治疗。追踪观察6个月。观察了手术前后的角膜新生血管、混浊度、水肿度、排斥反应指数、假性胬肉和患者的自觉症状及视力。结果术后的角膜新生血管、混浊度、水肿度、排斥反应指数、假性胬肉和自觉症状评分均比术前的显著下降,术后各眼RI值均小于9,术后的视力有显著改善。结论异体角膜缘干细胞移植联合丝裂霉素和血管抑素治疗严重眼表碱烧伤在临床上收到良效,是一种安全、有效的重建严重碱烧伤眼表的治疗方法。     

Amniotic membrane transplantation in ocular surface disorders

In this prospective study, 81 eyes of 70 patients diagnosed with various ocular surface disorders were enrolled to document the use of amniotic membrane transplantation in various ocular surface disorders. Detailed history and ocular examination was done. Ocular photographs and consent from all patients were taken. Fluorescein staining and impression cytology was done preoperatively and postoperatively in selected cases. Amniotic membrane was prepared from the placenta of a donor (consent taken and negative for infectious disorders), after separating amnion from chorion. It was washed with antibiotic solutions, transferred over nitrocellulose paper and stored in Dulbecco's modified Eagle's minimum essential medium at -80 degrees C. Recipient bed was prepared by removing the fibrovascular pannus and necrosed conjunctiva. Amniotic membrane was transplanted with the epithelial side up and sutured. Sixty-four eyes had good result by clinical evaluation or impression cytology findings, 5 eyes later required limbal stem cell culture and transplantation. All the 3 eyes had failure of the fornix reconstruction and 5 eyes had recurrence of the pterygium. Amniotic membrane provides lower recurrence rate in cases of recurrent pterygium. Alkali injuries are more dangerous but showed good response to amniotic membrane transplantation combined with limbal autografting or ex-vivo expansion and later transfer. Initial proper assessment of limbal involvement, conjunctival necrosis and corneal involvement is the key to the management of acute cases. Contracted sockets showed no improvement. Shield ulcers and persistent epithelial defect and ocular surface defects secondary to tumour excision showed excellent results.     

Transplantation of human limbal cells cultivated on amniotic membrane for reconstruction of rat corneal epithelium after alkaline burn

Background The transplantation of limbal epithelial cells cultivated on amniotic membrane is a newly developed treatment for limbal stem cell deficiency. The purpose of our study was to investigate the biological characteristics of limbal epithelial cells and evaluate the effect of transplantation of cultivated human limbal epithelial cells on ocular surface reconstruction in limbal stem cell deficiency rat model.Methods Human limbal cells were isolated and cultivated in vitro. Cytokertins 3, 12, and 19 (K3, K12 and K19) and p63 were detected by immunofluorescent staining or RT-PCR. BrdU labelling test was used to identify the slow cycling cells in the cultures. Limbal stem cell deficiency was established in rat cornea by alkali burn.Two weeks after injury, the rats received transplants of human limbal stem cells cultivated on amniotic membrane carrier. The therapeutic effect was evaluated by slit lamp observation, Hemotoxin and Eosin (HE) staining and immunofluorescent staining.Results On day 7 in primary culture, p63 and K19 were strongly expressed by most cells but only a few cells expressed K3. On days 14 and 21, p63 and K19 were still expressed by a majority of cells, but the expressive intensity of p63 decreased in a number of cells, while the proportion of K3 positive cells increased slightly and some cells coexpressed p63 and K3. RT-PCR showed that gene expression of both p63 and K12 were positive in cultivated limbal cells, but in mature superficial epithelial cells, only K12 was detected. BrdU labelling test showed that most cells were labelled with BrdU after 7 days‘ labelling and BrdU label retaining cells were observed after chasing for 21 days with BrdU free medium. For in vivo test, slit lamp observation, HE staining and immunofluorescent staining showed that the rats receiving transplant of human limbal stem cells cultivated on amniotic membrane grew reconstructed corneas with intact epithelium, improved transparency and slight or no neovascularization. A majority of epithelial cells of the reconstructed cornea were positive to antihuman nuclear antibody and cells expressing K3 were found mainly in superfacial epithelium.Conclusions Limbal stem cells can be cultivated in vitro: the cells are characterized by high proliferation and slow cycling and identified as p63/K19 positive and K3/K12 negative. During culture, some stem cells can proliferate and differentiate into mature cornea epithelial cells. Amniotic membrane is a suitable carrier for limbal stem cells. Transplantation of human limbal stem cells cultivated on amniotic membrane can functionally reconstruct rat cornea with limbal stem cell deficiency.     

Characterization and safety assessment of bioengineered limbal epithelium

Transplantation of cultivated limbal epithelium on substrates such as amniotic membrane is an established treatment for severe ocular surface disease with limbal stem cell deficiency. In this study, we adapted an established method to generate sheets of limbal epithelium on amniotic membrane and characterized the cells contained in these sheets and tested them for safety with regard to microbial contamination. Human limbal biopsies were cultivated on denuded amniotic membranes. After three weeks of culture, the phenotypes of cultivated cells were analyzed by immunohistochemistry and real-time RT-PCR for the expression of a panel of specific markers. Cultivated limbal epithelial cell sheets were also analyzed by scanning (SEM) and transmission (TEM) electron microscopy. Sterility tests and mycoplasma assays were conducted for the safety of product. A confluent layer of polygonal cells was formed in 2 weeks and 1-3 stratified layer of cells were observed after three weeks of culture. Cultivated cells were positive for p63, K3, K19, and involucrin but negative for K14, integrin alpha9 and ABCG2 when analyzed by immunohistochemistry. Expression of molecular markers was detectable with real-time RT-PCR. SEM showed multilayer of flat squamous polygonal epithelial cells. Desmosomal and hemidesmosomal attachments were evident. Our study showed that cultivated limbal epithelium consists of limbal progenitors as well as differentiated corneal epithelial cells. SEM and TEM analysis showed cultivated cells demonstrated typical features of corneal epithelium. The risk of contamination is low and can be prevented by culturing the cells in a clean room facility complying to Good Manufacturing Practice standard.     

自体角膜缘干细胞移植与新鲜羊膜移植治疗翼状胬肉临床比较

目的比较自体角膜缘干细胞移植和羊膜移植两种手术方法治疗翼状胬肉的临床效果。方法1组35例胬肉切除联合自体角膜缘干细胞移植,2组26例胬肉切除联合新鲜羊膜移植,术后随访3~24月,比较两种术式术后移植片反应和复发情况。结果1组病例术后7例(20%)1~2周内有结膜水肿增厚,1例(2·8%)12月内复发;2组病例术后12(46%)例出现结膜充血水肿,3例(11·8%)6月内复发;全部患者无术后睑球粘连。结论胬肉切除联合自体角膜缘干细胞移植或新鲜羊膜移植是治疗翼状胬肉的有效方法,可降低复发率,减少术后并发症。自体角膜缘干细胞移植效果更佳。     

玻璃体切割术治疗严重眼外伤14例

目的:探讨玻璃体切割术对严重眼外伤的治疗效果。方法:总结这类病人的临床资料例,对术后临床效果进行14分析、评价。结果:例病人均保住了眼球,且视力有不同程度的提高,最低为,最高达,无感染及交感性眼炎14FC/30cm0.3发生。结论:玻璃体切割术是治疗严重眼外伤的重要手段。     

人胎儿角膜缘干细胞的体外培养及鉴定

目的 探讨人胎儿角膜缘干细胞的体外培养及鉴定方法。方法 应用消化培养法对人胎儿角膜缘上皮组织进行体外培养并观察记录培养细胞的生长特性；对胎儿角膜缘组织及培养细胞采用一系列单克隆抗体进行免疫酶细胞化学染色检测；并用扫描电镜观察原代细胞的表面状况。结果 胎儿角膜缘上皮细胞原代培养时生长旺盛，传代培养时保持高增殖速率。免疫酶细胞化学染色见培养前胎儿角膜缘组织基底层有多量AE5阴性、AE1和PCNA阳性细胞，并见较多HLA-DR阳性细胞；原代细胞绝大部分AE5和HLA-DR染色阴性，AEI和PCNA阳性；传代培养至第3代时AE5阴性细胞仍占大部分；扫描电镜观察原代细胞以球形或短圆柱形为主，表面多突起和微绒毛。结论 采用消化培养法可成功培养出具有高增殖力和低抗原性的人胎儿角膜缘干细胞．为临床眼表重建开辟了新的思路。     

用新鲜羊膜与生物羊膜移植治疗眼表热烧伤的疗效研究

目的探讨新鲜羊膜和成品生物羊膜移植治疗眼表热烧伤的治疗效果。方法收集眼表烧伤50例（60眼）患者随机分成新鲜羊膜组和生物羊膜组2组,术后随访5~12个月。观察其临床疗效、并发症。结果新鲜羊膜组32眼,角膜上皮修复29,眼视力〉0.1;生物羊膜组28眼,角膜上皮修复24眼,视力〉0.1。结论在眼表修复与重建过程中新鲜羊膜与生物羊膜的疗效、预后无明显差别。