带角膜缘的深板层角膜移植联合自体结膜移植和羊膜移植治疗严重眼表烧伤

目的探讨带异体角膜缘干细胞的深板层角膜移植联合自体球结膜移植和羊膜移植治疗严重眼烧伤的临床疗效。方法陈旧性热烧伤和化学腐蚀伤的患者19例19眼。无睑球粘连6眼,轻度睑球粘连5眼,重度睑球粘连8眼。采用带巩膜环的全厚角膜移植,缺失的结膜组织用羊膜和自体结膜修补,随访时间为9m～26m,观察视力、角膜上皮修复情况、角膜透明性、睑球粘连复发情况及排斥反应的发生情况。结果术后半年角膜植片透明率为89.5%,睑球粘连复发率为23.1%(3/13);术后矫正视力在0.3以上的为52.6%,在0.1-0.3为89.5%,2例出现持续上皮缺损和排斥反应,术后一年以上植片透明率为57.8%,睑球粘连复发率为38.5%(5/13);术后最佳矫正视力在0.3以上的为36.8%,在0.1-0.3为57.9%,有8例(42.2%)患者出现排斥反应同时有角膜上皮的缺损。结论带异体角膜缘干细胞的深板层角膜移植手术是治疗重症眼烧伤角膜混浊的有效方法,具有良好的光学性能;自体结膜联合羊膜移植是治疗重症睑球粘连的有效方法,而羊膜在这类病例中只能起到暂时支架的作用;排斥反应是异体角膜缘移植失败的主要原因。     

羊膜加亲属活体角膜缘／结膜同种异体移植在Stevens-Johnson综合征眼表重建中的应用

目的：报道应用保存羊膜加亲属活体角膜缘／结膜移植治疗Stevens-Johnson综合征继发完全性角膜缘干细胞和结膜缺乏。设计：前瞻性，非对照性，干预性的一组病例。患者和方法：10位Stevens-Johnson综合征继发完全性角膜缘干细胞和结膜缺乏的患者的10只眼接受了瘢痕组织切除，随后进行羊膜加亲属活体角膜缘／结膜移植。主要测量结果：角膜上皮的重建(外观透明没有上皮缺损，正常的荧光素渗透性，印迹细胞学检查无结膜来源的杯状细胞)。角膜血管化减轻。视力改善。结果：在平均16．7个月的随访过程中，2例(20％)取得了令人满意的眼表重建效果，炎症和血管化减轻，平均上皮化的时间是3周。4例(40％)手术失败，4例(40％)出现并发症(感染)。4例(40％)视力改善，5例(50％)维持稳定，1例(10％)下降。结论：羊膜加亲属活体角膜缘／结膜移植治疗Stevens-Johnson综合征继发的完全性角膜缘干细胞和结膜缺乏的严重病例的成功率达20％。手术后高比例的并发症特别是感染的发生可能是其原因。     

自体带结膜瓣角膜缘干细胞移植联合羊膜移植治疗复发性翼状胬肉

目的:降低翼状胬肉术后复发率,减少术后并发症。方法:采用自体带结膜瓣的角膜缘干细胞移植联合羊膜移植治疗26例(26眼)复发性翼状胬肉患者。结果:在26眼中显效25眼,复发1眼,有效率为96%。结论:采用自体带结膜瓣角膜缘干细胞移植联合羊膜移植,可有效维持角膜上皮与结膜上皮之间的屏障途径,阻止结膜上皮长入和新生血管形成,并使结膜下成纤维增殖抑制,睑球粘连松解,从而降低翼状胬肉术后复发率,减少术后并发症。     

深低温保存异体角膜缘干细胞羊膜移植治疗重度眼表疾病

目的 应用以羊膜为载体的深低温保存的角膜缘干细胞同种异体移植术治疗重度眼表疾病。方法 19例19眼重度眼表疾病患者采用羊膜移植联合深低温保存的角膜缘干细胞同种异体移植，2例术后半年行穿透性角膜移植术，随访观察术后疗效。结果 14例平均随访超过一年，眼表结构恢复正常，整个角膜上皮未见结膜杯状细胞，保存了眼球，视力均有不同程度的提高；1例植片脱落；4例发生角膜自溶的免疫排斥反应。结论 羊膜移植联合深低温保存的角膜缘干细胞同种异体移植术对于重度眼表疾病患者能快速、稳定的促进角膜表面重新上皮化，是一项可采纳的技术。     

不同移植手术治疗角膜缘干细胞缺损的实验研究

目的探讨以人羊膜为载体培养的角膜缘干细胞，自体及异体移植治疗全角膜缘干细胞缺损。方法制作兔眼角膜缘干细胞完全缺损3个月的模型。实验动物随机分为自体移植组和异体移植组，前者取对侧眼角膜缘组织，后者取异体兔眼角膜缘组织，均以去除上皮细胞的羊膜基底膜为载体，培养12d后行角膜缘干细胞羊膜移植术。术后观察3个月，以角膜上皮染色、角膜浑浊和新生血管3项指标进行临床疗效评定，通过病理检查评估术后角膜上皮修复情况，印迹细胞学检查移植前后角膜上皮的细胞表型。结果体外培养的兔角膜缘干细胞可在羊膜上粘附生长并增生，体外培养12d可形成复层。自体移植组和部分异体移植组术后角膜上皮逐渐愈合，透明度提高，基质细胞浸润减轻，新生血管减退或消失。印迹细胞学检查显示：移植前角膜上皮细胞PAS阳性，而移植后转为阴性；组织病理学显示：移植前角膜上皮大部分缺损，移植后呈现角膜上皮结构。部分异体移植组术后出现了免疫排斥反应。结论兔自体角膜缘干细胞羊膜移植术可重建眼表；免疫排斥反应仍是异体角膜缘干细胞羊膜移植术失败的主要原因。     

烧伤后角膜新生血管性浑浊的手术治疗--异体角膜缘联合保存人羊膜移植

目的 观察异体角膜缘联合保存人羊膜移植治疗烧伤后角膜新生血管性浑浊的效果。方法 对12例15眼烧伤后半年以上，眼部行亲属角膜缘组织联合保存人羊膜移植，其中3例4眼行单纯病灶切除术，9例11眼联合板层角膜移植术。结果 全部病例术后2-4周，新生上皮覆盖眼表面，1年后眼表面上皮稳定，角膜透明或半透明，睑球粘连解除。术后矫正视力≥0.05。结论 双眼严重表面损伤，采用亲属的角膜缘组织联合保存人羊膜移植可有效提高眼表重建术的成功率，是一种有推广价值的术式。     

角膜缘干细胞与结膜移植治疗翼状胬肉

目的:观察角膜缘干细胞与结膜移植治疗翼状胬肉的疗效。方法:采用自体角膜缘干细胞与结膜移植术,对38例43眼初发和复发性翼状胬肉患者进行治疗,术后随访3～30月,平均10月。结果:38例43眼中34例38眼上皮愈合稳定,角膜恢复正常光滑、透明,胬肉无复发。4例5眼失访。结论:自体角膜缘上皮移植为病变区角膜和结膜提供新的干细胞来源,是治疗翼状胬肉的理想方法。眼科学报1999;15:89—90。     

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。     

羊膜培养兔角膜缘上皮细胞及自体移植--角膜缘干细胞缺损的治疗

目的　探讨以人羊膜为载体培养兔角膜缘上皮细胞及其自体移植治疗全角膜缘干细胞缺损。方法　在8只兔右眼用正庚醇脱上皮和角膜缘环切的方法构建全角膜缘干细胞缺损模型2月。其中6只兔为实验组,活体取左眼角膜缘浅层小块,置羊膜上常规和气_液培养42天后进行自体移植治疗右眼角膜缘干细胞缺损;2只兔为对照组,直接用解冻无细胞人羊膜移植治疗右眼角膜缘干细胞缺损。进行细胞和术眼活体观察、组织学观察和电镜观察。结果　角膜缘上皮细胞在羊膜上生长良好,形成复层,细胞间的联结结构存在,细胞与羊膜组织粘附牢固。实验组移植手术后角膜迅速上皮化,恢复角膜表面光滑和透明,组织学观察和电镜观察呈现生理角膜上皮层的结构特点。但眼睑闭合不全可导致手术失败。对照组术后出现角膜缘干细胞缺损导致的角膜病变。结论　以羊膜为载体培养角膜缘上皮细胞后自体移植可有效地治疗角膜缘干细胞缺损导致的角膜病变。     

自体带结膜瓣角膜缘干细胞移植治疗翼状胬肉

目的降低翼状胬肉术后复发率,减少术后并发症。方法采用自体带结膜瓣角膜缘干细胞移植治疗63例(68眼) 翼状胬肉患者。结果68眼中显效62眼,有效4眼,复发2眼,总有效率97％。结论采用自体带结膜瓣角膜缘干细胞移植, 可有效维持角膜上皮与结膜上皮之间的屏障作用,阻止异常结膜源性组织的再侵入,从而降低翼状胬肉术后复发率。     

Amniotic membrane transplantation or conjunctival limbal autograft for limbal stem cell deficiency induced by 5-fluorouracil in glaucoma surgeries

PURPOSE: To determine if human amniotic membrane transplantation or limbal stem cell transplantation is effective to restore the corneal surface with partial or total limbal stem cell deficiency, respectively, caused by 5-fluorouracil (5-FU) toxicity after glaucoma surgeries. METHODS: Partial and total limbal stem cell deficiency was confirmed by impression cytology as the cause of reduced vision and corneal surface breakdown in a 69-year-old man and a 67-year-old man, respectively, who both had received a total of 105 mg 5-FU injections. Amniotic membrane transplantation or conjunctival limbal autograft was performed for corneal surface reconstruction, respectively. RESULTS: For a period of 15 months of follow-up, the visual acuity improved, and their corneal surfaces remained avascular, smooth, and without recurrence of limbal stem cell deficiency. CONCLUSION: Limbal stem cell deficiency can occur as a late complication for patients receiving 5-FU after glaucoma filtering surgeries. Partial limbal stem cell deficiency can be treated with amniotic membrane transplantation alone, whereas limbal transplantation must be considered as an alternative for total limbal stem cell deficiency to restore the corneal surface integrity and vision.     

Amniotic membrane transplantation for partial and total limbal stem cell deficiency secondary to chemical burn

PURPOSE: To evaluate the surgical outcome of preserved amniotic membrane transplantation (AMT) for ocular surface reconstruction in chemical burn with limbal stem cell deficiency. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: Twenty eyes of 20 consecutive patients with limbal stem cell deficiency secondary to ocular chemical injury. INTERVENTION: AMT with or without adjunctive limbal transplantation using limbal tissue from either the healthy contralateral eye (CLAU) or a living related donor (lr-CLAL). MAIN OUTCOME MEASURES: Reconstruction of corneal epithelium (clear appearance without epithelial defect, normal fluorescein permeability and the absence of conjunctiva-derived goblet cells on impression cytology), decrease in corneal vascularization and improvement in visual acuity. RESULTS: With a mean follow-up time of 19 months (range, 8-27 months), satisfactory ocular surface reconstruction was obtained in 15 eyes (75%), with reduced inflammation and vascularization of the ocular surface and a mean epithelialization time of 3.3 weeks. Success was observed in all cases of partial limbal stem cell deficiency (PLD) and in 68.75% (11 eyes) of cases of total limbal stem cell deficiency (TLD). Surgical failure was observed in five severe cases (31.25%). A significant visual improvement was observed in all cases after surgery, except for 2 eyes that maintained preoperative visual acuity. CONCLUSIONS: AMT seems to be an efficient adjunct for ocular surface reconstruction in chemical burns with PLD. When performed in conjunction with limbal stem cell transplantation, it is also effective in most cases of TLD.     

Sectorial conjunctival epitheliectomy and amniotic membrane transplantation for partial limbal stem cells deficiency

CASE REPORT: A 74-year-old man developed a partial limbal stem cell deficiency secondary to a chemical injury in the left eye. He was treated with a sectorial conjunctival epitheliectomy and amniotic membrane transplantation. After 3 weeks there was an intact, smooth and stable corneal epithelium. DISCUSSION: This case report emphasizes that conjunctival epitheliectomy and amniotic membrane transplantation appears to be a safe and effective method of restoring a stable corneal epithelium for cases of partial limbal stem cell deficiency.     

Amniotic membrane transplantation for partial limbal stem cell deficiency

AIM: To examine the efficacy, safety, and long term outcomes of amniotic membrane transplantation for corneal surface reconstruction in cases of partial limbal stem cell deficiency. METHODS: 17 eyes of 15 patients with partial limbal stem cell deficiency underwent superficial keratectomy of the conjunctivalised corneal surface followed by amniotic membrane transplantation. Cases were followed up for at least a year. RESULTS: All eyes exhibited a stable, intact corneal epithelial surface after a mean follow up period of 25.8 months with no eyes developing recurrent erosion or persistent epithelial defect. The mean time to re-epithelialisation was 22.8 days. Overall improvement in visual acuity was observed in 92.9% of 14 eyes with visual potential. Of those, five eyes gained six or more lines, two eyes gained between four and five lines, six eyes gained between one and three lines, and one eye lost three lines of Snellen acuity. Pain and photophobia were abolished in 86% of cases and substantially reduced in 14%, with all eyes exhibiting decreased vascularisation and inflammation at final follow up. CONCLUSIONS: Amniotic membrane transplantation appears to be a safe and effective method of restoring a stable corneal epithelium for cases of partial limbal stem cell deficiency and can be considered as an alternative to limbal autograft or allograft.     

培养兔自体角膜缘干细胞移植的实验研究

目的 观察以羊膜为载体的兔角膜缘于细胞膜片移植治疗兔角膜缘于细胞缺损的效果。方法 制造兔角膜缘干细胞缺损的动物模型,以右眼为实验眼,从兔左眼取角膜缘组织,将兔角膜缘干细胞消化下来,接种于铺有羊膜的无菌六孔培养板中,待细胞形成多层角膜上皮细胞后,对兔角膜缘干细胞缺损动物模型行角膜缘干细胞羊膜移植术,并对治疗后的角膜进行裂隙灯及病理学检查。结果 临床和组织病理学染色证实：体外培养的兔角膜缘干细胞可在羊膜上继续增生、分化为多层角膜上皮细胞;角膜缘干细胞移植术后兔角膜缘轻度充血、角膜上皮完整、基质细胞浸润减轻、新生血管减少或消失。结论 应用角膜缘干细胞羊膜移植术可恢复其角膜上皮结构的完整性,减少角膜新生血管的形成,维持角膜缘的细胞屏障功能。     

Amniotic membrane transplantation with conjunctival limbal autograft for total limbal stem cell deficiency

PURPOSE: To evaluate the outcomes of corneal surface reconstruction with conjunctival limbal autograft when combined with amniotic membrane transplantation on both the donor and recipient eyes. DESIGN: Retrospective, noncomparative, interventional small case series. PARTICIPANTS: Five eyes of five patients with total limbal stem cell deficiency (LSCD) resulting from pseudopemphigoid (n = 1), chemical burns (n = 3), and extensive removal of conjunctival intraepithelial neoplasia (n = 1) were operated on by one surgeon (SCGT). INTERVENTION: After the removal of fibrovascular pannus from the corneal surface, two conjunctival limbal free grafts were harvested from the fellow eyes in all five patients with unilateral LSCD. Amniotic membrane, with the basement membrane side up, was grafted onto the defect created at the donor site and onto the recipient corneal and limbal sclera before placement of conjunctival limbal grafts. MAIN OUTCOME MEASURES: Symptomatic relief, improvement in visual acuity, fornix deepening, and rapid healing and restoration of normal cornea and limbus in the recipient and donor eyes were assessed. RESULTS: During the mean follow-up of 22 months (range, 11-48 months), all eyes experienced symptomatic relief. All recipient eyes had a mean improvement in visual acuity of nine lines (range, 7-12). The three eyes with stromal vascularization showed regression, and all recipient eyes had marked improvement in corneal clarity. Three eyes receiving simultaneous symblepharon lysis and fornix reconstruction successfully regained deep, stable fornices. The donor eyes showed rapid healing and restoration of the normal limbal landmark, even in one eye where nearly the entire limbus was removed. CONCLUSIONS: Limbal conjunctival transplantation is an effective procedure for restoring the corneal surface integrity in eyes with total LSCD. The additional use of amniotic membrane may contribute to a higher rate of success in the recipient eye and a lower rate of complications in the donor eye, as well as allow the simultaneous correction of concomitant cicatricial abnormalities.     

Amniotic membrane transplantation in acute chemical burns

PURPOSE: To evaluate the outcome of fresh amniotic membrane transplantation (AMT) for ocular surface reconstruction in acute chemical burns. METHODS: A prospective study of 15 consecutive eyes with acute chemical burns was performed. In all, 10 eyes had lime burns and five eyes had acid burns. There were three eyes of grade II, four eyes of grade III and eight eyes of grade IV burns. AMT was performed within 3 weeks of injury. RESULTS: Patients were followed up for 10.14 +/- 4.41 months. All patients had immediate relief of pain postoperatively. Of 15 eyes, nine (60%) showed epithelialization within 1-4 weeks (15.33 +/- 9.91 days). The final visual acuity improved in 10 of 15 eyes (66.66%). Eyes with burns of grade II and III showed more visual improvement than those with grade IV burns. None of the eyes showed perforation. Symblepharon was seen in nine of 15 eyes (60%). Of 15 eyes, 12 (80%) experienced limbal stem cell deficiency and showed superficial corneal vascularization. CONCLUSIONS: Amniotic membrane transplantation with fresh amniotic membrane increases patient comfort and reduces inflammation. In mild burns, AMT alone restores corneal and conjunctival surfaces. In moderate to severe burns, it probably reduces conjunctival scarring sequelae, but does not prevent the sequelae of limbal stem cell deficiency that requires further limbal stem cell transplantation. In the acute stage, amniotic membrane transplantation probably has a protective role against the progressive melting and perforation.     

Survival analysis of conjunctival limbal grafts and amniotic membrane transplantation in eyes with total limbal stem cell deficiency

PURPOSE: To evaluate the survival of conjunctival limbal grafts and amniotic membrane transplantation (AMT) for total limbal stem cell deficiency (LSCD) and the influence of several parameters as cause of LSCD, dry eye, keratinization, eyelid abnormalities, HLA compatibility, systemic immunosuppression, and keratoplasty (PKP) on surgical outcome. DESIGN: Prospective, noncomparative, interventional case series. METHODS: Thirty-three eyes of 31 patients with total LSCD that underwent conjunctival limbal grafts and AMT at the Department of Ophthalmology, Federal University of S?o Paulo were included in this study. Cumulative graft survival as well as the influence of several variables on surgical outcome was analyzed. RESULTS: Ten eyes (30%) underwent conjunctival limbal autograft and 23 (70%) underwent conjunctival limbal allograft from living HLA-matched donor. Graft survival was seen in 13 eyes (40%) at 1 year and in 11 eyes (33.3%) at 2 years, with a cumulative survival of 33% after a mean follow-up time of 33 months. Increase in postoperative visual acuity was observed in 20 eyes (60.6%) during this period. Marked impact on graft survival was observed for patients with Stevens-Johnson syndrome, dry eye, keratinization, eyelid abnormalities, and allogeneic conjunctival limbal transplantation (independently of HLA compatibility) (P < .05). Preoperative dry eye was the most important prognostic parameter for surgical outcome (P < .001). CONCLUSIONS: Conjunctival limbal grafts associated with AMT are useful for restoring corneal epithelium phenotype in eyes with total LSCD. However, the cumulative survival declines substantially over a 2-year period. Considering all investigated variables, dry eye was the most important prognostic parameter.     

羊膜移植与自体角膜缘干细胞移植治疗原发性翼状胬肉的临床疗效比较

目的羊膜移植与自体角膜缘干细胞移植治疗原发性翼状胬肉的临床疗效比较。方法将114例（131眼）随机分成A和B两组。A组53例（59眼）行翼状胬肉切除＋羊膜移植术,B组61例（72眼）行翼状胬肉切除＋自体角膜缘干细胞移植术,术后随访6-24个月,观察两组疗效并进行回顾性分析。结果 A组10眼复发,复发率为16.95%,B组6眼复发,复发率为8.33%,两组比较差异具有统计学意义（P〈0.05）,A组复发率明显高于B组。结论翼状胬肉切除＋自体角膜缘干细胞移植术,此手术取材方便、成功率高;而翼状胬肉切除＋羊膜移植术,手术耗材、成功率明显较自体角膜缘干细胞移植术低。     

Use of autologous cultured limbal and conjunctival epithelium in a patient with severe bilateral ocular surface disease induced by acid injury: a case report of unique application

PURPOSE: Reconstruction of the ocular surface in a case of severe bilateral partial limbal stem cell deficiency (LSCD) with extensive symblephara using autologous cultured conjunctival and limbal epithelium. CASE REPORT: A 31-year-old woman presented with severe bilateral ocular surface disease with partial limbal stem cell deficiency, symblephara, lid and facial scarring, with a vision of 20/400 and counting fingers at 1 m in both eyes. Limbal and conjunctival tissue was harvested from the healthy-appearing left eye and used to generate two sheets of composite epithelium consisting of central limbal and peripheral conjunctival cells. The limbal tissues were explanted in the central region while the conjunctival tissues were explanted on the periphery of the deepithelialized human amniotic membrane (HAM) and nurtured using human corneal epithelial cell medium. After successful generation of a monolayer from both tissues had been confirmed, the composite of cultivated limbal and conjunctival epithelium with HAM was transplanted in each eye after excision of fibrous tissue and release of symblephara. One year postoperatively, the patient had a best spectacle-corrected visual acuity of 20/40 in the right eye (preoperative acuity 20/400) and counting fingers at 1 m in the left eye (same as preoperative) with a stable ocular surface. CONCLUSIONS: Autologous cultured epithelial transplantation is as an excellent option in selected patients with bilateral partial LSCD with small area(s) of healthy limbus in either eye and avoids the attendant risk of rejection and cost and potential toxicity of immunosuppression in allogeneic tissue transplantation. This case also highlights the feasibility of generating a composite culture of limbal and conjunctival epithelium using a single amniotic membrane.