角膜缘部上皮移植治疗眼表疾病

角膜缘部上皮基底层的干细胞具有细胞更新和组织再生的功能,而眼部表面疾病则是干细胞缺乏的结果。采用自体角膜缘部上皮移植治疗15例。全部病例在术后5～15天上皮迅速愈合。术后随访半年以上,12例稳定不再脱落,11例无基质血管长入,11例矫正视力>0.05,由于明显改善眼部表面,11例联合板层角膜移植术的病例,全部获得透明成功。     

Factors influencing outcomes in cultivated limbal epithelial transplantation for chronic cicatricial ocular surface disorders

PURPOSE: To analyze factors influencing clinical outcomes in cultivated limbal epithelial transplantation (CLET). DESIGN: Retrospective, observational case series. METHODS: Twenty-seven eyes of 27 patients that had CLET for severe chronic cicatricial ocular surface disorders were studied. Two different cultivation methods were used to prepare epithelial sheets. Method 1 used the explant technique and neither feeder cells nor air-lifting were used. In Method 2, cell suspension technique and 3T3 feeder cells were used, and air-lifting was applied after cultivated cells became confluent. Clinical outcomes including corneal surface epithelialization and incidence of postoperative complications were studied. The relationship between the clinical outcome and type of cultivation method, original diseases, tear function, or preoperative ocular surface status was also studied. RESULTS: Both cultivation methods produced transplantable epithelial sheets with corneal phenotype. With a mean follow-up period of 127 weeks, corneal epithelialization was achieved in 16 eyes (59.3%). Eyes that had CLET with Method 1 suffered more severe postoperative complications such as infection, ulceration, and perforation (P = .053). Eyes with Stevens-Johnson syndrome (SJS) had poor final corneal epithelialization compared with other diseases (P = .034). CLET was more successful when performed to conjunctivalized corneas compared with eyes with persistent epithelial defects or with dermalized corneas. CONCLUSIONS: CLET offers new treatment modalities to chronic cicatricial ocular surface disorders with moderate success rates. The treatment is feasible for eyes with non-immune-mediated disorders with stable ocular surface conditions. Epithelial sheets with better structural integrity seem to be superior to obtain early postoperative epithelialization and to avoid serious postoperative complications.     

Cultivated corneal epithelial stem cell transplantation in ocular surface disorders

PURPOSE: To investigate the outcome of cultivated corneal epithelial transplantation for severe stem cell deficiencies using denuded amniotic membrane (AM) as a carrier. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Thirteen eyes of 11 patients were studied. These consisted of five eyes with acute Stevens-Johnson syndrome (SJS), two with chronic SJS, one with an acute chemical injury, two with chronic chemical injuries, two with ocular cicatricial pemphigoid, and one with drug-induced pseudopemphigoid. All of these eyes had total stem cell deficiencies. MAIN OUTCOME MEASURES: Adaptation of the cultivated corneal epithelium onto the host corneal surface was confirmed 48 hours after surgery. The reconstruction of the ocular surface and visual acuity were measured. METHODS: Corneal limbal epithelium from donor corneas was cultivated for 4 weeks on a denuded AM carrier, with 3T3 fibroblast coculture and air lifting. The cultivated corneal epithelium showed four to five layers of stratification and was well differentiated. After conjunctival tissue removal from the cornea up to 3 mm outside the limbus and subconjunctival tissue treatment with 0.04% mitomycin C, cultivated allocorneal epithelium, including the AM carrier, was transplanted onto the corneal surface up to the limbus. Lamellar keratoplasty, using preserved donor graft without epithelium, was performed simultaneously for five chronic-phase patients showing corneal stromal scarring. Systemic immunosuppression was used to prevent allograft rejection. RESULTS: In all 13 eyes, the entire corneal surface, on which cultivated allocorneal epithelium had been placed, was free from epithelial defects 48 hours after surgery, indicating complete survival of the transplanted corneal epithelium. Visual acuity improved in all eyes after surgery, and 10 of the 13 eyes were restored to good vision (postoperative visual acuity improved two or more lines) 6 months after the operation. During the follow-up period (mean +/- standard deviation, 11.2 +/- 1.3 months), the corneal surfaces were clear, although three eyes experienced epithelial rejection. CONCLUSIONS: Cultivated corneal epithelial transplantation using denuded AM as a carrier can be used for severe stem cell deficiencies.     

Transplantable cultivated mucosal epithelial sheet for ocular surface reconstruction

Ocular surface reconstruction by tissue engineering using somatic stem cells is a second-generation therapeutic modality. In view of future treatment of bilaterally affected, severe ocular surface disorders, two types of transplantable cultivated mucosal epithelial sheets can be used for reconstruction. One is an allogeneic corneal epithelial stem cell sheet, and the other is an autologous oral mucosal epithelial cell sheet. We first investigated the feasibility of amniotic membrane as an epithelial carrier, and found that denuded amniotic membrane was the most appropriate substrate for this purpose. Thus, cultivated corneal epithelial stem cell sheets were created by co-culturing with 3T3 fibroblast and air-lifting on amniotic membrane. These epithelial sheets demonstrated positive keratin 3 and 12 specific to in vivo corneal epithelium, light junction related proteins and proliferative activity. The transplanted allogeneic human corneal epithelial sheets existed successfully on the corneal surface, and were quite effective in achieving ocular surface stability in severe ocular surface disorders. A few cases, however, developed immunological reactions or opportunistic infections, etc. Secondly, we established transplantable autologous cultivated oral mucosal epithelial sheets in rabbits. The in vitro oral mucosal epithelial sheets showed histological characteristics similar to those of in vivo corneal epithelial sheets; for example, positive keratin 3 expression. Based on the fact that, the transplanted autologous oral mucosal epithelial sheets resembled corneal epithelium and that we achieved the recovery of corneal transparency in rabbits, we propose that cultivated oral mucosal epithelium may become the substitute for corneal epithelium in ocular surface reconstruction.     

Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation

PURPOSE: To perform a midterm assessment of the integrity and reproducibility of cultivated autologous oral mucosal epithelial sheets, and to evaluate the clinical efficacy of their transplantation in ocular surface. DESIGN: Observational case series. METHODS: Cultivated autologous oral mucosal epithelial sheets were created using amniotic membrane and buccal mucosal epithelium from 12 patients with Stevens-Johnson syndrome, chemical and thermal injury, pseudo-ocular cicatricial pemphigoid, and idiopathic ocular surface disorder. They were transplanted onto 15 eyes from these patients who were then followed up for a mean of 20 months; with the longest follow-up being 34 months. We assessed their clinical outcomes with special reference to neovascularization. RESULTS: Cultivated autologous oral mucosal epithelial sheets could be generated from all patients. On the second postoperative day, 14 of 15 sheets transplanted demonstrated total re-epithelialization on the cornea. During the follow-up, the ocular surface was stable and transparent without any major complications in 10 of 15 eyes (67%), and the transplanted epithelium survived for at least 34 months. There were five eyes (33%) with small but long-standing epithelial defects, three of these healed spontaneously, and two (13%) required reoperation. In 10 eyes, postoperative visual acuity was improved by more than 2 lines. All eyes manifested some peripheral corneal vascularization. CONCLUSIONS: We established a successful tissue-engineering technique to generate cultivated autologous oral mucosal epithelial sheets and succeeded in reconstructing the ocular surface. We suggest that this surgical modality may be both safe and useful, especially in younger patients with the most severe ocular surface disorders.     

Limbal autograft transplantation for ocular surface disorders

Limbal autograft transplantation is presented in 26 consecutive cases comprising both acute and chronic chemical injury (20 cases), thermal burns (2 cases), contact lens-induced keratopathy (3 cases), and ocular surface failure after multiple surgical procedures (1 case), with follow-up ranging from 2 to 45 months (mean, 18 months). The operative technique usually involved transfer of two free grafts of limbal tissue from the uninjured or less injured donor eye to the severely injured recipient eye, the latter having been prepared by limited conjunctival research and superficial dissection of fibrovascular pannus without keratectomy. Clinical results in 21 patients with follow-up of 6 months or more have consistently shown improved visual acuity (17 cases), rapid surface healing (19 cases), stable epithelial adhesion without recurrent erosion or persistent epithelial defect (20 cases), arrest or regression of corneal neovascularization (15 cases), and probable increased success for lamellar or penetrating keratoplasty (8 cases). No intraoperative complications were encountered, and no adverse reactions developed in donor eyes. Impression cytology in selected cases showed restoration of the corneal epithelial phenotype and regression of goblet cells from the recipient cornea. Therefore, limbal autograft transplantation is recommended for treatment of widespread ocular surface damage with loss of limbal epithelial stem cells and, specifically, for chemical or thermal burns, contact lens-induced keratopathy, and selected persistent corneal epithelial defects.     

羊膜移植治疗眼表疾病临床观察

目的　探讨羊膜移植治疗眼表疾病的临床效果。方法　采用新鲜羊膜移植治疗几种眼表疾病 2 5例 (2 7眼 ) ,术后随访观察 3月～ 1年。结果　复发性翼状胬肉 2眼 3月后复发 ;1眼 3 60°蚕蚀性角膜溃疡伴穿孔 ,行多层羊膜移植 ,1月后 ,部分羊膜脱落 ,再次给予羊膜加巩膜、结膜覆盖后治愈。结论　羊膜移植重建眼表是目前较理想的治疗手段。     

Transplantation of cultivated autologous oral mucosal epithelial cells in patients with severe ocular surface disorders

BACKGROUND/AIMS: To determine outcomes of transplants of cultivated autologous oral epithelial cells in patients with severe ocular surface disorders. METHODS: The eyes (n = 6) of four patients with Stevens-Johnson syndrome (three eyes) or chemical burns (three eyes) were studied. Autologous oral epithelial cells, grown for 2-3 weeks on a denuded amniotic membrane carrier in the presence of 3T3 fibroblasts, were air lifted. The resultant sheet was transplanted onto the damaged eye, and acceptance of the sheet by the corneal surface was confirmed 48 hours after surgery. The success of ocular surface reconstruction, graft survival, changes in visual acuity, and postoperative complications were assessed and the quality of the cultivated oral epithelial sheet was evaluated histologically. RESULTS: At 48 hours after transplant, the entire corneal surface of all six eyes was free of epithelial defects indicating complete survival of the transplanted oral epithelium. Visual acuity was improved in all eyes. During follow up (mean 13.8 (SD 2.9) months), the corneal surface remained stable, although all eyes manifested mild peripheral neovascularisation. CONCLUSIONS: Autologous oral epithelial cells grown on denuded amniotic membrane can be transplanted to treat severe ocular surface disorders.     

眼外伤及眼表疾病自体角膜缘干细胞移植术

目的 探讨用自体角膜缘干细胞移植术治疗眼外伤及眼表疾病新方法。方法 应用自体角膜缘干细胞移植术治疗42例48眼，包括角膜裂伤、角膜碱烧伤、翼状胬肉及角膜缘鳞癌等眼表疾病等。随访7月-2年，回顾性评价疗效。结果 术眼移植片成活良好，无新生血管长人，48眼均无复发，视力增进，无并发症发生。结论 自体角膜缘干细胞移植术，疗效可靠，无排斥反应。     

Limbal cell autograft transplantation for severe ocular surface disorders

· Background: Limbal cell transplantation may improve the visual outcome after chemical trauma and ocular surface diseases. · Methods: Nine eyes of nine consecutive patients (eight males and one female, age 9–60 years), underwent limbal autograft transplantation (LAUT). In five cases LAUT was done for severe chemical burns in the acute stage (group 1). In four patients with old chemical trauma LAUT was performed years after the trauma (group 2). Penetrating keratoplasty (PKP) was carried out within 6 months after LAUT in three patients of group 2. Preoperatively, the visual acuity in all the patients except one was counting fingers. Postoperatively, patients were treated with topical antibiotics, topical corticosteroids and oral steroids. Oral cyclosporin was used after penetrating keratoplasty. · Results: No complications were observed during the surgical procedure. Postoperatively, the epithelialization was complete between days 7 and 12. The inflammatory response subsided within 3 months and the stromal neovascularization decreased. Visual acuity improved in all the nine cases, ranging from 6/6 to 6/30. The decreased visual acuity was due to corneal haze, scars and vascularization. Following PKP, the three grafts remained clear with intact epithelium. No complications were observed during the follow-up period from 7 to 60 months. · Conclusions: Limbal cell transplantation is an efficacious procedure for rehabilitation of visual acuity after severe chemical trauma. Received: 19 March 1997 Revised version received: 22 July 1997 Accepted: 29 September 1997     

自体角膜缘上皮移植治疗眼表疾病

目的 评估角膜缘上皮移植治疗眼表疾病的疗效。方法 对59例患者(眼部化学伤9例、热烧伤2例、初发翼状胬肉36例、复发翼状胬肉12例)采用自体角膜缘上皮移植进行治疗，并对其疗效进行评估。结果 11例烧伤(化学伤和热烧伤)中9例一次手术成功，另2例二次手术后治愈。所有翼状胬肉患者术后2周角膜表面光滑透明。术后随访3～12个月。结论 自体角膜缘上皮移植治疗眼表疾病是一种方便、安全和有效的方法。     

体外培养口腔黏膜上皮细胞移植重建眼表的研究

采用体外培养的同种异体角膜缘干细胞重建眼表已在临床应用获得成功,而采用体外培养的自体口腔黏膜上皮重建眼表,国内尚鲜见报道。本文就体外培养口腔黏膜上皮移植重建眼表的研究进展,作一较为详尽的综述。