Sutureless amniotic membrane transplantation following excision of ocular surface neoplasia

【正】Dear Sir,S uccess of surgical management of conjunctival neoplasias primarily depends on the complete excision of the mass and eradication of tumor cells from the ocular surface[1-3].Especially in cases that extensive resections are required,primary closure may not be feasible;so ocular surface reconstruction can be a challenging problem for the surgeons.For conjunctival tissue defects different surgical

Amniotic membrane transplantation with topical interferon alfa-2b after excision of ocular surface squamous neoplasia

To evaluate the outcome of amniotic membrane transplantation（AMT） after tumor excision followed by topical interferon alfa-2 b（IFNα2 b） drops for primary ocular surface squamous neoplasia（OSSN）. Twelve eyes of 12 patients with a mean age of 66±10 y were included. The average followup was 23±10 mo. All 12 patients had limbal involvement. Smooth ocular surface and transparent cornea were achieved in all cases. No sign of inflammation, neovascularization, symblepharon or recurrence was noted at the last follow-up. We conclude that AMT with topical IFNα2 b drops restores a healthy ocular surface in OSSN without recurrence.

Medical and surgical approach to ocular surface reconstruction

Ocular surface disease(OSD)can have a severe impact on patients as it can lead to visual impairment and persistent discomfort.Ocular surface reconstruction(OSR)is an approach to the management of ocular diseases that cause structural damage to the ocular surface.OSR encompasses both medical and surgical treatment options.In this review,we discuss the medical and surgical strategies used in OSR.Medical management often aims to treat tear insufficiency,inflammation,and keratinization.Surgical treatments may be employed for a variety of reasons,including failure of medical management.This may include improving the oculo-palpebral structures in order to improve lid positioning and tear film.Additional therapies focus on improving tear production,such as through salivary gland transplantation.In situations where the ocular surface is so severely damaged that there is loss of limbal stem cells,limbal stem cell transplant(LSCT)may be indicated.Other surgeries such as amniotic membrane transplant(AMT)and conjunctival flaps(CFs)can help promote corneal healing.Finally,in severe situations where the cornea is beyond salvage,corneal transplantation,such as a penetrating keratoplasty(PKP),can be considered.OSR often requires a combination of medical and surgical approaches targeted to each specific patient’s presentation in order to achieve optimal outcomes.

Effect of autologous serum after amniotic membrane transplantation for persistent corneal ulcers

AIM:To investigate the effect of adding autologous serum eye drops to the postoperative regime after amniotic membrane transplantation for severe persistent corneal ulcers.METHODS:Forty eyes of 40 patients with persistent corneal ulcers were randomly assigned to artificial tears(sodium hyaluronate 0.2%,ATs group,n=20)or autologous serum eye drops(ASEDs,n=20)following treatment with amniotic membrane transplantation.Digital slit lamp images were acquired from all patients before and 30d post treatment.The area with fibrovascular tissue was calculated using Image J.Central corneal sensitivity was assessed by Cochet-Bonnet aesthesiometry before and one month after treatment.Scar tissue transparency was assessed with a novel optical densitometry.RESULTS:Mean age of patients was 61.65±16.47y and 57.3±19.11y in the ATs group and ASEDs group,respectively.Twenty-two male and 18 female patients were included in the study.The improvement in visual acuity was significantly greater in the ASEDs group(0.14±0.04)than the ATs(0.08±0.04;P=0.00046).Cochet-Bonnet aesthesiometry improved significantly after treatment with a similar rate between groups.There were no statistically significant differences in the area of postoperative fibrovascular tissue between the two groups(P=0.082).The success rate in the two groups was similar.The difference in densitometry between the ATs and ASEDs group was statistically significant(P=0.042)with greater reduction from baseline in the ASEDS group.CONCLUSION:Autologous serum eye drops can lead to better visual acuity,more stable results and improved densitometry and should be considered in the postoperative care following amniotic membrane transplantation.

Fresh Amniotic Membrane Transplantation for Conjunctival Surface Reconstruction

Purpose:To determine whether fresh human amniotic membrane can be used to reconstruct the conjunctiveal defect creates during symblepharon lysis.Methods:Forth-two eyes of 39 consecutive patients with eye burns and Stevens-Johnson syndrome were randomized to accept fresh or preserved human amniotic membrane transplantation(AMT) during the period of severe scarring.Impression cytology was performed in 12 eyes with normal tear secretion which received fresh AMT.Results:During a mean follow-up of 11 months (range,6 to 18 months),thirty-five patients (37 eyes)showed successful ocular surface reconstruction and resolution of motility restriction while four patients (2 eyes with fresh AMT,3 eye with preserved AMT)with minimal recurrence of symblepharon.There was no significant difference statistically between two groups(Chi-square test).Amniotic epithelial cells can survive about three months after being transplanted onto ocular surfaces with normal tear secretion.Conclusion:Both fresh and preserved human amniotic membrane can be considered an ideal alternative substrate for conjunctival surface reconstruction during removal of severe symblepharon.

Amniotic membrane transplantation: an updated clinical review for the ophthalmologist

Although amniotic membrane transplantation(AMT)has long been used as an essential surgical technique for ocular surface reconstruction,its role continues to evolve and expand.In the management of numerous ocular surface disorders,ranging from inflammatory to infectious,traumatic to neoplastic,the ability to perform AMT is a valuable addition to the skillset of any ophthalmologist.The purpose of this paper is to provide ophthalmologists with an updated,evidence-based review of the clinical indications for AMT in corneal and conjunctival reconstruction,reviewing its common and even experimental applications known to date.The methods of amniotic membrane preservation,the available commercial amniotic membrane products to date,and future directions for amniotic membrane use,including amniotic membrane extract eye drops(AMEED),are also discussed.It is paramount for ophthalmologists to stay up-to-date on the applications of AMT so as to effectively incorporate this versatile treatment modality into their practice,both in the operating room and in the clinic.By familiarizing the general ophthalmologist with its diverse applications,we hope to motivate general ophthalmologists to incorporate the use of AMT into their clinical practice,or provide guidance on how to recognize when referral to a corneal specialist for amniotic membrane application is prudent.

Human amniotic membrane transplantation: Different modalities of its use in ophthalmology

The amniotic membrane(AM) is the inner layer of the fetal membranes and consist of 3 different layers: the epithelium, basement membrane and stroma which further consists of three contiguous but distinct layers: the inner compact layer, middle fibroblast layer and the outermost spongy layer. The AM has been shown to have anti-inflammatory, anti-fibrotic, anti-angiogenic as well as anti-microbial properties. Also because of its transparent structure, lack of immunogenicity and the ability to provide an excellent substrate for growth, migration and adhesion of epithelial corneal and conjunctival cells, it is being used increasingly for ocular surface reconstruction in a variety of ocular pathologies including corneal disorders associated with limbal stem cell deficiency, surgeries for conjunctival reconstruction, as a carrier for ex vivo expansion of limbal epithelial cells, glaucoma surgeries and sceral melts and perforations. However indiscriminate use of human AM needs to be discouraged as complications though infrequent can occur. These include risk of transmission of bacterial, viral or fungal infections to the recipient if the donors are not adequately screened for communicable diseases, if the membrane is not processed under sterile condi-tions or if storage is improper. Optimal outcomes can be achieved only with meticulous case selection. This review explores the ever expanding ophthalmological indications for the use of human AM.

Effects of amniotic membrane transplantation on cytokines expression in chemically burned rat corneas

AIM: To investigate the effect of amniotic membrane transplantation (AMT) on the expressions of inflammatory-related, angiogenic-related and growth-related cytokines in rat corneas after chemical injury. METHODS: Alkali wounds were inflicted on the central corneas of rats by applying a round filter paper soaked in 1mol/L NaOH for 40 seconds. One week after alkali burn, 12 rats were randomly divided into 2 groups: the AMT group and the control group, and AMT was performed on the rats in the AMT group. Corneal opacity and neovascularization were observed by slit-lamp microscopy. The protein levels of interleukin (IL)-2, interferon (IFN)-gamma, IL-10 and transforming growth factor (TGF)-beta were determined by enzyme-linked immunosorbent assay 2 weeks after AMT. The mRNA levels of matrix metalloproteinase-2 (MMP-2), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) were evaluated by real-time quantitative PCR. RESULTS: In the AMT group, the corneal opacity was improved (P =0.011) and the area of corneal neovascularization was significantly decreased (P=0.005) compared with the control group. The amount of IL-2 and IFN-gamma secreted by Th1 cells were decreased after AMT, whereas the amount of IL-10 and TGF-beta secreted by Th2 cells were increased ( P <0.05). The level of MMP-2 was significantly down-regulated (P=0.013) at the nnRNA level in the AMT group, while the expression of EGF was significantly higher (P= 0.022) compared with the control. CONCLUSION: AMT may suppress corneal neovascularization after chemical injury by modulating the expressions of soluble factors.

角膜基质细胞诱导分化的脂肪间充质干细胞羊膜片移植治疗兔角膜碱烧伤的疗效及其机制

背景角膜化学烧伤是致盲眼病之一，先前的各种疗法在临床上的应用均受到一定的限制，而脂肪间充质干细胞（ADSCs）移植疗法受到密切关注。目的观察诱导分化的ADSCs羊膜片移植治疗兔角膜碱烧伤的疗效及其机制。方法无菌条件下剪取兔角膜，用悬浮基质片法分离并培养兔角膜基质细胞（CSCs）。取兔腹股沟脂肪组织，以酶消化法（质量分数0．25％胰蛋白酶）分离及培养兔ADSCs，并用流式细胞术鉴定ADSCs表面抗原的表达。将CSCs与ADSCs共培养，采用免疫荧光技术和逆转录PCR（RT-PCR）鉴定CSCs诱导后ADSCs向角膜基质样细胞分化的表达，分别将诱导或未诱导的ADSCs种植于羊膜上制备成细胞羊膜片。选择60只新西兰白兔，右眼用浸有质量分数1％NaOH的滤纸贴敷于角膜中央50S制备兔角膜碱烧伤模型，按随机数字表法将模型眼随机分为诱导ADSCs＋羊膜移植组、未诱导ADSCs＋羊膜移植组、单纯羊膜移植组及模型组。分别于术后1周、2周和1个月在裂隙灯显微镜下观察各组兔模型眼角膜的混浊程度和新生血管形成情况，并对角膜炎症进行评分，采用ELISA法检测术后1个月角膜组织匀浆中CD45、叮干扰素（IFN-γ）、白细胞介素．10（IL-10）蛋白及房水中血管内皮生长因子（VEGF）蛋白的质量浓度。结果从脂肪组织分离培养的ADSCs表面抗原CDl05、CD29、CD44的阳性表达率分别为90．23％、88．56％、98．88％；分离培养的CSCs波形蛋白呈阳性表达；CSCs与ADSCs共培养后多数细胞双标记染色阳性。苏木精一伊红染色显示，ADSCs种植于羊膜后生长良好。术后1个月，模型组角膜混浊呈瓷白色，新生血管面积最大，而诱导ADSCs＋羊膜移植组角膜透明，新生血管最少。术后1个月，诱导ADSCs＋羊膜移植组、未诱导ADSCs＋羊膜移植组、单纯羊膜移植组和模型组兔角膜混浊程度评分分别为1．65±0．18、2．05±0．17、2．68±0．25和2．904-0．18，新生血管面积分别为（10．59±1．78）、（22．58±1．63）、（37．98±1．90）和（45．37±1．65）mm^2，术后1周、2周及1个月4个组间角膜混浊程度评分和角膜新生血管（CNV）面积的差异均有统计学意义（F=280．826、330．172、465．707，均P=0．000；F=60．020、670．811、1510．231，均P=0．000），其中诱导ADSCs＋羊膜移植组角膜炎症评分睨显低于其他3个组，CNV面积小于其他3个组，差异均有统计学意义（均P〈0．01）。术后1个月，4个组间角膜组织匀浆中CD45、IL-10、IFN-γ质量浓度的差异均有统计学意义（F=916．545、1739．358、462．134，均P=0．000），其中与未诱导ADSCs＋羊膜移植组、单纯羊膜移植组和模型组比较，诱导ADSCs＋羊膜移植组CD45、IFN-γ质量浓度明显降低，而IL-10质量浓度明显升高，差异均有统计学意义（均P〈0．01）。4个组兔房水中VEGF质量浓度的总体比较差异有统计学意义（F=129．126，P=0．000），其中诱导ADSCs＋羊膜移植组兔房水中VEGF质量浓度明显低于未诱导ADSCs＋羊膜移植组、单纯羊膜移植组和模型组，差异均有统计学意义（均P〈0．01）。结论CSCs诱导的ADSCs羊膜片移植法治疗兔角膜碱烧伤有较好的疗效，其作用机制可能是ADSCs在CECs诱导下分化为具有角膜上皮细胞特征的细胞，同时羊膜可减轻免疫炎症反应，抑制新生血管形成。

保存人羊膜联合自体角膜缘移植治疗陈旧性眼部烧伤

目的 观察羊膜移植 (AMT)在重建眼表面 ,治疗陈旧性眼部烧伤中的应用价值。方法 采用保存人羊膜联合自体角膜缘移植 (LT)治疗 15例 ( 15只眼 )。结果 15只眼中 ,12只眼眼表面重建成功 ,10只眼角膜透明或半透明 ,8只眼脱盲。结论 保存人羊膜无抗原性 ,具有抗炎、抑制新生血管及纤维化 ,阻止结膜源性组织再侵入的作用 ,采用保存人羊膜联合自体LT治疗陈旧性眼部灼伤 ,是一种有价值的方法 。

保存羊膜移植在眼表重建方面的临床应用

目的:观察和探讨保存人羊膜移植在眼表重建方面的临床应用和作用机制。方法:采用保存人羊膜移植治疗各种原因造成的眼表疾患共144例(154眼),术后随访观察4mo～1a。结果:术后患者眼部充血、刺激症状均较轻,约在1～2wk后移植区水肿消退。111眼(72.1%)视力有提高;147眼(95.5%)一次性达到治疗效果。结论:保存人羊膜移植能够安全、有效地重建眼表疾患患者的眼表,值得广泛应用。

保存羊膜移植治疗中、重度眼化学烧伤和热烧伤急性期的作用

目的讨并分析保存羊膜在眼中、重度化学烧伤和热烧伤急性期的治疗作用以及对晚期并发症的预防作用。方法选择兰州医学院第二附属医院急性化学烧伤和热烧伤的住院患者19例(21眼)根据病情分为中度组(14眼烧伤为Ⅱ～Ⅲ度)和重度组(7眼烧伤为Ⅳ度),入院1周内分别行单纯羊膜移植术和羊膜移植术+自体(异体)角膜缘上皮干细胞移植术。观察早期眼表重建时间,晚期角膜血管化,睑球粘连,视力改善情况。结果中度组的眼表重建时间较重度组短,视力提高较后者明显,差异有显著性意义(P<0.001)。在角膜血管化,睑球粘连等方面也较后者轻。结论羊膜可用于眼中、重度化学烧伤和热烧伤的急性期治疗,其预后效果同眼部损伤程度有直接关系。

新鲜羊膜移植治疗早期重症眼部烧伤

目的 探讨新鲜羊膜移植联合结膜、角膜移植对早期重症眼部烧伤的治疗效果。方法 急性重症眼部烧伤患者21例22眼,分别行羊膜移植联合结膜移植12例12眼和羊膜、结膜移植联合板层或穿透性角膜移植9例10眼。术后观察羊膜、结膜及角膜植片情况,术后随访2～16个月,平均7 个月。结果 19眼羊膜植片愈合良好;20眼穹隆部结膜植片全部存活;羊膜移植联合结膜移植中10眼术后未见角膜融解,角膜缘均未见新生血管长入。羊膜、结膜移植联合板层或穿透性角膜移植,1例2眼出现轻度排斥反应,另8眼角膜植片均恢复透明。结论 新鲜羊膜移植可有效地促进上皮细胞分化移行、抑制局部的炎症反应、阻止新生血管和瘢痕形成。

深低温保存羊膜治疗严重睑球粘连的临床观察

目的:探讨深低温保存羊膜移植治疗严重睑球粘连的临床价值。方法:对43例46眼有严重睑球粘连及假性胬肉的患者,进行睑球粘连松解后,应用羊膜移植重建球结膜和角膜表面,观察其临床效果。结果:在随访期内(平均13±3.5mo),24例患者疗效为优,眼睑球粘连均得到明显改善,眼表基本恢复正常;15例疗效为良,睑球粘连部分复发,但较术前有所改善;7例因上下睑球全粘连、结膜囊闭锁愈后效果较差。结论:对于严重的睑球粘连,深低温保存羊膜移植重建眼表是较理想的治疗方法。

羊膜移植治疗眼烧伤疗效的观察

目的 评价羊膜移植治疗眼部烧伤的疗效。 方法 2 0 0 0年 10月～ 2 0 0 2年 5月 ,对 2 0例患者 (2 8眼 )采用羊膜移植治疗眼烧伤 ,其中碱烧伤 6例 (10眼 ) ,酸烧伤 8例 (10眼 ) ,热烧伤 6例 (8眼 )。年龄 8～ 4 5岁 ,平均 2 8.6岁。烧伤部位包括角膜、结膜。受伤至手术时间 1～ 2 1天 ,平均 10天。 结果 2 8眼术后 3～ 7天眼部炎症控制 ,角、结膜均未见继续溶解 ,无穿孔 ,无虹膜萎缩 ,视力有不同程度提高 ;16眼角膜恢复透明 ,4眼角膜白斑 ,8眼全角膜浑浊。全部患者均保存了眼球 ,眼表稳定 ,无免疫排斥反应及继发感染发生。 结论 羊膜移植可以减轻炎性反应 ,减少新生血管的生成 ,抑制纤维组织增生 ,对重建眼表功能及保存眼球有较高的价值 ,可有效地用于眼部烧伤的治疗 ,烧伤后手术愈早效果愈好。

羊膜移植治疗严重眼表面疾病的临床观察

目的：研究羊膜移植治疗严重眼表面疾病的临床疗效。方法：对严重眼表面疾病29例31眼，其中急性眼烧伤炎症期6眼，陈旧性眼烧伤睑球粘连及其它眼表疾病25眼，施行单纯羊膜移植（15眼）和羊膜移植联合板层角膜移植术（16眼），随访3～18个月，平均8.4个月。结果：急性眼烧伤病变中4眼的炎症显著减轻，角膜未继续溶解，上皮修复；慢性病变中有20只眼的眼表面获得重建，睑球粘连得到治愈或显著改善，视力不同程度提高。结论：对于急性严重的眼烧伤，羊膜移植能有效地阻止炎症和角膜溶解的发展，防止或减轻疤痕；对于陈旧性眼烧伤和其它眼表面疾病，羊膜是一种良好的生物学材料移植后能有效的重建眼表面。

新鲜和保存羊膜移植重建结膜眼表的临床对比研究

目的:比较新鲜羊膜和保存羊膜治疗睑球粘连的疗效差异,分析影响疗效的因素。方法:共51例55只眼因陈旧性化学伤、热烧伤或Stevens-Johnson综合征而发生睑球粘连的临床连续病例接受睑球粘连分离联合新鲜羊膜移植(22只眼)或保存羊膜移植(33只眼)。其中男30例32只眼,女21例23只眼。年龄4～51岁,平均(34.2±4.3)岁。其中11只眼在烧伤后5～11个月,平均(7.4±1.6)个月时接受手术,40只眼在烧伤后1～8年,平均(2.0±0.7)年进行手术。结果:术后随访12～32个月,平均(19.3±4.1)个月。所有移植在结膜眼表面的羊膜(包括新鲜羊膜)植片在术后早期均未见溃烂和溶解,周边对合良好。31/55(56.4%)只眼形成了足够深的穹窿部且恢复了眼球运动功能。9/55(16.4%)只眼发生部分睑球粘连,眼球运动轻度受限,但其面积远较术前为小。15/55(27.3%)眼术后发生中度以上的睑球粘连。新鲜羊膜和保存羊膜重建眼结膜表面的效果相似(χ2=0.466,P=0.797);不同程度睑球粘连的患者其羊膜移植术后的效果不同(新鲜羊膜,χ2=27.995,P=0.000;保存羊膜,χ2=33.610,P=0.000);在眼表烧伤后1年内手术比1年以上进行羊膜移植的效果也不同(χ2=4.243,P=0.039)。结论:新鲜羊膜和保存羊膜一样可以有效地用于重建睑球粘连解除后的结膜眼表。患眼术前睑球粘连程度以及烧伤后其眼表炎症是否处于安静状态等因素都会直接影响羊膜移植重建眼结膜表面的远期疗效。

角膜缘上皮联合羊膜移植治疗眼表疾病的研究

目的 探讨角膜缘上皮联合羊膜移植治疗眼表疾病的效果。方法 复发性翼状胬肉、睑球粘连、陈旧性碱烧伤、角膜缘外伤、角巩膜大面积皮样肿 ,采用甘油保存的人羊膜联合自体或异体角膜缘移植进行治疗。结果 复发性翼状胬肉 6例 6眼 ,术后随访 4月～ 2年 ,上皮愈合稳定 ,角膜光滑透明 ,无复发。视力提高或保持不变。陈旧性碱烧伤 3例 4眼 ,随访 3、8、12月 ,表面保持安静 ,周边部有少许的新生血管 ;一眼手术失败 ,出现血管生长 ,基质溶解。角巩膜大范围皮样肿 1眼 ,术后角膜半透明 ,外观正常。视力除 1例失败外 ,均有不同程度的提高。从术前光感～ 0 0 5提高至 0 0 2～ 0 8。结论 保存的人羊膜无抗原性 ,具有抗炎、抑制新生血管、阻止结膜源组织再侵入的作用。

新鲜羊膜移植治疗眼表疾病的临床评价

目的 评价新鲜人羊膜在眼表重建手术中的临床价值。方法 用新鲜羊膜移植 ,治疗陈旧性睑球粘连、结膜囊缺失、复发性翼状胬肉、蚕蚀性角膜溃疡、热烧伤后角膜和结膜溶解等眼表疾病的连续病例19例 ( 2 1只眼 )。结果 19例患者的羊膜移植手术均为一次成功。平均随访 12 .5个月 ,羊膜移植片未见免疫排斥反应或溶解吸收。手术后 ,患者的睑球粘连松解 ,结膜囊形成 ,蚕蚀性角膜溃疡和复发性翼状胬肉在随访期间未见复发 ,角膜和结膜溶解未继续进展。结论 新鲜羊膜和保存的羊膜一样 ,可有效地重建角膜和结膜表面。新鲜羊膜具有取材简单 ,使用方便 ,费用低廉等优点 ,更适合于基层医院在眼表重建手术中使用。

羊膜移植在眼表疾病中的应用

目的 探讨羊膜移植治疗眼表疾病的疗效，寻找有效降低翼状胬肉术后复发率的新方法。方法 对３４例翼状胬肉、１例硷烧伤致睑球粘连患者进行保存羊膜移植手术，并在术后３月、６月分别取患眼部分羊膜植片进行组织细胞学检查，随访３～１８月。结果 羊膜移植片生长良好。术后复发率为５７％。无感染现象。细胞学检查表明，羊膜能够在免疫条件下成活后适应生长。结论 羊膜移植可用于治疗多种眼表疾病，尤其可以有效降低翼状胬肉术后复发率，是一种能够重建正常眼表，值得推广的新手术。