Surgical Management of Bilateral Limbal Stem Cell Deficiency.

At the age of 43 years-old, a man was left with bilateral limbal stem cell deficiency after an ocular alkaline burn with lime, which resulted in corneal opacification. After multiple unsuccessful surgical attempts to restore vision, including penetrating keratoplasties and Boston keratoprosthesis, visual acuity was counting fingers in the left eye. At 73 years of age, the patient underwent another surgery in his left eye. Cauterization of neovessels and removal of the vascular pannus were followed by partial excision of Tenon's capsule. Penetrating keratoplasty was followed by an intrastromal injection of anti-VEGF (vascular endothelial growth factor), and the ocular surface was covered with amniotic membrane. Postoperatively, the graft was clear with no signs of inflammation; vision improved to 20/50 and remained stable throughout the following two years. Herein we describe some adjunctive procedures that might have delayed failure and rejection of the corneal graft. This case demonstrates the difficulties in treating bilateral limbal stem cell deficiency in a tertiary eye care center with no capacity to perform stem cell therapy.

[Research progress on limbal stem cell transplantation].

Limbal Stem Cell Deficiency (LSCD) is an ocular surface disease caused by the decrease of the quantity and dysfunction of limbal stem cell, which is characterized by conjunctivalization and other signs of epithelial dysfunction. For sever LSCD, surgery is the main treatment way. Recently, plenty of researches published the outcomes of different operation methods. This article summarized five major operations, including conjunctival limbal autograft (CLAU), simple limbal epithelial transplantation (SLET), limbal allograft, cultivated limbal stem cell transplantation (CLET) and cultivated oral mucosal epithelial transplantation (COMET). (Chin J Ophthalmol, 2020, 56:956-960).

Severe corneal burn due to the accidental application of salicylic acid packed in a plastic dropper bottle / Quemadura corneal grave debida a la aplicación accidental de ácido salicílico envasado en un frasco gotero plástico

Eye burns due to the accidental application of pharmacological or nonpharmacological substances packaged in plastic dropper bottles have been described for more than three decades and continue to occur. These burns can cause potentially serious corneal injuries. We report the case of a patient who mistakenly applied salicylic acid to the right eye after confusing it with an eye lubricant, which caused him a severe corneal burn. Fortunately, after aggressive medical and surgical management (including oxygen therapy and amniotic membrane grafting), the visual results were good. We suggest conducting educational campaigns and taking legislative measures in our country to avoid packaging corrosive substances in this type of dropper bottle to reduce the risk of accidental burns.

Las quemaduras oculares por aplicación accidental de sustancias farmacológicas o no farmacológicas envasadas en frascos goteros plásticos, se han descrito desde hace más de tres décadas y siguen ocurriendo. Estas quemaduras pueden causar lesiones potencialmente graves de la córnea. Se presenta el caso de un paciente que se aplicó ácido salicílico en el ojo derecho al confundir el envase con el de un lubricante ocular, lo que le causó una grave quemadura corneal. Afortunadamente, tras un manejo médico y quirúrgico agresivo (incluida oxigenoterapia e injerto de membrana amniótica) los resultados visuales fueron buenos. Se sugiere hacer campañas educativas y adoptar normas en el país para evitar el envase de sustancias corrosivas en este tipo de frascos goteros, con el fin de disminuir el riesgo de quemaduras accidentales.

Clinical features and corneal optical anisotropies in a rabbit model of limbal stem cell deficiency / Fenótipos clínicos e anisotropias ópticas da córnea em coelhos com deficiência de células tronco limbais

ABSTRACT Purposes: To investigate the intra-laboratory reproducibility of clinical features and to evaluate corneal optical anisotropies in a rabbit model of limbal stem cell deficiency. Methods: Limbal injury was induced in the right eye of 23 adult New Zealand White rabbits using a highly aggressive protocol that combined 360 degrees limbal peritomy, keratolimbectomy, alkaline chemical burn, and mechanical removal of the epithelium. Clinical evaluation of the injured eyes was performed for 28 days and included corneal impression cytology. Corneas with a severe clinical outcome set typical of limbal stem cell deficiency were then collected, subjected to a histopathological examination, and examined for optical anisotropies. Corneas from healthy rabbit eyes were used as controls. Differences in optical path due to stromal collagen birefringence, as well as linear dichroism related to the expression and spatial orientation of glycosaminoglycan chains from proteoglycans, were measured from cross-sections under a quantitative polarized light microscope. Results: One eye showed signs of hypopyon and was excluded. Signs of ocular inflammation were observed in all eyes studied (n=22). Corneal impression cytology did not detect goblet cells. Twelve of the 22 corneas presented a clinical outcome set typical of limbal stem cell deficiency, which is characterized by the presence of epithelial defects, inflammatory cells, moderate-to-severe opacity, and neovascularization. Microscopic studies under polarized light revealed that relative to controls, limbal stem cell deficiency caused a 24.4% increase in corneal optical path differences. Further, corneas with limbal stem cell deficiency were less dichroic than controls. Conclusions: These results suggest that rabbit models of limbal stem cell deficiency must be rigorously screened for use in preclinical studies to ensure experimental homogeneity because protocols used to create limbal stem cell deficiency could be not associated with good intra-laboratory reproducibility of clinical features. Limbal stem cell deficiency, as induced herein, altered the optical anisotropic properties of the corneal stroma. Such alterations are indicative of changes in collagen packing and the spatial orientation of glycosaminoglycan chains from proteoglycans. Knowledge of these changes is important to potentiate strategies aimed at restoring the morphofunctional integrity of the corneal stroma affected by limbal stem cell deficiency.

RESUMO Objetivos: Investigar a reprodutibilidade intra-la­boratorial dos fenótipos clínicos e avaliar anisotropias ópticas em córneas de coelhos com deficiência de células tronco limbais. Métodos: Lesões ao limbo foram feitas no olho direito de 23 coelhos adultos da Nova Zelândia Branco, usando um protocolo altamente agressivo, que envolveu peritomia limbal em 360 graus, ceratolimbectomia, cauterização por álcali, e remoção mecânica de epitélio remanescente. Os olhos foram clinicamente avaliados por 28 dias, inclusive por citologia de impressão corneal. As córneas que manifestaram um conjunto de alterações típicas de deficiência de células tronco limbais foram coletadas e submetidas à estudos em histopatologia e em anisotropias ópticas. Córneas saudáveis foram usadas como controles. Diferenças de caminho óptico de birrefringência relacionada à organização do colágeno estromal, e dicroísmo linear relacionado à expressão e à orientação das cadeias de glicosaminoglicanos dos proteoglicanos estromais, foram quantificados por microscopia de luz polarizada. Resultados: Um olho apresentou hipópio e foi excluído do estudo. Todos os olhos estudados (n=22) apresentaram sinais de inflamação ocular. A citologia de impressão não detectou células caliciformes na superfície corneal. Doze de 22 córneas manifestaram alterações clínicas típicas de deficiência de células tronco limbais, caracterizado por defeitos epiteliais, infiltrados inflamatórios, opacidade de moderada à severa, e neovascularização. Estudos por microscopia de luz polarizada mostraram que a deficiência de células tronco limbais aumentou a diferenças de caminho óptico corneal em 24,4% (versus controles). As cór­neas com deficiência de células tronco limbais foram menos dicroicas do que as córneas controle. Conclusões: Coelhos com deficiência de células tronco limbais, para aplicações em estudos pré-clínicos, devem ser rigorosamente selecionados para assegurar homogeneidade experimental, pois há evidências de que protocolos utilizados para indução de deficiência de células tronco limbais não estão associados com boa reprodutibilidade intra-laboratorial de fenótipos clínicos. A deficiência de células tronco limbais, como induzida aqui, alterou as propriedades ópticas anisotrópicas do estroma corneal. Tais alterações são indicativas de mudanças no empacotamento de colágeno e na orientação das cadeias de glicosaminoglicanos dos proteoglicanos. Conhecimentos nessas alterações são importantes para potencializar estratégias que visam a restabelecer a integridade morfofuncional do estromal corneal acometido pela deficiência de células tronco limbais.

Lamellar keratoplasty using femtosecond laser intrastromal lenticule for limbal dermoid: case report and literature review.

Small incision refractive lenticule extraction (SMILE) is a femtosecond laser technique to correct myopia and myopic astigmatism. Herein, we report a technique where intrastromal lenticule obtained from the SMILE procedure served as a graft for lamellar keratoplasty in the management of a limbal dermoid. An 18-year-old woman presented to the clinic with a corneal-limbal mass in the right eye. Slit-lamp examination revealed a vascularized circular mass of approximately 6 mm × 5 mm, which was attached at 7 o'clock in the inferotemporal region of the corneal limbus; this suggested limbal dermoid. Anterior segment optical coherence tomography revealed superficial involvement of the cornea. The patient was treated with excision and lamellar keratoplasty by using femtosecond intrastromal lenticule. The lenticule was sutured over the cornea with 10-0 interrupted nylon sutures. On postoperative follow-up, best-corrected visual acuity was 20/20; there was no corneal neovascularization and no sign of rejection. This case of limbal dermoid was managed by simple surgical excision and lamellar keratoplasty with a SMILE-extracted lenticule. This method may serve as an alternative surgical approach for management of limbal dermoid.

Pigmentation Is Associated with Stemness Hierarchy of Progenitor Cells Within Cultured Limbal Epithelial Cells.

Ex vivo cultured human limbal epithelial stem/progenitor cells (hLESCs) are the main source for regenerative therapy of limbal stem cell deficiency (LSCD), which is worldwide one of the major causes of corneal blindness. Despite many stemness-associated markers have been identified within the limbal niche, the phenotype of the earliest hLESCs has not been hitherto identified. We sought to confirm or refute the use of tumor protein p63 (p63) and ATP binding cassette subfamily B member 5 (ABCB5) as surrogate markers for hLESCs early within the limbal differentiation hierarchy. Based on a robust fluorescence-activated cell sorting and subsequent RNA isolation protocol, a comprehensive transcriptomic profile was obtained from four subpopulations of cultured hLESCs. The subpopulations were defined by co-expression of two putative stem/progenitor markers, the p63 and ABCB5, and the corneal differentiation marker cytokeratin 3. A comparative transcriptomic analysis yielded novel data that indicated association between pigmentation and differentiation, with the p63 positive populations being the most pigmented and immature of the progenitors. In contrast, ABCB5, either alone or in co-expression patterns, identified more committed progenitor cells with less pigmentation. In conclusion, p63 is superior to ABCB5 as a marker for stemness. Stem Cells 2018;36:1411-1420.

A supramolecular look at microenvironmental regulation of limbal epithelial stem cells and the differentiation of their progeny / Uma visão supramolecular sobre a regulação microambiental de células tronco epiteliais limbais e a diferenciação de sua progênie

ABSTRACT Various approaches have been taken to improve our knowledge of the microenvironmental regulation of limbal epithelial stem cells. Researchers have extensively investigated the roles of growth factors, survival factors, cytokines, enzymes, and permeable molecules secreted by the limbal cells. However, recent evidence suggests that stem cell fate (i.e., self-renewal or differentiation) can also be influenced by biophysical and mechanical cues related to the supramolecular organization and the liquid crystalline (mesophase) nature of the stromal extracellular matrix. These cues can be sensed by stem cells and transduced into intracellular biochemical and functional responses, a process known as mechanotransduction. The objective of this review is to offer perspectives on the supramolecular microenvironmental regulation of limbal epithelial stem cells and the differentiation of their progeny.

RESUMO Muitas abordagens têm sido utilizadas para ampliar entendimentos sobre a regulação microambiental das células tronco epiteliais limbais. Neste contexto, pesquisadores têm exaustivamente investigado a participação de fatores de crescimento, fatores de sobrevida, citocinas, enzimas e moléculas permeáveis secretadas pelas células limbais. Entretanto, evidências recentes sugerem que o destino (ie. autorrenovação ou recrutamento para a via de diferenciação) das células tronco também sofre influência de estímulos biofísicos ou mecânicos relacionados à organização supramolecular e à natureza liquido-cristalina (mesofases) da matriz extracelular estromal. Esses estímulos podem ser percebidos e traduzidos pelas células tronco em sinais bioquímicos que geram respostas funcionais, através de um processo designado de mecanotransdução. Objetiva-se, com a presente revisão, oferecer ao leitor perspectivas supramoleculares sobre a regulação microambiental das células tronco epiteliais limbais e a diferenciação de sua progênie.

[Allogeneic cultured limbal epithelial transplantation: a clinical observation of its treatment effects on large conjunctival lesions].

Objective: To introduce the surgical methods and investigate the treatment effects of conjunctival lesion resection combined with transplantation of allogeneic limbal epithelia cultured in vitro for treatment of varied large conjunctival lesions. Methods: A retrospective analysis of case data of the patients receiving transplantation of allogeneic limbal epithelia cultured in vitro for large conjunctival lesions at Qingdao Eye Hospital from January 2013 to December 2015 was conducted. Fresh donor corneal limbal epithelial tissues were obtained for cellular digestion and isolation. The cells were cultured on de-epithelized amniotic membrane to form a stratified cell sheet with 3 to 5 layers of cells for reservation. Conjunctival lesions were removed, and corneal limbal epithelial cell sheets corresponding to the size of the conjunctival defects were covered on the scleral flaps, before the conjunctivae were para sutured with corneal limbal epithelial cell sheets. The stitches were removed 7 days after surgery. The treatment effects were evaluated. Results: There were 11 cases (12 eyes) in this study, including 6 cases (6 eyes) of recurrent pterygium combined with symblepharon, 3 cases (3 eyes) of conjunctival compound nevus, 1 case (1 eye) of conjunctival primary acquired melanosis, and 1 case (2 eyes) of dermoid lipoma. The patients were 6 to 69 years old, with an average age of (46±23) years old. There were 5 male patients and 6 female patients. The follow-up time was 6 to 39 months, with an average time of (24.8±12.4) months. The conjunctival lesions in all patients were more than 2 quadrants. No patients had intraoperative and postoperative complications. No obvious irritation symptoms were observed after surgery. There were a few small pieces of epithelial defects on the limbal epithelial cell sheets at the end of the operation due to operative injury. With postoperative cellular proliferation and repair, the epithelium gradually healed and tended to become firm, and the boundaries with original conjunctival epithelium gradually disappeared. The healing time of the epithelium after surgery was 3 to 7 days, with an average time of (4.5±1.2) days. No immunological rejection occurred after surgery. No recurrence of original diseases was observed among all the patients at the last follow-up. Conclusions: Transplantation of allogeneic cultured limbal epithelial cells, which was carried out for the treatment of large conjunctival lesions, witnessed rapid epithelization at the areas of lesion resection. It could reduce the risk of recurrence of original diseases and effectively avoid the occurrence of postoperative symblepharon. The postoperative inflammatory reaction was also minor. (Chin J Ophthalmol, 2017, 53:172-176).

Autologous method for ex vivo expansion of human limbal epithelial progenitor cells based on plasma rich in growth factors technology.

PURPOSE: Develop an autologous culture method for ex vivo expansion of human limbal epithelial progenitor cells (LEPCs) using Plasma Rich in Growth Factors (PRGF) as a growth supplement and as a scaffold for the culture of LEPCs. METHODS: LEPCs were cultivated in different media supplemented with 10% fetal bovine serum (FBS) or 10% PRGF. The outgrowths, total number of cells, colony forming efficiency (CFE), morphology and immunocytochemistry against p63- α and cytokeratins 3 and 12 (CK3-CK12) were analyzed. PRGF was also used to elaborate a fibrin membrane. The effects of the scaffold on the preservation of stemness and the phenotypic characterization of LEPCs were investigated through analysis of CK3-CK12, ABCG-2 and p63. RESULTS: LEPCs cultivated with PRGF showed a significantly higher growth area than FBS cultures. Moreover, the number of cells were also higher in PRGF than FBS, while displaying a better morphology overall. CFE was found to be also higher in PRGF groups compared to FBS, and the p63-α expression also differed between groups. LEPCs cultivated on PRGF membranes appeared as a confluent monolayer of cells and still retained p63 and ABCG-2 expression, being negative for CK3-CK12. CONCLUSIONS: PRGF can be used in corneal tissue engineering, supplementing the culture media, even in a basal media without any other additives, as well as providing a scaffold for the culture.

Comparison of Toric Foldable Iris-Fixated Phakic Intraocular Lens Implantation and Limbal Relaxing Incisions for Moderate-to-High Myopic Astigmatism.

PURPOSE: To compare the effectiveness of toric foldable iris-fixated phakic intraocular lens (pIOL) implantation and non-toric foldable iris-fixated pIOL implantation with limbal relaxing incisions (LRIs) for correcting moderate-to-high astigmatism in myopic eyes. MATERIALS AND METHODS: The medical records of 146 patients (195 eyes) with myopic astigmatism who underwent toric foldable iris-fixated pIOL implantation (toric group; 94 eyes) or non-toric foldable iris-fixated pIOL implantation with concurrent LRIs (LRI group; 101 eyes) were retrospectively reviewed. For subgroup analysis, the two groups were subdivided according to preoperative astigmatic severity [moderate, 2.00 to <3.00 diopters (D); high, 3.00-4.00 D]. Visual and astigmatic outcomes were compared 6 months postoperatively. RESULTS: The uncorrected distance visual acuity was at least 20/25 in 100% and 98% of the toric and LRI group eyes, respectively. The toric group had lower mean residual cylindrical error (-0.67±0.39 D vs. -1.14±0.56 D; p<0.001) and greater mean cylindrical error change (2.17±0.56 D vs. 1.63±0.72 D; p<0.001) than the LRI group, regardless of the preoperative astigmatic severity. The mean correction index (1.10±0.16 vs. 0.72±0.24; p<0.001) and success index (0.24±0.14 vs. 0.42±0.21; p<0.001) also differed significantly between the groups. CONCLUSION: Both surgical techniques considerably reduced astigmatism and had comparable visual outcomes. However, toric foldable iris-fixated pIOL implantation was more reliable for correcting moderate-to-high astigmatism in myopic eyes.

Assessment of surgical outcomes of limbal transplantation using simple limbal epithelial transplantation technique in patients with total unilateral limbal deficiency / Avaliação dos resultados cirúrgicos do transplante de limbo utilizando a técnica SLET (simple limbal epithelial transplantation), em pacientes com deficiência límbica total unilateral

ABSTRACT This study aimed to evaluate the effectiveness of the novel simple limbal epithelial transplantation (SLET) technique, which reduces the risk of iatrogenic limbal stem cell deficiency in the donor eye. Four patients with total unilateral limbal stem cell deficiency received a limbal graft, measuring 4 mm × 2 mm, from the contralateral healthy eye in a single surgical procedure. The graft was divided into 10-20 pieces and distributed on the corneal surface. At 6-month follow-up, a completely avascular corneal epithelial surface was obtained in two patients, and there was improvement in visual acuity in one patient. The limbal grafts did not adhere to the cornea in one patient. No serious complications related to the surgery were observed in this study.

RESUMO Este trabalho tem como objetivo avaliar a eficácia de uma nova técnica cirúrgica denominada SLET (simple limbal epithelial transplantation), um procedimento promissor que reduz os riscos de indução de deficiência límbica iatrogênica no olho doador. Quatro pacientes com deficiência límbica total unilateral, secundária a queimadura química, receberam um enxerto de células límbicas, medindo 4 mm x 2 mm, do olho contralateral sadio, em apenas um tempo cirúrgico. Este foi divido em 10 a 20 fragmentos e distribuído sobre a superfície da córnea. Após 6 meses de cirurgia, superfície corneana totalmente epitelizada e avascular foi obtida em dois pacientes. Houve melhora da acuidade visual em um paciente. Não houve aderência dos enxertos de limbo na córnea em um paciente. Nenhum paciente apresentou efeitos colaterais graves decorrentes do procedimento cirúrgico.

Comparison of Toric Foldable Iris-Fixated Phakic Intraocular Lens Implantation and Limbal Relaxing Incisions for Moderate-to-High Myopic Astigmatism

PURPOSE: To compare the effectiveness of toric foldable iris-fixated phakic intraocular lens (pIOL) implantation and non-toric foldable iris-fixated pIOL implantation with limbal relaxing incisions (LRIs) for correcting moderate-to-high astigmatism in myopic eyes. MATERIALS AND METHODS: The medical records of 146 patients (195 eyes) with myopic astigmatism who underwent toric foldable iris-fixated pIOL implantation (toric group; 94 eyes) or non-toric foldable iris-fixated pIOL implantation with concurrent LRIs (LRI group; 101 eyes) were retrospectively reviewed. For subgroup analysis, the two groups were subdivided according to preoperative astigmatic severity [moderate, 2.00 to <3.00 diopters (D); high, 3.00–4.00 D]. Visual and astigmatic outcomes were compared 6 months postoperatively. RESULTS: The uncorrected distance visual acuity was at least 20/25 in 100% and 98% of the toric and LRI group eyes, respectively. The toric group had lower mean residual cylindrical error (-0.67±0.39 D vs. -1.14±0.56 D; p<0.001) and greater mean cylindrical error change (2.17±0.56 D vs. 1.63±0.72 D; p<0.001) than the LRI group, regardless of the preoperative astigmatic severity. The mean correction index (1.10±0.16 vs. 0.72±0.24; p<0.001) and success index (0.24±0.14 vs. 0.42±0.21; p<0.001) also differed significantly between the groups. CONCLUSION: Both surgical techniques considerably reduced astigmatism and had comparable visual outcomes. However, toric foldable iris-fixated pIOL implantation was more reliable for correcting moderate-to-high astigmatism in myopic eyes.

Comparison of Toric Foldable Iris-Fixated Phakic Intraocular Lens Implantation and Limbal Relaxing Incisions for Moderate-to-High Myopic Astigmatism

PURPOSE: To compare the effectiveness of toric foldable iris-fixated phakic intraocular lens (pIOL) implantation and non-toric foldable iris-fixated pIOL implantation with limbal relaxing incisions (LRIs) for correcting moderate-to-high astigmatism in myopic eyes. MATERIALS AND METHODS: The medical records of 146 patients (195 eyes) with myopic astigmatism who underwent toric foldable iris-fixated pIOL implantation (toric group; 94 eyes) or non-toric foldable iris-fixated pIOL implantation with concurrent LRIs (LRI group; 101 eyes) were retrospectively reviewed. For subgroup analysis, the two groups were subdivided according to preoperative astigmatic severity [moderate, 2.00 to <3.00 diopters (D); high, 3.00–4.00 D]. Visual and astigmatic outcomes were compared 6 months postoperatively. RESULTS: The uncorrected distance visual acuity was at least 20/25 in 100% and 98% of the toric and LRI group eyes, respectively. The toric group had lower mean residual cylindrical error (-0.67±0.39 D vs. -1.14±0.56 D; p<0.001) and greater mean cylindrical error change (2.17±0.56 D vs. 1.63±0.72 D; p<0.001) than the LRI group, regardless of the preoperative astigmatic severity. The mean correction index (1.10±0.16 vs. 0.72±0.24; p<0.001) and success index (0.24±0.14 vs. 0.42±0.21; p<0.001) also differed significantly between the groups. CONCLUSION: Both surgical techniques considerably reduced astigmatism and had comparable visual outcomes. However, toric foldable iris-fixated pIOL implantation was more reliable for correcting moderate-to-high astigmatism in myopic eyes.

Limbal transplantation at a tertiary hospital in Brazil: a retrospective study / Transplante de limbo em hospital terciário no Brasil: estudo retrospectivo

ABSTRACTPurpose:Epidemiological analysis of limbal transplantation surgeries performed in the Ophthalmologic Hospital of Sorocaba.Methods:Retrospective medical records review of 30 patients who underwent limbal stem cell transplants between January 2003 and March 2008. Cases involving conjunctival limbal autograft were classified as group I, and those involving conjunctival limbal allograft as group II.Results:Two patients were excluded due to incomplete data during postoperative follow-up. Of the total sample of 28 patients, 53.6% constituted group I, whereas 46.4% were included in group II. Males were predominant (67.9%), and right eyes were the most prevalent (67.9%). The mean age was 40.3 years. Unilateral cases accounted for 60.7%. The most frequent pathology causing limbal system failure was chemical burns (53%). The mean length of time from diagnosis to surgery was 11.18 years. The limbal graft and amniotic membrane were associated in 75% of all cases, and tarsorrhaphy in 57.1%. The average follow-up period was 24.84 months. The uncorrected visual acuity improved in 38% of the cases, was unchanged in 28.5%, and deteriorated in 33.3%. There was no persistent epithelial defect in 75% of the patients. The conjunctivalization rate was similar between the groups (53.3% and 58.3%, respectively). The transparency improved in only 38.4% of the cases, and 28.5% of the surgeries performed were successful. The most prevalent complication was persistent epithelial defect, which occurred in 25% of the patients, followed by corneal melting in 14.2%. Other complications observed included infectious ulcers, limbal graft necrosis or ischemia, perforation, and descemetocele.Conclusion:Chemical burns remain the main cause of limbal stem cell deficiency. In these cases, limbal transplantation is the standard procedure to restore the ocular surface even though the success rate is low.

RESUMOObjetivo:Análise epidemiológica dos transplantes de limbo realizados no Hospital Oftalmológico de Sorocaba.Método:Foi realizada uma análise retrospectiva através de revisão de prontuário de trinta casos de transplante de limbo realizados no período de janeiro de 2003 a março de 2008. Casos de transplante de limbo autólogo foram classificados como grupo I e alogênicos como grupo II.Resultados:Dois pacientes foram excluídos da análise por insuficiência de dados nos registros. Da amostra completa de 28 pacientes, 53,6% constituíram o grupo I enquanto 46,4% o grupo II. O olho direito (68%) e o sexo masculino (68%) foram mais acometidos, com uma média de idade de 40,3 anos. Casos unilaterais contabilizaram 60,7%. A patologia de base causadora da deficiência límbica mais prevalente foi a queimadura química (53%). A média do tempo de doença até a cirurgia foi de 11,18 anos. Na maioria dos casos o transplante foi associado a membrana amniótica (75%) e tarsorrafia (57%). O tempo médio de seguimento foi de 24,84 meses. Foi observado melhora da acuidade visual não corrigida em 38% dos casos enquanto 28,5% permaneceram inalteradas e 33,3% pioraram. Evolução sem defeito epitelial persistente ocorreu em 75% dos pacientes. A taxa de conjuntivalização foi semelhante nos 2 grupos (53,3% e 58,3%). Em apenas 38% dos casos houve melhora da transparência. A taxa de sucesso foi de cerca de 28%. A complicação mais prevalente foi defeito epitelial persistente (25%) seguida de melting (14,2%). Outras complicações observadas foram úlceras infecciosas, necrose ou isquemia do enxerto, perfuração e descemetocele.Conclusões:A queimadura química permanece como principal patologia causadora de deficiência límbica. Nestes casos o transplante de limbo é atualmente a técnica de eleição para restauração da superfície ocular, porém com baixa taxa de sucesso.

Impression cytology in the evaluation of ocular surface tumors: review article / Citologia de impressão na avaliação de tumores da superfície ocular: artigo de revisão

Impression cytology (IC) has been widely used as a method for evaluating the ocular surface and superficial cells layers in the diagnosis and follow-up after treatment of several ocular surface tumors of both epithelial and melanocytic origin. Information regarding this can be found in the English-language literature since 1992. Using either cellulose acetate or Biopore membranes for specimen collection, a high correlation has been found between IC and tissue histology. Compared with exfoliative cytology with spatula, IC is less traumatic to the patient’s eye, provides a precise location of the area being studied, and allows accurate observation of the cells the way they exist in vivo. The additional advantage of IC is the preservation of limbal stem cells responsible for continuous corneal epithelium renewal; these can be affected after incisional or excisional biopsy at the corneoscleral limbus, which is the most frequent site of appearance of tumors in the stratified epithelium. Treatment for ocular surface squamous neoplasia has historically included surgery, but nonsurgical interventions have also been adopted. Hence, in certain cases, ophthalmologists may prefer interventions less invasive than surgical biopsy such as of impression cytology for both initial diagnosis and therapeutic monitoring of treatment for ocular surface lesions. Nevertheless, it should be considered that IC may be less helpful if the results conflict with the clinical picture or if the clinical diagnosis is uncertain and results are negative. In such cases, surgical biopsy is required for accurate diagnosis. The purpose of this review is to examine the published literature on the utilization of IC for the diagnosis and management of ocular surface tumors and to discuss the requirement for further investigation on the subject.

A citologia de impressão (CI) tem sido amplamente utilizada como um método de avaliação da superfície ocular e das camadas de células superficiais no diagnóstico e no seguimento após tratamento de vários tumores da superfície ocular de origem epitelial ou melanocítica. As informações podem sem encontradas na literatura em língua inglesa desde 1992. Utilizando-se de membranas de acetato de celulose ou Biopore na coleta dos espécimes, uma alta correlação tem sido encontrada entre a CI e a histologia do tecido. Comparando-se com a citologia esfoliativa, a citologia de impressão é menos traumática para o olho do paciente, fornece uma localização precisa da área estudada e permite ver as células da forma como elas organizam-se in vivo. A vantagem adicional da citologia de impressão é a preservação das células- tronco germinativas responsáveis pela renovação contínua do epitélio da córnea. Elas podem ser afetadas após biópsia cirúrgica na região do limbo que é o sítio mais frequentemente acometido pelos tumores do epitélio estratificado. O tratamento para a neoplasia escamosa da superfície ocular tem sido historicamente a cirurgia, mas intervenções não cirúrgicas também foram adotadas. Por esta razão, em certos casos, oftalmologistas podem recorrer a formas menos invasivas que a biópsia cirúrgica (como a citologia de impressão) tanto para o diagnóstico inicial quanto para o monitoramento terapêutico das lesões da superfície ocular. No entanto, deve-se ter em mente que a citologia de impressão deixa de ser útil quando seu resultado não coincide com o quadro clínico ou quando o diagnóstico clínico é incerto e o resultado da citologia de impressão negativo. Nesses casos, a biópsia cirúrgica deve ser realizada para o diagnóstico. O objetivo desta revisão é examinar a literatura sobre a utilização da citologia de impressão no diagnóstico e tratamento dos tumores da superfície ocular bem como discutir a necessidade de uma investigação mais aprofundada sobre o assunto.

Co-transplantation of HLA-matched limbal stem cells and amniotic membrane for treatment of pterygium / 中国组织工程研究

BACKGROUND:Co-transplantation of HLA-matched limbal stem cels and amniotic membrane is a new concept for the treatment of pterygium, which is characterized as few complications, fast recovery, and low relapse rate. This treatment has been widely used because of reduced rejections. OBJECTIVE:To investigate the therapeutic effect of HLA-matched limbal stem cels and amniotic membrane co-transplantation in the treatment of pterygium. METHODS:Forty-seven patients with pterygium (47 eyes) who underwent the co-transplantation of HLA-matched limbal stem cels and amniotic membrane from March 1, 2013 to April 30, 2014 were enroled as test group. Another 40 patients with pterygium (47 eyes) who received traditional surgical method acted as control group. Therapeutic effectiveness and relapse were compared between the two groups. RESULTS AND CONCLUSION: The 47 cases in the test group were successfuly treated with no complications and rejections, and only one case suffered from incision infection but discharged from hospital after symptomatic treatment. After treatment, pterygium relapsed in 2 of 47 eyes in the test group and 15 of 47 eyes in the control group, and the relapse rates were 4.3% and 31.9%, respectively. Results from this study show that the co-transplantation of HLA-matched limbal stem cels and amniotic membrane for treatment of pterygium has fewer complications and lower relapse rate, which is an effective method for the treatment of pterygium.

Use of impression cytology for the detection of unsuspected ocular surface squamous neoplasia cells in pterygia / Uso da citologia de impressão para a detecção de células de neoplasia escamosa da superfície ocular insuspeita em pterígios

Purpose: To evaluate the agreement between the methodologies of impression cytology (IC) and histopathology regarding epithelial lesions clinically diagnosed as pterygium and also regarding the detection of unsuspected and associated ocular surface squamous neoplasia (OSSN). Methods: Thirty-two Brazilian patients were included and IC was performed on all pterygia before excision. Histopathogical examination was considered the gold standard and was performed by two experienced ocular pathologists in which consensus existed regarding pterygia diagnosis. IC accuracy was assessed by sensitivity and specificity with a 95% confidence interval. Results: From the 32 primary lesions studied, histopathological examination confirmed the diagnosis of pterygium without atypical cells in 19 cases (60%) and showed unsuspected and associated OSSN cells in 13 cases (40%). IC demonstrated one false-negative and one false-positive result for atypia. Statistical analysis showed an estimated sensitivity of 92%, specificity of 94%, positive predictive value of 92%, and negative predictive value of 94%. Conclusion: IC demonstrated high agreement with histopathological analysis in the detection of atypical epithelial cells in unsuspected OSSN in Brazilian pterygia patients. .

Objetivo: Avaliar a concordância entre a citologia de impressão (CI) e os achados histopatológicos de lesões epiteliais clinicamente diagnosticadas como pterígio, no que tange a detecção de células de neoplasia escamosa da superfície ocular (NESO) insuspeita associada. Métodos: Trinta e dois pacientes brasileiros foram incluídos e a CI foi obtida de todos os pterígios antes da excisão. O exame histopatológico foi realizado por dois patologistas oculares experientes em consenso de opinião e considerado o padrão-ouro para o diagnóstico. A acurácia da CI foi avaliada pela sensibilidade e especificidade com intervalo de confiança de 95% Resultados: Das 32 lesões estudadas, o exame histopatológico confirmou o diagnóstico de pterígio sem atipia em 19 casos (60%) e mostrou células de NESO insuspeita associada em 13 espécimes (40%). A CI demonstrou um resultado falso-positivo e um falso-negativo para atipia. A análise estatística mostrou uma sensibilidade estimada de 92%, especificidade de 94%, valor preditivo positivo de 92% e valor preditivo negativo de 94%. Conclusão: A CI apresentou alta concordância com o estudo histopatológico na detecção de células epiteliais atípicas de NESO insuspeita em pterígios do Brasil. .

Efficacy of three different methods for side port incision wound sealing

PURPOSE: To evaluate wound leakage and bacteria-sized particle influx in differently corneal sealed side port incisions. METHODS: Four 1.5mm tunnel squared incisions were created in each of four cadaveric human eyes. In each cornea, one incision was left unsealed, whereas the other three incisions were sealed using a 10-0 nylon suture, cyanoacrylate glue, or stromal hydration, respectively. A Seidel and an India ink test were performed on each eye. During each Seidel test, flourescein was applied, the IOP increased from 15 to 80mmHg, and the IOP at which each incision started to leak recorded. During each India ink test, ink was placed on the eye and rinsed out with balanced salt solution (BSS). Ink penetration was then measured by planimetry at physiologic conditions and after an IOP plunge from 80mmHg to 0mmHg. RESULTS: Regardless of IOP variations, no leakage or ink inflow was observed through the glued incisions. In contrast, leakage did occur in the other three sealing methods, albeit at significantly different IOP levels in each one (p=0.013). Ink inflow occurred in these sealing methods at physiologic IOP and, to a significantly greater extent, after the IOP challenge (p<0.05). At both of these IOP conditions, the differences in ink influx among these three incision-sealing methods were deemed statistically insignificant. CONCLUSION: This study showed that glue was more effective at preventing wound leakage and bacteria-sized particle influx than other commonly used methods especially hydrosealing.

OBJETIVO: Avaliar o sinal de Seidel positivo e a penetração de tinta da Índia em incisões corneanas acessórias seladas por diferentes métodos. MÉTODOS: Quatro incisões de 1,5 x 1,5mm foram criadas em cada um dos quatro olhos provindos do banco de olhos. Em cada córnea, uma incisão foi mantida sem manipulação, enquanto que as outras três incisões foram seladas respectivamente com Nylon 10-0, cola de cianoacrilato e hidratação estromal. Foram realizados dois testes: Sinal de Seidel positivo e penetração da tinta da Índia. No primeiro, foi aplicado fluoresceína gotas e a pressão intraocular (PIO) elevada de 15 para 80mmHg. No segundo, a tinta da Índia foi aplicada sobre o olho estudado em duas situações: sob PIO fisiológica e sob variação súbita da pressão, de 80 para 0mmHg. RESULTADOS: Na incisão selada com cola, apesar da variação da PIO, não houve vazamento e nem penetração de partículas de tinta. Por outro lado, o sinal de Seidel foi positivo nas outras três incisões em diferentes níveis de PIO (p=0,013). A penetração da tinta da Índia ocorreu nestas três incisões sob pressão fisiológica e com maior extensão após a variação da PIO (p<0,05). Esta diferença, no entanto não foi considerada estatisticamente significante quando comparadas as incisões entre si. CONCLUSÃO: No presente estudo, a cola foi mais eficaz em prevenir Seidel e entrada de partículas do que outro método comumente usado especialmente, hidratação estromal.

Impression cytology and in vivo confocal microscopy in corneas with total limbal stem cell deficiency / Citologia de impressão e microscopia confocal in vivo em córneas com deficiência total das células-tronco do limbo

PURPOSES: To describe corneal changes seen on in vivo confocal microscopy in patients with total limbal stem cell deficiency and to correlate them with cytological findings. METHODS: A prospective case series including 13 eyes (8 patients) with total limbal deficiency was carried out. Stem cell deficiency was diagnosed clinically and by corneal impression cytology. Confocal images of the central cornea were taken with the Heidelberg Retina Tomograph II, Rostock Corneal Module (Heidelberg Engineering, Heidelberg, Germany). RESULTS: Impression cytology of the cornea revealed conjunctival epithelial cells and goblet cells in all cases. In vivo confocal microscopy showed disruption of normal layers of the corneal epithelium in all eyes. Confocal images showed cells with characteristics of conjunctival epithelium at the cornea in 76.9% of the total. These findings on confocal microscopy were compatible to limbal stem cell deficiency. Additionally, goblet cells, squamous metaplasia, inflammatory cells and dendritic cells were observed. The sub-basal nerve plexus was not identified in any of the corneas. Corneal neovessels were observed at the epithelium and stroma. All cases showed diffuse hyper-reflective images of the stroma corresponding to opacity of the tissue. CONCLUSIONS: Limbal stem cell deficiency had been confirmed by impression cytology in all cases, and 76.9% of the cases could also be diagnosed by in vivo confocal microscopy through the conjunctival epithelial cell visualization on the corneal surface. Frequent confocal microscopy findings were abnormal cells at the cornea (conjunctival epithelial, goblet and inflammatory cells), corneal neovessels and diffuse hyper-reflection of the stroma.

OBJETIVOS: Descrever as alterações corneais observadas na microscopia confocal in vivo e relacioná-las aos achados da citologia de impressão em pacientes com deficiência total das células-tronco do limbo. MÉTODOS: Série de casos prospectiva incluindo 13 olhos (8 pacientes) com deficiência total das células-tronco do limbo. A deficiência límbica foi diagnosticada clinicamente e mediante citologia de impressão da córnea. Imagens confocais da córnea central foram obtidas com o Heidelberg Retina Tomograph II, Rostock Corneal Module (Heidelberg Engineering, Heidelberg, Alemanha). RESULTADOS: A citologia de impressão da córnea demonstrou células epiteliais da conjuntiva e células caliciformes em todos os casos. A microscopia confocal da área central da córnea mostrou alteração da estrutura normal das camadas do epitélio corneal em todos os casos. As imagens confocais da córnea mostraram a presença de células com características do epitélio conjuntival em 76,9%. Esses achados no exame confocal são compatíveis com deficiência das células-tronco do limbo. Adicionalmente, células caliciformes, metaplasia escamosa, células inflamatórias e dendríticas foram observadas. O plexo nervoso sub-basal não foi identificado em nenhum dos casos. Neovasos corneais foram observados no epitélio e no estroma. Em todos os casos havia imagens difusamente hiperreflectivas no estroma, correspondendo à opacidade do tecido. CONCLUSÕES: A deficiência das células-tronco do limbo, previamente confirmada por citologia de impressão, pôde ser demonstrada pela microscopia confocal in vivo em 76,9% dos casos através da visualização de células epiteliais da conjuntiva na córnea. Achados frequentes na microscopia confocal foram células anormais na córnea (células conjuntivais, caliciformes e inflamatórias), neovasos corneais e hiperreflexão difusa do estroma.

Alpins and thibos vectorial astigmatism analyses: proposal of a linear regression model between methods / Análises vetoriais de alpins e thibos para o astigmatismo - proposta de modelo de regressão linear entre os métodos

PURPOSE: To determine linear regression models between Alpins descriptive indices and Thibos astigmatic power vectors (APV), assessing the validity and strength of such correlations. METHODS: This case series prospectively assessed 62 eyes of 31 consecutive cataract patients with preoperative corneal astigmatism between 0.75 and 2.50 diopters in both eyes. Patients were randomly assorted among two phacoemulsification groups: one assigned to receive AcrySof®Toric intraocular lens (IOL) in both eyes and another assigned to have AcrySof Natural IOL associated with limbal relaxing incisions, also in both eyes. All patients were reevaluated postoperatively at 6 months, when refractive astigmatism analysis was performed using both Alpins and Thibos methods. The ratio between Thibos postoperative APV and preoperative APV (APVratio) and its linear regression to Alpins percentage of success of astigmatic surgery, percentage of astigmatism corrected and percentage of astigmatism reduction at the intended axis were assessed. RESULTS: Significant negative correlation between the ratio of post- and preoperative Thibos APVratio and Alpins percentage of success (%Success) was found (Spearman's ρ=-0.93); linear regression is given by the following equation: %Success = (-APVratio + 1.00)x100. CONCLUSION: The linear regression we found between APVratio and %Success permits a validated mathematical inference concerning the overall success of astigmatic surgery.

OBJETIVO: Estabelecer modelos de regressão linear entre os índices propostos por Alpins com os vetores astigmáticos de Thibos, avaliando a validade e a força dessas correlações. MÉTODOS: Série de casos na qual foram avaliados prospectivamente 62 olhos de 31 pacientes de catarata com astigmatismo corneano pré-operatório entre 0,75 e 2,50 dioptrias para ambos os olhos. Os pacientes foram aleatoriamente distribuídos entre 2 grupos: um submetido a implante de lente intraocular (LIO) AcrySof ToricTM em ambos os olhos e outro grupo, no qual seriam implantadas LIO AcrySof NaturalTM complementada por incisões relaxantes limbares, também em ambos os olhos. Todos os pacientes foram reavaliados aos 6 meses de pós-operatório, sendo feitas análises do astigmatismo refracional tanto pelo método vetorial proposto por Alpins, quanto pelo proposto por Thibos. A razão entre os vetores astigmáticos pós e pré-operatórios de Thibos (VApós/pré), bem como a correlação linear com o percentual de sucesso da cirurgia do astigmatismo, o percentual de astigmatismo corrigido e o percentual de redução do astigmatismo no eixo pretendido, propostos por Alpins, foram avaliados. RESULTADOS: Foi encontrada correlação negativa significativa entre a VApós/pré de Thibos e o percentual de sucesso da cirurgia do astigmatismo (%Successo), de Alpins (ρ de Spearman=-0.93); regressão linear dada pela seguinte equação: %Successo = (-VApós/pré + 1,00)x100. CONCLUSÃO: A regressão linear encontrada entre VApós/pré e %Successo permite uma inferência matemática validada a respeito do sucesso de cirurgia do astigmatismo em termos gerais.