Uso autólogo e xenólogo da fibrina rica em plaquetas em úlceras de córnea em coelhos (Oryctolagus cuniculus)
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Data
2022-02-25
Autores
Gonçalves, Lenise Garbelotti [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A membrana de fibrina rica em plaquetas e leucócitos (L-PRF) é um biomaterial com potencial regenerativo proveniente de preparo rápido e simples do sangue. Este trabalho descreveu o uso de membrana autóloga e xenóloga de L-PRF em úlceras corneais profundas induzidas em coelhos; correlacionando achados clínico-oftalmológicos, histomorfométricos e imuno-histoquímico. Foram selecionados 28 coelhos adultos sadios, e o olho direito submetido à indução de úlcera de córnea profunda e tratamento com membrana L-PRF xenóloga de cães, fixada na conjuntiva bulbar. Constituíram-se dois grupos experimentais (n=14), designados: grupo um (G1), tratado apenas com a membrana L-PRF xenólogo; e o grupo dois (G2), cujos olhos receberam adicionalmente membrana autóloga de L-PRF previamente no leito da úlcera. Os animais foram avaliados por exame oftalmológico antes da cirurgia e aos quatro (M4), sete (M7), 14 (M14) e 30 (M30) dias de pós-operatório, bem como a análise histomorfométrica da córnea no M30. Cinco animais de cada grupo em M4, foram submetidos apenas à avaliação histomorfométrica e imunohistoquímica para fator de crescimento transformador β1 (TGF- β1) e fator de crescimento de fibroblastos básico (FGF 2). Os resultados das avaliações clínico-oftalmológicas, histomorfométrico e imunohistoquímico foram similares entre os grupos na maioria das variáveis analisadas. Inflamação discreta e ausência de vascularização corneal foram observadas; a reparação total da lesão verificou-se no M14 com discreta opacidade corneal nos dois grupos. No G2, notou-se absorção mais lenta das membranas na fase inicial, bem como, maior expressão de TGF-β1 no epitélio. No M30, na área da lesão, o número de camadas epiteliais e espessura do epitélio foram significativamente maiores, similar à córnea íntegra nos dois grupos. Verificou-se, entretanto, que a redução da área da lesão foi superior no G1 no período inicial (M7). A metodologia do G2 poderá favorecer o uso clínico da membrana autóloga; considerando necessidade de menor volume de sangue autólogo coletado, promovendo sua instituição em animais de pequeno porte. A aplicação cirúrgica da membrana de L-PRF xenóloga e/ou associada à autóloga é exequível e prática, promove a cicatrização de úlceras profundas, e apresenta se como alternativa viável na clínica oftalmológica.
Platelet and leukocyte-rich fibrin (L-PRF) is a biomaterial made from blood that presents regenerative potential and quick and simple preparation. This study describes the use of autologous and/or xenologous L-PRF membrane in deep induced corneal ulcers in rabbits; correlating clinical-ophthalmological, histomorphometric, and immunohistochemical findings. Twenty-eight healthy adult rabbits were selected. A deep corneal ulcer was induced and the animals were divided into two groups (n = 14): overlay group (G1) was treated with an overlay xenologous L-PRF membrane made from dog blood, fixed in the bulbar conjunctiva; inlay group (G2) received autologous L-PRF inlay and an overlay xenologous L-PRF membrane. The animals were evaluated on M0, before surgery, and on M4, M7, M14, and M30 days postoperatively as well the histomorphometric analysis of the cornea in the M30. In M4, five animals from each group were submitted to the histomorphometric and immunohistochemical evaluation of transforming growth factor-b (TGF-β1) and fibroblast growth factor (FGF-2). The results of the clinical-ophthalmological, histomorphometric, and immunohistochemical evaluations were similar between the groups in most of the variables analyzed. Mild inflammation and low corneal vascularization were observed; complete repair of the lesion was observed in M14 with a minor corneal opacity in both groups.In G2, slower absorption of membranes was observed in the initial phase, as well as greater expression of TGF-β1 in the epithelium. In M30, area of the lesion, the number of epithelial layers and the thickness of the epithelium were significantly higher, similar to the intact cornea in both groups. However, the reduction of the lesion area was superior in G1 in the initial period (M7). The G2 methodology may favor the clinical use of the autologous membrane; considering the need for a smaller volume of autologous blood collected, promoting its institution in small animals. The surgical application of xenologous and/or associated with autologous plug L-PRF membrane is feasible and practical, promotes the healing of deep ulcers, and presents itself as a viable alternative in the ophthalmic clinic.
Platelet and leukocyte-rich fibrin (L-PRF) is a biomaterial made from blood that presents regenerative potential and quick and simple preparation. This study describes the use of autologous and/or xenologous L-PRF membrane in deep induced corneal ulcers in rabbits; correlating clinical-ophthalmological, histomorphometric, and immunohistochemical findings. Twenty-eight healthy adult rabbits were selected. A deep corneal ulcer was induced and the animals were divided into two groups (n = 14): overlay group (G1) was treated with an overlay xenologous L-PRF membrane made from dog blood, fixed in the bulbar conjunctiva; inlay group (G2) received autologous L-PRF inlay and an overlay xenologous L-PRF membrane. The animals were evaluated on M0, before surgery, and on M4, M7, M14, and M30 days postoperatively as well the histomorphometric analysis of the cornea in the M30. In M4, five animals from each group were submitted to the histomorphometric and immunohistochemical evaluation of transforming growth factor-b (TGF-β1) and fibroblast growth factor (FGF-2). The results of the clinical-ophthalmological, histomorphometric, and immunohistochemical evaluations were similar between the groups in most of the variables analyzed. Mild inflammation and low corneal vascularization were observed; complete repair of the lesion was observed in M14 with a minor corneal opacity in both groups.In G2, slower absorption of membranes was observed in the initial phase, as well as greater expression of TGF-β1 in the epithelium. In M30, area of the lesion, the number of epithelial layers and the thickness of the epithelium were significantly higher, similar to the intact cornea in both groups. However, the reduction of the lesion area was superior in G1 in the initial period (M7). The G2 methodology may favor the clinical use of the autologous membrane; considering the need for a smaller volume of autologous blood collected, promoting its institution in small animals. The surgical application of xenologous and/or associated with autologous plug L-PRF membrane is feasible and practical, promotes the healing of deep ulcers, and presents itself as a viable alternative in the ophthalmic clinic.
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Palavras-chave
Biomaterial, Cicatrização corneal, Concentrado de plaqueta, Fator de crescimento de fibroblastos (FGF), Fator de crescimento transformador (TGF), PRF, Corneal healing