Avaliação do desempenho de colírio usando soro alogênico e fatores de crescimento derivados de plaquetas em cultura de células do anel córneo-escleral

• Autor(a) principal: Martins, Juliana Ravelli Baldassarre [UNESP]
• Data de Publicação: 2014
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/110668
• topic: Celulas - Cultura e meios de cultura; Síndromes do olho seco; Soluções oftálmicas; Imuno-histoquímica; Tratamento oftalmologico; Dry eye syndromes

Introduction: Many diseases can affect cornea, including dry eye syndrome and related diseases. Innovative treatments for dry eye syndrome have emerged , among them the use of autologous serum eye drops produced with the use of eye serum, umbilical cord blood, conditioned medium obtained by expansion of limbal epithelial stem cells (ex vivo model) among others. Patients with probable indication of autologous serum eyedrops may be contraindicated in production originating immune status for viral diseases such as HIV, hepatitis B and C, syphilis , Chagas disease and HTLV I and II, from the point of view of eye drops´s production, have barriers . For these cases, eye drops from healthy donors allogeneic serum can be an alternative. Objectives: Evaluate in vitro effect of the action of allogeneic eye drops in corneal-scleral cell culture, established the culture and expansion of adherent cells obtained from the corneal- scleral region, compare the culture of adherent cells in culture medium containing eye drops and platelets derived growth factors, evaluating the cells by immunohistochemistry and analyze medical data of eye drops´s patients. Materials and Methods: The corneal- scleral rings were obtained in the operating room after surgery of tissue donor, and these rings were discarded after surgery. Then, tissue was sent to the Laboratory of Cell Engineering of Botucatu Blood Center, which was fragmented into small pieces and digested with collagenase type I. Cells were plated in 25 cm² flasks with DMEM Knockout in the first part of the experiment, and Keratinocyte in the second part of the experiment. After 80% confluence, cells passed through the trypsinization procedure for detachment of the cells from the culture flask. Mounting plaque experiment, cells were placed to control, or untreated cells, treated cells with eye drops and cells treated with platelets derived growth factors was performed. After 5 days, immunohistochemistry ...