title: Histology and immunochemistry evaluation of autologous translocation of retinal pigment epithelium-choroid graft in porcine eyes.
abstract: OBJECTIVE
  To evaluate structure and cellular functionality of retinal pigment epithelium (RPE)-choroid grafts after autologous translocation in porcine eyes.
  
  
  METHODS
  Retinal pigment epithelium-choroid grafts were obtained from the nasal midperiphery donor site and translocated to the central area in 12 pigs (12 eyes). Grafts were placed under the central retina through a retinotomy. Ophthalmoscopic and pathological evaluations were performed immediately (n = 1) and at 15 (n = 3) and 30 (n = 3) days after surgery. Untranslocated nasal RPE-choroid grafts were obtained at time of surgery and used as controls. Specimens were evaluated by standard histology and by immunochemical studies of RPE65, CRALBP and GFAP.
  
  
  RESULTS
  Five animals were lost to follow-up owing to surgery or anaesthesia complications. Ophthalmoscopic examination revealed that the grafts remained in place at all time-points studied. Fifteen and thirty days postsurgery, some areas of the transplanted RPE maintained a monolayered structure. Retinal pigment epithelium cells were firmly attached to Bruch's membrane and predominantly preserved polarity and pigment distribution. However, RPE65, CRALBP and GFAP patterns of expression and distribution were diminished and modified during follow-up. Ophthalmoscopic retinal detachment and proliferative vitreoretinopathy (PVR), confirmed by microscopic evaluation, complicated all cases at 30 days of follow-up.
  
  
  CONCLUSIONS
  Autologous RPE-choroid grafts survived up to 30 days in porcine eyes. Histological and immunochemical evaluation revealed preserved transplanted RPE cells morphology accompanied by alterations in the immunoreactivity expression of functional proteins, and development of significant PVR. The data presented in this manuscript provide insights into the fate, viability and cellular functionality of the transplanted RPE-choroid graft, serving as foundation for further knowledge and improvement of this technique.
authors.affiliation: Universitary Institute of Applied Ophthalmobiology (IOBA), University of Valladolid, Valladolid, Spain. ifernandezb@ioba.med.uva.es
authors.forename: Ivan
authors.initials: I
authors.lastname: Fernandez-Bueno
authors.forename: Enrique
authors.initials: E
authors.lastname: Rodriguez de la Rua
authors.forename: Denise
authors.initials: D
authors.lastname: Hileeto
authors.forename: Maria Luisa
authors.initials: ML
authors.lastname: Parrado
authors.forename: Marta
authors.initials: M
authors.lastname: Regueiro-Purriños
authors.forename: Anna
authors.initials: A
authors.lastname: Sala-Puigdollers
authors.forename: Girish K
authors.initials: GK
authors.lastname: Srivastava
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Gonzalo-Orden
authors.forename: Jose Carlos
authors.initials: JC
authors.lastname: Pastor
mesh_terms: D000818:Animals; D016570:Bruch Membrane / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D002352:Carrier Proteins / Q000378:metabolism; D002829:Choroid / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D058450:Endotamponade; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006085:Graft Survival; D008268:Macular Degeneration / Q000601:surgery; D023421:Models, Animal; D012160:Retina / Q000378:metabolism / Q000473:pathology / Q000601:surgery*; D055213:Retinal Pigment Epithelium / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D012827:Silicone Oils / Q000008:administration & dosage; D034421:Sus scrofa; D014182:Transplantation, Autologous; D018630:Vitreoretinopathy, Proliferative / Q000601:surgery; D019745:cis-trans-Isomerases / Q000378:metabolism; D000099178:Retinoid Isomerohydrolase
chemical_list: D002352:Carrier Proteins; D005904:Glial Fibrillary Acidic Protein; D012827:Silicone Oils; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase; C020932:11-cis-retinal-binding protein