RESUMO
Through the paracrine effects of stem cells, including the secretion of neurotrophic, immunomodulatory, and anti-apoptotic factors, cell-based therapies offer a new all-encompassing approach to treatment of neurodegenerative diseases. In this study, we used physically separated co-cultures of porcine neuroretina (NR) and human mesenchymal stem cells (MSC) to evaluate the MSC paracrine neuroprotective effects on NR degeneration. NR explants were obtained from porcine eyes and cultured alone or co-cultured with commercially available MSCs from Valladolid (MSCV; Citospin S.L.; Valladolid, Spain), currently used for several approved treatments. Cultures were maintained for 72â¯h. MSC surface markers were evaluated before and after co-culture with NRs. Culture supernatants were collected and the concentration of brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), and glial-derived neurotrophic factor (GDNF) were determined by enzyme-linked immunosorbent assays. NR sections were stained by haematoxylin/eosin or immunostained for terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL), glial fibrillary acidic protein, ß-tubulin III, and neuronal nuclei marker. NR morphology, morphometry, nuclei count, apoptosis rate, retinal ganglion cells, and glial cell activation were evaluated. Treatment effects were statistically analysed by parametric or non-parametric tests. The MSCs retained stem cell surface markers after co-culture with NR. BDNF and CNTF concentrations in NR-MSCV co-cultures were higher than other experimental conditions at 72â¯h (pâ¯<â¯0.05), but no GDNF was detected. NR general morphology, total thickness, and cell counts were broadly preserved in co-cultures, and the apoptosis rate determined by TUNEL assay was lower than for NR monocultures (all pâ¯<â¯0.05). Co-cultures with MSCV also protected retinal ganglion cells from degenerative changes and reduced reactive gliosis (both pâ¯<â¯0.05). In this in vitro model of spontaneous NR degeneration, the presence of co-cultured MSCs retarded neuroglial degeneration. This effect was associated with elevated concentrations of the neurotrophic factors BDNF and CNTF. Our data suggest that the paracrine secretion of these, and possibly other molecules, are a potential resource for the treatment of several neuroretinal diseases.
Assuntos
Células-Tronco Mesenquimais/citologia , Neuroproteção/fisiologia , Comunicação Parácrina/fisiologia , Retina/citologia , Degeneração Retiniana/prevenção & controle , Animais , Biomarcadores/metabolismo , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Sobrevivência Celular/fisiologia , Fator Neurotrófico Ciliar/metabolismo , Técnicas de Cocultura , Ensaio de Imunoadsorção Enzimática , Técnica Indireta de Fluorescência para Anticorpo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Humanos , Marcação In Situ das Extremidades Cortadas , Células-Tronco Mesenquimais/metabolismo , Microglia/metabolismo , Retina/metabolismo , Degeneração Retiniana/metabolismo , Células Ganglionares da Retina/metabolismo , Suínos , Tubulina (Proteína)/metabolismoRESUMO
SIGNIFICANCE: The present study provides quantitative data regarding the change of indications of fluorescein angiography in a tertiary hospital. Exhaustively compiled data over more than 10 years concerning all the angiographic studies including diagnosis, first-visit/follow-up, possible relation with antiangiogenics approval, and optic coherence tomography (OCT) are presented. PURPOSE: The aim of this study was to determine the frequency and indications of fluorescein angiography before and after OCT and anti-vascular endothelial growth factor (anti-VEGF) treatment implantation. METHODS: This was a retrospective and descriptive study of all fluorescein angiographies performed in a tertiary hospital between 2005 and 2016. Demographic data, diagnosis, follow-up, and type of angiograms were analyzed in relation with the implantation of time domain OCT (2006), spectral domain OCT (SD-OCT) (2013), and anti-VEGF (2007). RESULTS: Of 3263 angiograms (2342 patients) analyzed, 72% were baseline angiograms, and 28% were follow-up angiograms. After anti-VEGF initiation, the number of angiograms per year increased progressively with peaks that matched with the approval of anti-VEGF for wet age-related macular degeneration (2007) (164.2%), macular edema secondary to retinal vein occlusions (2010) (102.2%), and diabetic macular edema (2011) (123.8%). After using SD-OCT, fluorescein angiographies decreased up to 57%. CONCLUSIONS: Anti-VEGF introduction led to an increase in the indication of angiograms, which did not vary significantly after time domain OCT. Nevertheless, since SD-OCT became available, the indication of fluorescein angiography has halved in the hospital of reference.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Retinopatia Diabética/diagnóstico , Angiofluoresceinografia/estatística & dados numéricos , Edema Macular/diagnóstico , Oclusão da Veia Retiniana/diagnóstico , Tomografia de Coerência Óptica , Degeneração Macular Exsudativa/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Bevacizumab/uso terapêutico , Retinopatia Diabética/tratamento farmacológico , Feminino , Fundo de Olho , Humanos , Injeções Intravítreas , Edema Macular/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Oclusão da Veia Retiniana/tratamento farmacológico , Estudos Retrospectivos , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Degeneração Macular Exsudativa/tratamento farmacológicoRESUMO
PURPOSE: To evaluate the feasibility, safety, and biocompatibility of intravitreal injection of human mesenchymal stem cells (MSCs) in immunocompetent pigmented rabbits. MATERIALS AND METHODS: Thirty-two pigmented rabbits (24 females, 8 males; Chinchilla-New Zealand White) were divided into 8 groups of 4 animals. Commercially prepared human MSCs were injected (0.05 ml) into the post-lens vitreous of the right eyes. Groups 1 and 4 received isotonic medium (Ringer lactate-based), groups 2, 5, 7, and 8 received a low dose of 15 × 106 cells/ml. Groups 3 and 6 received a high dose of 30 × 106 cells/ml. Clinical signs were evaluated and scored before MSCs injection and weekly for 2 or 6 weeks. Animals were sacrificed at 2 or 6 weeks after injection. Eyes, liver, spleen, and gonads were assessed by histology and by fluorescent in situ hybridization to evaluate survival and extraocular migration of MSCs. RESULTS: There were no relevant clinical findings between control and MSC-injected rabbit eyes at any time point. There were also no relevant histological findings between control and MSC-injected rabbits related to ocular, liver, spleen, or gonad tissues modifications. MSCs survived intravitreally for at least 2 weeks after injection. Extraocular migration of MSCs was not detected. CONCLUSIONS: MSCs are safe and well-tolerated when administered intravitreally at a dose of 15 × 106 cells/ml in pigmented rabbits. These findings enable future research to explore the intravitreal use of commercially prepared allogenic human MSCs in clinical trials of retinal diseases.
Assuntos
Teste de Histocompatibilidade/métodos , Hospedeiro Imunocomprometido , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Doenças Retinianas/cirurgia , Animais , Modelos Animais de Doenças , Eletrorretinografia , Estudos de Viabilidade , Feminino , Humanos , Hibridização in Situ Fluorescente , Injeções Intravítreas , Masculino , Células-Tronco Mesenquimais/imunologia , Coelhos , Retina/patologia , Retina/fisiopatologia , Doenças Retinianas/diagnóstico , Doenças Retinianas/imunologiaRESUMO
Retinal and optic nerve diseases are degenerative ocular pathologies which lead to irreversible visual loss. Since the advanced therapies availability, cell-based therapies offer a new all-encompassing approach. Advances in the knowledge of neuroprotection, immunomodulation and regenerative properties of mesenchymal stem cells (MSCs) have been obtained by several preclinical studies of various neurodegenerative diseases. It has provided the opportunity to perform the translation of this knowledge to prospective treatment approaches for clinical practice. Since 2008, several first steps projecting new treatment approaches, have been taken regarding the use of cell therapy in patients with neurodegenerative pathologies of optic nerve and retina. Most of the clinical trials using MSCs are in I/II phase, recruiting patients or ongoing, and they have as main objective the safety assessment of MSCs using various routes of administration. However, it is important to recognize that, there is still a long way to go to reach clinical trials phase III-IV. Hence, it is necessary to continue preclinical and clinical studies to improve this new therapeutic tool. This paper reviews the latest progress of MSCs in human clinical trials for retinal and optic nerve diseases.