RESUMO
BACKGROUND: The World Health Organization (WHO) estimates that the number of individuals who lose their vision due to retinal degeneration is expected to reach 6 million annually in 2020. The retinal degenerative diseases affect the macula, which is responsible for central and detailed vision. Most macular degeneration, i.e., age-related macular degeneration (AMD) develops in the elderly; however, certain hereditary diseases, such as the Stargardt disease, also affect young people. This degeneration begins with loss of retinal pigmented epithelium (RPE) due to formation of drusen (atrophic) or abnormal vessels (exudative). In wet AMD, numerous drugs are available to successful treat the disease; however, no proven therapy currently is available to treat dry AMD or Stargardt. Since its discovery, human embryonic stem cells (hESCs) have been considered a valuable therapeutic tool. Some evidence has shown that transplantation of RPEs differentiated from hESCs cells can result in recovery of both RPE and photoreceptors and prevent visual loss. METHODS: The human embryonic WA-09 stem cell lineage was cultured under current Good Manufacturing Practices (cGMP) conditions using serum-free media and supplements. The colonies were isolated manually and allowed to spontaneously differentiate into RPE cells. RESULTS: This simple and effective protocol required minimal manipulation and yielded more than 10e8 RPE cells by the end of the differentiation and enrichment processes, with cells exhibiting a cobblestone morphology and displaying cellular markers and a gene expression profile typical of mature RPE cells. Moreover, the differentiated cells displayed phagocytic activity and only a small percentage of the total cells remained positive for the Octamer-binding transcriptions factor 4 (OCT-4) pluripotency cell marker. CONCLUSIONS: These results showed that functional RPE cells can be produced efficiently and suggested the possibility of scaling-up to aim at therapeutic protocols for retinal diseases associated with RPE degeneration.
RESUMO
STUDY DESIGN: Animal experimental study. OBJECTIVE: Evaluate the effect of physical activity and overtraining condition on glycosaminoglycan concentration on the intervertebral disc (IVD) using a rat running model. SUMMARY OF BACKGROUND DATA: Some guidelines recommend the implementation of a physical exercise program as treatment for low back pain; however, cyclic loading impact on the health of the IVD and whether there is a dose-response relationship is still incompletely understood. METHODS: Thirty-two rats ages 8 weeks were divided into four groups with eight animals each. The first 8 weeks were the adaptive phase, the overtraining phase was from the ninth to the eleventh week, which consisted of increasing the number of daily training sessions from 1 to 4 and the recovery phase was represented by the 12th and 13th weeks without training. Control group 1 (CG1) did not undergo any kind of training. Control group 2 (CG2) completed just the adaptive phase. Overtraining group 1 (OT1) completed the overtraining phase. Overtraining group 2 (OT2) completed the recovery phase. Running performance tests were used to assess the "overtraining" status of the animals. IVD glycosaminoglycans were extracted and quantified, and identified by electrophoresis. RESULTS: Glycosaminoglycans showed a distribution between chondroitin sulfate and dermatan sulfate. Glycosaminoglycans quantification showed decreasing concentration at the following order: OT1â>âCG2â>âOT2â>âCG1. Increased expression of dermatan sulfate was verified at the groups submitted to any training. CONCLUSION: Overtraining condition, as assessed by muscle and cardiovascular endurance did not lessen glycosaminoglycan concentration in the IVD. In fact, physical exercise increased glycosaminoglycan concentration in the IVD in proportion to the training load, even at overtraining condition, returning to normal levels after the recovery phase and glycosaminoglycan production is a reversible acute positive response for mechanical stimulation of the IVD. LEVEL OF EVIDENCE: N/A.