Banca de DEFESA: GESSICA SABRINA DE ASSIS SILVA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
DISCENTE : GESSICA SABRINA DE ASSIS SILVA
DATA : 01/02/2021
HORA: 14:00
LOCAL: online
TÍTULO:

THERAPEUTIC POTENTIAL OF EXTRACELLULAR VESICLES DERIVED FROM MESENQUIMAL CELLS IN DIABETIC SENSORY NEUROPATHY IN MICE

PALAVRAS-CHAVES:

neuropathic pain, chronic pain, mesenchymal stem cells, extracellular vesicles, opioid system, endogenous analgesia, cell therapy, cell-free therapy.

PÁGINAS: 94
GRANDE ÁREA: Ciências da Saúde
ÁREA: Farmácia
RESUMO:

INTRODUCTION: Sensory neuropathy is one of the most frequent and debilitating complications of diabetes and manifests itself through paradoxical symptoms such as loss of sensation. Its pharmacological treatment is not very effective, produces a series of adverse effects and has no curative action, acting only in the relief of symptoms. Mesenchymal stem cells have been considered promising for the treatment of painful neuropathies. The therapeutic properties of acquired mesenchymal cells are attributed to their ability to release soluble bioactive molecules and contained in extracellular vesicles (EV). OBJECTIVE: To investigate the therapeutic potential of LV derived from human bone marrow mesenchymal cells (BMMCs) in a murine model of sensory diabetic neuropathy (SDN). METHODS: BMMCs were slid from the human bone marrow and characterized by morphological analysis, flow cytometry and in vitro differentiation. The EV were adjusted by ultracentrifugation of the BMMCs secretome and characterized by their size, shape and quantity by transmission electron microscopy (TEM) and by the suspended particle analysis technique (NTA - Nanoparticle Tracking Analysis). The streptozotocin NDS model (ETZ; 80 mg / kg / ip) was induced in male C57Bl / 6 mice (22-25g), from the IGM Biotery (FIOCRUZ-BA) and the nociceptive threshold was performed throughout the period . experimental (92 days), with von Frey filaments. Thirty days after the model was induced, the mice received intravenous administration of saline (100 µL), EV-BMMCs (100 µL) or BMMCs (1x106). The contribution of the opioid system to the analgesic effect of cell therapy was investigated in an antagonist reversal assay, using naloxone (0.4 mL / kg i.p.) administered 7 and 60 days after transplantation. An expression of preproencephalin in the spinal cord and no supernatant from the BMMCs culture was analyzed by quantitative Real-Time PCR (RT-qPCR). RESULTS: Through the differentiation assay and flow cytometry characterization, it was confirmed that the expanding cells were mesenchymal. The characterization of EV-BMMCs by NTA describes an EV population with a size of 52.4 to 450 nm, corresponding to microvesicles and exosomes. The average total number of vesicles recovered from 1x106 BMMCs was 4.4x108. TEM data confirmed the presence of structures with morphology similar to that of EV. A single intravenous administration of BMMCs or EV-BMMCs was able to reverse the behavioral signs of painful diabetic neuropathy throughout the experimental period (p <0.05). The administration of naloxone (3 mg / kg, i.p.), a non-selective opioid receptor antagonist, reversed an antinociception induced by BMMCs and EV-BMMCs (p <0.05). Analyzes of RT-qPCR performed 7 days after treatments with BMMCs and EV-BMMCs indicate an increase in the expression of preproencephalin in neuropathic animals when compared to saline (p <0.001). Already 60 days after the treatments, it was observed that neuropathic animals treated with less saline expression of preproencephalin in the spinal cord compared to non-neuropathic animals (p <0.05). In addition, the administration of EV-BMMCs, but not BMMCs, increased the expression of preproencephalin in the spinal cord of mice with diabetic neuropathy when compared to neuropathies treated with saline (p <0.01). In addition, analyzes of BMMCs in culture showed that these cells express preprooencephalin. CONCLUSION: the results of the present study show that EV derived from human EV-BMMCs preserve the antinociceptive properties of the cells of origin, and that this effect seems to involve the activation of the endogenous opioid system. This study highlights the potential of cell-free therapy to control painful diabetic neuropathy.

MEMBROS DA BANCA:
Interno(a) - 2055299 - ANA LEONOR PARDO CAMPOS GODOY
Presidente - 1536719 - CRISTIANE FLORA VILLARREAL
Externo(a) à Instituição - LUIZ FERNANDO FERRARI