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TISSUE REGENERATION: IN VITRO EVALUATION OF A PROTOTYPE OF BIOCURATIVE FOR SLOW RELEASE OF STEM CELLS IN CHRONIC WOUNDS.

Grant number: 23/02643-8
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2023
Effective date (End): May 31, 2024
Field of knowledge:Health Sciences - Medicine - Surgery
Principal Investigator:Matheus Bertanha
Grantee:Lorena Amorim Tiritan
Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Introduction: Peripheral Arterial Disease (PAD) has become a worrying situation, especially for developed countries, and its condition is extremely debilitating. This disease is caused by the formation of atheroma plaques in the intimal layer of the peripheral arteries, affecting mainly the lower limbs, its development and worsening, leading to the formation of ulcerated wounds and even gangrene. For the treatment of this condition, it is essential to control risk factors, such as platelet aggregation, and to take care of the etiology. Sometimes, when the wound is already in an ischemic condition, surgical intervention is occasionally necessary to revascularize the limb, which can lead to ulcers that are difficult to heal. Objective: It is proposed to carry out an in vitro test of the stability of a new dressing with slow release of stem cells for the future treatment of ulcers. Materials and Methods: To develop the experiment, autologous mesenchymal stem cells from adipose tissue (MSC) will be obtained from 5 Norvegicus rats, Wistar lineage (rW), which will be collected after euthanasia, from the interscapular region of donors in aseptic manner, in addition to from whole blood collection to fractionation. MSCs will be expanded in the laboratory and phenotypically characterized as multipotent. A spongy hemostatic matrix of sterile absorbable gelatin (Gelfoam®) will serve as a framework for the study. 24-well plates with sterilized cover slips will be used at the bottom of the wells, with 5 repetitions for each observation, with observation times of 2 days, 4 days and 7 days: Control group: Gelfoam®; Group1: Gelfoam® + 5x105 CTM; Group2: Gelfoam® + 5x105 CTM + PRP + platelet-poor plasma cryoprecipitate (PPP). At the end of these observations, the culture medium will be aspirated, the product will be fixed with 10% buffered formaldehyde and the coverslips will be removed for histological analysis in HE. This experiment will allow the choice of the best protocol, from the point of view of stability, containing MSC, to perform a new biodressing.

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