Evaluation of the integration of rabbit mesenchymal stem cells spread on biological scaffolds - in vitro model

• Grant number: 19/03785-5
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2019
• End date: July 31, 2020
• Field of knowledge: Health Sciences - Medicine - Surgery
• Principal Investigator: Matheus Bertanha
• Grantee: Barbara Esteves Silva
• Host Institution: Faculdade de Medicina (FMB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

Cardiovascular diseases are the main cause of mortality in the western adult population and the Arterial Occlusive Disease (AOD), as a part of this group of diseases, is present in 5% of this population. As the disease advances, 25% of the symptomatic patients evolve to the amputation of the affected limb, despite the treatment techniques available now a day. Therefore, researches have been developed aiming to produce new arterial substitutes, to be used on patients who can't find a compatible replacement. The cellular engineering may represent a promising perspective to the provision of blood vessels for these diseased, falling in line with the personalized medical practice that the near future bestows. Objective: Improve the technique of cultivating mesenchymal stem cells (MSC) from adipose tissue (AT) in 3D biological scaffolds, produced through the decellularization of inferior vena cava (IVC) from rabbits. Test if the addition of a new stage to the manufacturing of the IVC scaffolds, with their exposure to the detergent agent Triton X-100, can improve the integration of the MSC with the scaffold. Lastly, histomorphologically evaluate the product of the experiment. Methods: The IVC and the AT will be obtained from 10 rabbits, non-pregnant females, for decellularization of 1,5 cm each and the decellularization will be executed by 2 protocols that have been verified by previous studies: 1% sodium Dodecyl sulfate (SDS); and 2% sodium deoxycholate (DS). The scaffolds will be divided in 2, being that half will be used as produced and half will be exposed to 1% TritonX-100 (TX) for 10 min. All the scaffolds will be spread with 1X105 or 1X106 MSC, with the assistance of Puramatrix only to cover up its lumen. For the obtention of the MSC, 1g of AT will be subjected to enzymatic digestion with collagenase I and expanded until third passageway. This way, we'll have 8 test groups, with 3 repetitions for each group, which will be kept in the following cultures for 21 days: G1 - aVCI-SDS + 1X105CTM; G2 G1 - aVCI-SDS + 1X106CTM; G3 - aVCI-SDS-TX + 1X105CTM; G4 - aVCI-SDS-TX + 1X106CTM; G5 - aVCI-DS + 1X105CTM; G6 - aVCI-DS + 1X106CTM; G7 - aVCI-DS-TX + 1X105CTM; G8 - aVCI-DS-TX + 1X106CTM. Lastly, the samples will be collected and submitted to histomorphological analysis, and the cell count will be made on 10 fields within each slide, to determine which is or are the best protocols.