Speaker
Description
Introduction
Cardiovascular disease (CVD) is a general term for a group of disorders of the heart and blood vessels. According to statistical data of World Health Organization (WHO), CVD is one of the most prevalent disease and the leading cause of death globally in recent years. Therefore, it is justified to search for new, effective therapeutic methods such as stem cell therapy and tissue engineering. One of the most promising groups of stem cells are mesenchymal stem cells (MSCs). MSCs are multipotent stem cells characterized with high proliferative potential, paracrine activity and the ability to differentiate into many cell types. In order to effectively use the potential of MSCs for tissue regeneration, new biocompatible scaffolds for cell culture are still being sought for. An interesting material for biomedical applications due to their unique physicochemical properties are graphene oxide (GO) and reduced graphene oxide (rGO).
Methodology
The main goal of this study was to investigate the potential of GO and rGO substrates as scaffolds that can promote cardiomyogenic and angiogenic differentiation of MSCs in in vitro and in vivo studies.
Different types of solvents (water, ethanol), size of flakes, level of GO reduction and graphene layer thickness have been tested to obtain the most promising cell culture surfaces. The influence of the graphene-based scaffolds on the biological and functional properties of MSCs, such as morphology, viability, proliferative capacity, adhesion ability were investigated. The cytotoxicity of the graphene substrates was evaluated using Cytotoxicity Detection Kit (Sigma Aldrich). Based on the collected results, the substrates with the best biocompatibility were selected. Next, the chosen biomaterials were used to differentiate MSCs into heart tissue and blood vessel cells. In order to investigate the efficiency of the differentiation of MSCs, an analysis of gene expression was performed. Moreover, the effectiveness of angiogenic differentiation of MSCs on selected rGO surfaces was evaluated in murine model of hind limb ischemia.
Results
Obtained results showed that the biocompatibility of the tested substrates depends on the thickness of the graphene layer, the size of the flakes and the level of graphene reduction. The most appropriate graphene-based surfaces for MSCs culture were large flakes of GO and slightly reduced rGO. It was observed that in MSCs cultured on one type of slightly reduced rGO surface the expression of endothelial genes increased in comparison to the control conditions.
Conclusions
The conducted research indicates that graphene-based substrates, can have a different biological effect on MSCs, depending on the thickness and size of graphene flakes as well as the level of GO reduction. Moreover, selected rGO surfaces may promote angiogenic differentiation of MSCs, which could contribute to their future use in cardiovascular repair. However, further studies are required to analyze these phenomenon.
Acknowledgements:
This study was funded by NCN grant SYMFONIA 3 (2015/16/W/NZ4/00071) to EZS, NCBR grant STRATEGMED 3 (Strategmed3/ 303570/5/NCBR/2017) to EZS and NCN grant MAESTRO 11 (2019/34/A/NZ3/00134) to EZS.
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