The effect of RGD pertide on cell proliferation and vasculogenesis during microtissue formation

3D (Dimensional) microtissues are commonly used in regenerative medicine, medicine design, toxicology studies and advance cancer research. Microtissues which are constructed by this technique mimic natural in vivo condition and it stands for shifting form between in vitro studies and in vivo research. Scaffold-free microtissues produce own extracellular matrix and cells proliferate faster. However microtissue is used in vitro experiments, they mimic in vivo condition by producing own matrix.
Recently, RGD peptides which are commonly used biomaterials consist of GRGDS sequences and it has active role cell-cell and cell-matrix attachment. This peptide has been shown to enhance cell proliferation by increasing cellular interaction and cell-matrix connection in previous studies.
HUVEC (Human Umbilical Vein Endothelial Cell) is one of the most commonly used model cells to understand the endothelial cell behavior. Moreover, the inner layer of the vascular structure is composed of endothelial cells. HUVEC is one of the model cell types frequently used in vascularization studies with high vasculogenesis capacity and endothelial origin. VEGF, Tie-1, Tie-2, PECAM, and Ve-cadherin are also effective genes for expressing preliminary vascular model studies in microtissue studies.
This study was carried out to investigate the effect of RGD peptide on HUVEC scaffold-free 3D microtissue formation and early vasculogenesis. Microtissue size analysis, viability analysis, focal adhesion analysis and qRT-PCR (Real Time PCR) data were used. In the results of working, we found that RGD peptides increase HUVEC microtissue proliferation and contribute to constituting microtissue formation. Furthermore, it plays the effective role of HUVEC 3D microtissue formation during vascularization process. We experimented different RGD concentration on microtissue formation. The most effective amount of those peptides was shown as 2 mM RGD. 2 mM RGD microtissue formation as depicted from enhanced viability studies compared to 0 mM, 1 mM and 4 mM RGD concentration. microtissue diameter. Additionally, the morphology of cells described with focal adhesion analysis on microtissue. qRT-PCR data displays that RGD peptide helps to initiate vascular processing. Results of all data demonstrate that 2 mM RGD peptide has important tasks in 3D HUVEC microtissue formation and also regulated vascularization period. This study put forward the RGD peptides can be useful for constructing vascular formation. Therefore using optimal RGD peptide amount for vascular tissue may subvene in vivo to construct vascular networks.