Abstract
One of the most important goals of regenerative medicines is to generate alternative tissues with a developed vascular network. Endothelial cells are the most important cell type required in angiogenesis process, contributing to the blood vessels formation. The stimulation of endothelial cells to initiate angiogenesis requires appropriate extrinsic signals. The aim of this study was to evaluate the effects of M13 phage along with RGD peptide motif on in vitro and in vivo vascularization. The obtained results demonstrated the increased cellular proliferation, HUVECs migration, cells altered morphology, and cells attachment to M13 phage-RGD coated surface. In addition, the expression of Vascular Endothelial Growth Factor A (VEGF-A), VEGF Receptors 2 and 3, Matrix Metalloproteinase 9 (MMP9), and epithelial nitric oxide synthase (eNOS) transcripts were significantly upregulated due to the HUVECs culturing on M13 phage-RGD coated surface. Furthermore, VEGF protein secretion, nitric oxide, and reactive oxygen species (ROS) production were significantly increased in cells cultured on M13 phage-RGD coated surface.
Highlights
Generating new tissues that can be used as alternative organs for damaged tissues is one of the important aims in tissue engineering[1]
The results of Transmission electron microscopy (TEM) imaging showed that isolated phages represented the typical morphology of native M13 phage with a diameter of 6.6 nanometer (Fig. 1)
The mean DCF fluorescent intensity of HUVECs after 48 h culture on control, M13phage, M13 phage-RGD and gelatin surfaces was 20.92, 33.14, 30.20 and 23.72, respectively. These results showed that the M13 phage and M13 phage-RGD surfaces could increase the level of intracellular reactive oxygen species (ROS) in HUVECs compared to gelatin and control surfaces
Summary
Generating new tissues that can be used as alternative organs for damaged tissues is one of the important aims in tissue engineering[1]. The stimulation of ECs to initiate angiogenesis requires to an increase in the level of pro-angiogenic growth factors and appropriate extracellular matrix which subsequently plays an important role in this process[4,5]. It should be noted that the recent advances in tissue engineering field have been led to the design of biomaterials with proper physical characteristics of vascular tissue microenvironment. Previous studies indicated that indirect factors in an engineered tissue could stimulate the resident cells to create an angiogenic microenvironment. The advantage of this method is in stimulating in vivo responses of cells in order to regulate factor secretion based on the tissues requirement during the angiogenesis[7]. It is worth noting that the reproduction of www.nature.com/scientificreports/
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