Influence of Recombinant Codon-Optimized Plasmid DNA Encoding VEGF and FGF2 on Co-Induction of Angiogenesis

Abstract

Several methods for the stimulation of skin wound repair have been proposed over the last few decades. The most promising among them are gene and stem cell therapy. Our present experiments combined several approaches via the application of human umbilical cord blood mononuclear cells (hUCB-MC) that were transfected with pBud-VEGF165-FGF2 plasmid (gene-cell therapy) and direct gene therapy using pBud-VEGF165-FGF2 plasmid to enhance healing of full thickness skin wounds in rats. The dual expression cassette plasmid pBud-VEGF165-FGF2 encodes both VEGF and FGF2 therapeutic genes, expressing pro-angiogenic growth factors. Our results showed that, with two weeks post-transplantation, some transplanted cells still retained expression of the stem cell and hematopoietic markers C-kit and CD34. Other transplanted cells were found among keratinocytes, hair follicle cells, endothelial cells, and in the derma. PCNA expression studies revealed that transplantation of transfected cells terminated proliferative processes in regenerating wounds earlier than transplantation of untransfected cells. In the direct gene therapy group, four days post-operatively, the processes of flap revascularization, while using Easy LDI Microcirculation Camera, was higher than in control wounded skin. We concluded that hUCB-MC can be used for the treatment of skin wounds and transfection these cells with VEGF and FGF2 genes enhances their regenerative abilities. We also concluded that the application of pBud-VEGF165-FGF2 plasmids is efficient for the direct gene therapy of skin wounds by stimulation of wound revascularization.

Overexpression of VEGF in dermal fibroblast cells accelerates the angiogenesis and wound healing function: in vitro and in vivo studies

“…In another study, Salafutdinov et al evaluated the efficiency of human umbilical cord blood mononuclear cells (hUCB-MC) transfection with the pBudVEGF-FGF2 vector. Seventy-two hours after electroporation, they demonstrated a noteworthy change in the mRNA levels of the FGF2 and VEGF genes expression in modified cells by 19,000 ± 1186 and 57,000 ± 2250 times, respectively compared to unmodified UCB-MC 36 . Also, our previous study showed a significant enhancement in VEGF expression in fibroblasts cultured on the PU-CA/gelatin.PRGF/PU-CA scaffold 17 .…”

Altered hypoxia inducible factor regulation in hereditary haemorrhagic telangiectasia

“…Following several studies’ descriptions of the influence of immune cells on angiogenesis and the vascular abnormalities in HHT patients, we analysed HIF target genes influencing the angiogenesis 34 – 37 . Surprisingly, changes in angiogenesis associated genes were not detected in HHT patients’ leukocytes (Fig.…”

Assessment of the Tissue Response to Modification of the Surface of Dental Implants with Carboxyethylphosphonic Acid and Basic Fibroblastic Growth Factor Immobilization (Fgf-2): An Experimental Study on Minipigs

“…They chemically modified the rough surface of implants, discovering an upregulation of osteoblastic differentiation and the suppression of osteoclastogenesis regulating the RANKL/RANK/OPG transcriptional axis. Fibroblastic growth factors are pleiotropic growth factors; that is, they intervene in a multitude of biological processes, such as the regulation of cell proliferation, migration, adhesion and differentiation of different tissues, such as epithelial tissue, soft connective tissue, nervous tissue and bone tissue [ 17 , 29 , 30 , 31 ]. As previously described, FGF generally stimulates cell proliferation [ 32 ], while melatonin is capable of promoting cell differentiation and the mineralization of bone tissue [ 33 , 34 , 35 ].…”

“…Surface bioactivation is a biochemical method of surface modification, whose objective is based on the immobilization of proteins, enzymes or peptides that induce a specific cellular response at the bone–implant interface. To modify this type of surface, organic components are used, which are known to create a response in the bone and promote cell adhesion, such as identification of the Arg-Gly-Asp (RGD) sequence, a mediator of cell binding with plasma proteins and extracellular matrix proteins (fibronectin, vitronectin, type I collagen, osteopontin or bone sialoproteins) [ 14 , 15 , 16 , 17 ]. The development of biocompatible layers that attempt to mimic the adhesion of osteoblasts to obtain better and faster osseointegration is an ongoing investigation.…”