Osteogenic differentiation effects on rat bone marrow-derived mesenchymal stromal cells by lentivirus-mediated co-transfection of human BMP2 gene and VEGF165 gene

Abstract

We proposed a novel combined gene therapy of human vascular endothelial growth factor 165 gene (hVEGF165) and human bone morphogenetic protein 2 gene (hBMP2) for bone regeneration by lentivirus-mediated co-transfection of both genes into rat bone marrow-derived mesenchymal stromal cells (MSCs). Both genes were successfully co-expressed in MSCs confirmed by real-time PCR and ELISA. And the alkaline phosphatase activity of MSCs was significantly augmented by the co-transfection with both genes than any single gene transfection (P < 0.01). These results demonstrated the feasibility of the combined gene therapy by using MSCs lentivirally co-transfected with hVEGF165 and hBMP2 for bone regeneration.

Odontogenic differentiation of vascular endothelial growth factor-transfected human dental pulp stem cells in vitro

“…In this study, the expression of ALP and OCN was significantly enhanced in the DPSCs/VEGF as compared with that in the DPSCs/Vector. This result was consistent with that from other reports on cells with VEGF overexpression in vitro (11,13). The expression of odontogenic differentiation-specific markers, such as DSPP and DMP1 genes, was also enhanced among the DPSCs/VEGF in this study.…”

“…VEGF gene-transfected rat bone marrow stromal cells (BMSCs) were reported to have higher ectopic osteogenesis in vivo (12). Another study also confirmed the improved osteogenic differentiation capacity of rat BMSCs by lentivirus-mediated VEGF gene transfection (13). Due to the similarity between osteogenic and odontogenic differentiation among DPSCs (14), lentiviral vector-mediated stable transfection of the VEGF gene may be an effective strategy to improve the odontogenic differentiation capacity of DPSCs.…”

Locally administered lentivirus-mediated siRNA inhibits wear debris-induced inflammation in murine air pouch model

“…Lane 1, DNA ladder; Lanes 2-4, DNA extracted from pouches in TNF-a group; Lanes 5-7, DNA extracted from pouches in MS group; Lanes 8-10, DNA extracted from pouches in control group These results indicated that successful transfection in vivo could be achieved using lentiviral vector, and the LV-mediated TNF-a siRNA could inhibit TNF-a expression in high efficiency both in mRNA and protein levels. Furthermore, the silencing of TNF-a lasted at least 4 weeks, suggesting that the vector was stably integrated into the host genome and passed on to all progeny cells (Jiang et al 2008), leading to relatively long-term stable silencing activity rather than a single bolus or limited expression in only one generation. The findings in our study were in accordance with the results in Zhang et al's (2007) study.…”

Gene therapy of arthritis and orthopaedic disorders: current experimental approaches in China and in Canada

“…The results demonstrated that APA microcapsules could prevent encapsulated AdBMP-2 gene-transfected MSCs from initiating cellular immune responses, and gene products from the encapsulated BMP-2 cells could induce undifferentiated MSCs to become osteoblasts [53] . BMP-7 and human vascular endothelial growth factor gene have been tested in bone regeneration [54,55] . Arthritic chondrocytes and synoviocytes are receptive to transduction via various gene delivery methods.…”

“…In vitro [55] Spinal cord injury NT-3 Naked DNA Mouse [82] Osteonecrosis BMP-2 Adenovirus Goat [83] Mandibular defects BMP-2 Liposome Rabbit [84] Osteoarthritis IL-1Ra Naked DNA Rabbit [56] Bone defects BMP2 Adenovirus Rat [52] Wuhan Intervertebral disc regeneration hSox9 Baculovirus Rabbit [85] AAV: Adeno-associated virus; BMP: Bone morphogenetic protein; Cbfa1: Core binding factor alpha1; CTLA4Ig: Cytotoxic T lymphocyte-associated antigen 4 Ig; hIGF-1: Human IGF I; hSox9: Human sry-related high-mobility-group box 9; NT-3: Neurotrophin-3; Runx2: Runt-related transcription factor 2; siRNA: Short interfering ribonucleic acid; sTNF-R1: Soluble tumor necrosis factor receptor 1; sTNF-R-IgG [8,9] . Nevertheless, gene therapy is not only a promising treatment innovation in cancer but it also offers the opportunity to provide prolonged systemic or local drug delivery in non-lethal conditions, such as various orthopaedic problems and disorders.…”