Transplantation of bone marrow stromal cells overexpressing human vascular endothelial growth factor 165 enhances tissue repair in a rat model of radiation-induced injury.

BACKGROUND: The multilineage differentiation potential ability of bone marrow stromal cells (BMSCs) showed great potential in tissue engineering, while vascular endothelial growth factor 165 (VEGF165) promotes vasculogenesis and further promotes tissue regeneration. This study aimed to assess the ability of rat BMSCs expressing human VEGF A165 (hVEGF165) to promote tissue repair in rat model of radiation-induced injury. METHODS: Rat BMSCs were isolated from the tibia. Plasmid DNA expressing hVEGF165 was stably transfected into BMSCs using liposomes. The right hindlimb muscle of 40 rats was irradiated using a (60)Co γ source (total dose 30 Gy). The animals were divided into four groups (n = 10): not injected with BMSCs (control; group 1) or intramuscularly injected two times (once in 2 weeks) with pcDNA(TM)3.1-transfected BMSCs (group 2), untransfected BMSCs (group 3), or hVEGF165-transfected BMSCs (group 4). Angiography was performed 1 week after the last injection of BMSCs; samples of the hindlimb muscle were subjected to transmission electron microscopy, ultrastructural analysis, reverse transcription-PCR (RT-PCR), Western blotting, and immunohistochemistry. RESULTS: Rat BMSCs with multipotent differentiation capacity were isolated. hVEGF165-transfected BMSCs overexpressed hVEGF165 mRNA and protein. Injection of BMSCs (groups 2-4) increased the average vessel number, density, diameter, and cross-sectional area; mRNA expression of the myogenic markers including myoblast determination protein, myogenin, and a-smooth muscle actin; and CD31 protein expression; and promoted the repair of blood vessels and myofibers after radiation-induced injury compared to group 1; each of these parameters and hVEGF165 mRNA or protein expression were markedly improved in rats injected with hVEGF165-transfected BMSCs compared to groups 2 and 3. CONCLUSIONS: BMSCs expressing hVEGF165 enhanced the repair of radiation-induced tissue injury by promoting vasculogenesis and muscle fiber regeneration. BMSCs expressing hVEGF165 may have potential clinical applications.

[Effect of two different kinds of occlusal splints on temporomandibular disorders].

PURPOSE: To investigate the effect of relaxed splint and stabilized splint on the treatment of temporomandibular disorders (TMD). METHODS: A total of 68 TMD patients were divided into 2 groups (30 patients with acute pain and 38 patients with chronic pain) and treated with relaxed splint or stabilized splint. Visual analog scale (VAS) scores and electromyography (EMG) of bilateral anterior temporal muscle (TA) and masseter muscle (MM) were recorded before treatment and 1 month after treatment. The data was analyzed using variance analysis and student's t test with SPSS11.0 software package. RESULTS: At rest position, patients' EMG decreased remarkably (P<0.05) after both kinds of splints treatment. During maximal voluntary clench, the EMG of masticatory muscle (TA, MM) of patients with acute pain and EMG of MM of patients with chronic pain increased significantly after relaxed splint treatment (P<0.05), but only EMG of MM increased significantly after stabilization splint treatment (P<0.05). Patients' VAS scores decreased remarkably after both kinds of splints treatment (P<0.05), but during function, patients' acute pain eased remarkably after relaxed splint treatment (P<0.05). CONCLUSIONS: The relaxed splint and stabilized splint can relax the masticatory muscles and ease TMD pain, but relaxed splint has significant effect on the treatment of acute TMD patients. Supported by Natural Science Foundation of Hainan Province (310126).

Transfection of bone marrow mesenchymal stem cells using green fluorescence protein labeled hVEGF165 recombinant plasmid mediated by liposome.

OBJECTIVE: To study the role of bone marrow mesenchymal stem cells (BMSCs) in construction of vascularized engineered tissue. METHODS: hVEGF165 was amplified via RT-PCR before recombinant with pShuttle- green fluorescence protein;green fluorescent protein (GFP)-CMV. Then the recombinant shuttle plasmid was transfected into BMSCs with Lipofectamine(TM) 2000 for packaging and amplifying. hVEGF165 mRNA expression in BMSCs cells was tested. RESULTS: The sequence of hVEGF165 in pShuttle-GFP-hVEGF165 plasmid was confirmed by double-enzyme cleavage method and sequencing. hVEGF165 was highly expressed in BMSCs. CONCLUSIONS: The GFP/hVEGF165 recombinant plasmid vector was constructed successfully and expressed effectively in host cells, which may be helpful for discussing the possibility of the application of VEGF165-BMSCs in tissue engineering and ischemic disease cure.