Lentiviral­mediated Shh reverses the adverse effects of high glucose on osteoblast function and promotes bone formation via Sonic hedgehog signaling.

Patients with diabetes tend to have an increased incidence of osteoporosis, which may be associated with hyperglycemia; however, the pathogenic mechanisms governing this interaction remain unknown. The present study sought to investigate whether elevated extracellular glucose levels of bone mesenchymal stem cells (BMSCs) could influence osteoblastic differentiation and whether the intracellular Sonic hedgehog (Shh) pathway could adjust the effects. Furthermore, to verify the results in vivo, a rat tooth extraction model was constructed. BMSCs were incubated in eight types of culture medium, including low glucose (LG), LG + lentivirus (Lenti), LG + Lenti­small interfering RNA (Lenti­siRNA), LG + Lenti­Shh, high glucose (HG), HG + Lenti, HG + Lenti­siRNA and HG + Lenti­Shh. The lentiviral transfection efficiency was observed using a fluorescence microscope; protein and mRNA expression was detected by western blotting and reverse transcription­quantitative polymerase chain reaction (RT­qPCR). The matrix mineralization and alkaline phosphatase (ALP) activity of BMSCs were examined by Alizarin red staining and ALP activity assays, respectively. The expression of osteogenesis­related genes in BMSCs were quantified by RT­qPCR. The alveolar ridge reduction was measured and histological sections were used to evaluate new bone formation in the tooth socket. With high concentrations of glucose, Shh expression, matrix mineralization nodules formation, ALP activity and the levels of bone morphogenic protein 4 (BMP4), bone sialoprotein (BSP) and osteopontin (OPN) expression were greatly reduced compared with LG and corresponding control groups. Whereas activated Shh signaling via Lenti­Shh could increase the number of matrix mineralization nodules, ALP activity, and the expression levels of BMP4, BSP and OPN in BMSCs. Additionally, in vivo assays demonstrated that Lenti­Shh induced additional bone formation. Collectively, the results of the present study indicated that HG inhibited the Shh pathway in osteoblasts and resulted in patterning defects during osteoblastic differentiation and bone formation, while the activation of Shh signaling could suppress these deleterious effects.

Vascular endothelial growth factor expression and pathological changes in the local tissue of facial hemangiomas following injections with pure alcohol.

The aim of the present study was to investigate the association between the formation of hemangioma and the expression of vascular endothelial growth factor (VEGF) following local injections of pure alcohol in patients exhibiting hemangioma. Ten healthy subjects (control group) and 10 hemangioma patients (treatment group) were included in the study population, with the hemangioma patients receiving one injection of pure alcohol. The VEGF levels were evaluated in the treatment and control group subjects prior to and following the injection using enzyme-linked immunosorbent assay; furthermore, local tissue was excised to perform pathological analysis one week after the injections. The VEGF levels of the healthy group were identified to be significantly lower when compared with those of the treatment group prior to the injections (P<0.01) and one week after the injections (P<0.01), however, were not significantly different when compared with the treatment group one month after the injections (P>0.01). Therefore, serum VEGF concentrations in the peripheral blood may be a clinical indicator of the efficacy of clinical treatment and aid with determination of the prognosis.

Study of TNF-α, IL-1ß and LPS levels in the gingival crevicular fluid of a rat model of diabetes mellitus and periodontitis.

OBJECTIVE: In this study, we sought to investigate the dynamic changes in the levels of TNF-α, IL-1ß and LPS in the gingival crevicular fluid (GCF) in a rat model of diabetes mellitus (DM) and periodontitis (PD). Additionally, we evaluated alveolar bone loss and the histopathological response associated with experimental diabetes mellitus and experimental periodontitis. METHODS: DM and PD were induced together in 15 rats (group 1) by streptozotocin injection and ligature induction. Periodontitis alone was produced by ligature induction in 15 rats (group 2), diabetes alone was produced by streptozotocin injection in 15 rats (group 3), and fifteen systemically and periodontally healthy rats were used as controls (group 4). The gingival TNF-α, IL-1ß and LPS levels were measured by using ELISA method. Periodontal destruction was assessed by measuring the alveolar bone loss. Periodontal inflammation was quantified by histopathological grading in H&E stained samples. RESULTS: Higher levels of TNF-α, IL1-ß and LPS, increased alveolar bone loss and more serve histopathology were found in group 1 compared with group 2, group 3 and group 4 (p< 0.05). The quantities of TNF-α, IL1-ß and LPS, the amount of alveolar bone loss and the severity of the histopathological finding were greater in group 2 than group 3 and group 4 (p< 0.05). Group 3 demonstrated higher levels of TNF-α, IL1-ß and LPS, increased alveolar bone loss and more serve histopathology than group 4 (p< 0.05). Statistically significant differences were noted between all of the groups. CONCLUSIONS: These data indicate that DM may lead to enhanced TNF-α, IL1-ß and LPS production in the periodontal tissues. The resorption values of alveolar bone and the histological inflammation were more severe in rats with periodontitis and diabetes mellitus than in those with periodontitis alone, diabetes mellitus alone and control rats. Our findings are consistent with the hypothesis that hyperglycemia contributes to the heightened inflammatory response associated with periodontitis.