RESUMO
BACKGROUND AND OBJECTIVE: We previously reported that low-intensity laser irradiation stimulated bone nodule formation through enhanced cellular proliferation and differentiation. However, the mechanisms of irradiation are unclear. Thus, we attempted to determine the responsibility of insulin-like growth factor (IGF)-I for the action observed. STUDY DESIGN/MATERIALS AND METHODS: Osteoblast-like cells were isolated from fetal rat calvariae and cultured with rat recombinant (r) IGF-I, IGF-I-antibody (Ab), and/or the cells were irradiated once (3.75 J/cm(2)) with a low-intensity Ga-Al-As laser (830 nm). The number and area of bone nodules formed in the culture were analyzed, and IGF-I expression was also examined. RESULTS: Treatment with rIGF-I significantly stimulated the number and area of bone nodules. This stimulatory effect was quite similar to those by laser irradiation, and this stimulation was abrogated dose-dependently by treatment with IGF-I-Ab. Moreover, laser irradiation significantly increased IGF-I protein and gene expression. CONCLUSION: The stimulatory effect of bone nodule formation by low-intensity laser irradiation will be at least partly mediated by IGF-I expression.
Assuntos
Fator de Crescimento Insulin-Like I/biossíntese , Crânio/metabolismo , Crânio/efeitos da radiação , Animais , Anticorpos , Células Cultivadas , Dinoprostona/biossíntese , Feminino , Feto , Expressão Gênica , Terapia com Luz de Baixa Intensidade , Modelos Animais , Osteoblastos/metabolismo , Osteoblastos/efeitos da radiação , Osteogênese/fisiologia , Osteogênese/efeitos da radiação , Gravidez , Ratos , Ratos Wistar , Crânio/citologiaRESUMO
BACKGROUND: The mechanism of stimulation of human gingival epithelial cells (HGEC) by Porphyromonas gingivalis (Pg) has not been fully clarified yet. In order to investigate the possible activation of HGEC by Pg through Toll-like receptors (TLRs), we analyzed the production of chemotactic factors and the activated nuclear factor-kappa B (NF-kappaB). METHODS: The mRNA expression of TLRs and the protein expression of TLR2 and TLR4 in HGEC and gingival tissue were assessed using reverse transcription-polymerase chain reaction (RT-PCR) assay and immunohistochemical staining. Primary cultured HGEC (nHGEC) and HGEC transformed by simian virus 40 T antigen (OBA-9) were activated by a sonic extract (SE) of Pg to examine cytokine production and NF-kappaB activation using enzyme-linked immunosorbant assay (ELISA). In addition, Pg mediated activation of NF-kappaB in a TLR2-transfectant was also investigated. RESULTS: RT-PCR results revealed that HGEC expressed mRNA of TLR2, TLR4, TLR5, and TLR9, although the expression profiles of each cell line were slightly different. In addition, immunostaining revealed the prominent expression of TLR2 not only in nHGEC, but also in the gingival epithelium of the tissue specimen. Interestingly, nHGEC and OBA-9 secreted IL-8 and monocyte chemoattractant protein (MCP)-1 upon stimulation with Pg SE more efficiently than LPS and fimbriae of Pg. Furthermore, Pg SE increased the activated NF-kappaB not only in OBA-9, but also in 293T cells transfected with the human TLR2 gene. CONCLUSION: TLR2 participates, at least partly, in the signaling pathway to induce chemokine production in gingival epithelium as a reaction against Pg component(s), probably other than lipopolysaccharide and fimbriae.