RESUMO
OBJECTIVES: Stem cells from the dental apical papilla (SCAPs) can be induced to differentiate along both osteoblast and odontoblast lineages. However, little knowledge is available concerning their differentiation efficiency in osteogenic media containing additional KH2 PO4 . MATERIALS AND METHODS: Stem cells from the dental apical papilla were isolated from apical papillae of immature third molars and treated with two kinds of mineralization-inducing media, MM1 and MM2, differing in KH2 PO4 concentration. Proliferation and osteo/odontogenic differentiation capacity of MM1/MM2-treated SCAPs were investigated and compared both in vitro and in vivo. RESULTS: Cell counting and flow cytometry demonstrated that MM2 containing 1.8 mm additional KH2 PO4 significantly enhanced proliferative potential of SCAPs, compared to MM1. Osteo/odontogenic capacity of SCAPs was much better in MM2 medium than in MM1, as indicated by elevated alkaline phosphatase activity, increased calcium deposition and upregulated expression of osteo/odontoblast-specific genes/proteins (for example, runt-related transcription factor 2, osterix, osteocalcin, dentin sialoprotein, and dentin sialophosphoprotein). In vivo transplantation findings proved that SCAPs in MM2 group generated more mineralized tissues, and presented higher expression of osteo/odontoblast-specific proteins (osteocalcin and dentin sialoprotein) than those in the MM1 group. CONCLUSION: Mineralization-inducing media supplemented with 1.8 mm additional KH2 PO4 significantly enhanced cell proliferation and improved differentiation capacity of SCAPs along osteo/odontogenic cell lineages, compared to counterparts lacking additional KH2 PO4 .
Assuntos
Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Papila Dentária/citologia , Osteogênese , Fosfatos/farmacologia , Compostos de Potássio/farmacologia , Células-Tronco/citologia , Adolescente , Adulto , Fosfatase Alcalina/metabolismo , Animais , Cálcio/metabolismo , Linhagem da Célula , Células Cultivadas , Técnicas de Cocultura , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Meios de Cultura/metabolismo , Papila Dentária/metabolismo , Proteínas da Matriz Extracelular/genética , Proteínas da Matriz Extracelular/metabolismo , Regulação da Expressão Gênica , Humanos , Dente Serotino/citologia , Dente Serotino/metabolismo , Odontoblastos/citologia , Odontoblastos/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Ratos , Sialoglicoproteínas/genética , Sialoglicoproteínas/metabolismo , Transplante de Células-Tronco/métodos , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Calcificação de Dente , Adulto JovemAssuntos
Bioensaio/métodos , Polpa Dentária/inervação , Neurofarmacologia/métodos , Nociceptores/metabolismo , Células Receptoras Sensoriais/metabolismo , Analgésicos/farmacologia , Animais , Peptídeo Relacionado com Gene de Calcitonina/análise , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Polpa Dentária/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Dente Serotino/inervação , Dente Serotino/metabolismo , Dente Serotino/cirurgia , Receptores Opioides mu/análise , Receptores Opioides mu/metabolismo , Canais de Cátion TRPV/análise , Canais de Cátion TRPV/metabolismo , Odontalgia/tratamento farmacológico , Odontalgia/metabolismo , Odontalgia/fisiopatologiaRESUMO
The aim of the study was to determine immunoreactive Substance P (iSP) and immunoreactive calcitonin gene-related peptide (iCGRP) content in the human dental pulp and whether local anesthesia has an effect on the neuropeptide content. Dental pulps were obtained from patients, who underwent surgical extraction of all 4 impacted wisdom teeth under general isoflurane gas anesthesia. There was a very high interindividual variation in tissue content, with small variation in levels found in teeth from the same patient. Pulps obtained from lower teeth without local anesthesia contained an average of 131 +/- 62 fmol/mg protein of iCGRP and 15 +/- 9 fmol/mg iSP (n = 10). With additional mandibular block anesthesia the values were 194 +/- 71 fmol/mg iCGRP (statistically significant, p = 0.0356, Mann-Whitney-rank-sum-test) and 12 +/- 3.6 fmol/mg iSP. The results suggest that local anesthesia attenuates neuropeptide release in the human dental pulp during surgical extraction.