RESUMO
Hydroxyapatite was immobilized on poly(ethylene-co-vinyl alcohol) (EVA) by alternate soaking in aqueous CaCl(2) and Na(2)HPO(4) solutions, followed by carboxyl groups introduction through ozone exposure in order to investigate the nature of the gingival cells, to control their proliferation and properties and to develop a highly organized hybrid implant possessing periodontium. Human gingival cells were cultured on the ozone-exposed EVA, collagen-immobilized EVA, hydroxyapatite-immobilized EVA, and a conventional tissue culture dish. Cell proliferation was highest on the tissue culture dish and lowest on the hydroxyapatite-immobilized EVA. The results of RT-PCR of gingival cells on hydroxyapatite-immobilized EVA shows that mRNAs expressed in bone and periodontal ligament were determined. Furthermore, alkaline phosphatase activity and ELISA assay revealed that gingival cells acquired the osteoblastic properties when cultured on hydroxyapatite-immobilized EVA, suggesting that the periodontium might be regenerated around implants using gingival cells.
Assuntos
Materiais Biocompatíveis/farmacologia , Durapatita/farmacologia , Gengiva/citologia , Gengiva/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Sequência de Bases , Proliferação de Células , Células Cultivadas , Materiais Revestidos Biocompatíveis , Colágeno Tipo I/genética , Colágeno Tipo XII/genética , Cristalização , Primers do DNA/genética , Gengiva/metabolismo , Humanos , Teste de Materiais , Osteocalcina/genética , Osteonectina/genética , Periodonto/citologia , Periodonto/fisiologia , Polivinil , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Regeneração , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
INTRODUCTION: This study determined the gene expression profiles of the human coronal pulp (CP) and apical pulp complex (APC) with the aim of explaining differences in their functions. METHODS: Total RNA was isolated from the CP and APC, and gene expression was analyzed using complementary DNA microarray technology. Gene ontology analysis was used to classify the biological function. Quantitative reverse-transcription polymerase chain reaction and immunohistochemical staining were performed to verify microarray data. RESULTS: In the microarray analyses, expression increases of at least 2-fold were present in 125 genes in the APC and 139 genes in the CP out of a total of 33,297 genes. Gene ontology class processes found more genes related to immune responses, cell growth and maintenance, and cell adhesion in the APC, whereas transport and neurogenesis genes predominated in the CP. Quantitative reverse-transcription polymerase chain reaction and immunohistochemical staining confirmed the microarray results, with DMP1, CALB1, and GABRB1 strongly expressed in the CP, whereas SMOC2, SHH, BARX1, CX3CR1, SPP1, COL XII, and LAMC2 were strongly expressed in the APC. CONCLUSIONS: The expression levels of genes related to dentin mineralization, neurogenesis, and neurotransmission are higher in the CP in human immature teeth, whereas those of immune-related and tooth development-related genes are higher in the APC.
Assuntos
Polpa Dentária/crescimento & desenvolvimento , Expressão Gênica , Odontogênese/genética , Ápice Dentário/crescimento & desenvolvimento , Adolescente , Receptor 1 de Quimiocina CX3C , Calbindina 1/genética , Proteínas de Ligação ao Cálcio/genética , Adesão Celular/genética , Criança , Pré-Escolar , Colágeno Tipo XII/genética , Polpa Dentária/anatomia & histologia , Polpa Dentária/citologia , Polpa Dentária/diagnóstico por imagem , Proteínas da Matriz Extracelular/genética , Feminino , Perfilação da Expressão Gênica , Proteínas Hedgehog/genética , Proteínas de Homeodomínio/genética , Humanos , Imuno-Histoquímica , Laminina/genética , Masculino , Análise em Microsséries/métodos , Neurogênese/genética , Osteopontina/genética , Fosfoproteínas/genética , RNA/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Receptores de Quimiocinas/genética , Receptores de GABA-A/genética , República da Coreia , Transmissão Sináptica/genética , Ápice Dentário/anatomia & histologia , Ápice Dentário/citologia , Ápice Dentário/diagnóstico por imagem , Calcificação de Dente/genética , Fatores de Transcrição/genética , Adulto JovemRESUMO
The dental follicle is a mesenchymal tissue that surrounds the developing tooth germ. During tooth root formation, periodontal components, viz., cementum, periodontal ligament (PDL), and alveolar bone, are created by dental follicle progenitors. Here, we report the presence of PDL progenitors in mouse dental follicle (MDF) cells. MDF cells were obtained from mouse incisor tooth germs and immortalized by the expression of a mutant human papilloma virus type 16 E6 gene lacking the PDZ-domain-binding motif. MDF cells expressing the mutant E6 gene (MDF( E6-EGFP ) cells) had an extended life span, beyond 150 population doublings (PD). In contrast, normal MDF cells failed to proliferate beyond 10 PD. MDF( E6-EGFP ) cells expressed tendon/ligament phenotype-related genes such as Scleraxis (Scx), growth and differentiation factor-5, EphA4, Six-1, and type I collagen. In addition, the expression of periostin was observed. To elucidate the differentiation capacity of MDF( E6-EGFP ) cells in vivo, the cells were transplanted into severe combined immunodeficiency mice. At 4 weeks, MDF( E6-EGFP ) cell transplants had the capacity to generate a PDL-like tissue that expressed periostin, Scx, and type XII collagen and the fibrillar assembly of type I collagen. Our findings suggest that MDF( E6-EGFP ) cells can act as PDL progenitors, and that these cells may be a useful research tool for studying PDL formation and for developing regeneration therapies.