Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
1.
Odontology ; 109(4): 895-903, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34086131

RESUMO

Pulp regeneration with stem cells is a promising alternative for treating periapical and pulp diseases of young permanent teeth. The aim of this study was to characterize decellularized dental pulp extracellular matrix (dECM) and investigate whether bone morphogenetic protein 4 (BMP4) regulates dental pulp stromal cells (DPSC)-mediated pulp regeneration combined with dECM. Dental pulp isolated from healthy third molars was decellularized with 10% sodium dodecyl sulfate (SDS) and Triton X-100. H&E staining, DAPI staining and electron microscopy were used to observe the dECM structure. The Cell Counting Kit-8 assay was used to analyse cell proliferation. Recombinant adenovirus was used to overexpress BMP4 in DPSCs. The cells were cultured in dECM and dECM + three-dimensional (3D) Vitrogel systems, and bone/dentin/angiogenesis marker expression was evaluated by real-time polymerase chain reaction (RT-PCR) and ALP staining. DPSCs mixed with dECM and BMP4 were transplanted into nude mice, and pulp-like tissue formation was evaluated. The expression of osteogenic and angioblastic genes was increased, and pulp-like tissue formed in vivo. Thus, dECM promotes DPSC proliferation, BMP4 and dECM together accelerate pulp-like tissue formation by DPSCs in vitro.


Assuntos
Polpa Dentária , Regeneração , Animais , Proteína Morfogenética Óssea 4 , Diferenciação Celular , Humanos , Camundongos , Camundongos Nus , Células Estromais
2.
Int J Med Sci ; 16(4): 567-575, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31171908

RESUMO

Bone tissue engineering requires a combination of cells, efficient biochemical and physicochemical factors, and biocompatible scaffolds. In this study, we evaluated the potential use of injectable Matrigel as a scaffold for the delivery of rat dental follicle stem/precursor cells (rDFSCs) transduced by bone morphogenetic protein (BMP) 9 to enhance osteogenic differentiation in vitro and promote ectopic bone formation in vivo. Recombinant adenovirus was used to overexpress BMP9 in rDFSCs. Alkaline phosphatase activity was measured using a histochemical staining assay and a chemiluminescence assay kit. Quantitative real-time polymerase chain reaction was used to determine mRNA expression levels of bone-related genes including distal-less homeobox 5 (DLX5), osteopontin (OPN), osterix (Osx), and runt-related transcription factor 2 (Runx2). Matrix mineralization was examined by Alizarin Red S staining. rDFSCs proliferation was analyzed using the Cell Counting Kit-8 assay. Subcutaneous implantation of rDFSCs-containing Matrigel scaffolds was used, and micro-computed tomography analysis, histological evaluation, and trichrome staining of implants extracted at 6 weeks were performed. We found that BMP9 enhanced alkaline phosphatase activity and mineralization in rDFSCs. The expression of bone-related genes (DLX5, OPN, Osx, and Runx2) was also increased as a result of BMP9 stimulation. Micro-computed tomography analysis and histological evaluation revealed that the bone masses retrieved from BMP9-overexpressing rDFSCs were significantly more pronounced in those with than in those without Matrigel. Our results suggest that BMP9 effectively promote osteogenic differentiation of rDFSCs, and Matrigel facilitate BMP9-induced osteogenesis of rDFSCs in vivo.


Assuntos
Fator 2 de Diferenciação de Crescimento/genética , Osteogênese/efeitos dos fármacos , Transplante de Células-Tronco , Alicerces Teciduais , Animais , Diferenciação Celular/efeitos dos fármacos , Colágeno/farmacologia , Saco Dentário/citologia , Combinação de Medicamentos , Fator 2 de Diferenciação de Crescimento/farmacologia , Humanos , Laminina/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/genética , Proteoglicanas/farmacologia , Ratos , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Engenharia Tecidual , Microtomografia por Raio-X
3.
Front Bioeng Biotechnol ; 10: 1036061, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36324890

RESUMO

Apical periodontitis is a common clinical disease caused by bacteria; bacterial metabolites can cause an imbalance in bone homeostasis, bone mass reduction, and tooth loss. Bone resorption in apical periodontitis causes a concentration of stress in the tooth and periodontal tissues during occlusion, which aggravates the disease. Emerging evidence indicates that bone morphogenetic protein 9 (BMP9), also known as growth differentiation factor 2(Gdf2), may play an important role in tooth and dentoalveolar development. Herein, we investigated the role of BMP9 in the development of apical periodontitis and its effects on the biomechanics of dentoalveolar bone. Apical periodontitis models were established in five BMP9 knockout (KO) mice and five C57BL/6 WT (wild-type) mice. At baseline and 14, 28, and 42 days after modeling, in vivo micro-computed tomography analysis and three-dimensional (3D) reconstruction were performed to evaluate the apical lesion in each mouse, and confirm that the animal models were successfully established. Finite element analysis (FEA) was performed to study the stress and strain at the alveolar fossa of each mouse under the same vertical and lateral stress. FEA revealed that the stress and strain at the alveolar fossa of each mouse gradually concentrated on the tooth cervix. The stress and strain at the tooth cervix gradually increased with time but were decreased at day 42. Under the same lingual loading, the maximum differences of the stress and strain at the tooth root in KO mice were greater than those in WT mice. Thus, these findings demonstrate that BMP9 could affect the biomechanical response of the alveolar fossa at the tooth root in mice with apical periodontitis. Moreover, the effects of BMP9 on the biomechanical response of the alveolar bone may be site-dependent. Overall, this work contributes to an improved understanding of the pathogenesis of apical periodontitis and may inform the development of new treatment strategies for apical periodontitis.

4.
PeerJ ; 5: e4057, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29158986

RESUMO

Prophages have been considered genetic units that have an intimate association with novel phenotypic properties of bacterial hosts, such as pathogenicity and genomic variation. Little is known about the genetic information of prophages in the genome of Streptococcus mutans, a major pathogen of human dental caries. In this study, we identified 35 prophage-like elements in S. mutans genomes and performed a comparative genomic analysis. Comparative genomic and phylogenetic analyses of prophage sequences revealed that the prophages could be classified into three main large clusters: Cluster A, Cluster B, and Cluster C. The S. mutans prophages in each cluster were compared. The genomic sequences of phismuN66-1, phismuNLML9-1, and phismu24-1 all shared similarities with the previously reported S. mutans phages M102, M102AD, and ϕAPCM01. The genomes were organized into seven major gene clusters according to the putative functions of the predicted open reading frames: packaging and structural modules, integrase, host lysis modules, DNA replication/recombination modules, transcriptional regulatory modules, other protein modules, and hypothetical protein modules. Moreover, an integrase gene was only identified in phismuNLML9-1 prophages.

5.
J Periodontol ; 88(5): 473-483, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-27982724

RESUMO

BACKGROUND: Periodontitis is a chronic inflammatory disease initiated by bacteria and their virulence factors. Bortezomib (BTZ) is the first proteasome inhibitor for clinical treatment of malignancies. Its anticancer activity is accompanied by an anti-inflammatory effect. However, there are few reports about its anti-inflammatory effect and underlying mechanism in periodontal disease, especially on human periodontal ligament cells (hPDLCs), which are considered a promising cell-based therapy for treating periodontitis. METHODS: hPDLCs were treated with lipopolysaccharide (LPS) and pretreated with BTZ. mRNA and protein levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1ß, IL-6, and IL-8 were determined. The anti-inflammatory mechanism of BTZ was studied. Further, experimental rat periodontitis was induced with ligature and LPS injection, and simultaneously and locally treated with BTZ (three injections/week). Four weeks after treatment, microcomputed tomography, immunohistochemical, and histopathologic analyses were performed. RESULTS: Bortezomib administration at safe concentrations (≤1 nM) inhibited production of proinflammatory cytokines in LPS-stimulated hPDLCs via nuclear factor (NF)-kappa B, p38/extracellular signal-regulated kinase, and mitogen-activated protein kinase/activator protein-1 pathways. Moreover, in the LPS and ligature-induced periodontitis rat model, BTZ suppressed expression of TNF-α, IL-1ß, IL-6, and IL-8, reduced the ratio of receptor activator of NF-κB ligand/osteoprotegerin, and prevented alveolar bone absorption. CONCLUSION: These findings demonstrate the anti-inflammatory activity of BTZ against periodontal inflammatory response and present BTZ as a promising therapy for periodontal disease.


Assuntos
Bortezomib/uso terapêutico , Ligamento Periodontal/efeitos dos fármacos , Periodontite/tratamento farmacológico , Inibidores de Proteassoma/uso terapêutico , Adolescente , Animais , Criança , Modelos Animais de Doenças , Feminino , Humanos , Inflamação/tratamento farmacológico , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Masculino , Ligamento Periodontal/citologia , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Fator de Necrose Tumoral alfa/metabolismo
6.
Sci Rep ; 7(1): 6373, 2017 07 25.
Artigo em Inglês | MEDLINE | ID: mdl-28743897

RESUMO

Cells, scaffolds, and growth factors play important roles in bone regeneration. Bone morphogenetic protein 9 (BMP9), a member of BMP family, could facilitate osteogenesis by regulating growth factors and promoting angiogenesis. Similar to other stem cells, rat dental follicle stem cells (rDFCs), the precursor cells of cementoblasts, osteoblasts and periodontal ligament cells, can self-renew and exhibit multipotential capacity. Coralline hydroxyapatite (CHA) has good biocompatibility and conductivity required for bone tissue engineering. Here, we reported that BMP9 could enhance the osteogenic differentiation of rDFCs in cell culture. Moreover, our results suggested that BMP9 acted through the Smad1/5/8 signaling pathway. We also produced a novel scaffold that encompasses bio-degradable CHA seeded with recombinant adenoviruses expressing BMP9-transfected rDFCs (Ad-BMP9-transfected rDFCs). With this implant, we achieved more alveolar bone regeneration in the alveolar bone defect compared to blank group, CHA group and rDFCs group. Our results provided a novel bio-implants composed of Ad-BMP9-transfected rDFCs and CHA scaffolds and its mechanism is regarding the activation of Smad1/5/8 signaling pathway in BMP9-induced rDFCs osteogenesis.


Assuntos
Osso e Ossos/lesões , Cerâmica/farmacologia , Saco Dentário/citologia , Fator 2 de Diferenciação de Crescimento/genética , Hidroxiapatitas/farmacologia , Osteogênese/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Animais , Regeneração Óssea/efeitos dos fármacos , Diferenciação Celular , Linhagem Celular , Saco Dentário/metabolismo , Dependovirus/genética , Fator 2 de Diferenciação de Crescimento/farmacologia , Ratos , Transdução de Sinais , Proteínas Smad/metabolismo , Transplante de Células-Tronco , Células-Tronco/citologia , Células-Tronco/metabolismo , Engenharia Tecidual , Alicerces Teciduais/química
7.
J Biomed Nanotechnol ; 12(4): 602-18, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27301188

RESUMO

Enhancing the biocompatibility and osteogenic activity of nano-apatite for applications in bone graft substitutes and bone tissue engineering have been the current challenge in regeneration of lost bone. Inspired by mussels, here we have developed facile biomimetic approaches for preparation of two types of peptide-conjugated apatite nanocompsoties assisted by polydopamine (pDA). We exploited polydopamine chemistry for the modification of nano-apatite crystals: polydopamine coated apatite (HA-c-pDA) and polydopamine template-mediated apatite (HA-t-pDA), on which bone forming peptide was subsequently immobilized under weakly basic conditions to obtain peptide-conjugated apatite nanocomposites (HA-c-pep and HA-t-pep, respectively). TEM images revealed that HA-c-pDA displayed typically rod-like morphology, while HA-t-pDA was sponge-like structure where pDA sheets were decorated by needle-like apatite crystals with low degree of crystallinity. In the cell culture experiments, HA-t-pep nanocomposite exhibited higher cell proliferation, spreading, and alkaline phosphatase activity as well as calcium nodule-formation, compared with pristine nano-HA and HA-c-pep nanocomposite. We then implanted the peptide-decorated apatite into rabbit calvarial defects and analyzed bone formation after 2 months. The data revealed that HA-t-pep group exhibited remarkably enhanced bioactivity and bone formation in vivo. Based on these results, our biomimetic approach could be a promising tool to develop peptide-conjugated apatites for bone regeneration. Meanwhile, the excellent biocompatibility and high osteogenesis of the peptide-conjugated apatite nanocomposite might confer its great potentials in bone repair, bone augmentation, as well as coating of biomedical implants.


Assuntos
Proteína Morfogenética Óssea 7/administração & dosagem , Durapatita/química , Indóis/química , Nanocompostos/química , Osteoblastos/citologia , Osteogênese/fisiologia , Polímeros/química , Células 3T3 , Animais , Proteína Morfogenética Óssea 7/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/síntese química , Materiais Revestidos Biocompatíveis/toxicidade , Durapatita/toxicidade , Camundongos , Nanocompostos/toxicidade , Nanocompostos/ultraestrutura , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Osteogênese/efeitos dos fármacos
8.
Genes Dis ; 3(4): 263-276, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28491933

RESUMO

Tooth is a complex hard tissue organ and consists of multiple cell types that are regulated by important signaling pathways such as Wnt and BMP signaling. Serious injuries and/or loss of tooth or periodontal tissues may significantly impact aesthetic appearance, essential oral functions and the quality of life. Regenerative dentistry holds great promise in treating oral/dental disorders. The past decade has witnessed a rapid expansion of our understanding of the biological features of dental stem cells, along with the signaling mechanisms governing stem cell self-renewal and differentiation. In this review, we first summarize the biological characteristics of seven types of dental stem cells, including dental pulp stem cells, stem cells from apical papilla, stem cells from human exfoliated deciduous teeth, dental follicle precursor cells, periodontal ligament stem cells, alveolar bone-derived mesenchymal stem cells (MSCs), and MSCs from gingiva. We then focus on how these stem cells are regulated by bone morphogenetic protein (BMP) and/or Wnt signaling by examining the interplays between these pathways. Lastly, we analyze the current status of dental tissue engineering strategies that utilize oral/dental stem cells by harnessing the interplays between BMP and Wnt pathways. We also highlight the challenges that must be addressed before the dental stem cells may reach any clinical applications. Thus, we can expect to witness significant progresses to be made in regenerative dentistry in the coming decade.

9.
Biomaterials ; 39: 145-54, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25468367

RESUMO

Dental pulp/dentin regeneration using dental stem cells combined with odontogenic factors may offer great promise to treat and/or prevent premature tooth loss. Here, we investigate if BMP9 and Wnt/ß-catenin act synergistically on odontogenic differentiation. Using the immortalized SCAPs (iSCAPs) isolated from mouse apical papilla tissue, we demonstrate that Wnt3A effectively induces early osteogenic marker alkaline phosphatase (ALP) in iSCAPs, which is reduced by ß-catenin knockdown. While Wnt3A and BMP9 enhance each other's ability to induce ALP activity in iSCAPs, silencing ß-catenin significantly diminishes BMP9-induced osteo/odontogenic differentiation. Furthermore, silencing ß-catenin reduces BMP9-induced expression of osteocalcin and osteopontin and in vitro matrix mineralization of iSCAPs. In vivo stem cell implantation assay reveals that while BMP9-transduced iSCAPs induce robust ectopic bone formation, iSCAPs stimulated with both BMP9 and Wnt3A exhibit more mature and highly mineralized trabecular bone formation. However, knockdown of ß-catenin in iSCAPs significantly diminishes BMP9 or BMP9/Wnt3A-induced ectopic bone formation in vivo. Thus, our results strongly suggest that ß-catenin may play an important role in BMP9-induced osteo/ondontogenic signaling and that BMP9 and Wnt3A may act synergistically to induce osteo/odontoblastic differentiation of iSCAPs. It's conceivable that BMP9 and/or Wnt3A may be explored as efficacious biofactors for odontogenic regeneration and tooth engineering.


Assuntos
Papila Dentária/citologia , Fatores de Diferenciação de Crescimento/farmacologia , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Animais , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Sinergismo Farmacológico , Feminino , Gossipol/análogos & derivados , Gossipol/farmacologia , Células HeLa , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Odontogênese , Ratos , Transdução de Sinais/efeitos dos fármacos , Proteínas Wnt
10.
Stem Cells Dev ; 23(12): 1405-16, 2014 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-24517722

RESUMO

Dental pulp/dentin regeneration using dental stem cells combined with odontogenic factors may offer great promise to treat and/or prevent premature tooth loss. We previously demonstrated that bone morphogenetic protein 9 (BMP9) is one of the most potent factors in inducing bone formation. Here, we investigate whether BMP9 can effectively induce odontogenic differentiation of the stem cells from mouse apical papilla (SCAPs). Using a reversible immortalization system expressing SV40 T flanked with Cre/loxP sites, we demonstrate that the SCAPs can be immortalized, resulting in immortalized SCAPs (iSCAPs) that express mesenchymal stem cell markers. BMP9 upregulates Runx2, Sox9, and PPARγ2 and odontoblastic markers, and induces alkaline phosphatase activity and matrix mineralization in the iSCAPs. Cre-mediated removal of SV40 T antigen decreases iSCAP proliferation. The in vivo stem cell implantation studies indicate that iSCAPs can differentiate into bone, cartilage, and, to lesser extent, adipocytes upon BMP9 stimulation. Our results demonstrate that the conditionally iSCAPs not only maintain long-term cell proliferation but also retain the ability to differentiate into multiple lineages, including osteo/odontoblastic differentiation. Thus, the reversibly iSCAPs may serve as an important tool to study SCAP biology and SCAP translational use in tooth engineering. Further, BMP9 may be explored as a novel and efficacious factor for odontogenic regeneration.


Assuntos
Diferenciação Celular/genética , Papila Dentária/crescimento & desenvolvimento , Fator 2 de Diferenciação de Crescimento/genética , Odontogênese , Animais , Proliferação de Células/genética , Papila Dentária/citologia , Regulação da Expressão Gênica no Desenvolvimento , Fator 2 de Diferenciação de Crescimento/biossíntese , Camundongos , Odontoblastos/metabolismo , Regeneração , Células-Tronco/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA