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1.
Int J Mol Sci ; 22(4)2021 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-33672551

RESUMO

Paired box protein 5 (Pax5) is a crucial transcription factor responsible for B-cell lineage specification and commitment. In this study, we identified a negative role of Pax5 in osteoclastogenesis. The expression of Pax5 was time-dependently downregulated by receptor activator of nuclear factor kappa B (RANK) ligand (RANKL) stimulation in osteoclastogenesis. Osteoclast (OC) differentiation and bone resorption were inhibited (68.9% and 48% reductions, respectively) by forced expression of Pax5 in OC lineage cells. Pax5 led to the induction of antiosteoclastogenic factors through downregulation of B lymphocyte-induced maturation protein 1 (Blimp1). To examine the negative role of Pax5 in vivo, we generated Pax5 transgenic (Pax5Tg) mice expressing the human Pax5 transgene under the control of the tartrate-resistant acid phosphatase (TRAP) promoter, which is expressed mainly in OC lineage cells. OC differentiation and bone resorption were inhibited (54.2-76.9% and 24.0-26.2% reductions, respectively) in Pax5Tg mice, thereby contributing to the osteopetrotic-like bone phenotype characterized by increased bone mineral density (13.0-13.6% higher), trabecular bone volume fraction (32.5-38.1% higher), trabecular thickness (8.4-9.0% higher), and trabecular number (25.5-26.7% higher) and decreased trabecular spacing (9.3-10.4% lower) compared to wild-type control mice. Furthermore, the number of OCs was decreased (48.8-65.3% reduction) in Pax5Tg mice. These findings indicate that Pax5 plays a negative role in OC lineage specification and commitment through Blimp1 downregulation. Thus, our data suggest that the Pax5-Blimp1 axis is crucial for the regulation of RANKL-induced osteoclastogenesis.


Assuntos
Regulação para Baixo/genética , Osteogênese , Fator de Transcrição PAX5/metabolismo , Fator 1 de Ligação ao Domínio I Regulador Positivo/genética , Animais , Osso e Ossos/patologia , Linhagem da Célula , Regulação para Baixo/efeitos dos fármacos , Expressão Gênica , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mutação/genética , Osteoclastos/metabolismo , Osteogênese/genética , Osteopetrose/genética , Osteopetrose/patologia , Fator de Transcrição PAX5/genética , Fenótipo , Ligante RANK/farmacologia , Células RAW 264.7 , Transgenes
2.
Ecotoxicol Environ Saf ; 214: 112080, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33677380

RESUMO

Resveratrol (RES) is a natural polyphenolic compound with a broad range of physiological and pharmacological properties. Previous studies have shown that RES also plays an important role in protecting and promoting early bone metabolism and differentiation. The accumulation of cadmium (Cd), one of the world's most poisonous substances, can inhibit skeletal growth and bone maturation, thus causing osteoporosis. However, whether RES can prevent the Cd-induced inhibition of osteogenic differentiation remains unknown. In this study, we found that RES promoted the early maturity of osteoblastic MC3T3-E1 cells, as demonstrated by the significantly increased mRNA and protein expression of a range of differentiation markers, including alkaline phosphatase (ALP), collagen 1 (COL1), bone morphogenetic protein-2 (BMP-2), and runt-related transcription factor 2 (RUNX2). In contrast, we found that cadmium chloride (CdCl2) inhibited the viability and osteogenic maturity of MC3T3-E1 cells. We also demonstrated that RES pretreatment for 30 min provided significant protection against Cd-induced apoptosis and attenuated the inhibition of osteogenic differentiation induced by Cd by modulating ERK1/2 and JNK signaling. In conclusion, our results indicate that RES is a potential femoral protectant that not only enhance the viability and early differentiation of osteoblasts, but also protect osteoblasts from cadmium damage.


Assuntos
Cádmio/toxicidade , Substâncias Protetoras/farmacologia , Resveratrol/farmacologia , Fosfatase Alcalina/metabolismo , Animais , Proteína Morfogenética Óssea 2 , Cádmio/metabolismo , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Subunidade alfa 1 de Fator de Ligação ao Core , Sistema de Sinalização das MAP Quinases , Proteína Quinase 3 Ativada por Mitógeno , Osteoblastos/citologia , Osteogênese/genética
3.
Int J Nanomedicine ; 16: 725-740, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33542627

RESUMO

Purpose: As a dental material, polyetheretherketone (PEEK) is bioinert that does not induce cellular response and bone/gingival tissues regeneration. This study was to develop bioactive coating on PEEK and investigate the effects of coating on cellular response. Materials and Methods: Tantalum pentoxide (TP) coating was fabricated on PEEK surface by vacuum evaporation and responses of rat bone marrow mesenchymal stem (RBMS) cells/human gingival epithelial (HGE) were studied. Results: A dense coating (around 400 nm in thickness) of TP was closely combined with PEEK (PKTP). Moreover, the coating was non-crystalline TP, which contained many small humps (around 10 nm in size), exhibiting a nanostructured surface. In addition, the roughness, hydrophilicity, surface energy, and protein adsorption of PKTP were remarkably higher than that of PEEK. Furthermore, the responses (adhesion, proliferation, and osteogenic gene expression) of RBMS cells, and responses (adhesion and proliferation) of HGE cells to PKTP were remarkably improved in comparison with PEEK. It could be suggested that the nanostructured coating of TP on PEEK played crucial roles in inducing the responses of RBMS/HGE cells. Conclusion: PKTP with elevated surface performances and outstanding cytocompatibility might have enormous potential for dental implant application.


Assuntos
Células Epiteliais/citologia , Gengiva/citologia , Cetonas/farmacologia , Células-Tronco Mesenquimais/citologia , Nanoestruturas/química , Óxidos/farmacologia , Polietilenoglicóis/farmacologia , Tantálio/farmacologia , Adsorção , Fosfatase Alcalina/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/ultraestrutura , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/enzimologia , Nanoestruturas/ultraestrutura , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Ratos Sprague-Dawley , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de Superfície , Difração de Raios X
4.
Nat Commun ; 12(1): 496, 2021 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-33479228

RESUMO

Chronic inflammation during many diseases is associated with bone loss. While interferons (IFNs) are often inhibitory to osteoclast formation, the complex role that IFN and interferon-stimulated genes (ISGs) play in osteoimmunology during inflammatory diseases is still poorly understood. We show that mice deficient in IFN signaling components including IFN alpha and beta receptor 1 (IFNAR1), interferon regulatory factor 1 (IRF1), IRF9, and STAT1 each have reduced bone density and increased osteoclastogenesis compared to wild type mice. The IFN-inducible guanylate-binding proteins (GBPs) on mouse chromosome 3 (GBP1, GBP2, GBP3, GBP5, GBP7) are required to negatively regulate age-associated bone loss and osteoclastogenesis. Mechanistically, GBP2 and GBP5 both negatively regulate in vitro osteoclast differentiation, and loss of GBP5, but not GBP2, results in greater age-associated bone loss in mice. Moreover, mice deficient in GBP5 or chromosome 3 GBPs have greater LPS-mediated inflammatory bone loss compared to wild type mice. Overall, we find that GBP5 contributes to restricting age-associated and inflammation-induced bone loss by negatively regulating osteoclastogenesis.


Assuntos
Reabsorção Óssea/metabolismo , Proteínas de Ligação ao GTP/metabolismo , Interferons/metabolismo , Osteoclastos/metabolismo , Osteogênese/fisiologia , Fatores Etários , Animais , Reabsorção Óssea/genética , Diferenciação Celular/genética , Fusão Celular , Células Cultivadas , Proteínas de Ligação ao GTP/genética , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Osteoclastos/citologia , Osteogênese/genética , Transdução de Sinais/genética
5.
Int J Mol Sci ; 22(2)2021 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-33466904

RESUMO

Reconstruction of segmental bone defects by autologous bone grafting is still the standard of care but presents challenges including anatomical availability and potential donor site morbidity. The process of 3D bioprinting, the application of 3D printing for direct fabrication of living tissue, opens new possibilities for highly personalized tissue implants, making it an appealing alternative to autologous bone grafts. One of the most crucial hurdles for the clinical application of 3D bioprinting is the choice of a suitable cell source, which should be minimally invasive, with high osteogenic potential, with fast, easy expansion. In this study, mesenchymal progenitor cells were isolated from clinically relevant human bone biopsy sites (explant cultures from alveolar bone, iliac crest and fibula; bone marrow aspirates; and periosteal bone shaving from the mastoid) and 3D bioprinted using projection-based stereolithography. Printed constructs were cultivated for 28 days and analyzed regarding their osteogenic potential by assessing viability, mineralization, and gene expression. While viability levels of all cell sources were comparable over the course of the cultivation, cells obtained by periosteal bone shaving showed higher mineralization of the print matrix, with gene expression data suggesting advanced osteogenic differentiation. These results indicate that periosteum-derived cells represent a highly promising cell source for translational bioprinting of bone tissue given their superior osteogenic potential as well as their minimally invasive obtainability.


Assuntos
Células da Medula Óssea/metabolismo , Transplante Ósseo/métodos , Osso e Ossos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Biossíntese de Proteínas , Engenharia Tecidual/métodos , Adulto , Bioimpressão/métodos , Células da Medula Óssea/citologia , Osso e Ossos/citologia , Diferenciação Celular/genética , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/citologia , Osteogênese/genética , Impressão Tridimensional , Tecidos Suporte , Transplante Autólogo
6.
Nat Commun ; 12(1): 421, 2021 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-33462210

RESUMO

Multiple myeloma (MM) progression and myeloma-associated bone disease (MBD) are highly dependent on bone marrow mesenchymal stromal cells (MSCs). MM-MSCs exhibit abnormal transcriptomes, suggesting the involvement of epigenetic mechanisms governing their tumor-promoting functions and prolonged osteoblast suppression. Here, we identify widespread DNA methylation alterations of bone marrow-isolated MSCs from distinct MM stages, particularly in Homeobox genes involved in osteogenic differentiation that associate with their aberrant expression. Moreover, these DNA methylation changes are recapitulated in vitro by exposing MSCs from healthy individuals to MM cells. Pharmacological targeting of DNMTs and G9a with dual inhibitor CM-272 reverts the expression of hypermethylated osteogenic regulators and promotes osteoblast differentiation of myeloma MSCs. Most importantly, CM-272 treatment prevents tumor-associated bone loss and reduces tumor burden in a murine myeloma model. Our results demonstrate that epigenetic aberrancies mediate the impairment of bone formation in MM, and its targeting by CM-272 is able to reverse MBD.


Assuntos
Antineoplásicos/farmacologia , Doenças Ósseas/tratamento farmacológico , Metilação de DNA/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Mieloma Múltiplo/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Antineoplásicos/uso terapêutico , Doenças Ósseas/diagnóstico , Doenças Ósseas/genética , Doenças Ósseas/patologia , Medula Óssea/patologia , DNA (Citosina-5-)-Metiltransferases/antagonistas & inibidores , DNA (Citosina-5-)-Metiltransferases/metabolismo , Inibidores Enzimáticos/uso terapêutico , Epigênese Genética/efeitos dos fármacos , Feminino , Fêmur/diagnóstico por imagem , Fêmur/patologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Antígenos de Histocompatibilidade/metabolismo , Histona-Lisina N-Metiltransferase/antagonistas & inibidores , Histona-Lisina N-Metiltransferase/metabolismo , Humanos , Masculino , Células-Tronco Mesenquimais/patologia , Camundongos , Pessoa de Meia-Idade , Mieloma Múltiplo/complicações , Mieloma Múltiplo/genética , Mieloma Múltiplo/patologia , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Ensaios Antitumorais Modelo de Xenoenxerto
7.
Methods Mol Biol ; 2248: 109-119, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33185871

RESUMO

TNFα/TNFR signaling plays a critical role in the pathogenesis of various inflammatory and autoimmune diseases, and anti-TNFα therapies have been accepted as the effective approaches for treating several autoimmune diseases. Progranulin (PGRN), a multi-faced growth factor-like molecule, directly binds to TNFR1 and TNFR2, particularly to the latter with higher affinity than TNFα. PGRN derivative Atsttrin is composed of three TNFR-binding domain of PGRN and exhibits even better therapeutic effects than PGRN in several inflammatory disease models, including collagen-induced arthritis. Herein we describe the detailed methodology of using (1) ELISA-based solid phase protein-protein interaction assay to demonstrate the direct binding of Atsttrin to TNFR2 and its inhibition of TNFα/TNFR2 interaction; and (2) tartrate-resistant acid phosphatase (TRAP) staining of in vitro osteoclastogenesis to reveal the cell-based anti-TNFα activity of Atsttrin. Using the protocol described here, the investigators should be able to reproducibly detect the physical inhibition of TNFα binding to TNFR and the functional inhibition of TNFα activity by Atsttrin and various kinds of TNF inhibitors.


Assuntos
Progranulinas/química , Receptores Tipo II do Fator de Necrose Tumoral/química , Fator de Necrose Tumoral alfa/química , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Progranulinas/metabolismo , Ligação Proteica , Mapeamento de Interação de Proteínas/métodos , Receptores Tipo II do Fator de Necrose Tumoral/metabolismo , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/antagonistas & inibidores
8.
Methods Mol Biol ; 2230: 3-16, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33197005

RESUMO

Development of cartilage and bone, the core components of the mouse skeletal system, depends on coordinated proliferation and differentiation of skeletogenic cells, including chondrocytes and osteoblasts. These cells differentiate from common progenitor cells originating in the mesoderm and neural crest. Multiple signaling pathways and transcription factors tightly regulate differentiation and proliferation of skeletal cells. In this chapter, we overview the process of mouse skeletal development and discuss major regulators of skeletal cells at each developmental stage.


Assuntos
Desenvolvimento Ósseo/genética , Mesoderma/crescimento & desenvolvimento , Osteogênese/genética , Células-Tronco/citologia , Animais , Cartilagem/crescimento & desenvolvimento , Diferenciação Celular/genética , Condrócitos/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Camundongos , Crista Neural/crescimento & desenvolvimento , Osteoblastos/metabolismo , Transdução de Sinais/genética
9.
Methods Mol Biol ; 2230: 397-413, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33197028

RESUMO

This chapter describes the methods of isolation of mouse periosteal progenitor cells. There are three basic methods utilized. The bone grafting method was developed utilizing the fracture healing process to expand the progenitor populations. Bone capping methods requires enzymatic digestion and purification of cells from the native periosteum, while the Egression/Explant method requires the least manipulation with placement of cortical bone fragments with attached periosteum in a culture dish. Various cell surface antibodies have been employed over the years to characterize periosteum derived progenitor cells, but the most consistent minimal criteria was recommended by the International Society for Cellular Therapy. Confirmation of the multipotent status of these isolated cells can be achieved by differentiation into the three basic mesodermal lineages in vitro.


Assuntos
Transplante Ósseo/métodos , Técnicas de Cultura de Células/métodos , Periósteo/crescimento & desenvolvimento , Células-Tronco/citologia , Animais , Diferenciação Celular/genética , Proliferação de Células/genética , Células-Tronco Mesenquimais/citologia , Camundongos , Osteogênese/genética , Periósteo/citologia
10.
Methods Mol Biol ; 2230: 425-436, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33197030

RESUMO

This chapter describes the isolation and culture of neonatal mouse calvarial osteoblasts. This primary cell population is obtained by sequential enzymatic digestion of the calvarial bone matrix and is capable of differentiating in vitro into mature osteoblasts that deposit a collagen extracellular matrix and form mineralized bone nodules. Maturation of the cultures can be monitored by gene expression analyses and staining for the presence of alkaline phosphatase or matrix mineralization. This culture system, therefore, provides a powerful model in which to test how various experimental conditions, such as the manipulation of gene expression, may affect osteoblast maturation and/or function.


Assuntos
Calcificação Fisiológica/genética , Técnicas de Cultura de Células/métodos , Separação Celular/métodos , Osteogênese/genética , Animais , Animais Recém-Nascidos , Matriz Óssea/crescimento & desenvolvimento , Matriz Óssea/metabolismo , Diferenciação Celular/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Camundongos , Osteoblastos/metabolismo
11.
Methods Mol Biol ; 2230: 457-465, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33197033

RESUMO

The osteoclast is the unique type of cell that resorbs bone in vivo and it is required for normal skeletal development and postnatal homeostasis. Osteoclast deficiency impairs skeletal development during embryogenesis and results in osteopetrosis and impaired tooth eruption. In contrast, excessive osteoclast formation in adults results in bone loss in a number of conditions, including osteoporosis, rheumatoid arthritis, and metastatic bone disease. Osteoclasts are derived from monocytes/macrophages; they can be generated in vitro by treatment of these precursor cells with macrophage colony stimulating factor (M-CSF) and receptor activator of NF-κB ligand (RANKL). This chapter describes procedures for generating osteoclasts from mouse bone marrow cells in vitro using M-CSF and RANKL and assessing their ability to form resorption lacunae on thin bone slices.


Assuntos
Células da Medula Óssea/metabolismo , Técnicas de Cultura de Células/métodos , Osteoclastos/metabolismo , Osteogênese/genética , Animais , Células da Medula Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Camundongos , Osteoclastos/efeitos dos fármacos , Ligante RANK/farmacologia
12.
J Periodontal Res ; 56(2): 379-387, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33368310

RESUMO

BACKGROUND AND OBJECTIVE: Periodontal ligament stem cells (PDLSCs) have potential for osteogenic differentiation and show a great foreground in treating bone diseases. Histone three lysine 27 (H3K27) demethylase lysine demethylase 6A (KDM6A) is a critical epigenetic modifier and plays an important role in regulating osteogenic differentiation. Multiple microRNAs have been found to play important roles in osteogenesis. The aim of this study was to explore the mechanisms underlying the roles of miR-153-3p and KDM6A in PDLSC osteogenesis. METHODS: The levels of the osteogenic markers alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), and osteopontin (OPN) were measured by western blotting. Osteoblast activity and mineral deposition were detected by ALP and Alizarin red S (ARS) staining. The levels of miR-153-3p and KDM6A were measured by quantitative real-time PCR (qRT-PCR). A luciferase reporter assay was used to confirm the interaction between KDM6A and miR-153-3p. Gain-of-function and loss-of-function assays were performed to identify the roles of miR-153-3p and KDM6A in the osteogenic differentiation of PDLSCs. RESULTS: In osteogenic PDLSCs, the expression of KDM6A, ALP, Runx2, and OPN was upregulated, whereas that of miR-153-3p was downregulated. miR-153-3p downregulation or KDM6A overexpression promoted the osteogenic differentiation of PDLSCs, as demonstrated by increases in ALP activity, matrix mineralization, and ALP, Runx2, and OPN expression. KDM6A was confirmed to be a target of miR-153-3p, and KDM6A overexpression reversed the inhibitory effect of miR-153-3p mimic on PDLSC osteogenesis. KDM6A promoted ALP, Runx2, and OPN expression through the demethylation of H3K27me3 on the promoter regions of these genes. CONCLUSION: miR-153-3p inhibited PDLSC osteogenesis by targeting KDM6A and inhibiting ALP, Runx2, and OPN transcription. These findings provide latent hope for PDLSCs application in periodontal therapy.


Assuntos
MicroRNAs , Ligamento Periodontal , Diferenciação Celular , Células Cultivadas , Desmetilação , Histona Desmetilases/genética , Histonas , MicroRNAs/genética , MicroRNAs/metabolismo , Osteogênese/genética , Ligamento Periodontal/metabolismo , Células-Tronco/metabolismo
13.
Gene ; 764: 145106, 2021 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-32889059

RESUMO

BACKGROUND: Circular RNAs (circRNAs) are a new class of non-coding RNA with a stable structure formed by special loop splicing. Research increasingly suggests that circRNAs play a vital role in the pathogenesis and progression of various diseases. However, the roles of circRNAs in osteoblast differentiation under microgravity remain largely unknown. Here, we investigated the roles and mechanobiological response of circRNAs in osteoblasts under simulated microgravity. METHODS: Differential circRNA and mRNA expression profiles of MC3T3-E1 cells during exposure to microgravity were screened by RNA transcriptome sequencing technology (RNA-seq). The selected RNAs were validated using quantitative real-time polymerase chain reaction (qRT-PCR). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were applied for gene function analyses. RESULTS: A total of 427 circRNAs and 1912 mRNAs were differentially expressed along with osteogenic differentiation in the simulated microgravity group (SMG) compared to the control group (CON). Of these, 232 circRNAs and 991 mRNAs were upregulated, whereas 95 circRNAs and 921 mRNAs were downregulated (fold change ≥ 2, p < 0.05). The results showed that the parental genes of circRNAs and mRNAs were mainly enriched in anatomical structure morphogenesis, anchoring junction and protein binding. KEGG analysis results showed that the differentially expressed mRNAs were enriched in the regulation of the actin cytoskeleton, focal adhesion, and Ras signalling pathway. Subsequently, 9 core regulatory genes, including 6 mRNAs and 3 circRNAs, were identified based on their possible function in osteoblast differentiation. Based on this analysis, circ_014154 was selected as the target circRNA, which likely plays important roles in osteogenic differentiation processes under microgravity. The circRNA-miRNA-mRNA network showed that circRNAs might act as miRNA sponges to regulate osteoblast differentiation. CONCLUSION: By presenting a better understanding of the molecular mechanisms of genes and circRNAs in simulated microgravity, the present study will provide a novel view of circRNAs in the regulation of osteogenic differentiation and bone formation.


Assuntos
Diferenciação Celular/genética , Osteoblastos/fisiologia , Osteogênese/genética , RNA Circular/metabolismo , Simulação de Ausência de Peso/efeitos adversos , Animais , Linhagem Celular , Biologia Computacional , Conjuntos de Dados como Assunto , Redes Reguladoras de Genes/fisiologia , Camundongos , MicroRNAs/metabolismo , RNA Mensageiro/metabolismo
14.
Int J Mol Sci ; 21(24)2020 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-33322825

RESUMO

This article provides a brief review of the pathophysiology of osteoarthritis and the ontogeny of chondrocytes and details how physical exercise improves the health of osteoarthritic joints and enhances the potential of autologous chondrocyte implants, matrix-induced autologous chondrocyte implants, and mesenchymal stem cell implants for the successful treatment of damaged articular cartilage and subchondral bone. In response to exercise, articular chondrocytes increase their production of glycosaminoglycans, bone morphogenic proteins, and anti-inflammatory cytokines and decrease their production of proinflammatory cytokines and matrix-degrading metalloproteinases. These changes are associated with improvements in cartilage organization and reductions in cartilage degeneration. Studies in humans indicate that exercise enhances joint recruitment of bone marrow-derived mesenchymal stem cells and upregulates their expression of osteogenic and chondrogenic genes, osteogenic microRNAs, and osteogenic growth factors. Rodent experiments demonstrate that exercise enhances the osteogenic potential of bone marrow-derived mesenchymal stem cells while diminishing their adipogenic potential, and that exercise done after stem cell implantation may benefit stem cell transplant viability. Physical exercise also exerts a beneficial effect on the skeletal system by decreasing immune cell production of osteoclastogenic cytokines interleukin-1ß, tumor necrosis factor-α, and interferon-γ, while increasing their production of antiosteoclastogenic cytokines interleukin-10 and transforming growth factor-ß. In conclusion, physical exercise done both by bone marrow-derived mesenchymal stem cell donors and recipients and by autologous chondrocyte donor recipients may improve the outcome of osteochondral regeneration therapy and improve skeletal health by downregulating osteoclastogenic cytokine production and upregulating antiosteoclastogenic cytokine production by circulating immune cells.


Assuntos
Condrócitos/metabolismo , Exercício Físico/fisiologia , Células-Tronco Mesenquimais/metabolismo , Osteoartrite/fisiopatologia , Osteogênese , Condicionamento Físico Animal/fisiologia , Regeneração/genética , Animais , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/enzimologia , Cartilagem Articular/patologia , Citocinas/metabolismo , Glicosaminoglicanos/metabolismo , Humanos , Metaloproteases/metabolismo , Osteoartrite/enzimologia , Osteoartrite/imunologia , Osteoartrite/terapia , Osteogênese/genética , Osteogênese/imunologia , Osteogênese/fisiologia , Regeneração/imunologia , Regeneração/fisiologia , Transplante de Células-Tronco
15.
J Biomed Nanotechnol ; 16(6): 899-909, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33187585

RESUMO

A well-studied subject of epigenetics, the histone methylation located at lysine and arginine is overseen via methyltransferases and demethylases. Lysine-specific demethylase 4A (KDM4A) comprises a lysine demethylase and possesses specificity for H3K9me3 and H3K36me3, which is capable of being used in order to activate histone transcription. Our team examined the expression of KDM4A within Sprague Dawley (SD) rats and further investigated the mechanism via which this phenomena regulates osteogenic variation within the present study. The overexpression of KDM4A facilitated the process of osteoblast differentiation in bone mesenchymal stem cells (BMSC), while the knocking down differentiation via osteoblast was restrained via the suppression of the expression of Runx2, Osterix, alkaline phosphatase (ALP), and osteocalcin (OCN). Knocking down KDM4A lowered levels of the promoter expression of Runx2, osterix, and OCN, and raised levels of H3K27me3 expression. The results demonstrated that KDM4A possesses a crucial role within the differentiation of osteoblasts and furthermore regulates the expression of Runx2, Osterix, and OCN via H3K9me3. The present research may provide new insights into the treatment of bone healing.


Assuntos
Histona Desmetilases , Lisina , Osteogênese , Animais , Diferenciação Celular , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Histona Desmetilases/fisiologia , Osteoblastos/metabolismo , Osteocalcina/genética , Osteogênese/genética , Ratos , Ratos Sprague-Dawley , Receptores de Ocitocina , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
16.
Int J Nanomedicine ; 15: 8465-8478, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33149587

RESUMO

Introduction: Decellularized matrix from porcine small intestinal submucosa (SIS) endows scaffolds with an ECM-like surface, which enhances stem cell self-renewal, proliferation, and differentiation. Mesoporous bioactive glass (MBG) is extensively recognized as an excellent bio-ceramic for fabricating bone grafts. Materials and Methods: In the current study, SIS was doped on an MBG scaffold (MBG/SIS) using polyurethane foam templating and polydopamine chemistry method. To mimic the bony environment of a natural bone matrix, an ECM-inspired delivery system was constructed by coupling the BMP2-related peptide P28 to a heparinized MBG/SIS scaffold (MBG/SIS-H-P28). The release of P28 from MBG/SIS-H-P28 and its effects on the proliferation, viability, and osteogenic differentiation of bone marrow stromal stem cells were investigated in vitro and in vivo. Results: Our research indicated that the novel tissue-derived ECM scaffold MBG/SIS has a hierarchical and interconnected porous architecture, and superior biomechanical properties. MBG/SIS-H-P28 released P28 in a controlled manner, with the long-term release time of 40 d. The results of in vitro experiments showed improvements in cell proliferation, cell viability, alkaline phosphatase activity, and mRNA expression levels of osteogenesis-related genes (Runx-2, OCN, OPN, and ALP) compared to those of MBG/SIS or MBG/SIS-P28 and MBG/SIS-H-P28. The in vivo results demonstrated that MBG/SIS-H-P28 scaffolds evidently increased bone formation in rat calvarial critical-sized defect compared to that in controls. Conclusion: MBG/SIS-H-P28 scaffolds show potential as ideal platforms for delivery of P28 and for providing a bony environment for bone regeneration.


Assuntos
Ácido Aspártico/química , Materiais Biocompatíveis/farmacologia , Proteína Morfogenética Óssea 2/farmacologia , Osso e Ossos/efeitos dos fármacos , Cerâmica/farmacologia , Matriz Extracelular/metabolismo , Osteoblastos/efeitos dos fármacos , Peptídeos/farmacologia , Fator de Crescimento Transformador beta/farmacologia , Animais , Regeneração Óssea/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Cinética , Masculino , Camundongos , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Porosidade , Ratos Sprague-Dawley , Proteínas Recombinantes/farmacologia , Suínos , Tecidos Suporte/química
17.
Shanghai Kou Qiang Yi Xue ; 29(4): 343-349, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-33089279

RESUMO

PURPOSE: To investigate the biological characteristics of human periodontal stem cells (hPDLSCs) modified with platelet derived growth factor BB(PDGFBB) gene, and to explore its influence on proliferation, migration and osteogenic induction of hPDLSCs. METHODS: hPDLSCs were isolated and amplified, and immunofluorescence staining was performed to identify cell surface markers and osteogenic differentiation ability. hPDLSCs were transfected with PDGFBB gene by lentivirus vector, and the effects on cell proliferation and migration were detected by CCK-8 and scratch test after transfection. Real-time PCR was performed to analyze the mRNA expression levels of osteogenic and angiogenic genes in hPDLSCs cells transfected with PDGFBB gene. Statistical analysis was performed using SPSS 22.0 software package. RESULTS: hPDLSCs were successfully obtained by tissue mass culture and finite dilution method. Compared with the blank virus group and non-transfected group, the proliferation and migration ability of the cells in the transfection group were significantly increased, and the mRNA expression levels of OPN, COL-1 and VEGF were significantly up-regulated(P<0.05). CONCLUSIONS: Lentiviral vector can transfer PDGFBB gene into hPDLSCs in vitro and obtain continuous and stable expression. PDGFBB can promote proliferation and migration of hPDLSCs cells and up-regulate expression of osteogenic and angiogenic genes.


Assuntos
Becaplermina , Ligamento Periodontal , Diferenciação Celular , Humanos , Osteogênese/genética , Células-Tronco
18.
Shanghai Kou Qiang Yi Xue ; 29(4): 365-369, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-33089283

RESUMO

PURSPOSE: To investigate the role of P2X7 receptor in osteogenic differentiation of human periodontal ligament stem cells. METHODS: Human periodontal ligament stem cells obtained from primary culture were divided into 4 groups: control group, adenosine triphosphate group, osteogenic induction group, adenosine triphosphate + osteogenic induction group. The differences of RUNX2, OCN gene expression and P2X7 receptor mRNA expression between the four groups were compared. Statistical analysis was performed with SPSS 16.0 software package. RESULTS: One week after osteogenic formation and two weeks after osteogenic formation, the expression of RUNX2 and OCN mRNA in the adenosine triphosphate + osteogenic induction group was significantly higher than that in the osteogenic induction group (P<0.05). The expression of RUNX2 and OCN mRNA in the 1 week after adenosine triphosphate + osteogenic induction fluid was significantly higher than that 2 weeks after osteogenic formation, and the difference was statistically significant (P<0.05). The expression of P2X7 receptor mRNA in the adenosine triphosphate group and the adenosine triphosphate + osteogenic induction group was significantly higher than that in the control group and the osteogenic induction group 1 week after osteogenesis and 2 weeks after osteogenesis (P<0.05). The expression of P2X7 receptor mRNA in the adenosine triphosphate group was significantly higher than that in the adenosine triphosphate + osteogenic induction group 2 weeks after osteogenesis(P<0.05). The expression of P2X7 receptor mRNA was significantly higher than that of osteogenic induction 1 week after adenosine triphosphate composition(P<0.05). CONCLUSIONS: P2X7 receptor can significantly improve the osteogenesis of periodontal ligament stem cells, and adenosine triphosphate can activate the expression of P2X7 receptor.


Assuntos
Osteogênese , Receptores Purinérgicos P2X7 , Células Cultivadas , Humanos , Osteogênese/genética , Ligamento Periodontal , RNA Mensageiro/genética , Receptores Purinérgicos P2X7/genética , Células-Tronco
19.
Shanghai Kou Qiang Yi Xue ; 29(4): 370-374, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-33089284

RESUMO

PURPOSE: To investigate the role of P2X7 receptor (P2X7r) in osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). METHODS: hPDLSCs were isolated from the premolars collected in the First Affiliated Hospital of Guiyang University of Traditional Chinese Medicine, and divided into four groups. Group A was cultured in conventional medium, group B was cultured in osteogenic induction medium, group C was cultured in osteogenic induction medium + 100 nmol/L adenosine triphosphate (ATP) solution, and group D was cultured in osteogenic induction medium + 100 nmol/L P2X7 receptor specific antagonist KN-62. After 7 days, alizarin red staining was used to observe the osteogenic effect of hPDLSCs in each group. The mRNA expression of osteocalcin (OCN), RUNX2 and P2X7r in hPDLSCs was detected by real-time PCR reaction (RT-PCR). The data were processed by SPSS 22.0 software package. RESULTS: Alisarin red staining showed that the morphology of hPDLSCs cells in group B and group C was significantly changed. The pale calcified nodules in group C were significantly more than those in group B, while very few calcified nodules were found in group A and group D. The mRNA expression of OCN, RUNX2 and P2X7r in hPDLSCs were the highest in group C, followed by group B(P<0.05), and no difference was found between group A and group D(P>0.05). CONCLUSIONS: P2X7 receptor can promote osteogenic differentiation of human periodontal ligament stem cells after being activated by ATP, which may provide a new direction for clinical treatment of periodontitis.


Assuntos
Ligamento Periodontal , Receptores Purinérgicos P2X7 , China , Humanos , Osteogênese/genética , Receptores Purinérgicos P2X7/genética , Células-Tronco
20.
Int J Nanomedicine ; 15: 6761-6777, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32982232

RESUMO

Purpose: Guided bone regeneration (GBR) therapy, which is a widely used technique in clinical practice and is effective in improving the repair of alveolar bone defects or bone mass deficiency regeneration, requires the use of membrane materials with good biocompatibility, barrier function, rigidity matching the space maintenance ability, economic benefits and excellent clinical applicability. The aim of this study was to develop an electrospun attapulgite (ATT)-doped poly (lactic-co-glycolic acid) (PLGA) scaffold (PLGA/ATT scaffold) as a novel material for GBR applications. Methods and Results: Scanning electron microscopy (SEM) and X-ray diffraction (XRD) were used to determine the morphology and the crystalline structure of the PLGA/ATT scaffolds, respectively. Porosity and contact-angle measurements were also carried out to further characterize the physical properties of the PLGA/ATT scaffolds. The results of in vitro studies showed that bone marrow mesenchymal stem cells (BMSCs) attached more readily to and spread better over the PLGA/ATT scaffolds than the Bio-Gide membrane. Furthermore, in the in vitro osteoinductive experiments with BMSCs, the PLGA/ATT scaffolds were found to enhance the activity of alkaline phosphatase (ALP), promote the formation of mineralized bone nodules, and up-regulate the expression of several osteogenic markers-namely, runt-related transcription factor 2, alkaline phosphatase, osteopontin, and osteocalcin-which are similar to the effects of the Bio-Gide membrane. Further, in in vivo studies, the results of sequential fluorescent labeling, micro-computed tomography, and histological analysis suggest that using the PLGA/ATT scaffolds for repairing V-shaped buccal dehiscence on a dog's tooth root improved bone regeneration, which is not only similar to the result obtained using the Bio-Gide membrane but also much better than that obtained using PLGA scaffolds and the negative control. Conclusion: To achieve satisfactory therapeutic results and to lower the cost of GBR treatment, this study provided a promising alternative material of bio-degradable membrane in clinical treatment.


Assuntos
Perda do Osso Alveolar/terapia , Regeneração Óssea/fisiologia , Compostos de Magnésio/farmacologia , Compostos de Silício/farmacologia , Tecidos Suporte/química , Animais , Regeneração Óssea/efeitos dos fármacos , Calcificação Fisiológica , Colágeno , Cães , Expressão Gênica , Gengiva/citologia , Humanos , Compostos de Magnésio/química , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Osteogênese/fisiologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Porosidade , Ratos Sprague-Dawley , Compostos de Silício/química , Raiz Dentária/diagnóstico por imagem , Microtomografia por Raio-X
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