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1.
Int J Mol Med ; 47(4): 1, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33537804

RESUMO

Quercetin (Quer) is a typical antioxidant flavonoid from plants that is involved in bone metabolism, as well as in the progression of inflammatory diseases. Elevated levels of tumor necrosis factor­α (TNF­α), a typical pro­inflammatory cytokine, can affect osteogenesis. In the present study, TNF­α was used to establish an in vitro model of periodontitis. The effects of Quer on, as well as its potential role in the osteogenic response of human periodontal ligament stem cells (hPDLSCs) under TNF­α­induced inflammatory conditions and the underlying mechanisms were then investigated. Within the appropriate concentration range, Quer did not exhibit any cytotoxicity. More importantly, Quer significantly attenuated the TNF­α induced the suppression of osteogenesis­related genes and proteins, alkaline phosphatase (ALP) activity and mineralized matrix in the hPDLSCs. These findings were associated with the fact that Quer inhibited the activation of the NF­κB signaling pathway, as well as the expression of NLRP3 inflammation­associated proteins in the inflammatory microenvironment. Moreover, the silencing of NLRP3 by small interfering RNA (siRNA) was found to protect the hPDLSCs against TNF­α­induced osteogenic damage, which was in accordance with the effects of Quer. On the whole, the present study demonstrates that Quer reduces the impaired osteogenesis of hPDLSCs under TNF­α­induced inflammatory conditions by inhibiting the NF­κB/NLRP3 inflammasome pathway. Thus, Quer may prove to be a potential remedy against periodontal bone defects.


Assuntos
Inflamassomos/metabolismo , NF-kappa B/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Osteogênese/efeitos dos fármacos , Ligamento Periodontal/patologia , Quercetina/farmacologia , Células-Tronco/patologia , Fator de Necrose Tumoral alfa/toxicidade , Adolescente , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Regulação para Baixo/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Humanos , Mediadores da Inflamação/metabolismo , Substâncias Protetoras/farmacologia , Transdução de Sinais/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Adulto Jovem
2.
Int J Nanomedicine ; 16: 61-73, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33442250

RESUMO

Objective: Cell sheet technology (CST) is advantageous for repairing alveolar bone defects in clinical situations, and osteogenic induction before implantation may result in enhanced bone regeneration. Herein, we observed the effect of gold nanoparticles (AuNPs) on osteogenic differentiation of periodontal ligament stem cell (PDLSC) sheets and explored their potential mechanism of action. Methods: PDLSCs were cultured in cell sheet induction medium to obtain cell sheets. PDLSC sheets were treated with or without AuNPs. Alkaline phosphatase, alizarin red S, von Kossa, and immunofluorescence staining were used to observe the effects of AuNPs on the osteogenic differentiation of PDLSC sheets. Western blotting was performed to evaluate the osteogenic effects and autophagy activity. The cell sheets were transplanted into the dorsa of nude mice, and bone regeneration was analyzed by micro-CT and histological staining. Results: AuNPs could promote the osteogenic differentiation of PDLSC sheets by upregulating bone-related protein expression and mineralization. The 45-nm AuNPs were more effective than 13-nm AuNPs. Additional analysis demonstrated that their ability to promote differentiation could depend on activation of the autophagy pathway through upregulation of microtubule-associated protein light chain 3 and downregulation of sequestosome 1/p62. Furthermore, AuNPs significantly promoted the bone regeneration of PDLSC sheets in ectopic models. Conclusion: AuNPs enhance the osteogenesis of PDLSC sheets by activating autophagy, and 45-nm AuNPs were more effective than 13-nm AuNPs. This study may provide an AuNP-based pretreatment strategy for improving the application of CST in bone repair and regeneration.


Assuntos
Autofagia/efeitos dos fármacos , Regeneração Óssea/efeitos dos fármacos , Ouro/farmacologia , Nanopartículas Metálicas/química , Ligamento Periodontal/fisiologia , Células-Tronco/citologia , Fosfatase Alcalina/metabolismo , Animais , Fosfatos de Cálcio/farmacologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Humanos , Masculino , Nanopartículas Metálicas/ultraestrutura , Camundongos Nus , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Osteopontina/metabolismo , Osteoprotegerina/metabolismo , Ligamento Periodontal/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Microtomografia por Raio-X
3.
Arch Oral Biol ; 122: 105026, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33348207

RESUMO

OBJECTIVE: This study evaluated the gene expression and protein synthesis of sex hormone receptors in human periodontal ligament cells (PDLCs) in relation to donor- and tooth-specific factors with the aim to clarify the debate about sex hormone receptors in PDLCs. DESIGN: The expression patterns of estrogen receptors (genes: ESR1 and ESR2; proteins: ERα and ERß), androgen receptor (AR) and progesterone receptor (PR) were investigated in the context of immortalization status, previous orthodontic tooth movement (OTM), donor age, sex and hormonal stimulation in PDLCs from 14 healthy donors (male: n = 8, female: n = 6; adolescents: n = 8, adults: n = 6) using quantitative real-time polymerase chain reaction, Western blot and immunocytochemistry. RESULTS: For ERß, the full-length isoform ERß1 and truncated variants were detected. For ERα, the expected isoform ERα66 was not observed, but a novel isoform ERα36 was detected. Immortalization status, previous OTM and donor age had no impact on ESR1 and ESR2 expression. Estradiol stimulation for 24 h doubled the ratio of ESR2/ESR1 in PDLCs from female but not male donors, indicating sex-specific patterns of receptor expression. AR and PR demonstrated insufficient protein synthesis in PDLCs. CONCLUSIONS: The data revealed a pivotal role for and complex interplay between ERα and ERß in human PDLCs regardless of variable donor characteristics. Therefore, PDLC biology might be altered in patients of each age group and both sexes due to hormonal changes. This should be kept in mind during periodontic and orthodontic treatment of patients with special hormonal status.


Assuntos
Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Ligamento Periodontal/citologia , Receptores Androgênicos/genética , Receptores de Progesterona/genética , Adolescente , Adulto , Estradiol , Feminino , Humanos , Masculino , Fatores Sexuais , Doadores de Tecidos
4.
J Formos Med Assoc ; 120(1 Pt 2): 388-394, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32540310

RESUMO

BACKGROUND/PURPOSE: Among various dental lasers, the erbium-doped yttrium-aluminum-garnet (Er:YAG) laser has great potential for periodontal treatment including soft and hard tissue ablation with minimal thermal side effects under suitable energy densities and it has multiple effects on tissues for wound-healing benefits. In the present study, we sought to reveal the molecular mechanism underlying the impact of Er:YAG laser on PDL fibroblasts. METHODS: Cells were irradiated by a Er:YAG laser with various energy densities (3.6-6.3 J/cm2). MTT assay was used for cell proliferation, and the transwell system was employed for migration and invasion abilities. The wound healing capacity was evaluated by a scratch assay. After confirming these effects, qRT-PCR and western blotting analysis was applied to identify the differentially galectin-7 expression in the irradiated cells. Knockdown experiments were conducted to reveal the functional role of galectin-7 in the modulation of Er:YAG laser-mediated effects. RESULTS: 4.2 J/cm2 was the lowest energy density to induce the optimal cell proliferation, migration and invasion abilities. In the group of upregulated genes, galectin-7 was selected for further examination and its elevation after Er:YAG laser treatment was validated by RT-PCR and Western blot. We demonstrated that silence of galectin-7 abrogated the effects of Er:YAG laser on cell proliferation, migration ad invasion, suggesting the Er:YAG laser promoted these effects through induction of galectin-7. CONCLUSION: These findings indicated that Er:YAG laser may accelerate the regeneration process in periodontal tissues through enhancement of their proliferative and mobile activities. Additionally, the significance of galectin-7 in the Er:YAG laser-elicited benefits was demonstrated.


Assuntos
Terapia a Laser , Lasers de Estado Sólido , Proliferação de Células , Fibroblastos , Galectinas/genética , Humanos , Ligamento Periodontal , Cicatrização
5.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 38(6): 628-636, 2020 Dec 01.
Artigo em Chinês | MEDLINE | ID: mdl-33377338

RESUMO

OBJECTIVE: To explore the mechanism of Piezo1 protein in mediating the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) via the Notch signaling pathway. METHODS: In this study, young permanent teeth extracted from impacted teeth of 8-14-year-old children from January 1, 2016 to January 1, 2018 in the Department of Orthodontic, Beijing Children's Hospital were selected as cell sources. hPDLSCs were extracted by enzymatic digestion. Immunohistochemical staining was used to detect the expression of keratin and vimentin, and flow cytometry was used to identify the markers (CD146 and STRO-1) of hPDLSCs. The construction and screening of Piezo1 siRNA gene interference vector and Piezo1 gene overexpression plasmid were completed. Flexcell 4000T mechanical distraction stress instrument was used to construct hPDLSC cell model in vitro. According to the preliminary results, the experiment was divided into five groups: siRNA interference group, overexpression group, blank control group, stretch stress group, and negative control group. Real time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression of Piezo1, Notch1, alkaline phosphatase (ALP), Runt-related transcription factor 2 (Runx2), osteocalcin (OCN), and bone sialoprotein (BSP). Western blot was used to detect the expression of ALP and Runx2. Fluo-3 AM probe was used to detect intracellular calcium content. RESULTS: Vimentin staining of hPDLSCs was positive, and keratin staining was negative. Flow cytometry was used to detect the expression of STRO-1 and CD146, markers of hPDLSC. Empty viral vectors, siRNA-Piezo1 interference sequence, and Piezo1 overexpression vector sequence could be transfected into hPDLSC by lentivirus, and the transfection efficiency was high (approximately 90%). The reverse transcription-polymerase chain reaction (RT-PCR) results showed that there were significant differences in Piezo1 gene levels among the siRNA interference group, overexpression group, blank control group, stretch stress group, and negative control group (F=9.573, P<0.05). The level of Piezo1 in the overexpression group was significantly higher than that in the siRNA interference group (q=3.893, P<0.05). The level of Piezo1 in the stretch stress group was significantly higher than that in the blank control group (q=2.006, P<0.05). The expression of Notch1 and osteogenic genes ALP, Runx2, OCN, and BSP had the same trend. Western blot results showed that there were significant differences in the expression of ALP in the siRNA interference group, overexpression group, blank control group, stretch stress group, and negative control group (F=11.207, P<0.001). The expression level of ALP in the overexpression group was significantly higher than that in the siRNA interference group (q=2.991, P<0.05). The expression of ALP in the stretch stress group was significantly higher than that in the blank control group (q=3.007, P<0.05). The expression of Runx2 protein showed the same trend. The intracellular calcium fluorescence intensity of the overexpression group was significantly higher than that of the siRNA interference group, and the intracellular calcium fluorescence intensity of the stretch stress group was significantly higher than that of the siRNA interference group. CONCLUSIONS: Mechanical stretch stress can promote the expression of Piezo1 protein. Ca2+ is the second messenger, activates the Notch1 signaling pathway and the expression of ALP, Runx2, OCN, and BSP; and promotes the osteogenic differentiation of hPDLSC. The siRNA-Piezo1 interfering plasmid can block this process. On the contrary, the overexpression plasmid of Piezo1 can promote the osteogenic differentiation of PDLSCs.


Assuntos
Osteogênese , Ligamento Periodontal , Fosfatase Alcalina , Diferenciação Celular , Células Cultivadas , Criança , Humanos , Canais Iônicos , Transdução de Sinais , Células-Tronco
6.
Ned Tijdschr Tandheelkd ; 127(12): 691-698, 2020 Dec.
Artigo em Holandês | MEDLINE | ID: mdl-33367296

RESUMO

Apical root resorption is a biological process induced when orthodontic force is exerted on a tooth and local necrosis of the periodontal ligament occurs. Macrophages remove the necrotic tissue. In this way, differentiating osteoclasts can both attach to the now available dental surface and can then provoke root resorption. There is considerable uncertainty among dental practitioners on how to deal with clinically relevant apical root resorption (bigger/equal 2 mm) during or after orthodontic treatment. To increase understanding and to improve the quality of care, the Dutch Association of Orthodontists has developed a clinical practice guideline. Recommendations have been formulated for the diagnosis of apical root resorption, possible risk factors and treatment management in order to respond adequately to this problem in practice.


Assuntos
Reabsorção da Raiz , Odontólogos , Humanos , Ligamento Periodontal , Papel Profissional , Reabsorção da Raiz/etiologia , Técnicas de Movimentação Dentária , Raiz Dentária
8.
Angle Orthod ; 90(5): 702-706, 2020 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-33378474

RESUMO

OBJECTIVES: To explore the expression of miR-34a and its effect on expression of matrix metalloproteinases (MMPs) during orthodontic tooth movement (OTM). MATERIALS AND METHODS: Twenty patients, age 12-18 years old, who underwent orthodontic treatment were enrolled. The expression of miR-34a and MMPs (MMP-1, MMP-2, MMP-3, MMP-8, MMP-9, and MMP-14) were detected in gingival crevicular fluid by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction at different time points. The miR-34a mimics or inhibitors were transfected into human periodontal ligament (hPDL) cells, and the MMP expression was measured by ELISA. RESULTS: The miR-34 expression in GCF on both the tension and pressure sides after orthodontic treatment were significantly downregulated, while the levels of MMPs were significantly upregulated compared with baseline level. The levels of miR-34 and MMPs returned to baseline level 3 months after orthodontic treatment. The expression of miR-34 was negatively correlated with the expression of MMP-2, MMP-9, and MMP-14. After transfection with miR-34, the MMP-2, MMP-9, and MMP-14 expression by hPDL cells were significantly downregulated compared with miR-control and miR-34 inhibitor. CONCLUSIONS: Downregulated miR-34 expression was positively correlated with MMP-2, MMP-9, and MMP-14 expression. The miR-34a transfection into hPDL cells inhibited expression of MMPs. The results suggest that miR-34a is involved in expression of MMPs during OTM.


Assuntos
Líquido do Sulco Gengival , MicroRNAs , Adolescente , Criança , Humanos , Metaloproteinase 9 da Matriz/genética , MicroRNAs/genética , Ligamento Periodontal , Técnicas de Movimentação Dentária
9.
Head Face Med ; 16(1): 26, 2020 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-33190638

RESUMO

BACKGROUND: Periodontal ligament (PDL) cells initiate local immune responses, similar to microglia regulating primary host defense mechanisms in neuroinflammatory events of the central nervous system. As these two cell types manifest similarities in their immunomodulatory behavior, this study investigated the thesis that the immunological features of PDL cells might be modulated by the endocannabinoid system, as seen for microglia. METHODS: A human PDL cell line and an Embryonic stem cell-derived microglia (ESdM) cell line were grown in n = 6 experimental groups each, incubated with cannabinoid receptor agonists arachidonoylethanolamine (AEA) (50 µM) or Palmitoylethanolamide (PEA) (50 µM) and challenged with centrifugation-induced inflammation (CII) for 6 and 10 h. Untreated samples served as controls. Quantitative real-time polymerase chain reaction was applied for gene expression analyses of inflammatory cytokines, cannabinoid receptors and ionized calcium binding adaptor molecule 1 (IBA-1). Microglia marker gene IBA-1 was additionally verified on protein level in PDL cells via immunocytochemistry. Proliferation was determined with a colorimetric assay (WST-1 based). Statistical significance was set at p < 0.05. RESULTS: IBA-1 was inherently expressed in PDL cells both at the transcriptional and protein level. AEA counteracted pathological changes in cell morphology of PDL cells and microglia caused by CII, and PEA contrarily enhanced them. On transcriptional level, AEA significantly downregulated inflammation in CII specimens more than 100-fold, while PEA accessorily upregulated them. CII reduced cell proliferation in a time-dependent manner, synergistically reinforced by PEA decreasing cell numbers to 0.05-fold in PDL cells and 0.025-fold in microglia compared to controls. CONCLUSION: PDL cells and microglia exhibit similar features in CII with host-protective effects for AEA through dampening inflammation and preserving cellular integrity. In both cell types, PEA exacerbated proinflammatory effects. Thus, the endocannabinoid system might be a promising target in the regulation of periodontal host response.


Assuntos
Endocanabinoides , Ligamento Periodontal , Proliferação de Células , Células Cultivadas , Humanos , Microglia
10.
Stomatologija ; 21(2): 49-53, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33245062

RESUMO

OBJECTIVE: The aim of this article is to review the effect of enamel matrix derivate (EMD) on growth factors activation for periodontal regeneration. MATERIAL AND METHODS: Online databases, such as PubMed, Cochrane Library, PMC, Science Direct were searched by using the following keywords in various combinations: emdogain, periodontal regeneration, growth factors, transforming growth factor, bone morphogenetic protein, fibroblast growth factor and vascular endothelial growth factors. All studies fulfilling the selection criteria were carefully reviewed for the focused question: "Does enamel matrix derivate induces the activity of growth factors, important in periodontal regeneration?". RESULTS: 1378 articles were found in the databases using keywords. After duplicate citations screened, inclusion/exclusion criteria applied, excluded articles after titles, summaries and full-text reading 14 articles were included in the literature review. CONCLUSION: Enamel matrix derivate (EMD) was found to have a possitive effect on periodontal tissue regneration. By stimulating secretion and activating functions of growth factors, such as transforming growth factor-ß (TGF-ß), bone morphogenetic proteins (BMP), vascular endothelial growth factors (VEGF) and fibroblast growth factor-2 (FGF-2), EMD induces production of new alveolar bone, new root cementum and functionl periodontal ligament (PDL) and new blood vessels formation in periodontal area. Due to this production, the probing depth of periodontal pocket is being reduced.


Assuntos
Peptídeos e Proteínas de Sinalização Intercelular , Ligamento Periodontal , Regeneração Óssea , Regeneração , Fator de Crescimento Transformador beta
11.
Am J Orthod Dentofacial Orthop ; 158(6): e151-e160, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33139146

RESUMO

INTRODUCTION: The Wnt signaling pathway acts as a key regulator of skeletal development and its homeostasis. However, the potential role of Wnt1 in the mechanotransduction machinery of orthodontic tooth movement-initiated bone remodeling is still unclear. Hence, this study focused on the regulatory dynamics of the Wnt1 expression in both the periodontal ligament (PDL) and osteocytes in vivo and in vitro. METHODS: The Wnt1 expression in the orthodontically moved maxillary first molar in mice was assessed at 0, 1, and 5 days, on both the compression and tension sides. Primary isolated human PDL (hPDL) fibroblasts, as well as murine long-bone osteocyte-Y4 (MLO-Y4) cells, were exposed to continuous compressive force and static tensile force. RESULTS: The relative quantification of immunodetection showed that orthodontic tooth movement significantly stimulated the Wnt1 expression in both the PDL and alveolar osteocytes on the tension side on day 5, whereas the expression on the compression side did not change. This increase in the Wnt1 expression, shown in vivo, was also noted after the application of 12% static tensile force in isolated hPDL fibroblasts and 20% in MLO-Y4 cells. In contrast, a compressive force led to the attenuation of the Wnt1 gene expression in both hPDL fibroblasts and MLO-Y4 cells in a force-dependent manner. In the osteocyte-PDL coculture system, recombinant sclerostin attenuated Wnt1 in PDL, whereas the antisclerostin antibody upregulated its gene expression, indicating that mechanically-driven Wnt1 signaling in PDL might be regulated by osteocytic sclerostin. CONCLUSIONS: Our findings provide that Wnt1 signaling plays a vital role in tooth movement-initiated bone remodeling via innovative mechanotransduction approaches.


Assuntos
Mecanotransdução Celular , Técnicas de Movimentação Dentária , Animais , Remodelação Óssea , Humanos , Camundongos , Osteócitos , Ligamento Periodontal , Estresse Mecânico , Proteína Wnt1/genética
12.
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
13.
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
14.
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
15.
Am J Orthod Dentofacial Orthop ; 158(6): 816-823, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33082075

RESUMO

INTRODUCTION: Periodontal ligament cells (PDLC) respond to the application of mechanical forces by releasing various molecules that participate in bone remodeling. Whether these cellular reactions take place at the same rate in adolescent and adult patients is not known. In this study, we aimed to evaluate differences in genetic expression, if any, between the release of various inflammatory mediators from PDLC in adolescent and adult patients before and after the application of orthodontic forces. METHODS: Forty subjects with bimaxillary dentoalveolar protrusion requiring extraction of first premolars for orthodontic treatment were selected and divided into 2 groups. Group A included 20 adolescents (aged 12-20 years), and group B included 20 adults (aged 35-50 years). Then, 35-50 g of force were applied to the maxillary first premolars, and teeth were extracted at different periods: pretreatment (control group), 7 days, 14 days, and 28 days (experimental group). The periodontal ligament was scraped from the middle third of the root, and the beta-galactosidase assay was performed in the control group. RNA extraction, DNase treatment, quantitative polymerase chain reaction, and complementary DNA synthesis were performed in the experimental group. RESULTS: Adult PDLC exhibited senescent changes through increased beta-galactosidase activity. The increase in the inflammatory response and bone resorption in adult patients was evident by increased prostaglandin E2, IL1B, and acid phosphatase mRNA expression levels. Controlled bone formation response by adolescent PDLC was evident from increased ALP and BGLAP mRNA levels and a balanced receptor activator of nuclear factor kappa-Β ligand/OPG ratio. CONCLUSIONS: The study could identify the reasons behind the differential response of adolescent and adult PDLC to orthodontic mechanics.


Assuntos
Ligamento Periodontal , Técnicas de Movimentação Dentária , Adolescente , Adulto , Dente Pré-Molar , Remodelação Óssea , Criança , Humanos , Pessoa de Meia-Idade , Estresse Mecânico , Adulto Jovem
16.
J Contemp Dent Pract ; 21(7): 776-780, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33020362

RESUMO

AIM: To evaluate the ability of osteogenic culture media in comparison with regular growth culture media in enhancing the osteoblastic cell differentiation of human periodontal ligament stem cells (hPDLSCs). MATERIALS AND METHODS: In vitro cultures of commercially obtained hPDLSCs were seeded onto xenograft bone blocks in both regular and osteogenic media. Confocal laser microscope images were obtained for cellular differentiation and adhesion, and scanning electron microscopy (SEM) images were obtained to validate the osteogenic differentiation by showing the morphological characteristics of the newly formed cells. RESULTS: Confocal laser microscope analysis showed positive staining for new bone cells with an increased signal intensity when samples were cultured in osteogenic culture media compared with regular culture media. These findings indicate the effect of the active ingredients of the osteogenic culture media in enhancing the osteogenic differentiation hPDLSC. Scanning electron microscopy images validated the osteogenic differentiation showing a flattened, polygonal morphology with multiple extending cytoplasmic processes of new cells. CONCLUSION: Xenograft bone blocks are biocompatible scaffold for the osteogenic differentiation of seeded hPDLSCs. Osteogenic culture media enhances and increases the osteogenic differentiation of hPDLSCs into new bone cells more than regular growth culture media. Periodontal ligament stem cells are a predictable biological input as a cell-based tissue-engineered construct and biologically acceptable when it is cultured in a suitable growth media that mimics the intended environment. CLINICAL SIGNIFICANCE: Consideration of the clinical use of equine bone blocks and periodontal ligament stem cells in a suitable biological environment as a potential new option for bone regeneration techniques.


Assuntos
Osteogênese , Ligamento Periodontal , Animais , Células Cultivadas , Meios de Cultura , Cavalos , Humanos , Microscopia Eletrônica de Varredura , Células-Tronco
17.
J Contemp Dent Pract ; 21(7): 798-802, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33020366

RESUMO

AIM: To illustrate, with two clinical cases of endoperiodontal lesions, the clinical application of the new classification of periodontal and peri-implant diseases and conditions. BACKGROUND: The endodont and the periodont are two entities that communicate with each other through physiological communication channels (apical foramen, lateral and secondary canals, and dentinal tubules) resulting in close anatomical and functional interaction. An endoperiodontal lesion is defined by pathological communication between the endodontic and periodontal tissues in a given tooth, according to the definition given by the new classification of periodontal and peri-implant diseases and conditions from the work of the Chicago Consensus Conference in 2017. This new classification differentiates the lesions with and without root damage. Diagnosis and therapeutic strategy will be analyzed through two clinical cases. REVIEW RESULTS: The clinical cases we presented show that the treatment of these lesions must involve endodontic and periodontal management due to the intimate relationship between the tooth and periodontium. CONCLUSION: The classification of periodontal and peri-implant diseases and conditions provides a clinical focus on endoperiodontal lesions, based on signs and symptoms that have a direct effect on the prognosis and the treatment of the tooth. The pathological communication between the endodont and the periodontium complicates the management of the involved tooth. CLINICAL SIGNIFICANCE: Chicago's new classification of periodontal and peri-implant diseases and conditions offers an up-to-date vision of periodontal lesions management and highlights the intimate links between endodontic and periodontal tissues.


Assuntos
Peri-Implantite , Doenças Periodontais , Chicago , Humanos , Ligamento Periodontal , Periodonto
18.
Shanghai Kou Qiang Yi Xue ; 29(3): 225-230, 2020 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-33043336

RESUMO

PURPOSE: To investigate the effects of exendin-4(EX-4) on proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells(PDLSCs). METHODS: PDLSCs were isolated and cultured using limited dilution method in vitro. Colony formation assay, osteogenic and adipogenic differentiation were applied to identify the stem cells. Immunofluorescence staining was used to detect the expression of EX-4 receptor glucagon-like peptide-1 receptor (GLP-1R) on the surface of PDLSCs. PDLSCs were stimulated with 5, 10, 20 or 50 nmol/L EX-4 in vitro. CCK-8, Transwell assay and alkaline phosphatase(ALP) activity assay were used to determine the effects of EX-4 on PDLSCs proliferation, migration and osteogenic differentiation. Quantitative real-time polymerase chain reaction was used to determine the expression of osteogenic related genes ALP, runt-related transcription factor 2(Runx2) and osteocalcin (OCN). The data were analyzed by Graphpad Prims 6.0 software package. RESULTS: PDLSCs were successfully isolated and cultivated. GLP-1R positively expressed on the surface of PDLSCs. EX-4 exerted no significant effect on PDLSCs proliferation(P>0.05). EX-4 significantly promoted migration, ALP activity and osteogenic related genes expression of PDLSCs (P<0.05). CONCLUSIONS: 10 nmol/L EX-4 could promote migration and osteogenic differentiation of PDLSCs.


Assuntos
Exenatida , Ligamento Periodontal , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Exenatida/farmacologia , Humanos , Osteogênese , Células-Tronco
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(7): 942-948, 2020 Jul 30.
Artigo em Chinês | MEDLINE | ID: mdl-32895159

RESUMO

OBJECTIVE: To investigate the effect of periostin on hypoxia-induced oxidative stress and apoptosis in human periodontal ligament fibroblasts and the molecular mechanism involved. METHODS: In vitro cultured human periodontal ligament fibroblasts were placed in an anaerobic gas-producing bag for hypoxia treatment for 48 h followed by treatment with periostin at low (25 ng/mL), moderate (50 ng/mL) or high (100 ng/mL) doses. MTT assay was used to measure the cell viability, and the cell apoptosis rate was determined using flow cytometry. The contents of IL-1ß, IL-6 and TNF-α in the cells were determined with ELISA, and ROS levels were measured using a fluorescent plate reader. The intracellular SOD activity was detected using ELISA. The expressions of HIF-1α, P21, cyclin D1, Bax, cleaved caspase-3, Bcl-2, P38MAPK and p-p38 MAPK proteins in the cells were detected with Western blotting. RESULTS: Hypoxia treatment significantly reduced the cell viability (P < 0.05), increased P21, Bax, and cleaved caspase-3 protein levels (P < 0.05), promoted cell apoptosis (P < 0.05), and decreased cyclin D1 and Bcl-2 protein levels (P < 0.05) in the cells. Compared with the hypoxic group, the cells treated with periostin at different concentrations showed significantly increased cell viability (P < 0.05) with significantly lowered apoptotic rates (P < 0.05) and decreased expression levels of Bax and cleaved caspase-3 (P < 0.05) but significantly increased expression levels of cyclin D1 and Bcl-2 (P < 0.05). Hypoxic exposure of the cells resulted in significantly increased expression levels of HIF-1α and p-p38 MAPK (P < 0.05) and increased levels of IL-1ß, IL-6, TNF-α and ROS (P < 0.05) but decreased SOD activity (P < 0.05). Periostin treatment at different concentrations significantly lowered the expression levels of HIF-1α and p-p38 MAPK (P < 0.05) and the levels of IL-1ß, IL-6, TNF-α and ROS (P < 0.05) and significantly increased SOD activity in the hypoxic cells (P < 0.05). CONCLUSIONS: Periostin promotes the proliferation, inhibits apoptosis, enhances cellular antioxidant capacity, and reduces inflammatory damage in human periodontal ligament fibroblasts exposed to hypoxia possibly by inhibiting the activation of the p38 MAPK signaling pathway.


Assuntos
Apoptose , Ligamento Periodontal , Fibroblastos , Humanos , Hipóxia , Estresse Oxidativo , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno
20.
Cell Prolif ; 53(10): e12912, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32964544

RESUMO

OBJECTIVES: Mechanical force plays an important role in modulating stem cell fate and behaviours. However, how periodontal ligament stem cells (PDLSCs) perceive mechanical stimulus and transfer it into biological signals, and thereby promote alveolar bone remodelling, is unclear. MATERIALS AND METHODS: An animal model of force-induced tooth movement and a compressive force in vitro was used. After force application, tooth movement distance, mesenchymal stem cell and osteoclast number, and proinflammatory cytokine expression were detected in periodontal tissues. Then, rat primary PDLSCs with or without force loading were isolated, and their stem cell characteristics including clonogenicity, proliferation, multipotent differentiation and immunoregulatory properties were evaluated. Under compressive force in vitro, the effects of the ERK signalling pathway on PDLSC characteristics were evaluated by Western blotting. RESULTS: Mechanical force in vivo induced PDLSC proliferation, which was accompanied with inflammatory cytokine accumulation, osteoclast differentiation and TRPV4 activation; the force-stimulated PDLSCs showed greater clonogenicity and proliferation, reduced differentiation ability, improved induction of macrophage migration, osteoclast differentiation and proinflammatory factor expression. The biological changes induced by mechanical force could be partially suppressed by TRPV4 inhibition. Mechanistically, force-induced activation of TRPV4 in PDLSCs regulated osteoclast differentiation by affecting the RANKL/OPG system via ERK signalling. CONCLUSIONS: Taken together, we show here that TRPV4 activation in PDLSCs under mechanical force contributes to changing their stem cell characteristics and modulates bone remodelling during tooth movement.


Assuntos
Remodelação Óssea , Ligamento Periodontal/citologia , Células-Tronco/citologia , Canais de Cátion TRPV/metabolismo , Animais , Fenômenos Biomecânicos , Proliferação de Células , Células Cultivadas , Humanos , Masculino , Osteoclastos/citologia , Osteoclastos/metabolismo , Ligamento Periodontal/metabolismo , Ratos , Ratos Sprague-Dawley , Células-Tronco/metabolismo , Estresse Mecânico
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