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1.
Arch Oral Biol ; 162: 105964, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38582010

RESUMO

OBJECTIVE: This study aimed to explore the effects of small extracellular vesicles derived from lipopolysaccharide-preconditioned dental follicle cells (L-D-sEV) on periodontal ligament cells from periodontitis affected teeth (p-PDLCs) in vitro and experimental periodontitis in mice. DESIGN: In vitro, the biological function of p-PDLCs and the underlying molecular mechanism were investigated by flow cytometry, Western blot, and quantitative real-time PCR (qRT-PCR) analysis. Eighteen-eight-week-old male C57BL/6 mice were randomly divided into three groups: control (Con), periodontitis (Peri), and L-D-sEV groups. Mice periodontitis model was induced by placing the 5-0 silk thread (around the maxillary second molar) and P.gingivalis (1 ×107 CFUs per mouse). In vivo, the alveolar bone loss, osteoclast activity, and macrophage polarization were measured by micro-computed tomography and histological analysis. RESULTS: In vitro, the RANKL/OPG ratio and phosphorylation of JNK and P38 protein levels of p-PDLCs were significantly decreased after L-D-sEV administration. Besides, flow cytometry and qRT-PCR analysis showed that L-D-sEV reduced apoptosis of p-PDLCs, down-regulated apoptosis-related genes Caspase-3 and BCL-2-Associated X expression, and up-regulated B-cell lymphoma-2 gene levels. In vivo, L-D-sEV administration significantly reduced alveolar bone loss, inhibited osteoclast activity, and induced M2 polarization. The histological analysis showed that iNOS/CD206, RANKL/OPG, p-JNK/JNK, and p-P38/P38 ratios were significantly lower in the L-D-sEV group than in the Peri group. CONCLUSIONS: L-D-sEV administration alleviated alveolar bone loss by mediating RANKL/OPG-related osteoclast activity and M2 macrophage polarization, alleviating p-PDLCs apoptosis and proliferation via the JNK and P38 pathways.


Assuntos
Perda do Osso Alveolar , Periodontite , Camundongos , Masculino , Animais , Perda do Osso Alveolar/patologia , Lipopolissacarídeos/farmacologia , Microtomografia por Raio-X , Saco Dentário/metabolismo , Camundongos Endogâmicos C57BL , Periodontite/metabolismo , Apoptose , Modelos Animais de Doenças
2.
ACS Biomater Sci Eng ; 10(5): 3173-3187, 2024 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-38605468

RESUMO

The application of bioengineering techniques for achieving bone regeneration in the oral environment is an increasingly prominent field. However, the clinical use of synthetic materials carries certain risks. The liquid phase of concentrated growth factor (LPCGF), as a biologically derived material, exhibits superior biocompatibility. In this study, LPCGF was employed as a tissue engineering scaffold, hosting dental follicle cells (DFCs) to facilitate bone regeneration. Both in vivo and in vitro experimental results demonstrate that this platform significantly enhances the expression of osteogenic markers in DFCs, such as alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), and type I collagen (Col1a1). Simultaneously, it reduces the expression of inflammation-related genes, particularly interleukin-6 (IL-6) and interleukin-8 (IL-8), thereby alleviating the negative impact of the inflammatory microenvironment on DFCs. Further investigation into potential mechanisms reveals that this process is regulated over time by the WNT pathway. Our research results demonstrate that LPCGF, with its favorable physical characteristics, holds great potential as a scaffold. It can effectively carry DFCs, thereby providing an optimal initial environment for bone regeneration. Furthermore, LPCGF endeavors to closely mimic the mechanisms of bone healing post-trauma to facilitate bone formation. This offers new perspectives and insights into bone regeneration engineering.


Assuntos
Regeneração Óssea , Saco Dentário , Peptídeos e Proteínas de Sinalização Intercelular , Alicerces Teciduais , Regeneração Óssea/efeitos dos fármacos , Saco Dentário/citologia , Saco Dentário/metabolismo , Alicerces Teciduais/química , Animais , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Células-Tronco/metabolismo , Células-Tronco/citologia , Osteogênese , Humanos , Engenharia Tecidual/métodos
3.
J Cell Physiol ; 239(6): e31283, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38651182

RESUMO

The long noncoding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) plays a crucial role in tumorigenesis and is frequently employed as a prognostic biomarker. However, its involvement in the osteogenic differentiation of oral stem cells, particularly human dental follicle stem cells (hDFSCs), remains unclear. Our investigation revealed that the absence of SNHG1 enhances the osteogenic differentiation of hDFSCs. Furthermore, the downregulation of SNHG1 induces autophagy in hDFSCs, leading to a reduction in intracellular oxidative stress levels. Notably, this effect is orchestrated through the epigenetic regulation of EZH2. Our study unveils a novel function of SNHG1 in governing the osteogenic differentiation of hDFSCs, offering fresh insights for an in-depth exploration of the molecular mechanisms underlying dental follicle development. These findings not only provide a foundation for advancing the understanding of SNHG1 but also present innovative perspectives for promoting the repair and regeneration of periodontal supporting tissue, ultimately contributing to the restoration of periodontal health and tooth function.


Assuntos
Autofagia , Diferenciação Celular , Saco Dentário , Proteína Potenciadora do Homólogo 2 de Zeste , Osteogênese , Estresse Oxidativo , RNA Longo não Codificante , Células-Tronco , Humanos , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Autofagia/genética , Estresse Oxidativo/genética , Osteogênese/genética , Diferenciação Celular/genética , Células-Tronco/metabolismo , Saco Dentário/metabolismo , Saco Dentário/citologia , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Epigênese Genética , Células Cultivadas , Técnicas de Silenciamento de Genes
4.
Eur J Orthod ; 46(1)2024 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-38001047

RESUMO

OBJECTIVE: To assess if the dental follicle volume of palatally impacted canines (PICs) affects the relative root position of the adjacent lateral incisors (LIs) and first premolars (FPs). MATERIALS AND METHODS: A retrospective cross-sectional study of 49 patients with unilaterally PICs with dental follicles who had CBCT imaging previously taken. Four orthodontic centers in different countries provided the sample. A mean difference of 5° between the angular measurements (mesiodistal tip, buccolingual inclination, or mesiodistal rotation) of the LI and FP adjacent to the PIC and the controls was considered clinically relevant. A value of 0.05 was set for significance level and a power of 80%. The minimum sample size was determined to be 26 patients. These patients were further assigned to an LI sample (n = 49) and an FP sample (n = 23), dependent on the direct contact of the dental follicle to that adjacent tooth. A manual segmentation technique was used to obtain the volumetric measurements of the dental follicle. Angular measurements of adjacent teeth were then compared to the contralateral nonimpacted side, which acted as the control. A multivariant regression analysis was performed using IBM SPSS software, and statistical significance was set at α = 0.05. RESULTS: Adequate intra-rater reliability was accomplished. The multivariant regression analysis implied that there is no difference in the mean change in the tip, torque, and rotation of the LI and FP between the impacted and control sides when dental follicle volumes are considered (P = .509 for the LI sample and P = .804 for the FP sample). LIMITATIONS: CBCT imaging of dental follicle border delimitations, partial volume effect, and scattering are limitations. This is a convenience sample where the FP subsample is small. CONCLUSION: The dental follicle volume of the PICs does not seem to influence the relative position of the adjacent LI and FP mesiodistal tip, buccolingual inclination, and mesiodistal rotation. Early intervention could have been suggested to avoid certain malocclusion traits if significant displacements were demonstrated.


Assuntos
Saco Dentário , Dente Impactado , Humanos , Saco Dentário/diagnóstico por imagem , Estudos Retrospectivos , Reprodutibilidade dos Testes , Estudos Transversais , Dente Canino/diagnóstico por imagem , Dente Impactado/diagnóstico por imagem , Tomografia Computadorizada de Feixe Cônico/métodos , Maxila
5.
J Contemp Dent Pract ; 24(10): 809-812, 2023 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-38152915

RESUMO

AIM: To evaluate the cystic changes in the radiographically normal dental follicle associated with impacted mandibular third molar. MATERIALS AND METHODS: This study was conducted on 80 patients. Samples were selected using a convenient sampling technique from the patients who had impacted mandibular third molars in Pell and Gregory's positions B and C, with follicular space less than 2.5 mm in diameter. After surgical removal of an impacted tooth, the dental follicle was sent for histopathologic evaluation. RESULTS: Pathologic alterations were found in 19% of cases out of 80 samples. Odontogenic keratocystic and dentigerous cystic changes were found in 7% of cases. A statistically significant cystic alteration was found in female patients and distoangular impacted teeth. CONCLUSION: This study shows a significant cystic alteration in the radiologically normal dental follicles. Clinical and radiographic features alone may not be a reliable indicator of the absence of pathology. Early intervention of impacted teeth will help to reduce morbidity due to the development of pathology. CLINICAL SIGNIFICANCE: This study will help educate patients on the risks of retaining impacted teeth, based on scientific facts, in order to minimize the risks and to assess the correlation of pathologic alterations with the depth of impaction and angular position of the impacted tooth.


Assuntos
Dente Serotino , Dente Impactado , Humanos , Feminino , Dente Serotino/diagnóstico por imagem , Dente Serotino/patologia , Dente Impactado/diagnóstico por imagem , Dente Impactado/cirurgia , Saco Dentário/patologia , Dente Molar/patologia , Mandíbula/patologia
6.
Int J Mol Sci ; 24(22)2023 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-38003310

RESUMO

N6-methyladenosine (m6A) is the most abundant RNA modification, regulating gene expression in physiological processes. However, its effect on the osteogenic differentiation of dental follicle stem cells (DFSCs) remains unknown. Here, m6A demethylases, the fat mass and obesity-associated protein (FTO), and alkB homolog 5 (ALKBH5) were overexpressed in DFSCs, followed by osteogenesis assay and transcriptome sequencing to explore potential mechanisms. The overexpression of FTO or ALKBH5 inhibited the osteogenesis of DFSCs, evidenced by the fact that RUNX2 independently decreased calcium deposition and by the downregulation of the osteogenic genes OCN and OPN. MiRNA profiling revealed that miR-7974 was the top differentially regulated gene, and the overexpression of m6A demethylases significantly accelerated miR-7974 degradation in DFSCs. The miR-7974 inhibitor decreased the osteogenesis of DFSCs, and its mimic attenuated the inhibitory effects of FTO overexpression. Bioinformatic prediction and RNA sequencing analysis suggested that FK506-binding protein 15 (FKBP15) was the most likely target downstream of miR-7974. The overexpression of FKBP15 significantly inhibited the osteogenesis of DFSCs via the restriction of actin cytoskeleton organization. This study provided a data resource of differentially expressed miRNA and mRNA after the overexpression of m6A demethylases in DFSCs. We unmasked the RUNX2-independent effects of m6A demethylase, miR-7974, and FKBP15 on the osteogenesis of DFSCs. Moreover, the FTO/miR-7974/FKBP15 axis and its effects on actin cytoskeleton organization were identified in DFSCs.


Assuntos
MicroRNAs , Osteogênese , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Saco Dentário/metabolismo , Células Cultivadas , Diferenciação Celular/genética , MicroRNAs/metabolismo , Células-Tronco/metabolismo
7.
J Oral Biosci ; 65(4): 371-378, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37806337

RESUMO

OBJECTIVE: This study aimed to examine the therapeutic effects of curcumin against replicative senescence in dental follicle cells (DFCs). METHODS: Human DFCs were cultured in Dulbecco's Modified Eagle Medium with growth supplements. Replicative senescence in DFCs at different passages was assessed using ß-galactosidase activity assay. Cell proliferation and size of DFCs at different passages were determined by CCK-8 kit and microscopy method, respectively. In addition, curcumin's effect on replicative senescence, cell proliferation, and size of DFCs at different passages was analyzed. Using western-blot analysis and siRNA-mediated gene silencing, we determined the molecular mechanisms involved in curcumin's effect against replicative senescence and osteogenic differentiation in DFCs at different passages. RESULTS: We observed decreased proliferation and increased cell size and replicative senescence in cultured human DFCs at higher passages. Intriguingly, despite not showing any effect on cell size, curcumin (50 µM) significantly restored proliferation ability in DFCs and inhibited their replicative senescence. Concerning mechanisms, we found that curcumin inhibits replicative senescence in DFCs via down-regulation of senescence markers (P16 & P21) and restoration of proliferation markers (E2F1 & P53). Additionally, curcumin also rescued the osteogenic differentiation potential in higher-passage DFCs via restoration of osteogenic markers RUNX2 and OPN. CONCLUSION: Our findings reveal for the first time that curcumin could act as a potential anti-senescence therapeutic for DFCs via regulation of proliferation, senescence, and osteogenic differentiation markers.


Assuntos
Curcumina , Osteogênese , Humanos , Osteogênese/genética , Curcumina/farmacologia , Saco Dentário , Diferenciação Celular/genética , Senescência Celular
8.
Eur J Oral Sci ; 131(5-6): e12952, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37664892

RESUMO

Dental follicle cells (DFCs) are osteogenic progenitor cells and are well suited for molecular studies of differentiation of alveolar osteoblasts. A recent study examined the metabolism in DFCs during osteogenic differentiation and showed that energy metabolism is increased after 14 days of differentiation (mid phase). However, previous studies have examined proteomes at early (2 h, 24 h) or very late (28 days) stages of differentiation, but not during the phase of increased metabolic activity. In this study, we examined the phosphoproteome at the mid phase (14 days) of osteogenic differentiation. Analysis of DFC phosphoproteomes showed that during this phase of osteogenic differentiation, proteins that are part of signal transduction are significantly regulated. Proteins involved in the regulation of the cytoskeleton and apoptosis were also increased in expression. As osteogenic differentiation induced oxidative stress and apoptosis in DFCs, the oxidative stress defense protein, catalase, was also upregulated during osteogenic differentiation, which supports the biomineralization of DFCs. In summary, this study revealed that during the middle phase (14 days) of osteogenic differentiation, processes in DFCs related to the control of cell organization, apoptosis, and oxidative stress are regulated.


Assuntos
Osteogênese , Proteoma , Humanos , Osteogênese/fisiologia , Saco Dentário/metabolismo , Diferenciação Celular/fisiologia , Células-Tronco , Células Cultivadas
9.
Arch Oral Biol ; 153: 105737, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37320885

RESUMO

OBJECTIVE: This study aimed to explore the effect of periostin in the osteogenic abilities of dental follicle stem cells (DFSCs) and DFSC sheets in the inflammatory microenvironment. DESIGN: DFSCs were isolated from dental follicles and identified. A lentiviral vector was used to knock down periostin in DFSCs. 250 ng/ml lipopolysaccharide from Porphyromonas gingivalis (P.g-LPS) was used to construct the inflammatory microenvironment. Osteogenic differentiation was evaluated by alizarin red staining, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot. The formation of extracellular matrix was assessed by qRT-PCR and immunofluorescence. The expressions of receptor activator of nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG) were measured by western blot. RESULTS: Knockdown of periostin inhibited osteogenic differentiation and promoted adipogenic differentiation of DFSCs. In an inflammatory microenvironment, knockdown of periostin attenuated the proliferation and osteogenic differentiation of DFSCs. Knockdown of periostin inhibited the formation of extracellular matrix collagen I (COL-I), fibronectin, and laminin in DFSC sheets, but did not affect the expression of osteogenesis-related markers alkaline phosphatase (ALP) and osteocalcin (OCN). In the inflammatory microenvironment, knocking down periostin inhibited the expression of OCN and OPG in DFSC sheets, and promoted the expression of RANKL. CONCLUSION: Periostin played a key role in maintaining the osteogenic abilities of DFSCs and DFSC sheets in the inflammatory microenvironment and might be an important molecule in the process of DFSCs coping with inflammatory microenvironment and promoting periodontal tissues regeneration.


Assuntos
Saco Dentário , Osteogênese , Células Cultivadas , Células-Tronco , Diferenciação Celular , Osteocalcina/metabolismo , Ligamento Periodontal
10.
Front Biosci (Landmark Ed) ; 28(5): 104, 2023 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-37258464

RESUMO

BACKGROUND: Dental follicle cells (DFCs) are promising candidates for tissue engineering. However, the molecular mechanisms that regulate the biological characteristics of DFCs are still unclear. Transient receptor potential melastatin 7 (TRPM7) is a Ca2+- and Mg2+-permeable cation channel. The aim of this study was to determine the impact of TRPM7 on the proliferation, migration and osteogenic differentiation of DFCs. METHODS: PCR, Western blotting, Immunocytochemical staining and Patch clamp methods were used to identify the gene and protein expression of TRPM7 in DFCs. DFCs were infected with lentiviruses that expressed either TRPM7 specific shRNA or scrambled non-effective shRNA to investigate its functional role. Cell proliferation and migration were assessed using Cell Counting Kit-8 assays and transwell cell culture chambers separately. Cell osteogenic differentiation were determined by ALP assay kit and Alizarin Red staining. RESULTS: Gene and protein expression of TRPM7 were detected in DFCs, but not of TRPM6, which is a closely related channel with similar function. In the absence of Mg2+, typical whole cell TRPM7-like currents were recorded by patch clamp. These were inhibited by low concentrations of 2-APB, but activated by high concentrations of 2-APB. Functional studies demonstrated that suppression of TRPM7 expression inhibited the proliferation and migration of DFCs, and promoted their osteogenic differentiation. Furthermore, Mg2+ deficiency mimicked the effects of TRPM7 knockdown in terms of osteogenic differentiation of DFCs. CONCLUSIONS: These results demonstrate that TRPM7 is involved in regulating the proliferation, migration and osteogenic differentiation of DFCs.


Assuntos
Osteogênese , Canais de Cátion TRPM , Humanos , Osteogênese/genética , Magnésio/farmacologia , Magnésio/metabolismo , Canais de Cátion TRPM/genética , Saco Dentário/metabolismo , Diferenciação Celular/genética , Proliferação de Células/fisiologia , Células Cultivadas , RNA Interferente Pequeno/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
11.
Int J Mol Sci ; 24(7)2023 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-37047322

RESUMO

Dental follicle stem cells (DFSCs) have been verified to promote periodontal regeneration in an inflammatory microenvironment. When coping with inflammatory stimulation, DFSCs highly express periostin, a bioactive molecule closely related to periodontal homeostasis. It is worth exploring whether and how periostin plays a role in the promotion of periodontal regeneration by DFSCs. By tracking the fate of DFSCs, it was found that DFSCs significantly contributed to periodontal regeneration in rat periodontal defects while they had a low survival rate. They highly expressed periostin and improved the immune microenvironment in the defect area, especially via the recruitment and reprogramming of macrophages. Silencing periostin attenuated the effects of DFSCs in promoting periodontal regeneration and regulating macrophages. Recombinant human periostin (rhPeriostin) could not only directly promote macrophage reprogramming through the integrin αM/phosphorylated extracellular signal-regulated kinase (p-Erk)/Erk signaling pathway, but it also exhibited the potential to promote periodontal regeneration in rats when loaded in a collagen matrix. These results indicated that periostin is actively involved in the process by which DFSCs promote periodontal regeneration through the regulation of macrophages and is a promising molecular agent to promote periodontal regeneration. This study provides new insight into the mechanism by which DFSCs promote periodontal regeneration and suggests a new approach for periodontal regeneration therapy.


Assuntos
Moléculas de Adesão Celular , Saco Dentário , Periodonto , Regeneração , Transplante de Células-Tronco , Células-Tronco , Saco Dentário/citologia , Saco Dentário/fisiologia , Células-Tronco/metabolismo , Periodonto/efeitos dos fármacos , Periodonto/imunologia , Periodonto/fisiologia , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Moléculas de Adesão Celular/farmacologia , Humanos , Animais , Ratos , Proteínas Recombinantes/farmacologia , Periodontite/imunologia , Periodontite/terapia , Masculino , Ratos Sprague-Dawley
12.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 41(2): 197-202, 2023 Apr 01.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-37056186

RESUMO

OBJECTIVES: To summarize the open-eruption technique of impacted anterior maxillary teeth, this study reports a technically improved operation on surgical exposure based on dental follicles and evaluates post-treatment periodontal health considering the effect of dental follicles. METHODS: Patients who underwent open-eruption technique with unilateral labially impacted maxillary central incisors were selected. The impacted teeth were assigned to the experimental group, and the contralateral unimpacted maxillary central incisors were assigned to the control group. In the surgical exposure, the new technique makes use of dental follicles to manage the soft tissue, so as to preserve soft tissue for better aesthetic results and healthier periodontal tissue. Tooth length, root length, alveolar bone loss, and alveolar bone thickness were recorded after the therapy. RESULTS: A total of 17 patients with unilateral maxillary central incisor impaction were successfully treated. The tooth length and root length of the two groups showed a statistically significant difference between the impacted and homonym teeth, with a shorter length in the impacted tooth (P<0.05). More labial alveolar bone loss was found in the experimental group compared with that in the control group (P<0.05). The outcomes of the cementoenamel junction width, pa- latal alveolar bone loss, and alveolar bone thickness did not indicate statistical significance between the experimental and control groups (P>0.05). CONCLUSIONS: In the surgical exposure, the new technique uses dental follicles to manage the soft tissue and preserve it for better aesthetic results and healthier periodontal tissues.


Assuntos
Perda do Osso Alveolar , Dente Impactado , Humanos , Dente Impactado/cirurgia , Incisivo , Perda do Osso Alveolar/diagnóstico por imagem , Raiz Dentária , Saco Dentário , Maxila/cirurgia , Estética Dentária
13.
J Cell Physiol ; 238(7): 1542-1557, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37120836

RESUMO

Large bone defect reconstruction undergoes hypoxia and remains a major practical challenge. Bone tissue engineering with a more promising stem cell source facilitates the development of better therapeutic outcomes. Human dental follicle stem cells (hDFSCs) with superior multipotency, osteogenic capacity, and accessibility have been proven a promising cell source for bone regeneration. We previously identified a novel long noncoding RNA (lncRNA), HOTAIRM1, to be highly expressed in hDFSCs. Here we found that HOTAIRM1 overexpressed hDFSCs promoted bone regeneration in rat critical-size calvarial defect model. Mechanically, HOTAIRM1 was induced in hDFSCs under hypoxic conditions and activated HIF-1α. RNA-sequencing analysis indicated that HOTAIRM1 upregulated oxygen-sensing histone demethylases KDM6A/B and suppressed methyltransferase EZH2 via targeting HIF-1α. The osteogenic differentiation of hDFSCs was accompanied with demethylation of H3K27, and HOTAIRM1 overexpression decreased the distribution of H3K27me3 in osteogenic genes, including ALP, M-CSF, Wnt-3a, Wnt-5a, Wnt-7a, and ß-catenin, thus promoted their transcription. Our study provided evidence that HOTAIRM1 upregulated KDM6A/B and inhibited EZH2 in a HIF-1α dependent manner to enhance the osteogenesis of hDFSCs. HOTAIRM1-mediated hDFSCs may serve as a promising therapeutic approach to promote bone regeneration in clinical practice.


Assuntos
Regeneração Óssea , RNA Longo não Codificante , Animais , Humanos , Ratos , Diferenciação Celular , Saco Dentário , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Histonas/genética , Osteogênese , RNA Longo não Codificante/genética , Células-Tronco/metabolismo
14.
Arch Oral Biol ; 150: 105676, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36934664

RESUMO

OBJECTIVE: Short telomeres and genomic DNA damage are causes of cellular senescence in dental follicle cells (DFCs). DESIGN: This study examined the role of the DNA damage response (DDR) during cellular senescence of DFCs by ß-galactosidase activity and DNA damage by comet assay. Expression of genes/proteins was determined by Western Blots and reverse transcription-quantitative polymerase chain reaction, while glycolysis was enzymatically estimated. Cell cycle stages and reactive oxygen species (ROS) were investigated by flow cytometry. RESULTS: During the induction of cellular senescence gene expression of DDR genes were down-regulated, while DNA double-strand breaks occurred at the same time. Furthermore, inhibition of DNA protein kinase (DNA-PK) reduced senescence and ROS, both of which are associated with cellular senescence. In contrast, while these data suggest that inhibition of DDR is associated with the induction of cellular senescence, inhibition of DNA-PK did not result in renewal of DFCs, as inhibition resulted in typical features of depleted cells such as increased cell size and reduced cell proliferation rate. DNA-PK repression inhibited both osteogenic differentiation potential and glycolysis, which are typical features of cellular exhaustion. Moreover, DNA-PK affects cellular senescence via activation of AKT1 (protein kinase B). CONCLUSION: Our results suggest that DNA-PK promotes cellular senescence, but DFCs may control the induction of cellular senescence via down-regulation of DDR genes. However, we also showed that inhibition of DNA-PK cannot renew senescent DFCs.


Assuntos
Osteogênese , Proteínas Quinases , Proteínas Quinases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Polinucleotídeo 5'-Hidroxiquinase/genética , Polinucleotídeo 5'-Hidroxiquinase/metabolismo , Saco Dentário , Senescência Celular , Proteínas/metabolismo , Dano ao DNA , DNA
15.
Oral Dis ; 29(5): 2297-2309, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35509129

RESUMO

OBJECTIVE: It aims to explore the effect of dental follicle cells-derived small extracellular vesicles (D-sEVs) with or without lipopolysaccharides (LPS) pretreating on the pathogenicity of Porphyromonas gingivalis (P. gingivalis). METHODS: The antibacterial effects of D-sEV were evaluated by measuring the growth, biofilm formation, gingipains, and type IX secretion system (T9SS) expression of P. gingivalis. And the influence of D-sEV on P. gingivalis adhesion, invasion, cytotoxicity, and host immune response was examined in gingival epithelial cells (GECs). Then P. gingivalis treated with D-sEV was applied to investigate the pathogenicity in experimental periodontitis of mice. RESULTS: It showed that both D-sEV and P. gingivalis LPS-pretreated D-sEV (L-D-sEV) could target P. gingivalis, inhibit their growth and biofilm formation, and hinder the attachment and invasion in GECs, therefore remarkably decreasing P. gingivalis cytotoxicity and the expression of IL-1ß and IL-6 in GECs. In addition, they significantly reduced the expression of P. gingivalis virulence factors (gingipains and T9SS). In vivo, it showed that the bacteria in the gingiva were significantly decreased after sEV treatment. Meanwhile, less bone loss and fewer inflammatory cells infiltration and osteoclast formation in D-sEV and L-D-sEV groups. CONCLUSION: Both D-sEV and L-D-sEV were proven to inhibit the pathogenicity of P. gingivalis and thus prevented the development of periodontitis.


Assuntos
Vesículas Extracelulares , Periodontite , Animais , Camundongos , Porphyromonas gingivalis/metabolismo , Virulência , Cisteína Endopeptidases Gingipaínas/metabolismo , Lipopolissacarídeos/farmacologia , Saco Dentário , Periodontite/metabolismo , Gengiva
16.
J Dent Res ; 102(4): 431-439, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36515316

RESUMO

Root apical complex, including Hertwig's epithelial root sheath, apical papilla, and dental follicle (DF), is the germinal center of root development, wherein the DF constantly develops into periodontal tissue. However, whether DF development is regulated by the adjacent apical papilla remains largely unknown. In this study, we employed a transwell coculture system and found that stem cells from the apical papilla (SCAPs) inhibit the differentiation and maintain the stemness of dental follicle stem cells (DFSCs). Meanwhile, partial SCAP differentiation markers were upregulated after DFSC coculture. High-throughput RNA sequencing revealed that the Hedgehog (Hh) pathway was significantly downregulated in DFSCs cocultured with SCAPs. Upregulation or downregulation of the Hh pathway can respectively activate or inhibit the multidirectional differentiation of DFSCs. Osteoglycin (OGN) (previously known as mimecan) is highly expressed in the dental papilla, similarly to Hh pathway factors. By secreting OGN, SCAP regulated the stemness and multidirectional differentiation of DFSCs via the OGN-Hh pathway. Finally, Ogn-/- mice were established using the CRISPR/Cas9 system. We found that the root length growth rate was accelerated during root development from PN0 to PN30 in Ogn-/- mice. Moreover, the hard tissues (including dentin and cementum) of the root in Ogn-/- mice were thicker than those in wild-type mice. These phenotypes were likely due to Hh pathway activation and the increased cell proliferation and differentiation in both the apical papilla and DF. The current work elucidates the molecular regulation of early periodontal tissue development, providing a theoretical basis for future research on tooth root biology and periodontal tissue regeneration.


Assuntos
Raiz Dentária , Dente , Animais , Camundongos , Diferenciação Celular , Proliferação de Células , Células Cultivadas , Cemento Dentário , Papila Dentária , Saco Dentário , Proteínas Hedgehog , Osteogênese
17.
Exp Gerontol ; 172: 112071, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36563529

RESUMO

Dental Follicle Cells (DFCs) are somatic stem cells with a limited lifespan, but little is known about a possible mechanism of cellular senescence. Previous studies have shown that cellular senescence is associated with increased demand of glycolsis or the "glycolytic metabotype", which can be induced by activation of 5' adenosine monophosphate-activated protein kinase (AMPK), and decreased autophagy. This study examined the role of AMPK in inducing senescence in DFCs. During the induction of cellular senescence, AMPK activity was impaired, suggesting a negative impact on senescence induction. In line with this assumption, cellular senescence was induced upon inhibition of AMPK with a specific siRNA. In addition, after this inhibition, autophagy was also inhibited. Moreover, specific inhibition of autophagy promoted cellular senescence. However, inducers of AMPK such as metformin or AICAR surprisingly increased senescence in DFCs. Interestingly, autophagy was impaired after long-term induction of AMPK with AICAR and metformin. Moreover, activation of AMPK induces the consumption of glucose but decreases NAD/NADH ratio in DFCs that suggest not only "glycolytic metabotype" of DFCs but also Mitochondrial Dysfunction Associated Senescence (MiDAS). Both changes are highly associated with the induction of cellular senescence. Hence, both AMPK activation and inhibition promote the induction of cellular senecence of DFCs.


Assuntos
Proteínas Quinases Ativadas por AMP , Metformina , Humanos , Proteínas Quinases Ativadas por AMP/metabolismo , Saco Dentário/metabolismo , Senescência Celular , Metformina/farmacologia , Fosforilação , Autofagia
18.
Macromol Biosci ; 23(1): e2200347, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36353916

RESUMO

Nanofibrous microspheres (NFM) are emerging as prominent next-generation biomimetic injectable scaffold system for stem cell delivery and different tissue regeneration where nanofibrous topography facilitates ECM-like stem cells niches. Addition of osteogenic bioactive nanosilicate platelets within NFM can provide osteoconductive cues to facilitate matrix mediated osteogenic differentiation of stem cells and enhance the efficiency of bone tissue regeneration. In this study, gelatin nanofibrous microspheres are prepared containing fluoride-doped laponite XL21 (LP) using the emulsion mediated thermal induce phase separation (TIPS) technique. Systematic studies are performed to understand the effect of physicochemical properties of biomimicking NFM alone and with different concentrations of LP on human dental follicle stem cells (hDFSCs), their cellular attachment, proliferation, and osteogenic differentiation. The study highlights the effect of LP nanosilicate with biomimicking nanofibrous injectable scaffold system aiding in enhancing stem cell differentiation under normal physiological conditions compared to NFM without LP. The laponite-NFM shows suitability as excellent injectable biomaterials system for stem cell attachment, proliferation and osteogenic differentiation for stem cell transplantation and bone tissue regeneration.


Assuntos
Nanofibras , Osteogênese , Humanos , Gelatina/farmacologia , Gelatina/química , Microesferas , Nanofibras/química , Saco Dentário , Diferenciação Celular , Transplante de Células-Tronco , Alicerces Teciduais/química , Engenharia Tecidual/métodos
19.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-981112

RESUMO

OBJECTIVES@#To summarize the open-eruption technique of impacted anterior maxillary teeth, this study reports a technically improved operation on surgical exposure based on dental follicles and evaluates post-treatment periodontal health considering the effect of dental follicles.@*METHODS@#Patients who underwent open-eruption technique with unilateral labially impacted maxillary central incisors were selected. The impacted teeth were assigned to the experimental group, and the contralateral unimpacted maxillary central incisors were assigned to the control group. In the surgical exposure, the new technique makes use of dental follicles to manage the soft tissue, so as to preserve soft tissue for better aesthetic results and healthier periodontal tissue. Tooth length, root length, alveolar bone loss, and alveolar bone thickness were recorded after the therapy.@*RESULTS@#A total of 17 patients with unilateral maxillary central incisor impaction were successfully treated. The tooth length and root length of the two groups showed a statistically significant difference between the impacted and homonym teeth, with a shorter length in the impacted tooth (P<0.05). More labial alveolar bone loss was found in the experimental group compared with that in the control group (P<0.05). The outcomes of the cementoenamel junction width, pa- latal alveolar bone loss, and alveolar bone thickness did not indicate statistical significance between the experimental and control groups (P>0.05).@*CONCLUSIONS@#In the surgical exposure, the new technique uses dental follicles to manage the soft tissue and preserve it for better aesthetic results and healthier periodontal tissues.


Assuntos
Humanos , Dente Impactado/cirurgia , Incisivo , Perda do Osso Alveolar/diagnóstico por imagem , Raiz Dentária , Saco Dentário , Maxila/cirurgia , Estética Dentária
20.
BMC Oral Health ; 22(1): 642, 2022 12 26.
Artigo em Inglês | MEDLINE | ID: mdl-36567318

RESUMO

OBJECTIVES: The indication for removal of asymptomatic fully impacted third molars is still controversial. In this study, radiological and histological investigation of the dental follicle of asymptomatic impacted mandibular third molars was performed, aiming to provide a reference for clinical prophylactic extraction of these teeth. METHODS: Patients with impacted mandibular third molars were included and the maximum width of the dental follicle around the crown was measured in horizontal, sagittal and coronal sections by cone beam computed tomography. The dental follicles were stained with haematoxylin-eosin, analysed by a pathologist and classified as normal, inflammatory or cystic. A Chi-squared test was used to analyse the association of the incidence of inflammation and cysts with the clinical variables of the impacted mandibular third molars. RESULTS: Thirty-seven samples were normal dental follicles; 52 samples showed inflammatory infiltration with an incidence of 57.14%; 2 samples with a maximum dental follicle width of 2-3 mm were diagnosed as odontogenic cysts, and the incidence was 2.20%. There was no significant difference in the incidence of inflammatory and cystic dental follicles between males and females, or between different age groups (P > 0.05). With an increase of the maximum width of the dental follicle, there was a rise in the incidence and degree of infiltration of chronic nonspecific inflammation. CONCLUSION: Asymptomatic impacted mandibular third molars tend to be extracted, especially for teeth with a 2-3 mm maximum width of the dental follicle on radiological examination.


Assuntos
Dente Serotino , Dente Impactado , Masculino , Feminino , Humanos , Dente Serotino/diagnóstico por imagem , Saco Dentário , Dente Molar/patologia , Mandíbula/diagnóstico por imagem , Mandíbula/patologia , Dente Impactado/diagnóstico por imagem , Dente Impactado/patologia , Inflamação/patologia
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