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1.
Braz Dent J ; 33(2): 73-82, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35508039

RESUMO

This study was conducted to assess the in vitro response of human periodontal ligament stem cells (hPDLSCs) to bacterial lipopolysaccharide (LPS) activation and application of three calcium silicate-based materials (CSBM): Bio-C Sealer, MTA Fillapex and Cimmo HP. Characterization of the CSBM was performed by FTIR (n = 3). Extracts of Bio-C Sealer, MTA Fillapex and Cimmo HP were prepared and diluted (1:1, 1:4 and 1:16). Culture of hPDLSCs was established and treated or not with LPS from Escherichia coli (1 µg/mL) for 7 days. MTT assay was used to assess cell viability at 24, 48 and 72 h (n = 9). Alkaline phosphatase (ALP) activity was indirectly assayed at day 7 (n = 5). TNF-α and Il -1 0 cytokines were quantified by ELISA at 24h-cell supernatants (n = 6). Data were analyzed by ANOVA and Tukey's test (α = 0.05). The cell viability of the LPS-activated hPDLSCs were higher than untreated control (p < 0.05). The application of CSBM affected the cell viability of untreated and LPS-activated cells (p < 0.05). ALP activity was higher for Bio-C Sealer and Cimmo HP in untreated and LPS-activated cells, respectively (p < 0.05). Application of CSBM normalized the TNF-α secretion in the LPS-activated cells (p < 0.05). Only MTA Fillapex in untreated hPDLSCs presented higher values of Il -1 0 (p < 0.05). Taken collectively, the results suggests that the simulation of the inflammatory process by LPS affect the in vitro response the hPDLSCs to the application of the CSBM.


Assuntos
Ligamento Periodontal , Materiais Restauradores do Canal Radicular , Compostos de Cálcio/farmacologia , Células Cultivadas , Humanos , Lipopolissacarídeos/farmacologia , Pemetrexede , Silicatos/farmacologia , Células-Tronco , Fator de Necrose Tumoral alfa
2.
Comput Math Methods Med ; 2022: 4634925, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35509853

RESUMO

Objective: miR-141-3p has been demonstrated to be both anti-inflammatory and osteoprotective. This study is aimed at investigating the effect of miR-141-3p on osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) stimulated by Porphyromonas gingivalis lipopolysaccharide (PgLPS) and its mechanism. Methods: PgLPS was used to induce an inflammatory environment, and overexpression of miR-141-3p was done to assess its effect on hPDLSCs in an inflammatory environment. The level of miR-141-3p and EZH2 in hPDLSCs from each treatment group was detected via qRT-PCR, and the inflammatory factors IL-6 and IL-8 in the supernatant of each group were detected by ELISA. ALP staining and alizarin red staining were used to assess the effect of miR-141-3p on the osteogenic differentiation ability of hPDLSCs, and also, western blot was used to detect expression of osteogenic differentiation-related proteins. Further, dual-luciferase reporter assay examined whether miR-141-3p targeted EZH2. Results: PgLPS led to a significant decrease of miR-141-3p in hPDLSCs. Overexpression of miR-141-3p could enhance ALP activity and alizarin red staining intensity and increase Runx2, OPN and OCN protein expression levels in PgLPS-treated hPDLSCs. Additionally, miR-141-3p could reduce IL-6 and IL-8. miR-141-3p could target and negatively regulate EZH2, and overexpression of EZH2 reversed the promoting effect of miR-141-3p on osteogenic differentiation. Conclusion: miR-141-3p can attenuate PgLPS-induced inhibition of osteogenic differentiation and inflammation in hPDLSCs by negatively regulating EZH2.


Assuntos
MicroRNAs , Ligamento Periodontal , Diferenciação Celular , Células Cultivadas , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/farmacologia , Humanos , Inflamação/genética , Inflamação/metabolismo , Interleucina-6/genética , Interleucina-8/genética , Interleucina-8/metabolismo , Interleucina-8/farmacologia , Lipopolissacarídeos , MicroRNAs/genética , MicroRNAs/metabolismo , Osteogênese , Porphyromonas gingivalis/metabolismo , Células-Tronco/metabolismo
3.
Bioengineered ; 13(4): 9602-9612, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35435112

RESUMO

Periodontitis is a chronic inflammation caused by the deposition of dental plaque on the tooth surface. Human periodontal ligament stem cells (hPDLSCs) have the potential of osteogenic differentiation. Long non-coding RNAs (lncRNAs) are collectively involved in periodontitis. This study was designed to explore the roles of Linc01133 in osteogenic differentiation of hPDLSCs. hPDLSCs obtained from the periodontal ligament (PDL) of patients with periodontitis were used to collect Linc01133, microRNA-30c (miR-30c), and bone gamma-carboxyglutamate protein (BGLAP) expression data, and their expression changes were traced during osteogenic differentiation of hPDLSCs. Quantitative reverse-transcription polymerase chain reaction as well as western blotting were used to analyze the levels of RNAs and proteins. Dual-luciferase reporter and RNA pull-down assays demonstrated the relationship between Linc01133, miR-30c, and BGLAP. Furthermore, alkaline phosphatase (ALP) staining and alizarin red staining were applied to evaluate the degree of osteogenic differentiation. Linc01133 was downregulated in the PDL of patients with periodontitis. Upregulated Linc01133 promoted osteogenic differentiation of hPDLSCs. Linc01133 could inhibit miR-30c expression by sponging miR-30c. miR-30c suppressed osteogenic differentiation. Additionally, miR-30c targeted BGLAP. Knockdown of BGLAP abrogated the effects of decreased miR-30c on osteogenic differentiation of hPDLSCs. Linc01133 acted as a ceRNA to regulate osteogenic differentiation of hPDLSCs via the miR-30c/BGLAP axis. Therefore, Linc01133 may participate in the progress of periodontitis.


Assuntos
MicroRNAs , Periodontite , RNA Longo não Codificante , Ácido 1-Carboxiglutâmico/metabolismo , Diferenciação Celular/genética , Células Cultivadas , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Osteogênese/genética , Ligamento Periodontal , Periodontite/genética , Periodontite/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Células-Tronco
4.
Braz Oral Res ; 36: e048, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35442377

RESUMO

PAR1 is a G-coupled protein receptor that regulates several cellular metabolism processes, including differentiation and proliferation of osteogenic and cementogenic related cells and our group previously demonstrated the regenerative potential of PAR1 in human periodontal ligament stem cells (hPDLSCs). In this study, we hypothesized that PAR1 regulates the cementogenic differentiation of hPDLSCs. Our goal was to identify the intracellular signaling pathway underlying PAR1 activation in hPDSLC differentiation. hPDLSCs were isolated using the explant technique. Cells were cultured in an osteogenic medium (OST) (α-MEM, 15% fetal bovine serum, L-glutamine, penicillin, streptomycin, amphotericin B, dexamethasone, and beta-glycerophosphate). The hPDLSCs were treated with a specific activator of PAR1 (PAR1 agonist) and blockers of the MAPK/ERK and PI3K pathways for 2 and 7 days. The gene expression of CEMP1 was assessed by RT-qPCR. The activation of PAR1 by its agonist peptide led to an increase in CEMP1 gene expression when compared with OST control. MAPK/ERK blockage abrogated the upregulation of CEMP1 gene expression induced by PAR1 agonist (p < 0.05). PI3K blockage did not affect the gene expression of CEMP1 at any experimental time (p > 0.05). We concluded that CEMP1 gene expression increased by PAR1 activation is MAPK/ERK-dependent and PI3K independent, suggesting that PAR1 may regulate cementogenetic differentiation of hPDLSCs.


Assuntos
Sistema de Sinalização das MAP Quinases , Receptor PAR-1 , Diferenciação Celular , Células Cultivadas , Expressão Gênica , Humanos , Osteogênese , Ligamento Periodontal , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas , Receptor PAR-1/genética , Receptor PAR-1/metabolismo
5.
Int J Mol Sci ; 23(7)2022 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-35409397

RESUMO

Periodontal ligament derived stem cells (PDLSC) are adult multipotent mesenchymal-like stem cells (MSCs) that can induce a promising immunomodulation to interact with immune cells for disease treatment. Metabolic reconfiguration has been shown to be involved in the immunomodulatory activity of MSCs. However, the underlying mechanisms are largely unknown, and it remains a challenging to establish a therapeutic avenue to enhance immunomodulation of endogenous stem cells for disease management. In the present study, RNA-sequencing (RNA-seq) analysis explores that curcumin significantly promotes PDLSC function through activation of MSC-related markers and metabolic pathways. In vitro stem cell characterization further confirms that self-renewal and multipotent differentiation capabilities are largely elevated in curcumin treated PDLSCs. Mechanistically, RNA-seq reveals that curcumin activates ERK and mTOR cascades through upregulating growth factor pathways for metabolic reconfiguration toward glycolysis. Interestingly, PDLSCs immunomodulation is significantly increased after curcumin treatment through activation of prostaglandin E2-Indoleamine 2,3 dioxygenase (PGE2-IDO) signaling, whereas inhibition of glycolysis activity by 2-deoxyglucose (2-DG) largely blocked immunomodulatory capacity of PDLSCs. Taken together, this study provides a novel pharmacological approach to activate endogenous stem cells through metabolic reprogramming for immunomodulation and tissue regeneration.


Assuntos
Curcumina , Células-Tronco Mesenquimais , Diferenciação Celular/fisiologia , Proliferação de Células , Células Cultivadas , Curcumina/metabolismo , Curcumina/farmacologia , Imunomodulação , Células-Tronco Mesenquimais/metabolismo , Ligamento Periodontal
6.
Genes (Basel) ; 13(4)2022 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-35456465

RESUMO

Ligament-fibroblastic cells and cementoblasts, two types of progenitor cells that differentiate from periodontal ligament stem cells (hPDLSCs), are responsible for the formation of the adhesive tissues in the tooth root. Since one of the factors that determines the fate of stem cell differentiation is the change in the microenvironment of the stem/progenitor cells, this study attempted to compare and analyze the molecular differences in the membrane and ECM of the two progenitor cells. Single cells derived from hPDLSCs were treated with TGF-ß1 and BMP7 to obtain ligament-fibroblastic and cementoblastic cells, respectively. The transcriptome profiles of three independent replicates of each progenitor were evaluated using next-generation sequencing. The representative differentially expressed genes (DEGs) were verified by qRT-PCR, Western blot analysis, and immunohistochemistry. Among a total of 2245 DEGs identified, 142 and 114 DEGs related to ECM and cell membrane molecules were upregulated in ligament-fibroblastic and cementoblast-like cells, respectively. The major types of integrin and cadherin were found to be different between the two progenitor cells. In addition, the representative core proteins for each glycosaminoglycan-specific proteoglycan class were different between the two progenitors. This study provides a detailed understanding of cell-cell and cell-ECM interactions through the specific components of the membrane and ECM for ligament-fibroblastic and cementoblastic differentiation of hPDLSCs.


Assuntos
Cemento Dentário , Ligamento Periodontal , Diferenciação Celular/genética , Matriz Extracelular/genética , Matriz Extracelular/metabolismo , Humanos , Ligamentos , Ligamento Periodontal/metabolismo , Transcriptoma/genética
7.
J Appl Oral Sci ; 30: e20210359, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35384987

RESUMO

Regenerative approaches using mesenchymal stem cells (MSCs) have been evaluated to promote the complete formation of all missing periodontal tissues, e.g., new cementum, bone, and functional periodontal ligaments. MSCs derived from bone marrow have been applied to bone and periodontal defects in several forms, including bone marrow aspirate concentrate (BMAC) and cultured and isolated bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate the periodontal regeneration capacity of BMAC and cultured BM-MSCs in the wound healing of fenestration defects in rats. METHODOLOGY: BM-MSCs were obtained after bone marrow aspiration of the isogenic iliac crests of rats, followed by cultivation and isolation. Autogenous BMAC was collected and centrifuged immediately before surgery. In 36 rats, fenestration defects were created and treated with suspended BM-MSCs, BMAC or left to spontaneously heal (control) (N=6). Their regenerative potential was assessed by microcomputed tomography (µCT) and histomorphometry, as well as their cell phenotype and functionality by the Luminex assay at 15 and 30 postoperative days. RESULTS: BMAC achieved higher bone volume in 30 days than spontaneous healing (p<0.0001) by enhancing osteoblastic lineage commitment maturation, with higher levels of osteopontin (p=0.0013). Defects filled with cultured BM-MSCs achieved higher mature bone formation in early stages than spontaneous healing and BMAC (p=0.0241 and p=0.0143, respectively). Moreover, significantly more cementum-like tissue formation (p<0.0001) was observed with new insertion of fibers in specimens treated with BM-MSCs within 30 days. CONCLUSION: Both forms of cell transport, BMAC and BM-MSCs, promoted bone formation. However, early bone formation and maturation were achieved when cultured BM-MSCs were used. Likewise, only cultured BM-MSCs were capable of achieving complete periodontal regeneration with inserted fibers in the new cementum-like tissue.


Assuntos
Medula Óssea , Células-Tronco Mesenquimais , Animais , Células da Medula Óssea , Regeneração Óssea , Ligamento Periodontal , Ratos , Microtomografia por Raio-X
8.
J Orthop Surg Res ; 17(1): 221, 2022 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-35399081

RESUMO

BACKGROUND: An increasing number of studies have shown that dysregulated miR-589-3p is associated with multiple diseases. However, the role of miR-589-3p in osteogenic differentiation of periodontal ligament stem cells (PDLSCs) remains unknown. This study aimed to explore the biological function and potential molecular mechanism of miR-589-3p in osteogenic differentiation of PDLSCs. METHODS: GSE159508 was downloaded from Gene Expression Omibus (GEO, http://www.ncbi.nlm.nih.gov/geo/ ). Differentially expressed miRNAs between osteogenic induction PDLSCs versus non-induction PDLSCs were obtained by R software. miR-589-3p mimic and miR-589-3p inhibitor and corresponding negative control were obtained and to identify the role of miR-589-3p in osteogenic differentiation of PDLSCs. ALP staining and ARS were used to evaluate ALP activity and mineralization, respectively. The targeted binding relationship between miR-589-3p and ATF1 was predicted and verified by target prediction analysis and dual-luciferase assay. Furthermore, the functional mechanism based on miR-589-3p and ATF1 in osteogenic differentiation of PDLSCs was further investigated through rescue experiments. RESULTS: According to the cut-off criteria with log 2 FC > 1.0 and P < 0.05, 514 differentially expressed miRNAs were identified between osteogenic induction and non-induction PDLSCs, including 309 upregulated miRNAs and 205 downregulated miRNAs. Compared with control PDLSCs, miR-589-3p expression level was notably increased in PDLSCs that underwent osteogenic induction. The overexpression of miR-589-3p promoted the cell viability of PDLSCs, while the low expression of miR-589-3p had the opposite effect. The dual luciferase reporter assay verified that ATF1 was proved to be a direct target of miR-589-3p in PDLSCs. And overexpressed miR-589-3p reduced the expression of ATF1. Overexpression of miR-589-3p enhanced the osteogenic capacity of PDLSCs, as demonstrated by increases in ALP activity, matrix mineralization, and RUNX2, OCN and OSX expression. In addition, the rescue experiments confirmed that overexpressed ATF1 restored the effects of overexpressed miR-589-3p on cell proliferation and osteogenic differentiation of PDLSCs. CONCLUSION: miR-589-3p could down-regulate the expression of ATF1, thereby promote the proliferation and osteogenic differentiation of PDLSCs. This finding may provide a new therapeutic target for molecular therapy of periodontitis.


Assuntos
MicroRNAs , Ligamento Periodontal , Diferenciação Celular/genética , Células Cultivadas , MicroRNAs/metabolismo , Osteogênese , Ligamento Periodontal/metabolismo , Células-Tronco
9.
Cells ; 11(7)2022 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-35406691

RESUMO

Periodontal ligament stem cells (PDLCs) can be used as a valuable source in cell therapies to regenerate bone tissue. However, the potential therapeutic outcomes are unpredictable due to PDLCs' heterogeneity regarding the capacity for osteoblast differentiation and mineral nodules production. Here, we identify epigenetic (DNA (hydroxy)methylation), chromatin (ATAC-seq) and transcriptional (RNA-seq) differences between PDLCs presenting with low (l) and high (h) osteogenic potential. The primary cell populations were investigated at basal state (cultured in DMEM) and after 10 days of osteogenic stimulation (OM). At a basal state, the expression of transcription factors (TFs) and the presence of gene regulatory regions related to osteogenesis were detected in h-PDLCs in contrast to neuronal differentiation prevalent in l-PDLCs. These differences were also observed under stimulated conditions, with genes and biological processes associated with osteoblast phenotype activated more in h-PDLCs. Importantly, even after the induction, l-PDLCs showed hypermethylation and low expression of genes related to bone development. Furthermore, the analysis of TFs motifs combined with TFs expression suggested the relevance of SP1, SP7 and DLX4 regulation in h-PDLCs, while motifs for SIX and OLIG2 TFs were uniquely enriched in l-PDLCs. Additional analysis including a second l-PDLC population indicated that the high expression of OCT4, SIX3 and PPARG TFs could be predictive of low osteogenic commitment. In summary, several biological processes related to osteoblast commitment were activated in h-PDLCs from the onset, while l-PDLCs showed delay in the activation of the osteoblastic program, restricted by the persistent methylation of gene related to bone development. These processes are pre-determined by distinguishable epigenetic and transcriptional patterns, the recognition of which could help in selection of PDLCs with pre-osteoblastic phenotype.


Assuntos
Osteogênese , Ligamento Periodontal , Células Cultivadas , Cromatina/metabolismo , Proteínas de Homeodomínio/metabolismo , Humanos , Metilação , Osteogênese/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
10.
Cells ; 11(7)2022 03 30.
Artigo em Inglês | MEDLINE | ID: mdl-35406732

RESUMO

Periodontal disease can cause irreversible damage to tooth-supporting tissues such as the root cementum, periodontal ligament, and alveolar bone, eventually leading to tooth loss. While standard periodontal treatments are usually helpful in reducing disease progression, they cannot repair or replace lost periodontal tissue. Periodontal regeneration has been demonstrated to be beneficial in treating intraosseous and furcation defects to varied degrees. Cell-based treatment for periodontal regeneration will become more efficient and predictable as tissue engineering and progenitor cell biology advance, surpassing the limitations of present therapeutic techniques. Stem cells are undifferentiated cells with the ability to self-renew and differentiate into several cell types when stimulated. Mesenchymal stem cells (MSCs) have been tested for periodontal regeneration in vitro and in humans, with promising results. Human umbilical cord mesenchymal stem cells (UC-MSCs) possess a great regenerative and therapeutic potential. Their added benefits comprise ease of collection, endless source of stem cells, less immunorejection, and affordability. Further, their collection does not include the concerns associated with human embryonic stem cells. The purpose of this review is to address the most recent findings about periodontal regenerative mechanisms, different stem cells accessible for periodontal regeneration, and UC-MSCs and their involvement in periodontal regeneration.


Assuntos
Células-Tronco Mesenquimais , Humanos , Ligamento Periodontal , Periodonto , Engenharia Tecidual , Cordão Umbilical
11.
J Med Microbiol ; 71(4)2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35451943

RESUMO

Periodontitis is initiated by hyper-inflammatory responses in the periodontal tissues that generate dysbiotic ecological changes within the microbial communities. As a result, supportive tissues of the tooth are damaged and periodontal attachment is lost. Gingival recession, formation of periodontal pockets with the presence of bleeding, and often suppuration and/or tooth mobility are evident upon clinical examination. These changes may ultimately lead to tooth loss. Mesenchymal stem cells (MSCs) are implicated in controlling periodontal disease progression and have been shown to play a key role in periodontal tissue homeostasis and regeneration. Evidence shows that MSCs interact with subgingival microorganisms and their by-products and modulate the activity of immune cells by either paracrine mechanisms or direct cell-to-cell contact. The aim of this review is to reveal the interactions that take place between microbes and in particular periodontal pathogens and MSCs in order to understand the factors and mechanisms that modulate the regenerative capacity of periodontal tissues and the ability of the host to defend against putative pathogens. The clinical implications of these interactions in terms of anti-inflammatory and paracrine responses of MSCs, anti-microbial properties and alterations in function including their regenerative potential are critically discussed based on literature findings. In addition, future directions to design periodontal research models and study ex vivo the microbial-stem cell interactions are introduced.


Assuntos
Células-Tronco Mesenquimais , Periodontite , Comunicação Celular , Humanos , Células-Tronco Mesenquimais/fisiologia , Ligamento Periodontal/fisiologia , Células-Tronco
12.
Anesth Prog ; 69(1): 10-17, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35377934

RESUMO

OBJECTIVE: The purpose of this prospective randomized study was to assess using a periodontal ligament (PDL) injection as an aide to decrease palatal infiltration pain. METHODS: A total of 133 subjects randomly received a PDL injection and alternative palatal infiltration or a mock PDL injection and conventional palatal infiltration at 2 separate appointments. PDL injection was given in the mid-palatal sulcus of the maxillary first molar. Mock PDL injection consisted of only needle insertion. All subjects then received a palatal infiltration administered into the blanched gingival tissue 3 mm (alternative palatal infiltration) or 7 mm (conventional palatal infiltration) from the gingival collar. Subjects recorded needle insertion and solution deposition pain using a Heft-Parker visual analog scale (VAS). RESULTS: The combined PDL injection and alternative palatal infiltration had significantly decreased mean VAS ratings for needle insertion and solution deposition pain (P < .0001). Incidence of moderate/severe pain for needle insertion and solution deposition was reduced from 65% to 1% and from 65% to 2%, respectively. CONCLUSION: Providing PDL anesthesia into the mid-palatal sulcus of the maxillary first molar and then administering an alternative palatal infiltration into the blanched collar around the PDL molar site led to significant reductions in needle insertion and solution deposition pain compared with a mock PDL and conventional palatal infiltration.


Assuntos
Anestesia Dentária , Anestesiologia , Humanos , Dor , Ligamento Periodontal , Estudos Prospectivos
14.
Biomolecules ; 12(3)2022 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-35327627

RESUMO

Different approaches to develop engineered scaffolds for periodontal tissues regeneration have been proposed. In this review, innovations in stem cell technology and scaffolds engineering focused primarily on Periodontal Ligament (PDL) regeneration are discussed and analyzed based on results from pre-clinical in vivo studies and clinical trials. Most of those developments include the use of polymeric materials with different patterning and surface nanotopography and printing of complex and sophisticated multiphasic composite scaffolds with different compartments to accomodate for the different periodontal tissues' architecture. Despite the increased effort in producing these scaffolds and their undoubtable efficiency to guide and support tissue regeneration, appropriate source of cells is also needed to provide new tissue formation and various biological and mechanochemical cues from the Extraccellular Matrix (ECM) to provide biophysical stimuli for cell growth and differentiation. Cell sheet engineering is a novel promising technique that allows obtaining cells in a sheet format while preserving ECM components. The right combination of those factors has not been discovered yet and efforts are still needed to ameliorate regenerative outcomes towards the functional organisation of the developed tissues.


Assuntos
Ligamento Periodontal , Tecidos Suporte , Diferenciação Celular , Tecnologia , Engenharia Tecidual/métodos
15.
Ann Afr Med ; 21(1): 34-38, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35313402

RESUMO

Background: Replantation is a commonly performed method for avulsed tooth. A vital periodontal membrane (periodontal ligament [PDL]) is significant for the successful healing of replanted teeth. Hence, various storage media are used to preserve the viability of periodontal cells before replantation. Objectives: The present study was conducted to evaluate the efficacy of ViaSpan, Aloe vera, Gatorade solution, and propolis storage media for maintaining the PDL cell viability. Materials and Methods: The present study was conducted on 40 recently extracted teeth which were randomly divided into four study storage groups: Group I: ViaSpan, Group II: Aloe vera, Group III: Gatorade solution, and Group IV: Propolis. Later they were subjected to centrifugation, and the cells from supernatant were colored with 0.4% trypan blue for determination of viability. The obtained data were statistically evaluated with SPSS package (21.0 version, Inc.; Chicago, IL, USA) using analysis of variance, Mann-Whitney test, and Post hoc tests. Results: The mean viable periodontal cell in Group I was 30.2 cumm, in Group II was 24.6 cumm, Group III was 14.5 cumm, and Group IV in 31.4. The difference was significant (P < 0.01). Post hoc test between different groups revealed a significant difference in mean viable periodontal cells (P < 0.001). Propolis, ViaSpan, and Aloe vera had higher pH and osmolality values. Conclusion: This study found that propolis had higher periodontal cell viability followed by ViaSpan solution and Aloe vera and least in Gatorade solution. Propolis, ViaSpan, and Aloe vera media can be used as a storage media.


Résumé Contexte: la Replantation est une méthode couramment utilisée pour la dent avulsée. Une membrane parodontale vitale (ligament parodontal [PDL]) est important pour la guérison réussie des dents replantées. Par conséquent, divers supports de stockage sont utilisés pour préserver la viabilité du parodontal les cellules avant la réimplantation. Objectifs: la présente étude a été menée pour évaluer L'efficacité de ViaSpan, Aloe vera, solution de Gatorade, et des supports de stockage de propolis pour maintenir la viabilité des cellules PDL. Matériaux et Méthodes: la présente étude a été menée sur 40 récemment dents extraites qui ont été divisées au hasard en quatre groupes de stockage d'étude: Groupe I: ViaSpan, Groupe II: aloe vera, Groupe III: Gatorade solution, et groupe IV: Propolis. Plus tard, ils ont été soumis à une centrifugation et les cellules du surnageant ont été colorées avec 0,4% de trypan bleu pour la détermination de la viabilité. Les données obtenues ont été évaluées statistiquement avec le package SPSS (version 21.0, Inc. Chicago, ILLINOIS, états-unis) en utilisant l'analyse de la variance, le test de Mann-Whitney et les tests post hoc. Résultats: la cellule parodontale viable moyenne dans le Groupe I était de 30,2 cumm, dans Le groupe II était de 24,6 cumm, le Groupe III de 14,5 cumm et le Groupe IV de 31,4. La différence était significative (P < 0,01). Essai post hoc entre différents groupes ont révélé une différence significative dans les cellules parodontales viables moyennes (P < 0,001). Propolis, ViaSpan et aloe vera avaient plus valeurs de pH et d'osmolalité. Conclusion: Cette étude a révélé que la propolis avait une viabilité cellulaire parodontale plus élevée suivie D'une solution de ViaSpan et Aloe vera et moins dans la solution de Gatorade. La Propolis, le ViaSpan et L'aloe vera peuvent être utilisés comme support de stockage. Mots-clés: aloe vera, Gatorade, cellules parodontales, propolis, ViaSpan.


Assuntos
Aloe , Própole , Adenosina , Alopurinol , Sobrevivência Celular , Glutationa , Humanos , Insulina , Soluções Isotônicas , Soluções para Preservação de Órgãos , Ligamento Periodontal , Própole/farmacologia , Rafinose
16.
Cells ; 11(6)2022 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-35326406

RESUMO

The interrelationships between periodontal disease, obesity-related hyperlipidemia and mechanical forces and their modulating effects on the epigenetic profile of periodontal ligament (PdL) cells are assumed to be remarkably complex. The PdL serves as a connective tissue between teeth and alveolar bone and is involved in pathogen defense and the inflammatory responses to mechanical stimuli occurring during tooth movement. Altered inflammatory signaling could promote root resorption and tooth loss. Hyperinflammatory COX2/PGE2 signaling was reported for human PdL fibroblasts (HPdLFs) concomitantly stressed with Porphyromonas gingivalis lipopolysaccharides and compressive force after exposure to palmitic acid (PA). The aim of this study was to investigate the extent to which this was modulated by global and gene-specific changes in histone modifications. The expression of key epigenetic players and global H3Kac and H3K27me3 levels were quantitatively evaluated in dual-stressed HPdLFs exposed to PA, revealing a minor force-related reduction in repressive H3K27me3. UNC1999-induced H3K27me3 inhibition reversed the hyperinflammatory responses of dual-stressed PA cultures characterized by increased COX2 expression, PGE2 secretion and THP1 adhesion. The reduced expression of the gene encoding the anti-inflammatory cytokine IL-10 and the increased presence of H3K27me3 at its promoter-associated sites were reversed by inhibitor treatment. Thus, the data highlight an important epigenetic interplay between the different stimuli to which the PdL is exposed.


Assuntos
Dinoprostona , Ligamento Periodontal , Células Cultivadas , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Dinoprostona/metabolismo , Fibroblastos/metabolismo , Histonas/metabolismo , Humanos , Palmitatos/metabolismo
17.
Bioengineered ; 13(3): 6966-6977, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35249460

RESUMO

Periodontitis is a chronic infectious disease that causes inflammation and immune response and has an ultimate impact on the health of the whole body. Sirtuin6 (SIRT6) and Krüppel-like factor 5 (KLF5) have been reported to regulate the inflammatory response and play an important role in the development of periodontitis. LPS was adopted to induce periodontal ligament stem cells (PDLSCs) to construct a periodontitis cell model. SIRT6 expression was assayed through RT-qPCR and Western blot. Subsequently, after SIRT6 was overexpressed, CCK8 was to appraise cell viability. ELISA analysis was used to estimate inflammatory response. ALP staining, ARS staining, and Western blot were used to detect osteogenic differentiation. The JASPAR website then predicts the binding of transcription factor KLF5 to SIRT6 promoter. The interaction between KLF5 and SIRT6 was verified by a luciferase reporter and ChIP assays. Additionally, the osteogenic differentiation and inflammation in LPS-induced PDLSCs transfected with Ov-SIRT6 and si-KIF5 were also explored. Finally, the protein levels of the nuclear factor kappa-B (NF-κB) pathway-related factors were detected by Western blot to further explore the mechanism. There was a marked decrease in SIRT6 expression in LPS-induced PDLSCs. SITR6 overexpression prevented LPS-induced cell viability loss and inflammation, while promoting osteogenic differentiation. In addition, KLF5 could transcriptionally activate SIRT6. Further, KLF5 knockdown reversed the impacts of SIRT6 on the proliferation, inflammation, and osteogenic differentiation of LPS-induced PDLSCs via mediating NF-κB pathway. Overall, KLF5-mediated SIRT6 promoted the viability and osteogenic differentiation, while inhibiting the inflammatory response of LPS-induced PDLSCs by inhibiting NF-κB pathway.


Assuntos
Periodontite , Sirtuínas , Diferenciação Celular , Células Cultivadas , Humanos , Inflamação/genética , Inflamação/metabolismo , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/farmacologia , NF-kappa B/metabolismo , Osteogênese/genética , Ligamento Periodontal , Periodontite/genética , Periodontite/metabolismo , Transdução de Sinais , Sirtuínas/genética , Sirtuínas/metabolismo , Células-Tronco
18.
J Periodontal Res ; 57(3): 545-557, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35246839

RESUMO

BACKGROUND AND OBJECTIVES: Many studies have been conducted to better understand the molecular mechanism involved with periodontitis progression. There has been growing interest in the potential impact of obesity on periodontitis onset and progression, but the mechanisms involved remain to be elucidated. The present study was designed to determine the impact of obesity on experimentally induced periodontitis in rats and identify novel pathways involved. METHODS: Sixteen Holtzman rats were distributed into two groups (n = 8): ligature-induced periodontitis (P) and obesity plus ligature-induced periodontitis (OP). Obesity was induced by a high-fat diet for 70 days, whereas periodontitis was induced for 20 days, with a cotton thread placed around the upper first molars bilaterally. Alveolar bone loss was measured by microtomographic analysis and histologically by histometry on the hemimaxillae. The protein composition of the periodontal ligament was evaluated by proteomic analysis. RESULTS: Data analysis (body weight, adipose tissue weight, and blood test) confirmed obesity induction, whereas bone loss was confirmed by micro-CT and histologic analyses. Proteome analysis from the periodontal ligament tissues (PDL) identified 819 proteins, 53 exclusive to the P group, 28 exclusive to the OP group, and 738 commonly expressed. Validation was performed by immunohistochemistry for selected proteins (spondin1, vinculin, and TRAP). CONCLUSION: Histologically, it was found that obesity did not significantly affect bone loss resulting from periodontitis. However, the present study's findings indicated that obesity affects the proteome of PDL submitted to experimental periodontitis, allowing for identifying potential targets for personalized approaches.


Assuntos
Perda do Osso Alveolar , Periodontite , Perda do Osso Alveolar/patologia , Animais , Obesidade/complicações , Ligamento Periodontal/metabolismo , Periodontite/metabolismo , Proteoma , Proteômica , Ratos , Ratos Wistar
19.
Bioengineered ; 13(3): 7951-7961, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35297308

RESUMO

Lipopolysaccharide (LPS) induces inflammatory stress and apoptosis. This study focused on the effect of nuclear factor kappa B (NF-κB) signaling pathway on proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) after LPS induction and its mechanism. We first isolated hPDLSCs from human tooth root samples in vitro. Then, flow cytometry detected positive expression of cell surface antigens CD146 and STRO-1 and negative expression of CD45, suggesting the hPDLSCs were successfully isolated. LPS significantly induced increased apoptosis and diminished proliferation of hPDLSCs. The NF-κB pathway agonist phorbol 12-myristate 13-acetate (PMA) or p65 overexpression inhibited the proliferation of LPS-treated hPDLSCs and promoted apoptosis. PMA also promoted LPS-induced up-regulation of the expression of inflammatory factors TNF-α and IL-6 and down-regulation of the expression of anti-inflammatory factor IL-10. Additionally, LPS was confirmed to lead to a reduction of alkaline phosphatase (ALP) activity, calcium nodules, and expression of osteogenic markers Runt-related transcription factor 2 (Runx2) and osteopontin. This reduction could be promoted by PMA. Western blotting further indicated that PMA could promote LPS-induced decrease of expression of p65 (cytoplasm), and total cellular proteins IKKα and IKKß in hPDLSCs, while protein expression of p-IκBα (cytoplasm) and p65 (nucleus), and p-IκBα/IκBα ratio was elevated. By contrast, inhibition of the NF-κB pathway (PDTC) or small-interfering RNA targeting NF-κB/p65 (p65 siRNA) showed the opposite results. In conclusion, activation of NF-κB signaling in LPS-induced inflammatory environment can inhibit the proliferation and osteogenic differentiation of hPDLSCs. This study provides a theory foundation for the clinical treatment of periodontitis.


Assuntos
NF-kappa B , Ligamento Periodontal , Diferenciação Celular , Células Cultivadas , Humanos , Inflamação/metabolismo , Lipopolissacarídeos/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/metabolismo , Osteogênese , Transdução de Sinais , Células-Tronco
20.
J Periodontal Res ; 57(3): 568-577, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35297053

RESUMO

BACKGROUND AND OBJECTIVE: As one of the widely expressed cell surface receptors binding to collagen, the most abundant component of the extracellular matrix (ECM), knowledge of the expression, functions, and mechanisms underlying the role of discoidin domain receptor 1 (DDR1) in human periodontal ligament cells (hPDLCs) is incomplete. This study determined the expression of DDR1 in hPDLCs and the effect of DDR1 upon migration and adhesion to hPDLCs, as well as the related regulatory mechanisms. MATERIALS AND METHODS: The expression of DDR1 and the DDR1 isoforms in hPDLCs from six donors were tested. The migratory ability (horizontal and vertical) and adhesive capacity of hPDLCs with or without specific knockdown of DDR1 were evaluated. After treatment with MEK-ERK1/2 inhibitors (PD98059 and U0126) with or without RNAi, the migratory and adhesive capacity of hPDLCs were re-tested. Western blotting was performed to verify p-MEK1/2 and p-ERK1/2, the key factors of the MEK-ERK1/2 signaling pathways. RESULTS: DDR1 was detected in hPDLCs in the mRNA and protein level; DDR1b was the dominant isoform. Knockdown of DDR1 almost halved the migratory capacity and significantly downregulated the adhesive capacity of hPDLCs. The use of MEK-ERK1/2 inhibitors caused declined migratory and adhesive capacity of hPDLCs as well. After DDR1 was knocked down, the expression of p-MEK and p-ERK protein declined significantly while total MEK and ERK showed no obvious change, which means the ratio of p-MEK/MEK and p-ERK/ERK was markedly reduced. CONCLUSIONS: DDR1 plays an important role in the migration and adhesion of hPDLCs and might be regulated via the MEK-ERK1/2 signaling pathway.


Assuntos
Receptor com Domínio Discoidina 1 , Ligamento Periodontal , Células Cultivadas , Receptor com Domínio Discoidina 1/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases/genética , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo
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