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International Journal of Molecular... Aug 2024Naringenin (NAR) is a prominent flavanone that has been recognized for its capacity to promote the osteogenic differentiation of human periodontal ligament stem cells...
Naringenin (NAR) is a prominent flavanone that has been recognized for its capacity to promote the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The present study aimed to explore how NAR promotes the osteogenic differentiation of hPDLSCs and to assess its efficacy in repairing alveolar bone defects. For this purpose, a protein‑protein interaction network of NAR action was established by mRNA sequencing and network pharmacological analysis. Gene and protein expression levels were evaluated by reverse transcription‑quantitative and western blotting. Alizarin red and alkaline phosphatase staining were also employed to observe the osteogenic capacity of hPDLSCs, and immunofluorescence was used to examine the co‑localization of NAR molecular probes and AKT in cells. The repair of mandibular defects was assessed by micro‑computed tomography (micro‑CT), Masson staining and immunofluorescence. Additionally, computer simulation docking software was utilized to determine the binding affinity of NAR to the target protein, AKT. The results demonstrated that activation of the nitric oxide (NO)‑cyclic guanosine monophosphate (cGMP)‑protein kinase G (PKG) signaling pathway could promote the osteogenic differentiation of hPDLSCs. Inhibition of AKT, endothelial nitric oxide synthase and soluble guanylate cyclase individually attenuated the ability of NAR to promote the osteogenic differentiation of hPDLSCs. Micro‑CT and Masson staining revealed that the NAR gavage group exhibited more new bone formation at the defect site. Immunofluorescence assays confirmed the upregulated expression of Runt‑related transcription factor 2 and osteopontin in the NAR gavage group. In conclusion, the results of the present study suggested that NAR promotes the osteogenic differentiation of hPDLSCs by activating the NO‑cGMP‑PKG signaling pathway through its binding to AKT.
Topics: Humans; Osteogenesis; Flavanones; Proto-Oncogene Proteins c-akt; Signal Transduction; Cell Differentiation; Nitric Oxide; Cyclic GMP-Dependent Protein Kinases; Stem Cells; Cyclic GMP; Animals; Male; Cells, Cultured
PubMed: 38940332
DOI: 10.3892/ijmm.2024.5391 -
Frontiers in Cellular and Infection... 2024Recent studies have demonstrated a positive role of hyaluronic acid (HA) on periodontal clinical outcomes. This study aimed to investigate the impact of four different...
INTRODUCTION
Recent studies have demonstrated a positive role of hyaluronic acid (HA) on periodontal clinical outcomes. This study aimed to investigate the impact of four different HAs on interactions between periodontal biofilm and immune cells.
METHODS
The four HAs included: high-molecular-weight HA (HHA, non-cross-linked), low-molecular-weight HA (LHA), oligomers HA (OHA), and cross-linked high-molecular-weight HA (CHA). Serial experiments were conducted to verify the influence of HAs on: (i) 12-species periodontal biofilm (formation and pre-existing); (ii) expression of inflammatory cytokines and HA receptors in monocytic (MONO-MAC-6) cells and periodontal ligament fibroblasts (PDLF) with or without exposure to periodontal biofilms; (iii) generation of reactive oxygen species (ROS) in MONO-MAC-6 cells and PDLF with presence of biofilm and HA.
RESULTS
The results indicated that HHA and CHA reduced the bacterial counts in a newly formed (4-h) biofilm and in a pre-existing five-day-old biofilm. Without biofilm challenge, OHA triggered inflammatory reaction by increasing IL-1β and IL-10 levels in MONO-MAC cells and IL-8 in PDLF in a time-dependent manner, whereas CHA suppressed this response by inhibiting the expression of IL-10 in MONO-MAC cells and IL-8 in PDLF. Under biofilm challenge, HA decreased the expression of IL-1β (most decreasing HHA) and increased IL-10 levels in MONO-MAC-6 cells in a molecular weight dependent manner (most increasing CHA). The interaction between HA and both cells may occur via ICAM-1 receptor. Biofilm stimulus increased ROS levels in MONO-MAC-6 cells and PDLF, but only HHA slightly suppressed the high generation of ROS induced by biofilm stimulation in both cells.
CONCLUSION
Overall, these results indicate that OHA induces inflammation, while HHA and CHA exhibit anti-biofilm, primarily anti-inflammatory, and antioxidant properties in the periodontal environment.
Topics: Biofilms; Hyaluronic Acid; Humans; Reactive Oxygen Species; Fibroblasts; Cytokines; Monocytes; Periodontal Ligament; Cell Line; Interleukin-1beta; Interleukin-10
PubMed: 38938883
DOI: 10.3389/fcimb.2024.1414861 -
Journal of Periodontology Jun 2024Diabetes is one of the major inflammatory comorbidities of periodontitis via 2-way interactions. Cystathionine γ-lyase (CTH) is a pivotal endogenous enzyme synthesizing...
BACKGROUND
Diabetes is one of the major inflammatory comorbidities of periodontitis via 2-way interactions. Cystathionine γ-lyase (CTH) is a pivotal endogenous enzyme synthesizing hydrogen sulfide (HS), and CTH/HS is crucially implicated in modulating inflammation in various diseases. This study aimed to explore the potential role of CTH in experimental periodontitis under a hyperglycemic condition.
METHODS
CTH-silenced and normal human periodontal ligament cells (hPDLCs) were cultured in a high glucose and Porphyromonas gingivalis lipopolysaccharide (P.g-LPS) condition. The effects of CTH on hPDLCs were assessed by Cell Counting Kit 8 (CCK8), real-time quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assay (ELISA). The model of experimental periodontitis under hyperglycemia was established on both Cth and wild-type (WT) mice, and the extent of periodontal destruction was assessed by micro-CT, histology, RNA-Seq, Western blot, tartrate-resistant acid phosphatase (TRAP) staining and immunostaining.
RESULTS
CTH mRNA expression increased in hPDLCs in response to increasing concentration of P.g-LPS stimulation in a high glucose medium. With reference to WT mice, Cth mice with experimental periodontitis under hyperglycemia exhibited reduced bone loss, decreased leukocyte infiltration and hindered osteoclast formation, along with reduced expression of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in periodontal tissue. RNA-seq-enriched altered NF-κB pathway signaling in healthy murine gingiva with experimental periodontitis mice under hyperglycemia. Accordingly, phosphorylation of p65 (P-p65) was alleviated in CTH-silenced hPDLCs, leading to decreased expression of IL6 and TNF. CTH knockdown inhibited activation of nuclear factor kappa-B (NF-κB) pathway and decreased production of proinflammatory cytokines under high glucose and P.g-LPS treatment.
CONCLUSION
The present findings suggest the potential of CTH as a therapeutic target for tackling periodontitis in diabetic patients.
PubMed: 38937859
DOI: 10.1002/JPER.23-0811 -
STAR Protocols Jun 2024Periodontal ligament cells (PDLCs) and macrophages in bone marrow cells have been widely used to investigate novel therapeutic agents to treat periodontitis. Here, we...
Periodontal ligament cells (PDLCs) and macrophages in bone marrow cells have been widely used to investigate novel therapeutic agents to treat periodontitis. Here, we present a protocol for collecting primary mouse PDLCs and bone marrow cells. We detail steps for culturing and differentiation for both cell types and review data analysis for in vitro experiments using primary PDLCs and bone marrow cells. This protocol can be used to explore the impact of novel therapeutic agents using in vitro experiments. For complete details on the use and execution of this protocol, please refer to Sirisereephap et al..
PubMed: 38935507
DOI: 10.1016/j.xpro.2024.103162 -
Indian Journal of Dental Research :... Jan 2024The application of direct current can have a significant impact on the rate of tooth movement and surrounding periodontal ligament collagen turnover. This study aims to... (Comparative Study)
Comparative Study
An Immunohistochemical and Histological Study of the Animal Periodontal Ligament During Orthodontic Force Application with Concomitant Application of Electric Current - An Animal Study.
INTRODUCTION
The application of direct current can have a significant impact on the rate of tooth movement and surrounding periodontal ligament collagen turnover. This study aims to provide insight into the optimal characteristics of applied current to achieve enhanced tissue response.
METHOD
Eighteen male Wistar rats were divided into three groups (I, II, and III). Split mouth design was used, and each side was allocated into an experimental group or control group. Experimental sides of groups I, II, and III received 20, 10, and 15 μA of current (15 min, twice daily for 3 days). Both the experimental and control groups receive an orthodontic force via the NiTi closed coil spring. The amount of tooth movement was determined daily. Immunohistochemistry slides were scored using the immunoreactive scoring (IRS) system for collagen types I and III. One-way Analysis of Variance (ANOVA) and Tukey post hoc test were used to analyse the rate of tooth movement, while Mann-Whitney test was used to analyse IRS distribution between control and experimental groups.
RESULTS
Compared with the control group, there was a statistically significant difference in tooth movement in all the experimental groups, with group 3 showing the maximum rate on days 2 and 3. This was supported by immunoreactive scores for both collagen types I and III.
CONCLUSIONS
After 72 hours, the expression of collagen types 1 and 3 increased significantly for group III. This finding was in harmony with the rate of tooth movement, which was maximum for group 3 (15 μA) as compared to other groups.
Topics: Periodontal Ligament; Animals; Rats, Wistar; Tooth Movement Techniques; Male; Rats; Collagen Type I; Immunohistochemistry; Collagen Type III; Orthodontic Wires; Dental Alloys; Nickel; Stress, Mechanical; Titanium
PubMed: 38934753
DOI: 10.4103/ijdr.ijdr_905_22 -
Cell Biology International Jun 2024This study explores the potential role and mechanism of Ginsenoside Rb3 (Rb3) in modulating osteoclastogenesis induced by human periodontal ligament fibroblasts (hPLFs)...
This study explores the potential role and mechanism of Ginsenoside Rb3 (Rb3) in modulating osteoclastogenesis induced by human periodontal ligament fibroblasts (hPLFs) within the periodontitis microenvironment. We investigated the anti-inflammatory effects of Rb3 on hPLFs stimulated with Porphyromonas gingivalis lipopolysaccharide (P.g-LPS) utilizing quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay techniques. Moreover, the functional role of Rb3 in hPLFs-induced osteoclast formation was assessed by treating human bone marrow-derived macrophages (hBMMs) with conditioned medium from hPLFs, followed by analyses through qPCR, western blot analysis, and staining for tartrate-resistant acid phosphatase (TRAP) and phalloidin. The impact of Rb3 on the activation of the STAT3 signaling pathway was determined via western blot analysis. Results indicated that Rb3 treatment significantly suppressed the upregulation of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6, MCP-1, and IL-18) at both gene and protein levels in hPLFs induced by P.g-LPS. Furthermore, conditioned medium from Rb3 plus P.g-LPS treated hPLFs notably decreased the number of TRAP-positive cells, actin ring formations, and the expression of osteoclast marker genes (including CTSK, NFATC1, and ACP5). Rb3 also inhibited the P.g-LPS-induced activation of the STAT3 pathway, with the activation of STAT3 partially reversing the effects of Rb3 on inflammation and osteoclast differentiation. Collectively, Rb3 ameliorates inflammation in P.g-LPS-stimulated hPLFs and reduces hPLFs-induced osteoclastogenesis by inhibiting the STAT3 signaling pathway, suggesting its potential as a therapeutic agent for periodontitis.
PubMed: 38934258
DOI: 10.1002/cbin.12201 -
Cureus May 2024Periodontitis, a persistent inflammatory condition, results in the deterioration of both the hard and soft tissues in the periodontium, leading to the formation of...
INTRODUCTION
Periodontitis, a persistent inflammatory condition, results in the deterioration of both the hard and soft tissues in the periodontium, leading to the formation of intrabony defects. Restoring the lost tissues, particularly bone, is possible through tissue engineering techniques utilizing scaffolds made from different polymers. Consequently, this research focuses on creating and assessing a scaffold infused with alginate (Sigma Aldrich, Gillingham, UK) and carrageenan (Sigma Aldrich, Gillingham, UK) for the purpose of bone regeneration.
METHODS
An in vitro investigation was conducted to assess the characteristics of the recently formulated scaffold. Spectroscopic analysis, tensile strength testing, scanning electron microscopy (SEM) analysis, and degradation testing were carried out to evaluate both the physical and biological attributes of the scaffold.
RESULTS
IBM SPSS Statistics for Windows, V. 1.2 (IBM Corp., Armonk, NY, USA) was used for statistical analysis. A one-way ANOVA test was done to determine the significance of tensile strength, and a paired t-test was done to check the significance of the degradation test. The in vitro research unveiled notable distinctions in the physical and biological attributes between the scaffold infused with alginate and carrageenan and the PerioCol® (p<0.05).
CONCLUSION
The scaffold incorporating alginate and carrageenan demonstrated superior outcomes concerning parameters such as tensile stress and strain, degradation rate, percentage bone volume, and object surface density when contrasted with the conventional PerioCol®. Therefore, the scaffold infused with alginate and carrageenan emerges as a promising candidate for bone regeneration.
PubMed: 38933614
DOI: 10.7759/cureus.61139 -
Medicina (Kaunas, Lithuania) May 2024: The present systematic review and meta-analysis were conducted to evaluate and compare the long-term clinical outcomes of immediate implants placed into fresh sockets... (Meta-Analysis)
Meta-Analysis Comparative Study Review
: The present systematic review and meta-analysis were conducted to evaluate and compare the long-term clinical outcomes of immediate implants placed into fresh sockets with and without periapical pathology. : After the search and review of the literature in the electronic databases, 109 publications were achieved. The titles and abstracts of 66 publications were screened. After the evaluation of the full text of 22 publications, based on the inclusion criteria, six controlled clinical studies were included in this systematic review and meta-analysis. The statistical calculation showed no heterogeneity among the studies included. The implant survival was 99.6% in the test (socket with periapical pathology) and control (socket without periapical pathology) groups of all the clinical trials. The results of the meta-analysis showed no statistically significant difference between test and control groups regarding the marginal bone level and the width of keratinized mucosa in all the studies. Other parameters indicating plaque level, bleeding on probing, and gingival recession also did not differ between test and control groups at the final follow-up in nearly all studies. : Within the limitation of this systemic review and meta-analysis, the obtained data suggest that implants immediately placed into the extraction sockets of teeth exhibiting periapical pathology can be successfully osseointegrated for an extended period.
Topics: Humans; Tooth Socket; Dental Implantation, Endosseous; Dental Implants; Immediate Dental Implant Loading
PubMed: 38929509
DOI: 10.3390/medicina60060893 -
Medicina (Kaunas, Lithuania) May 2024Peri-implant soft tissue deficiency (PSTD) is a significant factor impacting aesthetics, particularly in the anterior zone, where labial bone resorption and thin...
Peri-implant soft tissue deficiency (PSTD) is a significant factor impacting aesthetics, particularly in the anterior zone, where labial bone resorption and thin peri-implant phenotypes are common. The occurrence of a gray color around the implant fixture due to PSTD can be aesthetically concerning in the esthetic zone. In cases involving natural teeth, autogenous soft tissue grafts such as subepithelial connective tissue grafts (SCTGs), free gingival grafts (FGGs), and coronally advanced flaps (CAFs) are commonly utilized. However, there are limited reports of using bone grafts in conjunction with these techniques for modifying the gingival phenotype around both teeth and implants. In the presented cases where PSTD resulted in visible gray coloration of the implant fixture in the esthetic zone, mechanical and chemical decontamination of the exposed implant surface was performed using a titanium brush and tetracycline (Tc) HCl. Subsequently, to enhance peri-implant mucosa thickness and mask the titanium color, simultaneous SCTG and bone grafting procedures were conducted. Within the limitations of these case reports, successful esthetic outcomes were achieved and maintained without recurrence for 3-6 years following the simultaneous subepithelial connective tissue graft and bone graft procedures. These findings suggest the potential efficacy of this combined approach in addressing PSTD and enhancing aesthetic results around dental implants, though further studies are needed to validate these outcomes.
Topics: Humans; Connective Tissue; Bone Transplantation; Female; Phenotype; Gingiva; Esthetics, Dental; Adult; Middle Aged; Male; Dental Implants
PubMed: 38929458
DOI: 10.3390/medicina60060841 -
International Journal of Molecular... Jun 2024The pathology of medication-related osteonecrosis of the jaw (MRONJ), often associated with antiresorptive therapy, is still not fully understood. Osteocyte networks are...
The pathology of medication-related osteonecrosis of the jaw (MRONJ), often associated with antiresorptive therapy, is still not fully understood. Osteocyte networks are known to play a critical role in maintaining bone homeostasis and repair, but the exact condition of these networks in MRONJ is unknown. On the other hand, the local application of E-coli-derived Recombinant Human Bone Morphogenetic Protein 2/β-Tricalcium phosphate (E-rhBMP-2/β-TCP) has been shown to promote bone regeneration and mitigate osteonecrosis in MRONJ-like mouse models, indicating its potential therapeutic application for the treatment of MRONJ. However, the detailed effect of BMP-2 treatment on restoring bone integrity, including its osteocyte network, in an MRONJ condition remains unclear. Therefore, in the present study, by applying a scanning electron microscope (SEM) analysis and a 3D osteocyte network reconstruction workflow on the alveolar bone surrounding the tooth extraction socket of an MRONJ-like mouse model, we examined the effectiveness of BMP-2/β-TCP therapy on the alleviation of MRONJ-related bone necrosis with a particular focus on the osteocyte network and alveolar bone microstructure (microcrack accumulation). The 3D osteocyte dendritic analysis showed a significant decrease in osteocyte dendritic parameters along with a delay in bone remodeling in the MRONJ group compared to the healthy counterpart. The SEM analysis also revealed a notable increase in the number of microcracks in the alveolar bone surface in the MRONJ group compared to the healthy group. In contrast, all of those parameters were restored in the E-rhBMP-2/β-TCP-treated group to levels that were almost similar to those in the healthy group. In summary, our study reveals that MRONJ induces osteocyte network degradation and microcrack accumulation, while application of E-rhBMP-2/β-TCP can restore a compromised osteocyte network and abrogate microcrack accumulation in MRONJ.
Topics: Animals; Bone Morphogenetic Protein 2; Osteocytes; Calcium Phosphates; Mice; Recombinant Proteins; Disease Models, Animal; Bisphosphonate-Associated Osteonecrosis of the Jaw; Humans; Bone Regeneration; Male; Tooth Extraction; Transforming Growth Factor beta; Alveolar Process
PubMed: 38928355
DOI: 10.3390/ijms25126648