Network pharmacological study and in vitro studies validation-Molecular dynamics simulation of Cistanche deserticola in promoting periodontitis and bone remodeling.

OBJECTIVE: Periodontitis is a chronic infectious disease, characterized by loss of alveolar bone and supporting tissues. Cistanche deserticola(Cd), a local medicinal herb in Xinjiang, possesses favorable biological characteristics and potential applications. Our aim is to investigate the remodeling properties of Cd extract and elucidate the specific mechanisms underlying its therapeutic effects on periodontitis, by employing a combination of basic experimental and network pharmacology approaches. METHODS: Firstly, UHPLC-QTOF-MS analysis was conducted on Cd extract to identify its main components, with several compounds were identified by standard. Subsequently, in vitro studies were performed using the Cd extract on MC3T3-E1 cells. Cell proliferation viability was assessed using CCK-8 and apoptosis assays, while ALP and ARS staining and quantitative experiments, qRT-PCR, and Western blot assays were employed to evaluate the osteogenic differentiation capability. Network pharmacology analysis was then carried out using the identified compounds to establish a database of Cd components and targets, along with a database of periodontitis. The intersection of these databases revealed the network relationship between Cd components-mapped genes-signaling pathways. KEGG/GO pathway analysis of the targets was performed to filter potential enriched pathways. PPI/CytoHubba protein interaction network analysis was utilized to identify hub genes. Molecular docking and molecular dynamics simulations were employed to analyze the docking and interaction between core gene and Cd components. RESULTS: We detected 38 major components in the Cd extract, with Echinacoside, Acteoside, Tubuloside A, and Cistanoside A undergoing standard substance verification. In vitro studies indicated that the Cd, at concentrations below 100 µg/ mL, did not affect cell proliferation and inhibited apoptosis. Osteogenesis assays demonstrated that Cd at concentrations of 1 µg/ mL, 10 µg/ mL, and 100 µg/ mL significantly promoted the osteogenic differentiation ability of MC3T3-E1 cells. It also notably upregulated the mRNA and protein levels of Alp, Bmp2, Runx2, and Opn, and the optimal concentration was 10 µg/mL. Network pharmacology results revealed the network relationship between Cd's components, crossed targets and signaling pathways. Combined with KEGG/GO pathway analysis and PPI/CytoHubba protein interaction network analysis. The key pathway and hub genes of Cd regulating periodontitis are both related to hypoxia pathway and HIF-1α. Molecular docking results showed a strong binding affinity between Cd compounds and hub genes, and molecular dynamics simulation results indicated the stability of the complexes formed between HIF-1α and several Cd compounds. CONCLUSION: Cistanche deserticola exhibits a notable capacity to promote bone regeneration, and its mechanism of action in regulating periodontitis is associated with the hypoxia signaling pathway. HIF-1α may serve as a potential core gene. Future research will focus on exploring the mechanism of Cd in intervene periodontitis and promoting bone remodeling in hypoxic environment.

HIF-1α and periodontitis: Novel insights linking host-environment interplay to periodontal phenotypes.

Periodontitis, the sixth most prevalent epidemic disease globally, profoundly impacts oral aesthetics and masticatory functionality. Hypoxia-inducible factor-1α (HIF-1α), an oxygen-dependent transcriptional activator, has emerged as a pivotal regulator in periodontal tissue and alveolar bone metabolism, exerts critical functions in angiogenesis, erythropoiesis, energy metabolism, and cell fate determination. Numerous essential phenotypes regulated by HIF are intricately associated with bone metabolism in periodontal tissues. Extensive investigations have highlighted the central role of HIF and its downstream target genes and pathways in the coupling of angiogenesis and osteogenesis. Within this concise perspective, we comprehensively review the cellular phenotypic alterations and microenvironmental dynamics linking HIF to periodontitis. We analyze current research on the HIF pathway, elucidating its impact on bone repair and regeneration, while unraveling the involved cellular and molecular mechanisms. Furthermore, we briefly discuss the potential application of targeted interventions aimed at HIF in the field of bone tissue regeneration engineering. This review expands our biological understanding of the intricate relationship between the HIF gene and bone angiogenesis in periodontitis and offers valuable insights for the development of innovative therapies to expedite bone repair and regeneration.

Mandibular residual ridge morphology in relation to complete dentures and implant overdentures-Part I: Predictors for perceived conventional denture stability.

BACKGROUND: There is lack of reliable predictors for success of conventional complete denture (CCD) therapy, which in turn might affect the effectiveness of subsequent implant-retained overdenture (IOD) therapy. PURPOSE: To investigate relationships between digitally obtained geometrical mandibular residual ridge measures and perceived CCD-stability. MATERIALS AND METHODS: 30 CCD wearing patients (67.9 ± 7.0 years) for whom a new set of CCDs was advised, were treated with new CCDs. Digitalized mandibular gypsum models were measured using the Geomagic Studio 2013 software. Data were obtained for (1) height, width, and cross-section surface area of the residual ridge at different locations (midline, premolar, and anterior edge of retromolar pad) and (2) denture base surface area. Scatter plots and multivariate regression analyses were used to investigate associations between the geometric data and denture base surface area, and correlated with denture stability scores (Spearman rank test). RESULTS: Scatter plots showed that best model fit for denture base surface area was mean ridge height (R2 = 0.906). Multivariate regression showed that height at premolar location (p = 0.001) had largest effect on denture base surface area (R2 = 0.796). Ridge morphology variables, except width at midline location, were significantly correlated with CCD-stability (p-values <0.05). CCD-stability was significantly correlated with denture base surface area (p ≤ 0.001). CONCLUSION: Residual ridge height at premolar location was most predictive for denture base surface area and perceived CCD-stability.

Mandibular residual ridge morphology in relation to complete dentures and implant overdentures-Part II: Predictors for effectiveness of implant overdenture therapy.

BACKGROUND: The significance of mandibular residual ridge height and satisfaction with conventional complete dentures (CCD) as predictors for the added value of implant-overdenture (IOD) therapy is unknown. PURPOSE: To investigate the predictive value of thresholds for (1) residual ridge height at premolar location (PRH), and (2) satisfaction with CCD-stability for the added value of two intraforaminal implants supporting the mandibular CCD. METHODS: Thirty CCD wearing patients (67.9 ± 7.0 years) for whom a new CCD was advised, received a new CCD. Mandibular gypsum models were digitally measured. After 3 months free of complaints (T1), perceived CCD-stability was evaluated, and participants received two intraforaminal implants. At T1 and T2 (3 months free of complaints after IOD therapy) participants completed OHIP14-CN, and denture satisfaction (VAS) questionnaires, and performed mixing ability tests. Participants were grouped according to PRH of ≥6.15 mm versus < 6.15 mm, and perceived CCD-stability satisfied vs. dissatisfied. Scores at T2 were compared to T1 (paired t-tests). Predictive values of PRH and CCD-stability were analyzed with logistic multivariate regression models. RESULTS: At T2, only participants with PRH of <6.15 mm or dissatisfied with CCD-stability had significant lower OHIP-total and domain scores for 'physical pain' and 'physical disability' and significantly higher VAS scores for perceived chewing function, denture retention and oral comfort. Regression analyses showed that participants with PRH of <6.15 mm, or dissatisfied with CCD-stability had significantly higher chance for lower OHIP-total and domain scores 'physical pain' and 'physical disability', and for higher VAS scores for perceived chewing function, denture retention and oral comfort at T2. Masticatory performance improved significantly after IOD therapy, but independent of PRH and CCD-stability. CONCLUSION: PRH and satisfaction with CCD-stability were adequate prognostic indicators for improvement of oral health-related quality of life and denture satisfaction by mandibular IOD therapy.

Hydrogen exerts neuroprotective effects on OGD/R damaged neurons in rat hippocampal by protecting mitochondrial function via regulating mitophagy mediated by PINK1/Parkin signaling pathway.

Cerebral ischemia/reperfusion injury (IRI) is a serious complication during the treatment of stroke patients with very few effective clinical treatment. Hydrogen (H2) can protect mitochondria function and have favorable therapeutic effects on cerebral IRI. Mitophagy plays an important role in eliminating damaged or dysfunctional mitochondria and maintaining mitochondria homeostasis. However, whether the protection of H2 on cerebral IRI is via regulating mitophagy is still unknown. In this study, OGD/R damaged hippocampal neurons were used to mimic cerebral IRI in vivo and we detected the effect of H2, Rap (autophagy activator) and 3-MA (autophagy inhibitor) on OGD/R neurons. The results of MTT indicated that H2 and RAP could increase cell viability after OGD/R treatment, while 3-MA further aggravated injury and inhibited the protection of H2 and RAP. Furthermore, the intracellular ROS and apoptosis ratio were determined, the results showed that ROS and apoptosis level significantly increased after OGD/R, H2 and RAP effectively restrained the increment of ROS level and apoptosis ratio but their protective effect can be weakened by 3-MA. Mitochondrial membrane potential (MMP) and mitophagy level were also determined, the data showed that H2 and RAP protected against the loss of MPP and increased the co-localization of mitochondria with GFP-LC3 while 3-MA exerted antagonistic effect. At last, the mitophagy-related factors LC3, PINK1 and Parkin expression were detected and analyzed. We found that the expression of LC3 was increased after OGD/R which can be further enhanced by H2 and RAP treatment, but treatment with 3-MA was opposite. The result revealed H2 and RAP could activate mitophagy while 3-MA inhibit mitophagy. In addition, the study found H2 and RAP could significantly induce the expression of PINK1 and Parkin in OGD/R neurons which was inhibited by 3-MA. Taken together, our findings demonstrated H2 had a neuroprotective effect on OGD/R damaged neurons by protecting mitochondrial function and the potential protection mechanism may closely related to enhancement of mitophagy mediated by PINK1/Parkin signaling pathway.

Antibacterial Activity and Bonding Ability of an Orthodontic Adhesive Containing the Antibacterial Monomer 2-Methacryloxylethyl Hexadecyl Methyl Ammonium Bromide.

Irreversible white spot lesion (WSL) occurs in up to 50% of patients during orthodontic treatment. Therefore, orthodontic adhesives need to be able to inhibit or reduce bacterial growth in order to prevent or minimize WSL. This study evaluated the antibacterial effect and shear bond strength (SBS) of a resin-based orthodontic adhesive containing the antibacterial monomer 2-methacryloxylethyl hexadecyl methyl ammonium bromide (MAE-HB). MAE-HB was added at three concentrations (1, 3, and 5 wt%) to a commercial orthodontic adhesive Transbond XT, while the blank control comprised unmodified Transbond XT. Their antibacterial effects on Streptococcus mutans were investigated after 0 and 180 days of aging. The SBS of metal brackets bonded to the buccal enamel surface of human premolars was assessed. Compared with the blank control, the MAE-HB-incorporated adhesive exhibited a significant contact inhibitory effect on the growth of S. mutans (P < 0.05), even after 180 days of aging. SBS and adhesive remnant index values revealed that the bonding ability of the experimental adhesive was not significantly adversely affected by the incorporation of MAE-HB at any of the three concentrations. Therefore, orthodontic adhesives with strong and long-lasting bacteriostatic properties can be created through the incorporation of MAE-HB without negatively influencing bonding ability.

Single nucleotide polymorphisms in interleukin-6 and their association with venous thromboembolism.

The aim of the present study was to reveal the contribution of single nucleotide polymorphisms of the interleukin­6 (IL­6) gene and the progression of venous thromboembolism (VTE). A case­control study composed of 246 VTE patients, including 160 from the Han population (76 males and 84 females, mean age 57.41±13.25 years), 86 from the Uyghur population (41 males and 45 females, mean age 51.61±13.73 years) and 292 gender and ethnicity­matched control participants, including 170 from the Han population (91 males and 79 females, mean age 55.82±11.83 years) and 122 from the Uyghur population (64 males and 58 females, mean age 53.52±13.64 years) were enrolled in the present study. The results demonstrated that the serum levels of IL­6, C­reactive protein (CRP), D­dimer, fibrinogen, plasminogen activator inhibitor­1 and leptin were significantly higher in the VTE group compared with the control group (P<0.05). The frequencies of the ­572C/G promoter polymorphisms of the IL­6 genotypes CC, CG and GG were identified to be 34, 48 and 18% in the Han population and 33, 47 and 20% in the Uyghur population, respectively. The allele frequency distributions of the C and G alleles were 58 and 42% in the Han population and 56 and 43% in the Uyghur population, respectively. Significant differences were identified in the ­572C/G promoter polymorphisms between the VTE group and the control group (P<0.05). For the ­597G/A polymorphism, all individuals carried the GG and GA genotype; AA genotypes were not detected. Logistic regression analysis was used to identify the risk factors for VTE, adjusting by confounding factors, the results of which demonstrated that the CC homozygote of the IL­6 ­572G/C, CRP, IL­6 and high­density lipoprotein­cholesterol were independent risk factors of VTE (P<0.05). In conclusion, the ­572G/C genotype of IL­6 may be a genetic marker of VTE in the Han and Uyghur populations.

Association of interleukin-6 and C-reactive protein genetic polymorphisms levels with venous thromboembolism.

BACKGROUND: Increased levels of interleukin-6 (IL-6) and C-reactive protein (CRP) have been reported in patients with venous thromboembolisms (VTE). However, prospective studies did not confirm an association between IL-6, CRP and their polymorphism with the risk of VTE. METHODS: One hundred and forty patients (including 66 males and 74 females, mean age (55.55 ± 17.11) years) and one hundred and sixty controls (including 74 males and 86 females, mean age (56.58 ± 12.24) years) were involved. An enzyme linked immunosorbent assay (ELISA) method was used for detecting the serum levels of inflammatory factors IL-6 and CRP in both groups. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for analyzing the distribution of polymorphisms at the -572C/G and -597G/A sites of the promoter of the IL-6 gene and at 1059G/C of the CRP gene. RESULTS: Serum levels of IL-6 and CRP were significantly higher in the VTE group than in the control group (P < 0.05). The frequencies of -572C/G promoter polymorphisms CC, CG, and GG in the IL-6 gene were found to be 34%, 48%, and 18%, respectively, and the derived allele frequencies for the C and G alleles were 58% and 42%. There was a significant difference in the -572C/G promoter polymorphisms between the VTE group and control group (P < 0.05). For the -597G/A polymorphism, individuals all carried the GG and GA type; AA genotypes were not detected. The frequency of the GG, GC, and CC genotypes at the CRP1059G/C promoter was 87.57%, 7.86% and 3.57% in VTE group, while 86.25%, 10%, and 3.75% in control group, respectively. The frequency of G and C alleles at CRP 1059G/C was 91.43% and 8.57% in VTE group and 91.56% and 8.44% in the control group. The results showed that there was no statistically significant difference of 1059G/C genotype and mutation frequency of the allele between the VTE group and control group (P > 0.05). Multiple Logistic regression analysis showed CC homozygotes of the IL-6 -572G/C, body mass index (BMI), and CRP, IL-6, and high-density lipoprotein cholesterol (HDL-C) were independent risk factors for VTE (P < 0.05). CONCLUSIONS: We found that VTE was associated with IL-6 and CRP levels, and there was an association of IL-6 and its promoter polymorphism at -572G/C with the risk of VTE. Thus far, a causal relationship between inflammation and VTE remains to be clarified and more prospective data are required.

[Association between gene polymorphisms of methylenetetrahydrofolate reductase and plasma homocysteine in Uygur patients with venous thromboembolism].

OBJECTIVE: To investigate methylenetetrahydrofolate reductase (MTHFR) gene C677T mutation and plasma homocysteine (Hcy) levels in Uygur patients with venous thromboembolism (VTE) in Xinjiang. METHODS: A total of 222 VTE patients including 74 Uygur and 148 Han ethnic patients were examined, and 86 Uygur ethnic and Han 134 ethnic healthy people were included as controls. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was applied to detect MTHFR gene C677T polymorphism and plasma Hcy levels were measured by fluorescence polarization immunoassay. RESULTS: The MTHFR gene C677T genotypes distribution in Uygur VTE patients and control groups were: TT [28.38% (35/86) vs. 12.79% (11/86), P < 0.05], CT [41.89% (31/74) vs. 52.33% (45/86), P > 0.05]and CC [29.73% (22/74) vs. 34.88% (30/86), P > 0.05], respectively; and in Han VTE patients and control groups were: TT[27.03% (40/148) vs. 14.92% (20/134), P < 0.05], CT [44.59% (66/148) vs. 52.99% (71/134), P > 0.05] and CC [28.38% (42/148) vs. 32.09% (43/134), P > 0.05], respectively. SNP genotyping distribution frequency in Uygur and Han ethnic population was similar between controls and between VTE patients (P > 0.05). Plasma levels of Hcy in MTHFR gene TT genotype were statistically higher than CT and CC genotype (P < 0.05). After adjusting for age, gender, smoking history, hyperlipidemia, hypertension, diabetes, and MTHFR genotype, multifactor logistic regression analysis showed that plasma Hcy level (OR = 1.025, 95%CI 1.003 - 1.046, P = 0.024) and obesity (OR = 4.660, 95%CI 1.417 - 15.324, P = 0.011) were independent risk factors for Uygur ethnic patients with VTE while plasma Hcy level (OR = 1.020, 95%CI 1.006 - 1.034, P = 0.004) and smoking (OR = 2.867, 95%CI 1.062 - 6.586, P = 0.024) were independent risk factors for Han ethnic patients with VTE. CONCLUSION: MTHFR C677T polymorphism (TT genotype carrier) and increased plasma levels of Hcy are risk factors for Uygur and Han ethnic patients with VTE in Xinjiang.