Attachment and proliferation of human gingival fibroblasts seeded on barrier membranes using Wharton's jelly-derived stem cells conditioned medium: An in vitro study.

The effect of Wharton's jelly mesenchymal stem cells conditioned medium (WJMSCs-CM) and zinc oxide nanoparticles (ZnO-NPs) on cultured human gingival fibroblasts on various barrier membranes was investigated in this study. In this study, human gingival fibroblasts were prepared and cultured on three membranes: collagen membrane, acellular dermal matrix (ADM) with ZnO-NPs, and ADM without ZnO-NPs. WJMSCs-CM was given to the testing groups, while control groups received the same membranes without WJMSCs-CM. Following 48 and 72 h, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tests were performed to assess cell survival. Cell proliferation on the membranes was also evaluated using 4',6-diamidino-2-phenylindole (DAPI) staining after 48 and 72 h. Field emission scanning electron microscopy was used to determine membrane surface structure and cell adhesion. Nanoparticles were also subjected to an energy-dispersive x-ray analysis to identify their chemical structure. Two-way analysis of variance was used to conduct the statistical analysis. The p-value ≤.05 was considered significant. When ADM-ZnO-NPs were combined with CM, fibroblast viability, and adhesion significantly differed from ADM-ZnO-NPs alone. DAPI results confirmed cell proliferation in all six groups on both experiment days. The abundance and concentrated distribution of cells during cell proliferation were found in CM-containing membranes, specifically the ADM-ZnO-NPs membrane, demonstrating the improved biocompatibility of the ADM-ZnO-NPs membrane for cell proliferation. The other groups did not significantly differ from one another. WJMSCs-CM positively affected the viability and proliferation of gingival fibroblasts, but only marginally. Under certain conditions, ZnO-NPs below a specific concentration increased the biocompatibility of the membranes.

Bone repair potential of collagen-poly(3-hydroxybutyrate)-carbon nanotubes scaffold loaded with mesenchymal stem cells for the reconstruction of critical-sized mandibular defects.

The poor structural stability of collagen (COL) upon hydration poses a significant challenge in tissue engineering (TE). To overcome this limitation, the incorporation of hydrophobic polymers such as poly(3-hydroxybutyrate) (PHB), and nanomaterials such as carbon nanotubes (CNTs) has been explored. In this study, we investigated the physical, chemical, and biological characteristics of COL-based scaffolds modified with PHB and CNTs for bone tissue engineering (BTE) applications. The tensile strength analysis revealed a substantial improvement in the ultimate tensile strength with the addition of 10 % PHB and 4 % CNTs. Scanning electron microscopy (SEM) images depicted a denser and more compact structure resulting from the presence of PHB and CNTs, enhancing the scaffold's mechanical properties. Fourier-transform infrared spectroscopy (FTIR) confirmed the successful incorporation of PHB and CNTs into the composite scaffold, maintaining the chemical integrity of COL. Stereological studies also conducted in a rat model with induced critical-sized bone defects in the mandibular bone further emphasize the substantial increase in bone formation and reduction in defect volume achieved by the scaffold loaded with stem cells. These findings underscore the promising approach to enhance bone healing, using COL-based scaffolds loaded with stem cells, and the favorable results obtained in this study can contribute to the advancement of BTE strategies.

Evaluation of the association between osteoporosis and periodontitis in postmenopausal women: A clinical and radiographic study.

Background: This study aimed to assess the relationship between the osteoporotic condition and periodontitis in postmenopausal women. Materials and Methods: Ninety-four women aged 50-80 years were studied in this cross-sectional study. Bone mineral density (BMD) of lumbar vertebra BMD (LBMD), total hip BMD (HBMD), and neck of femur BMD (NBMD) was assessed using standardized dual-energy X-ray absorptiometry (DXA) (normal: T-score ≥-1, osteopenic: -2.5 ≤ t-score <-1, osteoporotic: T-score <-2.5). Bleeding point index (BI), O'Leary plaque index (PI), and clinical attachment loss (CAL) were recorded. Cementoenamel junction, alveolar-crest distance (CEJ-AC) was measured from cone-beam computed tomography images. Periodontitis severity was represented by CAL and CEJ-AC distance. One-way analysis of variance followed by Post hoc Tukey was performed for examining differences among the groups for different variables. Pearson correlation coefficient® and backward regression analysis were used to investigate the effect of confounding variables on CEJ-AC as the dependent variable. Significance was considered at P < 0.05. Results: Mean CEJ-AC was significantly higher in the osteoporotic and osteopenic groups compared to the normal group (P = 0.001). There was a significant positive correlation between CEJ-AC and NBMD and LBMD (P < 0.001). The associations between LBMD and CEJ-AC existed even after adjusting for confounding factors (P = 0.002). The differences in BI, PI, and CAL were not statistically different between the groups (P > 0.05). Conclusion: Although osteoporosis is not the main cause of periodontitis, it can indirectly affect periodontal status by increasing CEJ-AC. Early diagnosis of osteoporosis followed by early referral to a dentist for the treatment of potential existing periodontal diseases is important to avoid complications among postmenopausal women.

Comparison of Osteogenic and Chondrogenic Differentiation Ability of Buccal Fat Pad Derived Mesenchymal Stem Cells and Gingival Derived Cells.

STATEMENT OF THE PROBLEM: One major goal of tissue engineering and regenerative medicine is to find an appropriate source of mesenchymal stem cells (MSCs) with higher differentiation ability. PURPOSE: In this experimental study, the osteogenic and chondrogenic differentiation ability of buccal fat pad derived MSCs (BFP-MSCs) with gingival derived cells (GDCs) were compared. MATERIALS AND METHOD: BFP-MSCs and GDCs were cultured enzymatically and expanded. The expanded cells were analyzed for membrane-associated markers, using flow cytometry. Then the ability of these cells to differentiate into osteocyte and chondrocyte was assessed morphologically and by mRNA expression of collagen I (COLL), BGLA and bone morphogenetic protein 2 (BMP2) using qRT-PCR. RESULTS: Flow cytometry analysis showed that both BFP-MSCs and GDCs expressed the characteristic stem cell markers such as CD73, CD44, and CD90, whereas they did not express hematopoietic markers. Mineralized calcium deposition was observed apparently in BFP-MSCs cultured in osteogenic medium but GDCs showed fewer mineralized nodules. The mRNA expression levels of BGLA and BMP2 showed 7×105 and 733-fold more mRNA expression in BFP-MSCs treated with differentiation media compared to the control group. In chondrogenic differentiation, BFP-MSCs transformed from a spindle to a cuboidal shape while GDCs showed only a slight transformation. In addition, mRNA expression of COLL showed 282-fold higher expression in BFP-MSCs in comparison to the control group. Such significant difference in mRNA expression of BGLA, BMP2, and COLL was not observed in GDCs compared to their corresponding controls. CONCLUSION: Based on the present results, BFP yields a greater proportion of stem cells compared to gingiva. Therefore, this tissue can be introduced as an easily available source for the treatment of periodontal defects and other maxillofacial injuries.

Screening the antigenic properties of Porphyromonas gingivalis and level of raised antibodies after antigenic injection into Balb/c Mice.

BACKGROUND: Different diagnostic screening tests have been developed to detect periodontal disease in the early stages. Despite these advances still, there is a need for a more practical and beneficial diagnostic test. AIM: The aim of this study was to investigate the possibility of developing such a kit based on the body immune response against Porphyromonas gingivalis. METHOD AND MATERIALS: This experimental study was conducted by culturing P. gingivalis and extracting its antigens. These antigens were injected into peritoneal cavity of four Balb/c mice. Finally, the pattern, type, and quantity of antibody response against P. gingivalis antigen were detected. Results of the study showed that 3.0 × 108 cells of P. gingivalis are an appropriate count for stimulating the immunization in Balb/c mice and the subsequent amount of antibody (IgG) production was 81.5 µg/ml. RESULT: The antigenic injections which were done in the current study could mimic the condition of periodontal disease and the raise of P. gingivalis in the body. CONCLUSION: The obtained data can be used in future attempts to develop practical and usable test kits against P. gingivalis.

Amnion membrane as a novel barrier in the treatment of intrabony defects: a controlled clinical trial.

PURPOSE: The purpose of this 6-month randomized, controlled, blinded, clinical trial was to evaluate and compare the efficacy of amnion membrane (AM) with deproteinized bovine bone mineral (BBM) and a collagen membrane (CM) with BBM in guided tissue regeneration (GTR) for the treatment of intrabony periodontal defects. MATERIALS AND METHODS: Ten chronic periodontitis patients with bilateral intrabony defects with radiographic evidence of intrabony component ≥ 4 mm and probing pocket depths (PPDs) ≥ 6 mm were treated with AM+BBM, and the control group was managed with CM+BBM. Periodontal clinical parameters were recorded at baseline and at 6 months after treatment. RESULTS: PPD, clinical attachment level (CAL), and probing bone (PB) showed significant improvements after 6 months in both the test and control groups. Gingival recession showed a significant increase in the control group but not in the test group. The changes in mean PPD, PB, and CAL preoperatively and postoperatively between the groups were not significant. There was no significant relationship between the depth of the baseline bony defect and CAL gain. CONCLUSION: Both AM and CM in conjunction with BBM provided improvement of clinical periodontal parameters. AM did not induce significant gingival recession and is suggested as a new barrier membrane in GTR treatment.

The Relationship Between Periodontal Disease and Public Health: A Population-Based Study.

BACKGROUND: Periodontal diseases, such as periodontitis, are considered the main cause of tooth loss in the elderly.The present study is aimed to determine the relationship between periodontal condition and quality of life. Quality of life consists of a range of people's objective needs related to the self-perception of well-being. METHODS: This study was done from January 2014 to June 2015 in a healthcare clinic in Zahedan, southeast Iran. Using the random sampling method, the researchers enrolled 700 individuals over 35 years of age. The participants initially completed a demographic questionnaire consisting of data, such as age, sex, educational level, and smoking habit. Then, the periodontal chart was completed. Moreover, patients, based on the number of their natural teeth, were divided into two groups (≥10 teeth in both maxillary and mandible arches and <10 teeth in at least one arch). The body mass index (BMI) was also measured.To assess the participants' general health, the WHO's quality of life questionnaire (WHOQOL-BREF) was used. RESULTS: Of the 700 enrolled individuals, 53.3% were womenand 47.7% were men. Moreover, most of the participants (63.71%) had BMI of less than 25 and 68% did not smoke.We found that as the people's periodontal status deteriorated, their quality of life also declined and the total mean score in all four health domains decreased (P<0.001).Moreover, people with more than 10 teeth in both arches scored higher with respect to life quality than those with less than 10 teeth in at least one arch (P<0.001). CONCLUSION: This studyindicates a decrease in the general quality of life in patients with periodontal disease.The authors suggest performing studies with larger sample sizes andcohort studies for more reliable results.

Evaluation of serum anti-cardiolipin antibodies after non-surgical periodontal treatment in chronic periodontitis patients.

The present study investigated the effect of non-surgical periodontal therapy on serum level of anti-cardiolipin antibodies (aCLA), which are potentially involved in the pathogenesis of cardiovascular diseases in periodontal patients. Twenty volunteers (11 females and 9 males) with the mean age of 40.55 years participated in this study. Generalized chronic periodontitis was diagnosed through clinical periodontal examination at baseline visit. This examination included measuring the probing pocket depth and clinical attachment loss. Plaque index and gingival index were also recorded. After baseline examination, all the subjects received full-mouth non-surgical periodontal treatment. Subjects returned for a final visit 6 weeks after the last session of scaling for reevaluation of the periodontal parameters. At baseline and final visits 2 ml of venous blood was collected from each patient and an available commercially enzyme-linked immunosorbent assay was used for analyzing aCLA (IgM and IgG). The collected data were analyzed using the paired sample t test. Mean levels of both forms of aCLA, before and after treatment, showed statistically significant difference (P = 0.003 for IgM and P = 0.001 for IgG). In addition, study results showed significant reductions in periodontal parameters after non-surgical periodontal therapy (P < 0.001). The results of this study suggested that successful periodontal therapy can improve the serum level of one of the inflammatory biomarkers involved in the cardiovascular problems.

mRNA expression of pattern recognition receptors and their signaling mediators in healthy and diseased gingival tissues.

BACKGROUND: Gingivitis and periodontitis are initiated by inflammation caused by microorganisms. Pathogen-associated molecular patterns (PAMPs) from these microorganisms are recognized through various toll-like receptors (TLRs) and NOD-like receptors (NLRs). In this study, we have chosen five TLRs and two NLRs as representatives taking part in the recognition and inflammation process, along with a few of their signaling mediators including CD14, MYD88, and TRIF to compare their mRNA expression levels between healthy and diseased gingival tissues. This will provide deeper insight into the mechanisms underlying gingivitis and periodontitis. Understanding the mechanisms involved in the onset and progression of the periodontal diseases could greatly help in establishing effective ways for prevention and treatment of these diseases besides decreasing the risk factor for relevant systemic disorders. MATERIALS AND METHODS: Gingival tissue samples for mRNA extraction and cDNA synthesis were taken from patients with gingivitis and periodontitis and from healthy control subjects. Messenger RNA expression of all genes was assessed using real-time polymerase chain reaction (PCR). RESULTS: Among the genes studied in different groups, only MYD88 mRNA expression was significantly higher in the periodontitis group compared to that of the controls. The expression level of this molecule was also significantly higher in patients with severe periodontitis compared to other patients and also compared to healthy individuals. In different tissues, positive significant correlations were observed between the mRNA expression levels of some genes. CONCLUSIONS: Elevated mRNA levels of MYD88 in periodontitis might have a key role in the pathogenesis of this disease. Therefore, MYD88 may be a useful target for the therapy of this disease.

Antibacterial susceptibility patterns of Porphyromonas gingivalis isolated from chronic periodontitis patients

Objectives: To test the antimicrobial sensitivity of Porphyromonas gingivalis to a panel of eight orally administrable antibiotics in chronic periodontal diseases and to evaluate factors associated with periodontitis in adultpatients.Study Design: A total of fifty strains of P. gingivalis were isolated from one hundred and twenty adult patients with chronic perio-dontitis. Identification of bacteria was carried out by anaerobic culture and biochemical tests.Selected colonies of P. gingivalis were used to evaluate the antibacterial activities of penicillin, metronidazole,amoxicillin, amoxicillin/clavulanic acid, clindamycin, doxy-cycline, ciprofloxacin and azithromycin.Results: Most of the patients were female, age ranging between 40 to 50 years. Majority of the patients frequently had scaling and depths of periodontal pockets in infected teeth were 5-8 mm and most of them had hemorrhageduring sampling. Susceptibility testing revealed a sensitivity of 100% of P. gingivalis to azithromycin, doxycyclineand amoxicillin/clavulanic acid but lower susceptibilities were found for the rest of antibiotic agents evaluated.Conclusions: Frequent scaling in women aged between 40-50 years had positive correlation with chronic periodontitis.The application of antibiotics in conjuction with mechanical debridation, may reflect in the level of resistance of P. gingivalis in patients with chronic periodontal infections. This could suggest periodical antibiotic susceptibility testing is necessary to determine the efficacy of antimicrobial agents if the perfect curing of chronicperiodontal diseases after mechanical debridation is meant. Further clinical studies are required to confirm thein vitro results. The only limitation in this study was identification of bacteria to species rather than subspecies level (AU)

Antibacterial susceptibility patterns of Porphyromonas gingivalis isolated from chronic periodontitis patients.

OBJECTIVES: To test the antimicrobial sensitivity of Porphyromonas gingivalis to a panel of eight orally administrable antibiotics in chronic periodontal diseases and to evaluate factors associated with periodontitis in adult patients. STUDY DESIGN: A total of fifty strains of P. gingivalis were isolated from one hundred and twenty adult patients with chronic perio-dontitis. Identification of bacteria was carried out by anaerobic culture and biochemical tests. Selected colonies of P. gingivalis were used to evaluate the antibacterial activities of penicillin, metronidazole, amoxicillin, amoxicillin/clavulanic acid, clindamycin, doxy-cycline, ciprofloxacin and azithromycin. RESULTS: Most of the patients were female, age ranging between 40 to 50 years. Majority of the patients frequently had scaling and depths of periodontal pockets in infected teeth were 5-8 mm and most of them had hemorrhage during sampling. Susceptibility testing revealed a sensitivity of 100% of P. gingivalis to azithromycin, doxycycline and amoxicillin/clavulanic acid but lower susceptibilities were found for the rest of antibiotic agents evaluated. CONCLUSIONS: Frequent scaling in women aged between 40-50 years had positive correlation with chronic periodontitis. The application of antibiotics in conjuction with mechanical debridation, may reflect in the level of resistance of P. gingivalis in patients with chronic periodontal infections. This could suggest periodical antibiotic susceptibility testing is necessary to determine the efficacy of antimicrobial agents if the perfect curing of chronic periodontal diseases after mechanical debridation is meant. Further clinical studies are required to confirm the in vitro results. The only limitation in this study was identification of bacteria to species rather than subspecies level.