Mesenchymal stem cells in PRP and PRF containing poly(3-caprolactone)/gelatin Scaffold: a comparative in-vitro study.

Scaffold design is one of the three most essential parts of tissue engineering. Platelet-rich plasma (PRP) and platelet-rich fibrin (PRF) have been used in clinics and regenerative medicine for years. However, the temporal release of their growth factors limits their efficacy in tissue engineering. In the present study, we planned to synthesize nanofibrous scaffolds with the incorporation of PRP and PRF by electrospinning method to evaluate the effect of the release of PRP and PRF growth factors on osteogenic gene expression, calcification, proliferation, and cell adhesion of human bone marrow mesenchymal stem cell (h-BMSC) as they are part of scaffold structures. Therefore, we combined PRP/PRF, derived from the centrifugation of whole blood, with gelatin and Polycaprolactone (PCL) and produced nanofibrous electrospun PCL/Gel/PRP and PCL/Gel/PRF scaffolds. Three groups of scaffolds were fabricated, and h-BMSCs were seeded on them: (1) PCL/Gel; (2) PCL/Gel/PRP; (3) PCL/Gel/PRF. MTS assay was performed to assess cell proliferation and adhesion, and alizarin red staining confirmed the formation of bone minerals during the experiment. The result indicated that PCL/Gel did not have any better outcomes than the PRP and PRF group in any study variants after the first day of the experiment. PCL/gelatin/PRF was more successful regarding cell proliferation and adhesion. Although PCL/gelatin/PRP showed more promising results on the last day of the experiment in mineralization and osteogenic gene expression, except RUNX2, in which the difference with PCL/gelatin/PRF group was not significant.

Effects of platelet-rich plasma local injection on dentin sialo protein and dentin matrix acidic phosphoprotein 1 levels of secretion in gingival sulcular fluid during orthodontic movement in animal samples.

Background: Prolonged orthodontic treatment causes complications such as root resorption, gingivitis, and caries. Platelet-rich plasma (PRP) can accelerate the dental movements of orthodontic treatments. The aim of the present study was to evaluate the effect of local PRP injection on the secretion of dentin sialoprotein (DSP) and dentin matrix acidic phosphoprotein (DMP1) biomarkers in gingival crevicular fluid during orthodontic movements in animal samples. Materials and Methods: In this experimental study, one maxillary quadrant of six beagle dogs was randomly selected as the experimental group, and the other side was considered as the control group. Afterward, maxillary of the first premolars was extracted, and then, a titanium nickel coil spring (150 g) was then used between the second premolar and the canine. PRP that was previously activated with CaCl 2-thrombin was injected intraligaman to the experimental side at days 0, 21, and 42. Moreover, the mixture of the CaCl 2-thrombin with placebo was injected in to the control side. The study period was 63 days. Sampling was performed on days 0, 1, 2, 7, 21, 42, and 63. Then, in all the samples, the DSP and DMP1 level was measured using special kits by ELISA method. Data were analyzed by the analysis of variance and t-test. Results: Upon the application of orthodontic forces, the mean DMP1 and DSP levels in the experimental group at different times were significantly increased compared to the control group (P < 0.001). Conclusion: PRP injection enhanced DMP1 and DSP level with probable rise in the rate of root resorption.

Evaluation and comparison number of gingival fibroblast and osteosarcoma cell (MG-63 cell line) adhesive to mocugraft, alloderm, and collagen membrane with or without advanced platelet-rich fibrin.

BACKGROUND: The tissue engineering has recently shown a significant progress in the fields of membranes and biosynthetic materials. Advanced platelet-rich fibrin (A-PRF) contains functional molecules that have newly shown great interest in regenerative therapies. The purpose of this study was to evaluate the effect of A-PRF on the adhesion of gingival fibroblast cells and osteosarcoma cells to different membranes. MATERIALS AND METHODS: In this experimental in vitro study, three collagen, alloderm, and mucograft membranes were studied, which were cut into four 5 mm × 5 mm pieces and placed in the bottom of a 24-well culture medium. One milliliter of A-PRF was added to two wells from each group and the other two wells remained without A-PRF. The gingival fibroblasts and osteosarcoma cells were individually added to each well. The cell adhesion was studied using an electron microscope after 24 h. The data were analyzed by independent t-test, one-way analysis of variance, and least significant difference test. RESULTS: In the presence of A-PRF, there was a significant higher osteoblast adhesion to collagen membrane compared to alloderm and mucograft membranes (P < 0.001). In the absence of A-PRF, adhesion of osteoblasts to collagen membrane was significantly higher than alloderm and mucograft (P = 0.019). Moreover, in the presence of A-PRF, fibroblast adhesion to collagen membrane was significantly higher than alloderm and mucograft membranes (P < 0.001). Furthermore, in the absence of A-PRF, no significant difference was found among the study groups (P = 0.830). CONCLUSION: A-PRF was effective on fibroblast adhesion to the collagen membrane, which is similar to its absence. A-PRF was also found to be very effective on the adhesion of fibroblast cells to the collagen membrane, and in its absence, even less adhesion was observed compared to the other membranes. The presence or absence of A-PRF showed no significant differences in both cells' adhesion for alloderm and mucograft membranes.

Maximum Insertion Torque, Stress, and Strain in Bone during Insertion of Three Dental Implants with Different Macroscopic Designs: A Dynamic Finite-Element Analysis.

Using finite element analysis, this study assesses maximum insertion torque, stress, and strain in bone during insertion of three dental implant types with different macroscopic designs. We model a bone block including cancellous and cortical bones and an osteotomy hole matching the size of the final implant drill. We then model three implants of the same length and diameter but with different thread and body designs. Model I has a conical body with greater apical region tapering and specifically designed threads, model II has wedge-shaped threads and a conical body, and model III is a conical implant with progressive threads. We place models into the bone block at 30 rounds/min and evaluate the implant insertion process in three equal phases of apical, middle, and coronal thirds. We record mean maximum von Mises stress, strain, and insertion torque at 10 points in each third of the osteotomy hole (total of 30 points). In all three implant models, increasing fixture insertion depth into the osteotomy hole results in augmented maximum von Mises stress, strain, and insertion torque in bone. Maximum values are recorded in model I, whereas minimum values are recorded in model III. The dental implant model with crestal microthread design shows maximum von Mises stress, strain, and insertion torque, whereas the model with progressive thread design and absence of coronal threads shows minimum stress, strain, and torque.

The effect of laser irradiation and doxycycline application on the production of matrix metalloproteinase-8 and collagen I from cultured human periodontal ligament cells.

BACKGROUND: Periodontal ligament fibroblasts (PDLF) play a key role in periodontal wound healing and tooth-supporting structures. Various approaches have been tried to enhance the fibroblastic activity such as laser irradiation or doxycycline application. The current study explored the influence of laser irradiation and doxycycline application on human PDLF. The aim of the study was the effect of low-level laser treatment and doxycycline application on the expression of collagen I and matrix metalloproteinase-8 (MMP8) from cultured human periodontal ligament cells. MATERIALS AND METHODS: In this experimental study After preparation of human PDLF in three replications, they were divided into five treatment groups. The first group was day 0, which was used for standardization. The second group was the control group, which received no treatment within 4 days of the study. The third group was treated with doxycycline 30, daily for 4 consecutive days. The fourth group was treated with diode laser 2 daily for 4 consecutive days. The fifth group was treated with both doxycycline and laser irradiation pertaining to the third and fourth groups. After 4 days of treatment, cells were tested for collagen I and MMP-8 secretion through real-time-polymerase chain reaction and ELISA reader. The data were analyzed using the ANOVA and least significant difference pair tests ( P < 0.05 ). RESULTS: Treatment of human PDLF either with diode laser or doxycycline reduced the secretion of MMP-8 significantly. The maximum reduction was related to doxycycline application. Regarding collagen, I, only doxycycline application significantly increased collagen I secretion. Other groups showed no significant increase in collagen I secretion. CONCLUSION: This study showed that treatment of human PDLF either with diode laser or doxycycline significantly reduced MMP-8. Treatment with doxycycline significantly increased the secretion of collagen I.

The effect of low-level laser radiation and doxycycline on the levels of osteoprotegerin and receptor activator of nuclear factor kappa-B ligand.

The present in vitro study was conducted to investigate the effect of low-level laser (LLL) radiation and doxycycline on the levels of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) derived from MG-63 osteosarcoma cell line. MG-63 cells were divided into four groups. In the first group, 2 mg/mL of doxycycline was injected into the cell culture medium. Diode laser (810 nm, 100 mw, 75 s) was radiated to the culture medium of the second group. The third group received both doxycycline and laser radiation. In the fourth group (control), the culture medium was replaced daily, similar to the above three groups. Mentioned interventions were performed once a day for 4 consecutive days. Then, on the sixth day, the levels of OPG and RANKL mediators were measured using real-time polymerase chain reaction by isolating the cells from the samples. OPG expression had the highest to lowest levels in the laser + doxycycline, doxycycline, laser, and control groups, respectively. The level of OPG was significantly different between all the study groups (p < 0.05) except in the doxycycline + laser and doxycycline groups (p = 0.061). The highest to lowest levels of RANKL was observed in the doxycycline, laser + doxycycline, control, and laser groups, respectively. The RANKL expression was not significantly different between all the study groups (p > 0.05). The results of this study revealed that LLL and doxycycline reduced the RANKL/OPG ratio derived from the MG-63 osteosarcoma cell line, which may result in the diminished activity of osteoclasts and osteoclastogenesis.

Interleukin - 17 and Interleukin-10 as Inflammatory and Prevention Biomarkers in Periimplant Diseases.

BACKGROUND: Periimplant diseases are inflammatory diseases. Thus, the level of preinflammatory cytokines which has important role in the inflammation processes can consider as biomchemical markers for early diagnosis and prevention of periimplant diseases. The aim of this study was to determine and compare the level of interleukin (IL)-17 and IL-10 in patients with periimplant mucositis and periimplantitis. METHODS: This case--control study was conducted on 51 patients with implants which were loaded at least 1 year previously, 17 patients with periimplant mucositis, 17 patients with periimplantitis, and 17 individuals with healthy implants. After clinical examination, gingival crevicular fluid sampling was carried out by paper point number 25 for 4 min and the mean value of IL-17, IL-10 in samples was measured using enzyme linked immunosorbent assay (ELISA), least square differences (LSD) reader in laboratory. The data was analyzed using statistical software SPSS 22. Quantitative analysis was done using One-way analysis of variance (ANOVA) test and LSD past test. RESULTS: The results of analysis showed that there was a significant difference in the mean value of IL-17 and IL-10 between the three study groups (P < 0.001). Individuals with healthy implants showed a significant lower level of IL-17 than patients with periimplantitis (P = 0.001) and for patients with periimplantitis, the level of IL-17 was significantly lower than that of patients with periimplant mucositis (P < 0.001) and IL-10 level was significantly lower in mucositis than periimplantitis (P < 0.001). CONCLUSIONS: The level of IL-17 and IL-10 increased in patients with periimplant compared to individuals with healthy periimplant tissues and the results showed that the highest concentrations of IL-17 and IL-10 were observed in patients with periimplant mucositis and periimplantitis, respectively.

Evaluation of Preventive Antibacterial Properties of a Glass-Ionomer Cement Containing Purified Powder of Salvia officinalis: An In vitro Study.

BACKGROUND: In this study, the anti-Streptococcus mutans and anti-Lactobacillus casei properties of a restorative glass-ionomer cement (GIC) modified with extract powder of Salvia officinalis as a safe and effective herbal extract at weight concentration levels of 0.5%, 0.75%, 1%, and 1.25% are investigated. METHODS: The S. officinalis extract powder is provided by doing a multistep laboratory procedure and is filtered to obtain particles smaller than 50 µ. The GIC powder is modified by adding extract powder in weight concentrations of 0.5% (Group II), 0.75% (Group III), 1% (Group IV), and 1.25% (Group V) to form experimental groups, each of 1 g. Five disk-shaped samples, 1 cm in diameter and 2 mm height, of each group (including control group [Group I]) are prepared for each bacterial category of S. mutans and L. casei. The twenty-five samples for each category are tested in vitro against strains of S. mutans and L. casei. Following Agar diffusion tests, the inhibition zone diameters are recorded. The data are tested for normality by means of Kolmogorov-Smirnov procedure. The Kruskal-Wallis and Mann-Whitney tests are used to perform a one-way ANOVA and to do pair-wise comparisons, respectively, at 5% significance level. RESULTS: The mean diameter of the inhibition zones are significantly different among the test groups and also the test groups and the control group except for the group with 0.5% in L. casei category which shows no significant difference with the control group. CONCLUSIONS: The present study revealed direct inhibitory activities of S. officinalis-containing GIC against S. mutans and L. casei in a dose-response manner.

Relationship between Quality of Life, Relationship Beliefs and Attribution Style in Infertile Couples.

BACKGROUND: Many infertile couples experience psychological distress and suffer from impaired quality of life. Generally, when couples are dealing with uncontrolled events such as infertility, it is important to manage it well and to use the suitable coping style; so this can represent an example of attribution style. The purpose of this study is to investigate the quality of life, relationship beliefs and attribution style in infertile couples. MATERIALS AND METHODS: This cross-sectional study consisted of 50 infertile couples, who were at least 18 years of age and could read and write in Persian. Participants provided demographic and general characteristics and completed the quality of life (SF-12), relationship belief inventory (RBI) and attribution style (ASQ) forms. Data was analyzed by the paired t test, Pearson correlation tests and multiple linear regression analysis, using SPSS version 22 statistical software. RESULTS: Overall, 50 infertile couples participated in our study. The males had a significantly higher score for quality of life compared to the females (P=0.019). In RBI subscales except "Disagreement is Destructive" all others significantly higher in wives than husbands. All subscales of RBI had a negative correlation with the quality of life. The quality of life had a significant correlation with positive internal (r=0.213, P=0.033). The adjusted regression model showed that the quality of life for males was higher than in females (ß=-3.098, P=0.024). CONCLUSION: The current data indicate that in infertile couples, the husbands have a higher quality of life in comparison to their wives. Also, all subscales of relationship beliefs have a negative correlation with the quality of life, but in attribution style, just internal attribution style for positive events is associated with the quality of life. In general, there is a correlation between relationship beliefs and the quality of life in infertile couples.

The Effect of Photodynamic Therapy and Diode Laser as Adjunctive Periodontal Therapy on the Inflammatory Mediators Levels in Gingival Crevicular Fluid and Clinical Periodontal Status.

STATEMENT OF THE PROBLEM: The presence of bacterial biofilms is the major cause of gingivitis and periodontitis, their mechanical removal is not often enough. Therefore, laser therapy and photodynamic therapy can be effective as adjunctive treatment. PURPOSE: This study aimed to evaluate the impact of these treatments on the level of gingival crevicular fluid (GCF), inflammatory mediators, and periodontal clinical status. MATERIALS AND METHOD: In this clinical trial, three quadrants were studied in 12 patients with chronic periodontitis aged 30-60 years. The clinical parameters were recorded and GCF samples were taken. After the first phase of periodontal treatment, one of the three quadrants was determined as the control group, one was treated by diode laser, and one underwent photodynamic therapy. The clinical parameters were recorded 2 and 6 weeks later. The data were statistically analyzed by using Friedman, ANOVA, and LSD post-test. RESULTS: Significant reduction was observed over time in the level of Interleukin-1ß (IL-1ß), Interleukin-17 (IL-17), clinical attachment loss, and pocket depth in the three treatment groups (p< 0.000). The three treatment methods significantly reduced the IL-1ß and IL-17 at the baseline, up to 2 weeks, and 2-6 weeks (p< 0.05). Diode laser and photodynamic therapy significantly decreased the average bleeding on probing over time (p< 0.000 and p< 0.002, respectively). CONCLUSION: Laser and photodynamic therapy reduced the inflammatory mediators (IL-1ß and IL-17) and improved the clinical symptoms.

The effect of chronic periodontitis on serum levels of tumor necrosis factor-alpha in Alzheimer disease.

BACKGROUND: Despite the outbreak in dental science, oral and dental complications in Alzheimer are of the unsolved problems. It is assumed that tumor necrosis factor-α, which is a key factor in Alzheimer, has a relation with periodontal complications in patients with Alzheimer disease. The present study evaluated the effect of chronic periodontitis on serum levels of tumor necrosis factor-α in Alzheimer disease. MATERIALS AND METHODS: This case-control study was performed on 80 patients with Alzheimer disease seeking medical care at Nour Hospital, Isfahan, Iran. Eighty patients with Alzheimer disease between 40 and 70 years old attended this study. Forty had chronic periodontitis (case group), and 40 patients had healthy periodontium (control group). Blood sample was taken, and serum levels of tumor necrosis factor-α were measured by means of an ELISA Reader device. Independent T-Test was used to analyze data, and P < 0.05 was considered significant. RESULTS: The mean of tumor necrosis factor-α was 749.1 ng/µL in case group and 286.8 ng/µL in control group. Independent t-test showed that the mean of tumor necrosis factor-α in patients with Alzheimer and periodontitis was approximately three folds higher than the patients only with Alzheimer, and this difference was statistically significant (P < 0.001). CONCLUSION: According to the results of this study, it seems that there is a difference between serum levels of tumor necrosis factor-α in patient with Alzheimer and chronic periodontitis and patients with Alzheimer disease and healthy periodontium. Tumor necrosis factor-α level in serum may act as a diagnostic marker of periodontal disease in patients with Alzheimer disease.

Emdogain effect on gingival fibroblast adhesion in bioabsorbable and non-resorbable barrier membranes: An in vitro study.

BACKGROUND: Tissue engineering represents very exciting advances in regenerative medicine; however, periodontal literature only contains few reports. Emdogain (EMD) consists of functional molecules that have shown many advantages in regenerative treatments. This study investigated EMD effect on gingival fibroblast adhesion to different membranes. MATERIALS AND METHODS: Two dense polytetrafluoroethylene membranes (GBR-200, TXT-200), Alloderm and a collagenous membrane (RTM Collagen) were used in this experimental study. Each membrane was cut into four pieces and placed at the bottom of a well in a 48-well plate. 10 µg/mL of EMD was added to two wells of each group. Two wells were left EMD free. Gingival fibroblasts were seeded to all the wells. Cell adhesion was evaluated by means of a Field Emission Scanning Electron Microscope after 24 hours incubation. Data was analyzed by independent t-test, one-way and two-way ANOVA and post hoc LSD test. P < 0.05 in independent t-test analysis and P < 0.001 in one-way ANOVA, two-way ANOVA and post hoc LSD analysis was considered statistically significant. RESULTS: Alloderm had the highest cell adhesion capacity in EMD+ group and the difference was statistically significant (P < 0.001). In EMD- group, cell adhesion to TXT-200 and Alloderm was significantly higher than GBR-200 and collagenous membrane (P < 0.001). CONCLUSION: This study showed that EMD may decrease the cell adhesion efficacy of GBR-200, TXT-200 and collagenous membrane but it can promote this efficacy in Alloderm. It also showed the composition of biomaterials, their surface textures and internal structures can play an important role in their cell adhesion efficacy.

The effect of adjunctive low-dose doxycycline and licorice therapy on gingival crevicular fluid matrix metalloproteinase-8 levels in chronic periodontitis.

BACKGROUND: This study compared the effect of adjunctive low dose doxycycline and licorice on gingival crevicular fluid (GCF) matrix metalloproteinase-8 (MMP-8) levels in patients with chronic periodontitis. MATERIALS AND METHODS: In this in vivo, experimental study 39 patients with mild to moderate chronic periodontitis were selected. Samples of GCF were collected from three deepest pockets and MMP-8 concentration was measured. Patients were divided into three groups (n = 13). Groups were treated with doxycycline, licorice and placebo. Sampling and measurement of MMP-8 was repeated after 6 weeks. Data was analyzed by t-paired and ANOVA test. P > 0.001 was considered significant. RESULTS: The decrease in mean of MMP-8 concentration was higher in doxycycline and licorice group in comparison with the placebo group and the difference was statistically significant (P value > 0.001). The decrease in mean of MMP-8 concentration was higher in licorice group than doxycycline group, but the difference was not statistically significant. CONCLUSION: The present study showed that licorice extract can prevent the production of MMPs by host cells and can be as useful as antibiotics like doxycycline to cure periodontal and other inflammatory diseases. It must be added that no side-effects were observed in usage of licorice extract.

Debris and smear layer removal efficacy and changes in morphology of dentinal tubules after using citric acid, tetracycline-hydrochloride and mixture of tetracycline and acid and detergent.

BACKGROUND: In periodontal diseases, contamination of roots with bacteria and their active biologic agents and smear layer formation following periodontal treatments, prevents reattachment of periodontal cells. As a matter of fact biomodification of root surface and removal of smear layer and debris is critical for regeneration of periodontal structures. The aim of this study was to compare the efficacy of smear layer removal by citric acid, tetracycline-hydrochloride (TTC-HCL) and mixture of tetracycline and acid and detergent (MTAD). MATERIALS AND METHODS: In this in vitro and experimental study, 12 impacted third molars were sectioned from cervical enamel junction (CEJ) to mid-root area into four dentinal specimens (n = 48). Three groups were conditioned by citric acid 3%, TTC-HCL 50 mg/mL and MTAD by means of cotton pellets, which were changed every 30 s for 3 min. The control group was conditioned by distilled water. Images from scanning electron microscope (SEM) were examined by two different analyzers. Data was analyzed by Kroskal Wallis and Mann-Whitney statistical analysis. RESULTS: The mean of dentinal tubules diameter was the most in MTAD group and the least in TTC-HCL and this difference was statistically significant (P = 0.05). CONCLUSION: According to the present study, TTC-HCL could remove debris and smear layer but citric acid and MTAD had better debris and smear layer removal efficacy and demineralization effect in comparison with TTC-HCL.