Anti-tuberculosis activity of morusin: a promising flavonoid from white mulberry.

BACKGROUND: TB has remained a significant public health concern from historical times to the present day. Each year, growing drug resistance problems necessitate the discovery of new drugs and drug precursors for TB treatment. Morusin is an important flavone found in the bark of white mulberry (Morus alba L.) with anti-oxidant, antimicrobial, anti-tumour, anti-inflammatory and antiallergic activity.OBJECTIVE: To determine the anti-TB efficacy of morusin on Mycobacterium tuberculosis strains.DESIGN: Anti-TB efficacy of morusin was tested on H37Ra (American Type Culture Collection [ATCC] 25177), H37Rv (ATCC 27294), ATCC 35822 (isoniazid [INH] resistant), ATCC 35838 (rifampicin [RIF] resistant), and ATCC 35820 (streptomycin [SM] resistant) standard strains and its efficacy was determined using nitrate reductase assay (NRA).RESULTS: The minimum inhibitory concentration (MIC) of morusin was tested in the range of 53.83â-"0.21 λg/ml. The MIC for H37Ra (ATCC 25177), H37Rv (ATCC 27294) and ATCC 35838 (RIF-resistant) strains were found to be 6.72 λg/ml, and this was 13.45 λg/ml for the ATCC 35822 (INHresistant) and ATCC 35820 (SM-resistant) strains.CONCLUSION: To consider morusin as a viable alternative or precursor drug for TB treatment, it is imperative to conduct an exhaustive examination of its mechanism of action and conduct in vitro studies using clinical isolates.

Porphyromonas gingivalis-Lipopolysaccharide induces cytokines and enzymes of the mouse cementoblasts.

Lipopolysaccharide is a potent virulence factor of Porphyromonas gingivalis and has been implicated predominant pathogen in the development and progression of periodontal diseases. The aim of this study was to determine the effect of Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) on cementoblasts. Cementoblast (OCCM-30) were evaluated proliferation using real-time cell analyzer. In addition, total RNA was isolated at 8, 16, 24 and 72 h from 1000 ng/mL Pg-LPS treated OCCM-30 cells and mRNA expressions of pro/anti-inflammatory cytokine mediators, extracellular matrix enzymes and their tissue inhibitors and of oxidative stress enzymes were studied by real-time polymerase chain reaction. Proliferation analysis indicated that Pg-LPS slightly decreased proliferation of OCCM-30. Pg-LPS had a time-dependent impact on the expression of cytokines and enzymes. There was statistically significant up-regulation of IL-1ß and IL-10 in response to Pg-LPS at 8, 16, 24, 72 h but IL-6 expression was reduced compared to control at 8 h. While IL-8 and IL-17 expressions were determined higher than control group at 16 and 24 h, their expressions were decreased compared to control groups at 72 h (p < 0.01). While MMP-1, MMP-2, MMP-3, TIMP-1, TIMP-2 expressions increased, MMP-9 expression reduced at time-points. Also, a time-dependent up-regulation in mRNA levels for oxidative stress enzymes was detected. These results indicated that up-regulation in the transcripts of inflammation-associated cytokines and degradation enzymes were noted in the cementoblasts exposed to Pg-LPS. Cementoblasts infected with the virulence factors of periodontopathogens might also involve to the induction of inflammation and degradation of the periodontal tissues.

Internalized stigma in acne vulgaris and its relationship with quality of life, general health, body perception, and depression.

BACKROUND: Internalized stigma, adoption of negative attitudes, and societal stereotypes regarding acne vulgaris (AV) have not been previously studied. OBJECTIVE: To investigate the internalized stigma state in AV and determine its association with quality of life, perceived health, body image, and depression. METHODS: A total of 77 AV patients (43 female, 34 male; aged 19.7 ± 2.3 years) were enrolled in this cross-sectional study. The scales used in the study were Acne Internalized Stigma Scale (AISS), Acne Quality of Life scale (AQOL), FDA Global score, Perceived Health Status (PHS), Body Image Scale (BIS), and Beck Depression Inventory (BDI). RESULTS: Mean AISS scores (53.68 ± 13.6) were significantly higher in males than in females (57.41 ± 14.37, 50.39 ± 12.25, P = 0.042). There was a significant positive correlation between mean values of AISS and AQOL (r = 0.816, P < 0.001), FDA Global grade (r = 0.391, P = 0.002) and BDI (r = 0.440, P < 0.001). Lower PHS (P = 0.027) was another determinant of high AISS scores. The mean AISS score of patients with a family history was significantly lower than those without a family history (P = 0.007). VAS was also found to be correlated with mean values of AISS and AQOL. Linear regression analysis revealed that the most important determinant of internalized stigma was AQOL (ß = 0,632; P < 0.001), followed by gender (ß = -0,229; P = 0.001), FDA Global score (ß = 0,193; P = 0.007), and BDI (ß = 0,177; P = 0.024). DISCUSSION: Significant and independent predictive factors for high internalized stigma state were the negative quality of life, male gender, the severity of the illness, and depression. Therefore, internalized stigma may be one of the major factors responsible for the psychosocial burden of AV.

Oral health related quality of life and disease severity in autoimmune bullous diseases.

BACKGROUND: There is an increased risk of long-term dental and periodontal disease in autoimmune bullous diseases (AIBD). AIMS: In this cross-sectional study, we aimed to determine whether the oral health-related quality of life status (OHRQoL) was associated with disease severity and activity in patients with AIBD. SUBJECTS AND METHODS: 67 patients with AIBD were enrolled in this study. Autoimmune Bullous Skin Disorder Intensity Score (ABSIS) was used to evaluate the disease severity. The score was categorized as a significant course (≥17) and moderate course (<17). Oral health impact profile-14 (OHIP-14) questionnaire was filled to assess the OHRQoL. Self-reported oral health status and oral lesion related pain score were also evaluated in the study group. RESULTS: OHIP-14 score was significantly higher in active patients (42.28 ± 13.66) than inactive patients (29.08 ± 12.25) (P = 0.004) and it was correlated with the pain score (6.33 ± 2.78; r = 0.409, P = 0.013). Furthermore, OHIP-14 score was higher in patients with a significant disease course (45.18 ± 15.08) (P = 0.010) than in patients with a moderate course (36.09 ± 9.73). CONCLUSIONS: OHRQoL may be useful in the disease management and treatment. Since it can be affected by both presence of oral erosions and disease severity, a collaboration between dermatologists and dentists could be crucial to the disease management in AIBD.

Langerhans Cell Histiocytosis: Single Center Experience of 25 Years.

OBJECTIVES: To review a single center outcome of patients with Langerhans Cell Histiocytosis diagnosed at a tertiary referral hospital from Turkey.Methods: The files between 1989 and 2015 of 80 patients with LCH were retrospectively analyzed. RESULTS: During the 25 years, 80 patients were diagnosed with LCH. The median age at diagnosis was 53 months (2-180 months) and the median follow-up time of patients was 10 years and 9 months (24 months-25 years). Bone was the most frequently affected organ (n:60, 75%). Initially, 43 patients (54%) had single system (SS) disease, 20 patients (25%) had multisystem (MS) disease without risk organ involvement (MS-RO-), and 17 patients (21%) had a multisystem disease with risk-organ involvement (MS-RO+). The overall survival (OS) rate was 91%, and event-free survival (EFS) rate was 67% at 10 years. 10-year OS rate was lower for patients with MS-RO+ (65%) when compared to those with, MS-RO-, and SS (100%, 97%, p value=<0.001). The overall survival rate was also lower in patients with lack of response to systemic chemotherapy on 12th week (p=<0.001), younger age (<2 years) at presentation (p=<0.02), skin involvement (<0.001) and lack of bone lesions at presentation (<0.001). DISCUSSION: In the group with MS-RO+, OS is significantly low compared to other groups. Further efforts are warranted to improve survival in MS-RO+ patients.

Recombinant amelogenin regulates the bioactivity of mouse cementoblasts in vitro.

Amelogenin (AMG) is a cell adhesion molecule that has an important role in the mineralization of enamel and regulates events during dental development and root formation. The purpose of the present study was to investigate the effects of recombinant human AMG (rhAMG) on mineralized tissue-associated genes in cementoblasts. Immortalized mouse cementoblasts (OCCM-30) were treated with different concentrations (0.1, 1, 10, 100, 1000, 10,000, 100,000 ng · mL-1) of recombinant human AMG (rhAMG) and analyzed for proliferation, mineralization and mRNA expression of bone sialoprotein (BSP), osteocalcin (OCN), collagen type I (COL I), osteopontin (OPN), runt-related transcription factor 2 (Runx2), cementum attachment protein (CAP), and alkaline phosphatase (ALP) genes using quantitative RT-PCR. The dose response of rhAMG was evaluated using a real-time cell analyzer. Total RNA was isolated on day 3, and cell mineralization was assessed using von Kossa staining on day 8. COL I, OPN and lysosomal-associated membrane protein-1 (LAMP-1), which is a cell surface binding site for amelogenin, were evaluated using immunocytochemistry. F-actin bundles were imaged using confocal microscopy. rhAMG at a concentration of 100,000 ng · mL-1 increased cell proliferation after 72 h compared to the other concentrations and the untreated control group. rhAMG (100,000 ng · mL-1) upregulated BSP and OCN mRNA expression levels eightfold and fivefold, respectively. rhAMG at a concentration of 100,000 ng · mL-1 remarkably enhanced LAMP-1 staining in cementoblasts. Increased numbers of mineralized nodules were observed at concentrations of 10,000 and 100,000 ng · mL-1 rhAMG. The present data suggest that rhAMG is a potent regulator of gene expression in cementoblasts and support the potential application of rhAMG in therapies aimed at fast regeneration of damaged periodontal tissue.

Comparison of Different Sources of Mesenchymal Stem Cells: Palatal versus Lipoaspirated Adipose Tissue.

OBJECTIVES: The purpose of this study was to compare the proliferation and differentiation potential of mesenchymal stem cells (MSCs) derived from palatal adipose tissue (PAT) and lipoaspirated adipose tissue (LAT). MATERIALS AND METHODS: PATs were obtained from 2 healthy female patients undergoing surgery for gingival recession, and LATs were obtained from 2 healthy female patients undergoing plastic surgery. LAT- and PAT-derived MSCs were confirmed by flow cytometry using MSC-specific surface markers. The multilineage differentiation capacity of the MSCs was analyzed. The expression of immunophenotyping, embryonic, and differentiation markers was compared between both MSC lines. The proliferation of PAT- and LAT-MSCs was evaluated using a real-time cell analyzer, and telomerase activity was determined using an ELISA-based TRAP assay. Stem cells isolated from PAT and LAT were analyzed by real-time PCR and whole genome array analysis. RESULTS: The cells isolated from PAT had MSC characteristics. In addition, PAT-MSCs had significantly higher alkaline phosphatase activity and osteogenic potential than LAT-MSCs. Although the proliferation and telomerase activities of LAT-MSCs were higher than those of PAT-MSCs, the difference was not statistically significant. The level of embryonic stem cell markers (Oct4 and Nanog) was higher in LAT-MSCs than in PAT-MSCs. The whole genome array analysis demonstrated that 255 gene sequences were differentially expressed, with more than a twofold change in expression. CONCLUSIONS: This is the first comparative analysis of the isolation and characterization of MSCs from PAT and LAT. PAT is an accessible source of MSCs, which could be used in periodontal and craniofacial tissue engineering.

HE4: really a novel biomarker for ovarian cancer?.

Ovarian cancer forms 4% of all cancers and approximately 23% of all gynecological cancers in women and is responsible for the 47% of deaths related to cancers of the genital tract of women. Tumor markers are the biochemical substances which can be detected in the presence of tumors. Generally they are either the products of tumoral tissues or secreted from the normal cells which are in the inter- action with tumoral ones. The present authors attempted to determine the efficacy of the tumor marker CA- 125 and HE4 to differentiate the malign cases from the benign adnexal masses. A total of 76 patients with the appropriate criteria were included in the study. They were divided into three groups; healthy control group (n=3 1), ones with benign masses (n=23), and ones with malign ovarian masses (n=22). In the study, when the cut-off values were accepted as 55I U/ml for CA-125 and 150 pM for HE4 in differentiation of benign and malign groups, the sensitivity was found as 59.09%, specificity 91.3%, PPV 86.67% and NPV 70% LR = +6.8. This combination gives one false positive result to every five positive cases which were detected as high. With the combination of CA-125 and HE4, the value of sensitivity was found decreased as expected, although the value of the specificity increased.

Porphyromonas gingivalis Lipopolysaccharide Induces a Pro-inflammatory Human Gingival Fibroblast Phenotype.

Human gingival fibroblasts (HGFs) are the major constituents of the gingival tissues responsible for the synthesis and degradation of the connective tissue while actively participating in immune reactions and inflammation. The aim of this study was to test the impact of lipopolysaccharide (LPS) from Porphyromonas gingivalis (P. gingivalis) on human gingival fibroblasts. Human gingival fibroblasts were treated with different P. gingivalis LPS concentrations. Cell survival rate was evaluated with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) after 24 h. Cell proliferation was determined by counting cells on days 3 and 12. Expression of matrix metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), and pro-inflammatory cytokine transcripts in HGFs was determined by quantitative PCR (Q-PCR) analysis on days 3 and 8. P. gingivalis LPS decreased cell proliferation on day 3 (p < 0.05) compared to the control group without significantly impacting the cell survival (p > 0.05).The experiments showed that P. gingivalis LPS dose-dependently and differentially modulated the expression of MMP-1, 2, and 3 and TIMP-1 and 2 on days 3 and 8. TIMP-1 expression was significantly induced in P. gingivalis LPS-treated cells while TIMP-2 was increased in response to 10 and 30 ng/ml of LPS on day 3. P. gingivalis LPS induced up-regulation of MMP-1/TIMP-1 ratio on day 3 and increased MMP-2/TIMP-2 ratio on day 8 dose-dependently. Expression of interleukin (IL)-6 and IL-8 was stimulated at higher concentrations (1000 and 3000 ng/ml) of LPS. These findings demonstrate that P. gingivalis LPS suppresses cell proliferation and leads to increased pro-inflammatory changes in HGFs, suggesting that P. gingivalis LPS-induced modification of phenotypic and inflammatory characteristics in HGF could potentially be a pathogenic mechanism underlying the tissue destruction.

Clinical Significance of Intraoperative Frozen Section Analysis of Pancreatic Cancer Surgical Margin at the Time of Pancreaticoduodenectomy.

BACKGROUND: Today, in the management of pancreas cancers, achieving an R0 resection is one of the most powerful independent predictors of long-term survival. The aim of this study is to assess the clinical significance of intraoperative frozen section analysis of the pancreatic surgical margin for pancreatic cancer during pancreaticoduodemectomy. MATERIAL AND METHODS: We conducted a retrospective analysis of prospectively collected data of 37 patients who were operated for pancreatic head cancer and who were evaluated for surgical margin by frozen section analysis intraoperatively, between September 2013 and August 2014 in our center. The intraoperative biopsy reports were compared with final pathological reports. RESULTS: The mean age of the patients was 64.55(19-82) years (range), the mean tumor size was 3.96(1.16-6.25) cm (range) and the mean harvested lymph node number was 18.52(9-45) (range). In the intraoperative frozen section, one patient was positive for surgical margin (%2.7) who underwent total or complementary pancreatectomy. CONCLUSION: To secure a tumor-free margin by frozen section, intraoperatively, may increase R0 resection rate in pancreas cancers. The preoperative estimation of tumor margin by endoscopic ultrasonography, computerized tomography or magnetic resonance imaging mostly correlate with intraoperative findings, however in suspected cases intraoperative frozen section for margin determination should be performed.

Evaluation of gingival crevicular fluid levels of tissue plasminogen activator, plasminogen activator inhibitor 2, matrix metalloproteinase-3 and interleukin 1-ß in patients with different periodontal diseases.

OBJECTIVES: The purpose of this study was to evaluate the gingival crevicular fluid levels of interleukin-1beta (IL-1ß), matrix metalloproteinases-3 (MMP-3), tissue type plasminogen activator (t-PA) and plasminogen activator inhibitor 2 (PAI-2) in patients with chronic periodontitis, aggressive periodontitis (AgP) and healthy individuals (controls). MATERIAL AND METHODS: Systemically healthy (21 chronic periodontitis, 23 AgP and 20 controls) subjects were included in this study. Plaque index, gingival index, probing pocket depth and clinical attachment level were recorded and gingival crevicular fluid samples were collected. Assays for IL-1ß, MMP-3, t-PA and PAI-2 levels in gingival crevicular fluid were carried out by an enzyme-linked immunosorbent assay. The one-sample Kolmogorov-Smirnov test, Mann-Whitney U test and Spearman correlation coefficient were used for data analyses. RESULTS: Gingival crevicular fluid levels of t-PA and IL-1ß were significantly higher in chronic periodontitis and AgP groups than in the control group (p < 0.001). MMP-3 levels in gingival crevicular fluid were detected as significantly higher in the chronic periodontitis and AgP groups compared with the control group (p < 0.05). The t-PA/PAI-2 rate of patients with chronic periodontitis and AgP were significantly higher than the control group (p < 0.05). The positive correlations were found among the PAI-2, t-PA, IL-1ß and MMP-3 levels in gingival crevicular fluid. The volume of the gingival crevicular fluid correlated with all of the clinical parameters (p < 0.001). There were positive correlations between the gingival crevicular fluid levels of PAI-2 and the probing pocket depth and between gingival crevicular fluid levels of PAI-2 and the clinical attachment level (p < 0.01). Similarly, significant correlations were found between t-PA levels and probing pocket depth and between t-PA levels and clinical attachment level measurements (p < 0.001). CONCLUSION: The present data showed that gingival crevicular fluid levels of IL-1 ß, MMP-3 and t-PA increased in periodontal disease regardless of the periodontitis type and played a part in tissue destruction.

Comparison of mesenchymal stem cells isolated from pulp and periodontal ligament.

BACKGROUND: Cell-based therapy using mesenchymal stem cells (MSCs) seems promising to obtain regeneration of dental tissues. A comparison of tissue sources, including periodontal ligament (PDL) versus pulp (P), could provide critical information to select an appropriate MSC population for designing predictable regenerative therapies. The purpose of this study is to compare the proliferation and stemness and the MSC-specific and mineralized tissue-specific gene expression of P-MSCs and PDL-MSCs. METHODS: MSCs were obtained from PDL and P tissue of premolars (n = 3) extracted for orthodontic reasons. MSC proliferation was evaluated using a real-time cell analyzer for 160 hours. Telomerase activity was evaluated by a telomeric repeat amplification protocol assay based on enzyme-linked immunosorbent assay. Total RNA was isolated from the MSCs on day 3. A polymerase chain reaction (PCR) array was used to compare the expression of MSC-specific genes. The expression of mineralized tissue-associated genes, including Type I collagen (COL I), runt-related transcription factor 2 (RunX2), bone sialoprotein (BSP), and osteocalcin (OCN) messenger RNA (mRNA), was evaluated using quantitative real-time PCR. RESULTS: Higher proliferation potential and telomerase activity were observed in the P-MSCs compared to PDL-MSCs of premolar teeth. Fourteen of 84 genes related to MSCs were expressed differently in the PDL-MSCs versus the P-MSCs. The expressions of bone morphogenetic protein 2 (BMP2) and BMP6; sex-determining region Y-box 9 (SOX9); integrin, alpha 6 (ITGA6); melanoma cell adhesion molecule (MCAM); phosphatidylinositol glycan anchor biosynthesis, class S (PIGS); prominin 1 (PROM1); ribosomal protein L13A (RPL13A); and microphthalmia-associated transcription factor (MITF) were higher in the P-MSCs compared to the PDL-MSCs, and higher expression of matrix metalloproteinase 2 (MMP2), interleukin (IL)-6, insulin (INS), alanyl (membrane) aminopeptidase (ANPEP), and IL-10 were observed in the PDL-MSCs. However, there was no statistically significant difference in the expression of mineralized tissue-associated genes, including BSP and RunX2, between the P-MSCs and the PDL-MSCs. Higher expression of COL I and lower expression of OCN mRNA transcripts were noted in the PDL-MSCs compared to the P-MSCs. CONCLUSIONS: The results of this study suggest that MSCs isolated from P and PDL tissues show different cellular behavior. To increase the predictability of MSC-based regenerative treatment, differences in dental tissue-derived MSCs and favorable aspects of cell sources should be further clarified.

Laparoscopic versus open appendectomy: where are we now?

PURPOSE: Although the advantages of laparoscopic procedures has been well studied over the last two decade, laparoscopic appendectomy could not to be a standard therapy due to some disadvantages such as longer operative time and higher cost.The objective of our study is to re-evaluate the outcomes of laparoscopic versus open appendectomy with current data. METHODS: Between January 2012 and July 2012, the data of the patients who had appendectomy were recorded prospectively. Patients' demographics, duration of procedure, length of hospital stay, need of analgesics, postoperative visual analogue scale scores and morbidity were assessed. RESULTS: Of 241 patients, 120 (49.8%) underwent open and 121(50.2%) laparoscopic appendectomy. The operating time was similar for both groups (p=0.855). The visual analog scale scores of 1st (p=0.001), 6th (p=0.001) and 12th (p=0.028) hours were higher in open the appendectomy group. The total need of analgesics significantly was higher in open group (p=0.001).There was no statistical difference in terms of total morbidity rate between open and laparoscopic appendectomy groups (p=0.617). CONCLUSION: Two operative techniques are similar in terms of length of hospital stay, operative time, and postoperative complications. Laparoscopic appendectomy reduces the need for analgesics and visual analog scale scores; therefore,it should be considered as the gold standard for surgical treatment of acute appendicitis.

A reliable pancreaticojejunal anastomosis with V-Loc 180 wound closure device for soft pancreatic stump.

BACKGROUND/AIMS: Although mortality rates decreased in recent years, pancreaticoduodenectomy is still associated with high morbidity rates. Pancreatic fistula is the leading cause of morbidity after pancreaticojejunal anastomosis and commonly occurs in soft pancreas. The objective of this study is to compare outcomes of conventional modified invaginated end to side pancreaticojejunostomy with a new practical method using V-Loc 'rM 180 wound closure device in soft pancreas. METHODOLOGY: Between December 2011 and August 2013, a total of 90 pancreaticoduodenectomy procedures were performed in our hospital. 28 of them were defined as soft pancreas according to attending surgeon and included in this study. Patients were divided into two groups consecutively and analysed for postoperative pancreatic fistula (POPF) rate, length of stay, operation time, cost and particular duration of anastomosis. Pancreatic fistulas were classified according to International Study Group on Pancreatic Fistula (ISGPF) definition. RESULTS: 1 grade A and 2 grade B fistulas appeared in V-Loc group (Group 1), whereas 1 grade A, 2 grade B and 1 grade C fistulas appeared in conventional anastomosis group (Group 2). CONCLUSIONS: Pancreaticojejunostomy with V-Loc suture is a convenient method in soft pancreas and can be performed safely.

On the number of spanning trees of graphs.

We establish some bounds for the number of spanning trees of connected graphs in terms of the number of vertices (n), the number of edges (m), maximum vertex degree (Δ1), minimum vertex degree (δ), first Zagreb index (M 1), and Randic index (R -1).

Efficacy of collagen membrane seeded with autologous gingival fibroblasts in gingival recession treatment: a randomized, controlled pilot study.

BACKGROUND: Gingival recession (GR) is one of the most common esthetic concerns associated with periodontal tissues. Recently, tissue engineering technology has been developed and applied in periodontology for the treatment of GR. The aim of this study is to compare the clinical efficacy of collagen membrane with or without autologous gingival fibroblasts under a coronally advanced flap for root coverage. METHODS: In this split-mouth, controlled clinical study, 22 sites are selected from 11 patients with Miller Class I recessions affecting canines or premolars in the maxillary arch. One tooth in each patient was randomized to receive either a collagen membrane (CM) (control group) or a collagen membrane seeded with autologous gingival fibroblasts (CM+GF) (test group) under a coronally advanced flap. Thickness of the gingiva, GR, and percentage of root coverage (PRC) were recorded by a calibrated examiner at baseline and 3, 6, and 12 months postoperatively. Furthermore, GR and PRC were evaluated using photogrammetric analysis at baseline and 3, 6, and 12 months. RESULTS: Both treatments resulted in a significant gain in root coverage compared with baseline. A statistically significant increase was detected in PRC in the test group compared with the control group. No significant difference was noted between the test and control sites regarding the thickness of the gingiva. CONCLUSIONS: The results indicated that CM+GF prepared by tissue engineering technology can be considered an alternative method for the treatment of Miller Class I recession defects.

Osteogenic differentiation of MC3T3-E1 cells on different titanium surfaces.

mRNA expressions related to osteogenic differentiation of MC3T3-E1 cells on electro-polished smooth (S), sandblasted small-grit (SSG) and sandblasted large-grit (SLG) surfaces of titanium alloys were investigated in vitro. Gene expression profiles of cells were evaluated using the RT2 Profiler PCR microarray on day 7. Mineralizing tissue-associated proteins, differentiation factors and extracellular matrix enzymes mRNA expressions were measured using Q-PCR. SLG surface upregulated 23 genes over twofolds and downregulated 3 genes when compared to the S surface. In comparison to the SSG surface, at least a twofold increase in 25 genes was observed in the SLG surface. BSP, OCN, OPN, COL I and ALP mRNA expressions increased in the SLG group when compared to the S and the SSG groups. BMP-2, BMP-6 and TGF-ß mRNA expressions increased in both the SSG and the SLG surfaces. MMP-2 and MMP-9 mRNA expressions increased as the surface roughness increased. This study demonstrated that surface roughness of titanium implants has a significant effect on cellular behavior and SLG surface apparently increased gene expressions related to osteogenesis when compared to the S and the SSG surfaces.

Effects of different setting of diode laser on the mRNA expression of growth factors and type I collagen of human gingival fibroblasts.

The aim of this study was to analyze the influence of non-surgical applications of diode laser (940 nm) on the cell proliferation and mRNA expressions of type I collagen and growth factors in human gingival fibroblasts (GF). Gingival fibroblasts were isolated from human gingival connective tissue of systemically healthy individuals. Cells were treated with different laser parameters as follows; (1) Infected pocket setting (power: 2 W, pulse interval: 1 ms, pulse length: 1 ms, 20 s/cm(2)); (2) Perio-pocket setting (power: 1.5 W, pulse interval: 20 ms, pulse length: 20 ms, 20 s/cm(2)); and (3) Biostimulation setting (power: 0.3 W in continuous wave, 20 s/cm(2)). Proliferation of GF was evaluated after different laser applications using a real-time cell analyzer. Total RNA was isolated on day 2 and cDNA synthesis was performed. Type I collagen, insulin-like growth factor (IGF), vascular endothelial growth factor (VEGF) and transforming growth factor-beta (TGF-ß) mRNA expressions were determined with quantitative RT-PCR. In a proliferation experiment, no significant differences were observed in the different laser applications when compared to the control group. Statistically significant increases in IGF, VEGF, and TGF-ß mRNA expressions were noted in the laser groups when compared to the untreated control group (p < 0.05). A significant increase in collagen type I mRNA expression was noted in only biostimulation set-up of diode laser (p < 0.05). The results of this study demonstrate that non-surgical laser applications modulate behavior of gingival fibroblasts inducing growth factors mRNA expressions and these applications can be used to improve periodontal wound healing.

Periodontal ligament fibroblast response to root perforations restored with different materials: a laboratory study.

AIM: To compare the effect of several materials on the attachment of periodontal ligament (PDL) fibroblasts to experimentally perforated root surfaces. METHODOLOGY: Root specimens (size 5 × 5 mm) were obtained from extracted human molar teeth and perforations with a 1 mm diameter were created. One group was kept as a control and the rest were repaired with the following materials: Amalgam, Dyract, IRM, Super Bond C&B and Mineral trioxide aggregate (MTA). PDL fibroblasts were placed at a density of 8 × 10(4) cells on the root specimens, incubated on tissue culture inserts (48 h) and then transferred to 48 well-plates. MTT assays were performed at 48 and 96 h for PDL fibroblast survival. Cell attachment was observed using confocal microscopy on days 2 and 5. Total RNAs from the root specimens were isolated on day 5 and type I collagen (COL I) and Runt-related transcription factor 2 (Runx2) mRNA expressions were checked using Quantitative-Polymerase Chain Reaction (QPCR). For the MTT assay and QPCR, one-way analysis of variance (anova) and Tukey HSD multiple comparison tests were used to compare the groups. RESULTS: Mineral trioxide aggregate resulted in a significantly higher cell density (P < 0.001). Dyract, IRM and Super Bond C&B groups had a lower cell density when compared with the control and MTA groups at 48 h (P < 0.001). Confocal microscopy revealed that, among the experimental groups, the MTA group had the largest viable cell population over the restoration site when compared with the other materials; however, reduced cell attachment was noted in all groups when compared with the control. Increased Runx2 mRNA expressions were noted in MTA (P < 0.001) and IRM (P < 0.01) groups when compared with control and other tested materials. COL I transcripts were increased in IRM (P < 0.01), D, C&B and MTA (P < 0.001) when compared with the control. CONCLUSION: Mineral trioxide aggregate provided a more favorable environment for PDL cell adhesion and growth.

Bone morphogenetic protein-7 enhances cementoblast function in vitro.

BACKGROUND: Bone morphogenetic protein (BMP)-7 is a potent bone-inducing factor and was shown to promote periodontal regeneration in vivo and in vitro; however, to our knowledge, the specific effect of BMP-7 on cementoblasts has not been defined. We aimed to investigate the effects of BMP-7 on cementoblasts, which are cells responsible for tooth root-cementum formation. We hypothesized that BMP-7 would regulate mineralized tissue-associated genes in cementoblasts and influence the expression profile of genes associated with cementoblast extracellular matrix (ECM) and cell adhesion molecules (CAMs). METHODS: A murine immortalized cementoblast cell line (OCCM.30) was cultured with and without 50 ng/ml BMP-7. After 72 hours, total RNA was isolated, and mRNA levels for bone/cementum markers, including bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor-2 (Runx2), were investigated by real-time quantitative reverse transcription-polymerase chain reaction (Q-PCR). In vitro mineral nodule formation was assayed on day 8 using von Kossa staining. A pathway-specific gene-expression array was used to determine BMP-7-responsive ECM and CAM genes in cementoblasts. RESULTS: Mineralized tissue markers were strongly regulated by BMP-7, with an almost three-fold increase in BSP and OCN transcripts and significant increases in OPN and Runx2 mRNA expressions. BMP-7 treatment markedly stimulated cementoblast-mediated biomineralization in vitro compared to untreated cells at day 8. BMP-7 treatment altered the OCCM.30 expression profile for ECM and CAM functional gene groups. BMP-7 tended to increase the expression of collagens and matrix metalloproteinases (MMPs), mildly decreased tissue inhibitors of MMPs (TIMPs), and had mixed regulatory effects on integrins. Using Q-PCR, selected array results were confirmed, including a significant BMP-7-induced increase in MMP-3 and a decrease in TIMP-2 mRNA expression. CONCLUSION: These results support the promising applications of BMP-7 in therapies aimed at regenerating periodontal tissues lost as a consequence of disease.