Bacterial adhesion and colonization differences between zirconium oxide and titanium alloys: an in vivo human study.

PURPOSE: The purpose of this study was to compare zirconium oxide and titanium alloys with respect to their tendency to adhesion and colonization of two periodontal pathogens on both hard surfaces and on soft tissues in vivo. MATERIALS AND METHODS: The present study was designed as a prospective stratified randomized controlled clinical trial. Patients were scheduled to receive two implants with different types of abutments in the posterior mandible. Three months after implant placement, titanium and zirconium abutments were connected. Five weeks after abutment connections, the abutments were removed, probing depth measurements were recorded, and gingival biopsy samples were obtained. Abutments and biopsy specimens were analyzed by reverse-transcriptase polymerase chain reaction to compare the DNA copy numbers of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and total bacteria. The surface free energy of the abutments was calculated by sesile water drop before replacement. RESULTS: No statistically significant differences were found between probing depths or DNA copy numbers of A actinomycetemcomitans, P gingivalis, and total bacteria both for both titanium alloys and zirconium oxide surfaces and the biops specimens obtained from their buccal gingival. With respect to the surface free energy of zirconium and titanium abutments, zirconium abutments showed lower surface free energy than titanium abutments. CONCLUSION: The results of this study showed that zirconium oxide surfaces have comparable properties to titanium alloy surfaces in their tendency to adhesion and colonization of two periodontal pathogens on both hard surfaces and in soft tissues.

Differentiation of human embryonic stem cells on periodontal ligament fibroblasts in vitro.

Human embryonic stem cells (hESCs) are pluripotent cells with unlimited proliferation potential and differentiation capacity to all types of somatic cells. Periodontal tissue engineering based on in vitro expanded cells holds the promise to overcome the limitations associated with contemporary regenerative techniques. The aim of this study was to investigate the differentiation patterns of hESCs under the influence of periodontal ligament cells in vitro. hESCs (HUES-9) were expanded and characterized for their pluripotency. Then they were transfected with green fluorescent protein-carrying plasmid, and cocultured with human periodontal ligament fibroblastic cells for 21 days. Two experimental groups were established with different medium constituents. Specimens were fixed at days 7, 14, and 21 and were analyzed morphologically under inverted light microscope, and by immunohistochemistry using antibodies against collagen types I and III, fibronectin, fibroblast surface protein, vimentin, and pancytokeratin. Our results demonstrate different patterns of cell differentiation between groups, with about one-fifth of cells in colonies acquiring characteristics similar to periodontal ligament fibroblastic progenitors while others proceed toward distinctive lineages. This indicates the feasibility to direct the differentiation of hESCs toward the periodontal ligament fibroblastic progenitors to some extent. These findings support the notion that hESCs may become a cell source with unlimited supply for periodontal tissue engineering applications.

Encapsulation and osteoinduction of human periodontal ligament fibroblasts in chitosan-hydroxyapatite microspheres.

Periodontal ligament cells play a crucial role in the regeneration of periodontal tissues and an undifferentiated mesenchymal cell subset is thought to exist within this population. The aim of this study was to assess the osteogenic differentiation potential of human periodontal ligament fibroblasts (hPDLFs) in three dimensional (3D)-osteogenic culture environment following encapsulation in chitosan-hydroxyapatite (C/HA) microspheres with the size range of 350-450 microm. Human PDLF cultures were established and three experimental groups were formed: (i) two-dimensional (2D)-culture as single cell monolayer, (ii) 3D-static culture of C/HA encapsulated hPDLFs, and (iii) 3D-dynamic culture of C/HA encapsulated hPDLFs in a rotating wall vessel bioreactor. The cells were cultured in standard culture medium supplemented with beta-glycerophosphate, dexamethasone, and ascorbic acid. After 21 days, immunohistochemistry was performed using antibodies against osteonectin, osteopontin, bone-sialoprotein, and osteocalcin as osteogenic differentiation markers. Phase-contrast and scanning electron microscopy observations were used for histological and morphological evaluation. The combined effects of osteoinductive medium and HA-containing composite microsphere material on encapsulated hPDLFs resulted in the transformation of a considerable portion of the cells into osteoblastic lineage at the end of the experiments. Results demonstrate the ability of hPDLFs to undergo osteogenic differentiation upon induction in vitro, both under 2D and 3D culture conditions. C/HA microspheres in microgravity bioreactor may serve as a suitable 3D environment to support the osteogenic differentiation of human PDLFs, in vitro.

Comparison of different methods to detect Helicobacter pylori in the dental plaque of dyspeptic patients.

The aim of this study was to compare different methods of detection of Helicobacter pylori (H. pylori) in the dental plaque of dyspeptic patients. After recording the clinical indices, culture and polymerase chain reaction (PCR) methods were performed on plaque samples, while rapid urease test in addition to these tests was carried on gastric samples from 67 dyspeptic patients who attended for an upper gastrointestinal endoscopy. Forty-seven of 67 patients were H. pylori-positive in gastric biopsy material whereas the microbial dental plaque from 19 patients demonstrated H. pylori positivity detected by PCR. Among the patients, 25.4% harbored H. pylori both in the stomach and in microbial dental plaque. No significant correlations were found among the presence of H. pylori in the stomach, in plaque, and clinical variables (P > 0.05). Although oral hygiene was observed optimal and the mean of pocket depth was not found to be higher, the prevalence of H. pylori was observed to be higher in dental plaque. According to our results, PCR technique gave the highest detection rate both in gastric biopsy and in dental plaque compared to the other methods used.

Gingival crevicular fluid alkaline phosphatase levels in postmenopausal women: effects of phase I periodontal treatment.

BACKGROUND: The aim of this study was to determine how estrogen status may possibly influence gingival crevicular fluid (GCF) alkaline phosphatase (ALP) levels in estrogen-deficient (ED) and -sufficient (ES) postmenopausal women at baseline (BL) and 1 year after periodontal phase I treatment (AT). METHODS: Thirty-six postmenopausal women on estrogen supplements (mean serum estradiol levels >30 pg/ml; estrogen sufficient) and 37 postmenopausal women not on estrogen supplements (mean serum estradiol levels <30 pg/ml; ED) were divided into two subgroups as chronic periodontitis and clinically healthy controls after clinical and radiographic examination. The ES group consisted of 19 control (ES/C) and 17 chronic periodontitis (ES/P) patients, and the ED group consisted of 20 control (ED/C) and 17 chronic periodontitis (ED/P) patients. Plaque (PI) and gingival (GI) indices, bleeding on probing (BOP), probing depths (PD), clinical attachment loss (CAL) scores, and GCF samples were recorded at BL and AT. The levels of ALP in the GCF were measured photometrically. The paired samples Student t and Wilcoxon tests were used to compare the ALP levels and clinical parameters between BL and AT. The correlation among the ALP and clinical parameters was analyzed using the Pearson correlation. RESULTS: The mean of all clinical parameters (PI, GI, BOP, PD, and CAL) was significantly (P <0.05) higher in periodontitis groups (ES/P and ED/P) than controls (ES/C and ED/C). Periodontitis groups (ES/P and ED/P) demonstrated significantly increased GCF volumes and GCF ALP levels (P <0.05) compared to controls (ES/C and ED/C). There were no significant differences in the concentrations of ALP between periodontitis and control groups (P >0.05). The BL GCF ALP total levels of the ED/P group were significantly higher than the ES/P group (P <0.05). The BL and AT serum ALP levels of the ED/P group were not significantly but were numerically higher than the ES/P group. One year after periodontal treatment, the GCF volume, GCF ALP total, and concentrations decreased significantly in both periodontitis groups (P <0.05). However, the GCF ALP levels were still numerically higher in the ED/P group. A positive statistical correlation was found between total ALP levels and PD (r = 0.621; P <0.05). CONCLUSION: These data suggest that the presence of ALP in GCF is not simply a reflection of the local inflammation state and that a patient's estrogen status may possibly influence local ALP levels in GCF.

Levels of interleukin-8 during tooth movement.

A host-derived neutrophil-activating cytokine interleukin-8 (IL-8) is secreted mainly by monocytes and is considered to be important in regulating alveolar bone resorption during tooth movement. The aim of this study was to evaluate the levels of IL-8 during mechanical forces on periodontal tissues at different stages of orthodontic therapy. Ten canine teeth of patients having different Angle classifications were selected for the study. After the premolars were extracted, the maxillary/mandibular canines were tipped distally. Gingival crevicular fluid was sampled from mesial and distal gingival crevices of each canine separately at baseline and one hour, 24 hours, six days, 10 days, and 30 days after the application of the force. An enzyme-linked immunosorbent assay for quantitative detection of IL-8 was used. Although there was an increase in the concentration of IL-8 at tension (mesial) sites after one hour, 24 hours, six days, and 10 days, a decrease was observed at 30 days. Pressure (distal) sites did not demonstrate such an increase at any period except at 10 days. However, the concentration of IL-8 at both sites showed a similar decrease and approached each other at day 30. We concluded that local host response toward the orthodontic forces might lead an increase in IL-8 and neutrophil accumulation, and this may be one of the triggers for bone remodeling processes.

Periodontal regenerative potential of autogenous periodontal ligament grafts in Class II furcation defects.

BACKGROUND: The aim of the present study was to evaluate the regenerative potential of autogenous periodontal ligament (PDL) grafts in the treatment of Class II furcation defects. METHODS: Twenty mandibular Class II furcation defects from 10 systemically healthy patients with chronic periodontitis were selected. In experimental defects, flaps were coronally positioned following placing autogenous PDL grafts that were obtained from third molars; in controls, coronally advanced flap procedure without graft was applied. Clinical measurements including plaque index, gingival index, probing depth (PD), vertical and horizontal clinical attachment level (CAL), and gingival recession (GR) were obtained at baseline and after 3 and 6 months postoperatively. Vertical and horizontal defect fill was evaluated with open clinical measurements at initial surgery and reentry after 6 months. Gingival biopsies from the experimental and control defects were obtained at reentry and evaluated histopathologically in order to examine the soft tissue response towards PDL grafts. RESULTS: Sites treated with PDL grafts demonstrated significant improvement in vertical and horizontal defect fill, PD, and CAL at 3 and 6 months compared to presurgical values. The difference determined for the PD values of both groups at a statistically significant degree in favor of grafted sites was maintained at all observation periods. No foreign body reaction was observed in PDL grafts. CONCLUSIONS: These short-term results point to the potential of PDL grafts in promoting healing of furcation lesions. This preliminary study suggests that the use of PDL grafts may have beneficial effects in the treatment of furcation defects.

Effectiveness of scaling and root planing versus modified Widman flap on nitric oxide synthase and arginase activity in patients with chronic periodontitis.

BACKGROUND: Nitric oxide (NO) is synthesized from the conversion of L-arginine to L-citrulline by NO synthase (NOS). Arginase, which is an arginine-depleting enzyme, can compete with NOS for the common substrate L-arginine and thus inhibit NO production. OBJECTIVES: In the present study, we aimed to examine the correlation between the arginase and NOS activity in patients with chronic periodontitis and to compare the effects of scaling and root planing and modified Widman flap procedures on enzyme activity. MATERIAL AND METHODS: The study included 13 patients diagnosed with chronic periodontitis. Using a split-mouth design, the defects showing>or=7 mm of attachment loss were treated either with scaling and root planing or with modified Widman flap. Gingival biopsies from both sites were obtained at baseline and 2 months after periodontal treatment. Immunohistochemical staining was performed for evaluating NOS expression and specific arginase activity was determined spectrophotometrically. RESULTS: Although inflamed periodontal tissues demonstrated a strong inducible NOS (iNOS) expression at baseline, immunostaining decreased after periodontal treatment. iNOS expression intensity and the number of inflammatory cells showing iNOS expression were found to be higher in the scaling and root planing group compared to the modified Widman flap group. The specific activity of arginase was measured as 0.18+/-0.07 IU/mg protein in the modified Widman flap group and 0.25+/-0.11 IU/mg protein in the scaling and root planing group at baseline. After periodontal therapy, the enzyme level was increased to 0.68+/-0.14 IU/mg protein in the modified Widman flap and to 1.10+/-0.23 IU/mg protein in the scaling and root planing group. CONCLUSION: This study was the first report of evaluating the involvement of the arginine-NO pathway in chronic periodontitis and this might be considered to be of value in understanding the periodontal disease mechanisms.

Comparative clinical study of connective tissue graft and two types of bioabsorbable barriers in the treatment of localized gingival recessions.

BACKGROUND: Localized buccal recessions occur in more than 60% of individuals; therefore, there is a need for predictable root coverage techniques. The objective of the present study was to evaluate the clinical effectiveness of the guided tissue regeneration (GTR) procedure versus connective tissue graft (CTG) in the treatment of localized gingival recessions over a 12-month postoperative period. METHODS: Thirty bilateral matched pairs of localized buccal recessions in 22 patients were treated with CTG and GTR in this study. For the GTR procedure, two types of bioabsorbable barriers, polylactide/polyglycolide acid (PLGA) and solvent dehydrated duramater allograft (SDDA) membranes, were used and CTG was obtained from the palatal mucosa. The selected pairs of teeth were randomly assigned to one of three groups (10 pairs per group): group 1, PLGA (10 recessions) or CTG (10 recessions); group 2, SDDA (10 recessions) or CTG (10 recessions); or group 3, PLGA (10 recessions) or SDDA (10 recessions). Statistical analysis evaluated both intra- and intergroup measurements. The height of gingival recession (GR), width of keratinized tissue (KT), clinical attachment level (CAL), and probing depth (PD) were assessed at baseline and at 6, 9, and 12 months following surgery. RESULTS: The amount of root coverage was 74.3%, 69.6%, and 86.3% with PLGA, SDDA, and CTG, respectively. The gain in KT was significant in the CTG group (P<0.05). No significant differences were observed among the three groups with respect to PD and CAL. CONCLUSIONS: Results of this study indicate that all techniques led to an improvement of all clinical parameters except PD from baseline. However, CTG increased KT considerably compared to GTR. The final esthetic results were similar for the two membranes and connective tissue graft.

Evaluation of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 levels in gingival fibroblasts of cyclosporin A-treated patients.

BACKGROUND: Cyclosporin A (CsA) is a potent immunosuppressant used to prevent organ transplant rejection and to treat various autoimmune diseases. CsA-induced gingival overgrowth (CsA GO) is the most widely seen side effect of this drug; its pathogenesis is not completely understood. The aim of this study was to identify and compare matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) levels in gingival fibroblast cultures of tissues derived from renal transplant patients receiving CsA and exhibiting gingival overgrowth and from periodontally healthy control subjects. METHODS: Gingival overgrowth samples were obtained from patients undergoing therapy with CsA, and control tissues were obtained from systemically healthy donors. Gingival fibroblasts were grown using explant cultures. Three different study groups were identified: 1) CsA GO fibroblast culture; 2) CsA-treated healthy gingival fibroblast culture (H+CsA); and 3) healthy gingival fibroblast culture (H). The levels of MMP-1 and TIMP-1 in these groups of gingival fibroblasts were analyzed by enzyme-linked immunoabsorbent assay (ELISA). RESULTS: The levels of TIMP-1 were significantly lower in CsA GO than H (P < 0.05). There was no statistically significant difference in the levels of MMP-1 between H and CsA GO (P = 0.505). The ratio of MMP-1 to TIMP-1 was significantly higher in CsA GO than H (P < 0.05). CONCLUSIONS: The results of this study indicate that CsA therapy does not have a significant effect on MMP-1 levels. However, low TIMP-1 levels can be an important factor in the pathogenesis of CsA GO, since the balance between MMP-1 and TIMP-1 levels was changed by CsA.

Effect of a collagen membrane enriched with fibronectin on guided tissue regeneration in dogs.

The present study was planned to assess the capacity of a resorbable collagen membrane enriched with fibronectin to prevent the apical migration of epithelium and to facilitate new attachment and new bone. Experimental osseous dehiscence defects were produced on the bilateral labial aspect of mandibular 2nd, 3rd and 4th premolar teeth in six mongrel dogs. Guided tissue regeneration therapy using collagen membranes, which were rehydrated with fibronectin solution, was performed on one quadrant (group A). In the contralateral quadrant, the same collagen membranes, but rehydrated only with saline (group B), were placed over the bony defects. The third premolar teeth, which were treated by open-flap debridement, served as control (group C). Flaps were positioned slightly coronally and sutured; sutures were removed 10 days later. The dogs were killed 30 days after reconstructive therapy. Tissue blocks containing the experimental and control teeth were excised, demineralized in EDTA, and embedded in paraffin. Histological and histometric evaluation revealed that all groups demonstrated similar effects on preventing the down-growth of epithelium and formation of new cementum and new bone. Collagen membranes were tolerated well within the tissues, and membrane remnants were identified at 30 days. In summary, this study indicated that in this dog model similar healing results could be achieved with a bovine type I collagen membrane with or without fibronectin solution and open-flap debridement.

Evaluation of cellular response to collagen membranes enriched with fibronectin light and transmission electron microscope study.

This study was undertaken to evaluate the biocompatibility, cellular reaction and resorption characteristics of a type I bovine collagen membrane material either enriched with or without fibronectin solution in vivo using light (LM) and transmission electron microscopy (TEM). Experimental osseous dehiscence defects were surgically produced bilaterally on the labial aspect of the mandibular 2nd, 3rd, and 4th premolar teeth in four mongrel dogs. Collagen membranes rehydrated with fibronectin solution (group FM) and membranes rehydrated with saline (group M) were placed over the bony defects. The third premolar teeth on which the flap operation was performed served as control (group C), with no membrane placed. Flaps were positioned slightly coronally and sutured. Gingival tissue samples and block biopsies were obtained from all experimental and control sites for LM and TEM evaluation at 7 days. For each group, morphometric analysis was performed and the numbers of macrophages in the most coronal area of the free gingiva were counted. Postoperative healing was uneventful during the experimental period, and all membranes remained covered. Light microscopic evaluation revealed similar resorption patterns in the most coronal area of the membranes both enriched with and without fibronectin solution within the first 7 days. The mean numbers of macrophages were higher in experimental groups than in the control group. In TEM evaluation, more excessive intracellular macrophage activity was observed in group M than group FM. As a result of these observations it may be concluded that similar resorption characteristics existed in the most coronal area in both experimental groups with LM evaluation, but with TEM it was observed that the membranes enriched with fibronectin solution were resorbed more slowly at the ultrastructural level.