Maxillary sinus augmentation with leukocyte and platelet-rich fibrin and deproteinized bovine bone mineral: A split-mouth histological and histomorphometric study.

OBJECTIVES: To evaluate the effect of leukocyte and platelet-rich fibrin (L-PRF) in combination with deproteinized bovine bone mineral (DBBM) on bone regeneration in maxillary sinus augmentation. MATERIAL AND METHODS: Thirteen patients (nine males and four females, mean age ± SD; 49.92 ± 10.37) were enrolled to the study. 26 maxillary sinus augmentation procedures were randomly performed using DBBM and L-PRF mixture (test) or DBBM alone (control) in a split-mouth design. The same surgical procedures were performed in both groups, and bone biopsies were harvested from the implant sites 6 months postoperatively for histological and histomorphometric evaluations as the primary outcome of the study. Implants were placed and then loaded in the augmented sites after 6 months. The secondary outcomes included clinical and radiographic data and were obtained pre- and postoperatively. RESULTS: There was no qualitative difference in histological analyses among the groups. In all samples, a newly formed bone was in direct contact with the residual material. The percentages of newly formed bone (test; 21.38 ± 8.78% and control; 21.25 ± 5.59%), residual bone graft (test; 25.95 ± 9.54% and control; 32.79 ± 5.89%), bone graft in contact with the newly formed bone (test; 47.33 ± 12.33% and control; 54.04 ± 8.36%), and soft tissue (test; 52.67 ± 12.53% and control; 45.96 ± 8.36%) were similar among the groups (p < .05). Similar radiographic bone height in the augmented area was observed, and implant survival rate was 100% for both groups. CONCLUSIONS: Both techniques were effective for maxillary sinus augmentation, and after 6 months of healing, the addition of L-PRF in DBBM did not improve the amount of regenerated bone or the amount of the graft integrated into the newly formed bone under histological and histomorphometric evaluation.

Levels of ll-37 antimicrobial peptide in the gingival crevicular fluid of young and middle-aged subjects with or without gingivitis.

PURPOSE: LL-37 is an antimicrobial peptide which plays an important role in the innate immunity. The aim of this study was to investigate the LL-37 levels in the gingival crevicular fluid (GCF) of middle-aged and young adults who have either gingivitis or healthy periodontal tissues. MATERIALS AND METHODS: Forty middle-aged adults (20 healthy controls and 20 with gingivitis) and 41 younger adults (20 healthy controls and 21 with gingivitis) were included in the present study. Probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. LL-37 levels in the GCF were analyzed by enzyme-linked immunosorbent assay. RESULTS: No significant differences were observed in the GCF LL-37 levels between young healthy and middle-aged healthy subjects. Also, there were no significant differences in GCF LL-37 levels between young and middle-aged gingivitis subjects. However, gingivitis groups had significantly higher GCF LL-37 levels than healthy groups (p<0.001). Correlation analysis demonstrated no significant correlation between age and GCF LL-37 levels neither in healthy nor in gingivitis groups. CONCLUSION: The levels of LL-37 in GCF increase in the presence of gingival inflammation, however, this does not vary according to subjects being young or middle-aged.

Gingival crevicular fluid and serum hCAP18/LL-37 levels in generalized aggressive periodontitis.

OBJECTIVES: hCAP18/LL-37 is an endogenous antibiotic having a role in innate immunity. The aim of the present study was to evaluate serum and gingival crevicular fluid (GCF) hCAP18/LL-37 levels in patients with generalized aggressive periodontitis (G-AgP). MATERIALS AND METHODS: Twenty-six G-AgP patients, 24 gingivitis patients, and 25 healthy subjects were included in this study. Periodontal parameters including probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. GCF and serum hCAP18/LL-37 levels were analyzed by enzyme-linked immunosorbent assay. RESULTS: GCF hCAP18/LL-37 level was significantly higher in G-AgP compared to others (p = 0.038, p < 0.001). Gingivitis patients had significantly higher GCF hCAP18/LL-37 levels than controls (p < 0.001). No significant differences were observed in serum hCAP18/LL-37 levels among the study groups (p = 0.524). While there were positive correlations between GCF hCAP18/LL-37 levels and periodontal parameters of sampling sites (p < 0.005), no significant correlation was observed between serum hCAP18/LL-37 levels and whole-mouth periodontal parameters (p > 0.05). CONCLUSION: Increased levels of GCF hCAP18/LL-37 in G-AgP might show that it is abundantly expressed in the presence of periodontal tissue destruction. Serum hCAP18/LL-37 levels do not seem to be related with the presence of G-AgP. CLINICAL RELEVANCE: hCAP18/LL-37 antimicrobial peptide might be associated with periodontal tissue destruction in the presence of aggressive periodontitis.

Monocyte chemotactic protein-1, RANTES and macrophage migration inhibitory factor levels in gingival crevicular fluid of metabolic syndrome patients with gingivitis.

OBJECTIVE: The aim of the present study was to determine gingival crevicular fluid (GCF) levels of monocyte chemotactic protein-1 (MCP-1), regulated on activation, normal T-cell expressed and secreted protein (RANTES) and macrophage migration inhibitory factor (MIF) in metabolic syndrome patients with gingivitis. DESIGN: Twenty metabolic syndrome patients with gingivitis (MSG), 20 MetS patients with clinically healthy periodontium (MSH), 20 systemically healthy subjects with gingivitis and 20 subjects who were both systemically and periodontally healthy were included. Periodontal and systemical parameters were recorded. GCF MCP-1, RANTES and MIF levels were assayed by enzyme-linked immunosorbent assay method. RESULTS: MSG and MSH groups had elevated blood pressure, triglyceride, waist circumference and fasting glucose values in comparison to gingivitis and healthy groups (P<0.0001). Clinical periodontal parameters were higher in MSG and gingivitis groups when compared to those of the MSH and healthy groups (P<0.0001). MCP-1 and RANTES levels (ng/mg total protein) of MSG group were higher than those of the MSH groups (P=0.005, P=0.0001, respectively). Also gingivitis group had higher MCP-1, RANTES and MIF levels compared to the healthy group (P=0.011, P=0.0001, P=0.011 respectively). The RANTES level of MSG group was significantly higher than those of the gingivitis group (P=0.01), but MCP-1 and MIF levels were similar in the MSG and gingivitis groups (P>0.05). CONCLUSION: Elevated levels of GCF RANTES in MetS patients with gingivitis might associate with the presence of increased gingival inflammation by MetS. Low-grade systemic inflammation associated with MetS and adipose tissue-derived RANTES might lead to altered GCF RANTES levels in the presence of gingival inflammation.

Evaluation of gingival crevicular fluid cyclophilin a and extracellular matrix metalloproteinase inducer levels in different periodontal diseases.

OBJECTIVE: Cyclophilin A (CypA) is able to regulate inflammatory responses and matrix metalloproteinase production via its interaction with extracellular matrix metalloproteinase inducer (EMMPRIN). EMMPRIN is the cell surface receptor of CypA. The aim of the present study was to evaluate the gingival crevicular fluid (GCF) CypA and EMMPRIN levels in patients with chronic periodontitis (CP), generalized aggressive periodontitis (G-AgP) and periodontally healthy controls. METHODS: Twenty CP patients, 19 G-AgP patients and 20 healthy control subjects were included in the present study. All study participants were non-smokers. Full mouth clinical periodontal parameters including probing depth, clinical attachment level, plaque index, and papilla bleeding index were recorded. GCF CypA and EMMPRIN levels were analyzed by enzyme-linked immunosorbent assay. Data were analyzed statistically with parametric and non-parametric tests. RESULTS: GCF CypA total amount was higher in the G-AgP group compared to healthy controls (p<0.05), whereas CypA total amounts were similar in CP and healthy controls (p>0.05). No significant difference in GCF CypA total amount between CP and G-AgP was observed (p>0.05). Also, there was no significant difference in GCF EMMPRIN total amounts among the study groups (p>0.05). CONCLUSION: Higher levels of GCF CypA in patients with G-AgP might demonstrate that CypA is associated with the inflammatory infiltrate and alveolar bone destruction of G-AgP. However, GCF CypA level does not seem to be affected by CP. Similar GCF EMMPRIN levels in diseased and healthy groups might suggest that EMMPRIN has role in the turn over of connective tissues in physiological conditions as well as pathological state.

Effect of centrifugation time on growth factor and MMP release of an experimental platelet-rich fibrin-type product.

Platelet-rich fibrin (PRF) has a controlled release of growth factors due to the fibrin matrix structure. Different centrifugation protocols were suggested for PRF preparation. Since the derivation method of PRF can alter its contents, in the present study it is aimed to investigate the cell contents and transforming growth factor beta-1 (TGF-ß1), platelet-derived growth factor (PDGF-AB), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-1 and-8 release from experimental PRF-type membranes obtained with different centrifugation times at 400 gravity. Three blood samples were collected from 20 healthy non-smoker volunteers. One tube was used for whole blood analyses. The other two tubes were centrifuged at 400 g for 10 minutes (group A) or 12 minutes (group B). Each experimental PRF-type membrane was placed in Dulbecco's Modified Eagle's Medium (DMEM)and at 1, 24 and 72 hours, TGF-ß1, PDGF-AB, VEGF, MMP-1 and -8 release amounts were analysed by enzyme-linked immunosorbent assay (ELISA). The blood cell count of membranes was determined by subtracting plasma supernatant and red blood cell (RBC) mixture from the whole blood cell counts. At 72 hours, the VEGF level of group B was statistically higher than that of group A (p = 0.040). The centrifugation time was not found to influence the release of other growth factors, enzymes and cell counts. Within the limits of the present study, it might be suggested that centrifugation time at a constant gravity has a significant effect on the VEGF levels released from experimental PRF-type membrane. It can be concluded that due to the importance of VEGF in the tissue healing process, membranes obtained at 12-minute centrifugation time may show a superior potential in wound healing.

Does smoking affect gingival crevicular fluid LL-37 levels following non-surgical periodontal treatment in chronic periodontitis?

OBJECTIVE: LL-37 contributes to maintaining the balance between health and disease. Smoking is a risk factor for periodontitis that impairs neutrophil functions. The aim of the present study was to comparatively evaluate gingival crevicular fluid (GCF) LL-37 levels in smoker and non-smoker chronic periodontitis (CP) patients and controls, as well as the effect of non-surgical periodontal treatment on GCF LL-37 levels. DESIGN: Thirty-one CP patients (16 smokers, 15 non-smokers) and thirty-one controls (16 smokers, 15 non-smokers) were included in the study. CP patients received non-surgical treatment. GCF LL-37 levels and periodontal parameters were assessed at baseline, 1 and 3 months after completion of non-surgical periodontal treatment. GCF LL-37 levels were analyzed by ELISA. RESULTS: No significant difference was observed in GCF LL-37 levels between smoker and non-smoker controls (p>0.05). Smoker CP group had significantly lower GCF LL-37 level than non-smoker CP group at baseline (p<0.05). GCF LL-37 levels significantly decreased in non-smoker CP group at first week, 1 and 3 months after completion of non-surgical periodontal treatment (p<0.05) although no significant decrease in GCF LL-37 levels was observed in smoker CP group (p>0.05). Periodontal parameters were correlated with GCF LL-37 levels in non-smoker CP group (p<0.05), but not in smoker CP group (p>0.05). CONCLUSIONS: GCF LL-37 levels do not seem to be affected from smoking in periodontal health. However, smoking might have a suppressive effect on GCF LL-37 levels in CP. Non-surgical treatment is effective in decreasing GCF LL-37 levels in non-smoker CP patients but not in smokers with CP.

Vascularization after treatment of gingival recession defects with platelet-rich fibrin or connective tissue graft.

OBJECTIVE: The aim of this study was to evaluate histologically the following treatment of bilateral localized gingival recessions with coronally advanced flap (CAF) combined with platelet-rich fibrin (PRF) or subepithelial connective tissue graft (SCTG). MATERIALS AND METHODS: Tissue samples were harvested from 14 subjects either 1 or 6 months after the surgeries. The 2-mm punch biopsies were obtained from the mid-portion of the grafted sites. Neutral buffered formalin fixed, paraffin-embedded 5-µm thick tissue sections were stained with hematoxylin eosin and Masson's trichrome in order to analyze the collagen framework, epithelium thickness and rete-peg length. Multiple sequential sections were cut from paraffin-embedded blocks of tissue and immunohistochemically prepared for detection of vascular endothelial growth factor, CD31 and CD34, for the assessment of vascularization. RESULTS: Rete peg formation was significantly increased in the sites treated with PRF compared to the SCTG group after 6 months (p < 0.05). On the contrary, the number of vessels was increased in the SCTG group compared to the PRF group after 6 months (p < 0.05). No statistically significant differences were observed in the collagen density. Staining intensity of CD31 increased in submucosal area of PRF group than SCTG group after 1 month. Higher staining intensity of CD34 was observed in the submucosal area of PRF group compared with SCTG group after 6 months. CONCLUSIONS: The results of the present study suggest that in histological evaluation because of its biological compounds, PRF results earlier vessel formation and tissue maturation compared to connective tissue graft. CLINICAL RELEVANCE: PRF regulated the vascular response associated with an earlier wound healing.

Evaluation of GCF MMP-1, MMP-8, TGF-ß1, PDGF-AB, and VEGF levels in periodontally healthy smokers.

BACKGROUND/AIM: The effect of smoking on inflammatory biomarkers in gingival crevicular fluid (GCF) is well established in the presence of periodontal inflammation. However, it is not clear if smoking has an influence on matrix metalloproteinase (MMP) and growth factor levels in the GCF of periodontally healthy subjects. The aim of this study was to investigate GCF levels of MMP-1, MMP-8, transforming growth factor (TGF)-ß1, platelet-derived growth factor (PDGF)-AB, and vascular endothelial growth factor (VEGF) in smoking versus nonsmoking periodontally healthy subjects. MATERIALS AND METHODS: Thirty-two periodontally healthy subjects were included in this study. Probing depths, bleeding on probing, and plaque index was assessed. GCF levels of MMP-1, MMP-8, TGF-ß1, PDGF-AB, and VEGF were analyzed by enzyme-linked immunosorbent assay. RESULTS: No significant differences were observed in the distribution of demographic data between study groups. GCF total amount of PDGF-AB was significantly lower in smokers compared to nonsmokers (P = 0.014). Total amount of GCF MMP-1, MMP-8, TGF-ß1, and VEGF levels were similar in both study groups (P = 0.022). CONCLUSION: Smoking has the effect of decreasing GCF PDGF-AB while it does not affect GCF MMP-1, MMP-8, TGF-ß1, and VEGF in periodontally healthy subjects. Since increased levels of these molecules are involved in periodontal breakdown, our findings may emphasize the importance for maintenance of periodontal health in smokers.

Platelet-rich fibrin in the treatment of localized gingival recessions: a split-mouth randomized clinical trial.

OBJECTIVES: The aim of this study is to evaluate the clinical efficacy of platelet-rich fibrin (PRF) in combination with coronally advanced flap (CAF) in the treatment of localized gingival recessions. MATERIALS AND METHODS: Twenty-two patients with localized gingival recession defects (Miller I, II) participated in this split-mouth trial. Forty-four defects received either CAF + PRF (test) or CAF with subepithelial connective tissue graft (SCTG) (control). Gingival recession depth (RD), gingival recession width (RW), keratinized tissue width (KTW), recession area (RA), probing depth (PD), clinical attachment level (CAL) and gingival thickness (GT) were evaluated at baseline and 6 months. RD, RW, RA and KTW were calculated on standardized photographs with a computer image analysis program. RESULTS: Percentage of root coverage in test group was 92.7 % and in control group was 94.2 % (p > 0.05). Percentage of complete root coverage of the test and control groups was 72.7 and 77.3 %, respectively (p > 0.05). KTW and GT were increased in both groups from baseline to 6 months (p < 0.001), but there was no statistically significant difference between treatment groups (p > 0.05). CONCLUSION: Within the limits of the present study, it can be concluded that localized gingival recessions could be successfully treated with CAF + PRF as well as CAF + SCTG. The digital measuring method provided high accuracy and precision in the evaluation of treatment outcomes after both surgical procedures. CLINICAL RELEVANCE: PRF might be suggested as an alternative to SCTG for the treatment of localized gingival recessions.