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Öğe Attachment, proliferation and collagen type I mRNA expression of human gingival fibroblasts on different biodegradable membranes(TAYLOR & FRANCIS INC, 2013) Hakki, Sema S.; Korkusuz, Petek; Purali, Nuhan; Bozkurt, Buket; Kus, Mahmut; Duran, IsmetThe purpose of this study was to investigate adhesion, proliferation and type I collagen (COL I) mRNA expression of gingival fibroblasts on different membranes used in periodontal applications. Collagen (C), acellular dermal matrix (ADM) and polylactic acid; polyglycolic acid; lactide/glycolide copolymer (PLGA) biodegradable membranes were combined with gingival fibroblasts in culture and incubated for 48 h. Cell adhesion was examined with scanning electron and confocal microscopy. MTT assay was used to measure proliferation. COL I mRNA expression was assessed using quantitative-polymerase chain reaction (QPCR). The PLGA group exhibited the lowest cell survival on day 5 and 10, and lowest cell proliferation on days 5, 10 and 14. While cell proliferation was similar in C and ADM groups, the C membrane showed a slightly greater increase in viable cells to day 10. Confocal and scanning electron microscopy confirmed the results of proliferation and MTT assays. The highest COL I mRNA expression was noted in the PLGA membrane group when compared to the C (p<0.01) and ADM (p<0.05) membrane groups. These data revealed that adherence and proliferation of primary gingival fibroblasts on collagen-based C and ADM membranes is better than that seen with PLGA membranes, and thus may be preferable in the treatment of gingival recession defects.Öğe Comparison of Er,Cr:YSGG Laser and Hand Instrumentation on the Attachment of Periodontal Ligament Fibroblasts to Periodontally Diseased Root Surfaces: An In Vitro Study(Amer Acad Periodontology, 2010) Hakkı, Sema S.; Korkusuz, Petek; Berk, Gizem; Dündar, Niyazi; Sağlam, Mehmet; Bozkurt, Buket; Purali, NuhanBackground: This study investigates the effects of erbium, chromium: yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation and hand instrumentation on the attachment of periodontal ligament (PDL) fibroblasts to periodontally involved root surfaces. Methods: Twenty-four single-rooted periodontally involved human teeth (test groups), and six healthy premolar teeth extracted for orthodontic reasons (control group) were included in this study. A total of 45 root slices were obtained from all selected teeth and assigned to the following five groups: 1) untreated healthy group (+control); 2) untreated periodontally diseased group ( control); 3) hand instrumentation group (scaled Gracey); 4) laser I, Er,Cr:YSGG laser irradiation setting-I (short pulse); and 5) laser II, Er,Cr:YSGG laser irradiation setting-II (long pulse). All of the root slices were autoclaved in phosphate buffered saline and slices were placed onto cell culture inserts. PDL fibroblasts were placed at the density of 80,000 cells on the root plate (5 x 6 mm) and incubated for 48 hours and transferred to 24-well plates. The attachment PDL fibroblasts on the root plates were observed using confocal microscopy (at 12 hours and on days 3 and 7) and scanning electron microscopy (at 12 hours and day 3). 3- (4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay was performed on day 5 for PDL fibroblast survival. Results: 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay shows that whereas laser-treated specimens showed a significantly higher cell density, the Gracey-treated group showed a lower cell density compared to the positive control group (P < 0.05). Based on confocal microscopy, apparent reduction was observed in the attachment of PDL cells to the periodontally diseased root surfaces. In the laser and Gracey groups, cells looked well-oriented to the root surfaces. Laser-treated groups provided suitable environment for cell adhesion and growth. Laser I treatment was more favorable for the attachment of PDL compared to scaled Gracey, laser II, and even healthy root surfaces. Conclusion: The results of the study indicate that short-pulse laser setup (laser I) looks more promising regarding the attachment, spreading, and orientation of PDL cells.Öğe Interaction of MC3T3-E1 cells with titanium implants(TURKISH JOINT DISEASES FOUNDATION, 2008) Korkusuz, Petek; Hakki, Sema S.; Purali, Nuhan; Gorur, Ilker; Onder, Ercuement; Nohutcu, Rahime; Koc, NursenObjectives The aim of the present study was to evaluate in-vitro MC3T3-E1 preosteoblastic cell osseointegration on surfaces of polished, sand-blasted (smooth and rough) and sodium titanate coated titanium alloys. Materials and methods MC3T3-E1 cell proliferation and mineralization was assessed comparatively on polished, sand-blasted (smooth and rough) and sodium titanate coated titanium alloys. Cell morphology, attachment and proliferation were also comparatively evaluated using confocal (CM) and scanning electron microscopy (SEM). Results All implants Used in this study were biocompatible. Cells started to attach on the surfaces of the implants following exposure to the in vitro medium for 3 days. The cells were viable and metabolically active as observed by CM. Cell Population increased exponentially from day 3 to day 22. Proliferation rate was highest on polished surfaces and lowest on sodium titanate-coated surfaces. In contrast, mineralized nodules were numerous on sand-blasted and sodium titanate-coated Surfaces when compared to the polished ones on day 30. Conclusion This study demonstrated that sand-blasting and sodium titanate coating provided by NaOH favored the attachment, mineralization and early differentiation of osteoblasts on titanium alloys.Öğe Periodontal ligament cell behavior on different titanium surfaces(TAYLOR & FRANCIS LTD, 2013) Hakki, Sema S.; Korkusuz, Petek; Purali, Nuhan; Korkusuz, Feza; Bozkurt, Buket S.; Hakki, Erdogan E.; Onder, M. ErcumentAim. The purpose of this study was to investigate proliferation, morphology, mineralization and mRNA expressions of mineralized tissue associated proteins of PDL cells on smooth (S), sandblasted small-grit (SSG), sandblasted large-grit (SLG) and sodium titanate (NaTi) coated titanium alloys, in vitro. Methods and materials: PDL cells were cultured with DMEM media containing 10% FBS on the S, SSG, SLG and NaTi titanium surfaces. PDL cell proliferation, mineralization and immunohistochemistry experiments for Bone Sialoprotein (BSP) were performed. The morphology of the PDL cells was examined using confocal and scanning electron microscopy (SEM). Gene expression profiles of cells were evaluated using a quantitative-polymerase chain reaction (Q-PCR) for type I collagen (COL I), Osteocalcin (OCN), osteopontin (OPN) and Runt-related transcription factor-2 (Runx2) on days 7 and 14. Results. Proliferation results on days 6 and 10 were similar in groups, while those of day 13 revealed a decrease in the NaTi group when compared to the S group. NaTi surface induced BSP mRNA expression which was correlated with mineralization tests and BSP immunostaining results. Increased Runx2 mRNA expression was also noted in the NaTi surface when compared to other surfaces. Conclusions. This study considers the NaTi surface as a potential alternative to SSG and SLG surfaces. This surface might provide a promising environment for PDL ligament-anchored implants.Öğe Recombinant amelogenin regulates the bioactivity of mouse cementoblasts in vitro(NATURE PUBLISHING GROUP, 2018) Hakki, Sema S.; Bozkurt, S. Buket; Tuerkay, Emre; Dard, Michel; Purali, Nuhan; Goetz, WernerAmelogenin (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.
