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Biomedical subjects

L Kuru

Publications and source records attributed to L Kuru.

11 recordsLinked to original sources

Oral health in patients on inhaled corticosteroid treatment.

OBJECTIVE: The aim of this study was to investigate the effects of long-term inhaled corticosteroids on bone mineral density (BMD) of the mandible in relation with the tooth loss. DESIGN: Cross sectional analytic study. SUBJECTS AND METHODS: Patients (n = 30) with chronic obstructive pulmonary disease under inhaled corticosteroid therapy for at least 1 year were compared with sex- and age-matched healthy controls (n = 30). BMD of the mandible was measured by dual-energy X-ray absorptiometry. The clinical examination included recording the number of teeth present together with periodontal condition. Levels of serum osteocalcin, alkaline phosphatase, calcium, phosphorus and cortisol were also assessed. RESULTS: BMD of the mandible in patients on corticosteroid treatment was significantly lower than that in the control group (P = 0.001). Patients under treatment had more missing teeth than the control group but the difference did not reach statistical significance. The two groups exhibited similar clinical parameters of periodontal condition. Significantly lower levels of osteocalcin (P < 0.0001), calcium (P = 0.004) and cortisol (P = 0.03) were observed in the patients on corticosteroid treatment. CONCLUSION: Long-term use of inhaled corticosteroids may impair bone metabolism and lead to a marked decrease in the mandibular BMD.

Absorptiometry, Photon↗

Expression of growth factors in the gingival crevice fluid of patients with phenytoin-induced gingival enlargement.

The mechanism underlying phenytoin (PHT)-induced gingival enlargement (GE) is not yet known. The aim of the present study was to investigate transforming growth factor-beta1 (TGF-beta1), platelet-derived growth factor-BB (PDGF-BB) and basic fibroblast growth factor (bFGF) profiles in the gingival crevice fluid (GCF) of patients with PHT-induced GE and to compare the results with healthy controls. Five PHT-treated patients and five healthy subjects with normal periodontal tissue were included in this study. GCF samples were collected from (i) enlarged gingival sites in patients receiving PHT (GE+); (ii) non-enlarged gingival sites in the same patients (GE-); (iii) normal gingival sites of healthy subjects (control). The levels of TGF-beta1, PDGF-BB and bFGF in the GCF samples were analysed by ELISA. The results showed that the total amounts of TGF-beta1 and PDGF-BB in the GE+ group were higher than in the GE- group and significantly higher than in the control group (P < 0.05). However, no significant differences were found between the groups when the concentrations of these growth factors were compared. bFGF levels were not compared as this growth factor could be detected in only 33, 41 and 44% of the GE+, GE- and control GCF samples, respectively. These results show that TGF-beta1 and PDGF-BB are readily detectable in GCF obtained from enlarged and non-enlarged sites of PHT recipients and suggest that since the amounts were markedly higher at the GE+ than the GE- sites, the systemic administration of PHT has a pronounced localised effect on the levels of these growth factors. Moreover, our findings provide evidence that both TGF-beta1 and PDGF-BB are closely associated with the clinical manifestation of PHT-induced GE.

Adolescent↗

Changes in transforming growth factor-beta1 in gingival crevicular fluid following periodontal surgery.

OBJECTIVES: Growth factors play a major part in wound healing, including in the periodontium. However, the presence of growth factors in gingival crevicular fluid (GCF) in humans during periodontal wound healing has not yet been determined. Our hypothesis is that such factors are present in GCF and that changes in their levels might be of value as a prognostic marker of wound-healing activity and therapeutic progress following periodontal surgery. The aim of this study was therefore to measure transforming growth factor-beta1 (TGF-beta1) in GCF collected from sites that have undergone guided tissue regeneration (GTR) and conventional flap (CF) surgery and to compare these with GCF collected from unaffected healthy sites. MATERIALS AND METHODS: GCF samples were collected, using filter paper strips, at baseline (pre-surgical) and then at intervals up to 26 weeks from 16 patients undergoing GTR and from 11 patients undergoing CF surgery. After elution and acid treatment, TGF-beta1 levels were measured by ELISA. RESULTS: Treatment of periodontal defect sites significantly reduced the mean probing pocket depth (PPD) and improved the mean lifetime cumulative attachment loss (LCAL). Average GCF volumes also significantly increased at all sites at 2 weeks post-surgery and thereafter declined to baseline levels, except at the GTR test sites that were still elevated at 7 weeks. TGF-beta1 could be detected in almost all GCF samples, and 2 weeks after surgery, the average levels increased two-fold at the surgically treated but not at the control sites, which remained unchanged. CONCLUSION: TGF-beta1 is readily detectable in GCF and increases transiently following periodontal surgery. This suggests that changes in the levels of this growth factor in GCF might be useful for monitoring the progress of periodontal repair and regeneration.

Adult↗

Expression of growth-factor receptors in normal and regenerating human periodontal cells.

Growth factors are biologically active mediators that bind to specific receptors on target cells and regulate genes involved in cell growth, wound healing and regeneration. The expression of these receptors is thus of fundamental importance for the response of the cells to the factors. The aim here was to examine, using immunohistochemistry and flow cytometry, the expression of growth factor receptors in normal gingiva, periodontal ligament and in cells derived from these tissues, and also in regenerated tissues following guided tissue regeneration (GTR). By immunocytochemistry platelet-derived growth factor receptor-alpha (PDGF-Ralpha) was not detected in any of the tissues, whereas the PDGF-Rbeta and transforming growth factor-beta receptor types I and II (TGF-beta RI, RII) appeared to be upregulated in regenerated tissues compared with gingival and periodontal ligament tissues. Epidermal growth factor receptor (EGF-R) was also notably elevated in the regenerated tissue and was strongly expressed in the gingival epithelium but not in the periodontal ligament. Neither were fibroblast growth factor receptor-I (FGF-RI) or insulin-like growth factor receptor (IGF-R) detected in the periodontal ligament, nor in the gingiva, but they sometimes stained weakly in the regenerated tissues. Flow cytometry (FCM) showed that all the cells derived from the normal gingiva and the periodontal ligament expressed the PDGF-Rbeta, whereas the TGF-beta RI and RII, FGF-RI and IGF-R were detected in only a proportion of the total cells. In contrast, none of the cells expressed the PDGF-Ralpha or the EGF-R. These observations show that the growth factor receptors are differentially expressed by the periodontal tissues and cells and suggest that the corresponding factors may also be differentially involved in periodontal wound healing and regeneration.

Epithelium↗

Flow cytometry analysis of guided tissue regeneration-associated human periodontal cells.

BACKGROUND: Expanded polytetrafluoroethylene (ePTFE) barrier membranes have been widely used for guided tissue regeneration (GTR) of the human periodontal ligament (PL). However, the precise cellular and molecular events involved in the re-growth of the new tissue are still unclear. METHODS: Retrieved membranes and the newly-regenerated soft tissue (RT) underlying the membranes were used to examine the cells associated with GTR compared with normal human PL and gingival cells. Flow cytometry (FCM) was used, for the first time, to analyze the spindle-shaped fibroblast-like cells which were adherent to these membranes and the cells which grew out of the RT. RESULTS: The results showed that the membrane-associated (M) cells had the lowest rate of proliferation and appeared to be larger and more granular than the other types of cell. Moreover, both the M- and RT-derived cells were found to express higher levels of the extracellular matrix (ECM) proteins collagen type 1, fibronectin, tenascin, and decorin. In addition, evidence based on FCM profiles identified distinct sub-populations of GTR cells in which fibronectin expression was markedly up-regulated compared with normal PL cells and which also differed in size and granularity. CONCLUSIONS: The results of this study show that cells associated with GTR barrier membranes and with the underlying tissue appear to have distinct phenotypic and functional activities consistent with the production of new periodontal connective tissue and periodontal regeneration.

Adult↗

Retroviral transduction of human periodontal cells with a temperature-sensitive SV40 large T antigen.

The periodontal ligament (PDL) is considered to contain subpopulations of cells responsible for the development, repair and regeneration of the periodontium. Cell cultures have been used as model systems in order to understand the complex cellular and biochemical events underlying these processes. In order to obtain long-term cultures of these cells that can be cloned and characterized, primary cultures of PDL and gingival cells were infected with an amphotropic retroviral construct encoding a temperature-sensitive SV40 large T antigen (tsT). After selection for drug resistance, the cells expressed the T antigen and proliferated at 34 degrees C for more than 40 passages. However, when the T antigen was inactivated by incubation at 39 degrees C, the cultures became growth-arrested and the granularity of the cells increased, possibly as a result of differentiation. Reverse transcribed-polymerase chain reaction and flow cytometry showed that the tsT-transduced cells expressed a number of soft and hard connective-tissue antigens, including osteocalcin, osteonectin, osteopontin, collagen type I and alkaline phosphatase. Moreover, incubation of the transduced PDL cells at 39 degrees C was found to upregulate the expression of osteocalcin, osteopontin and collagen type I, but downregulate osteonectin. At this temperature, the presence of the dexamethasone downregulated type I collagen, while vitamin D3 had no effect on the expression of any of the antigens examined. Under all culture conditions, antigen expression was far higher in the transduced PDL cells than the gingival cells. The findings thus show that growth of the tsT-transduced PDL and gingival cells is temperature-dependent and that the presence of the T antigen increases their lifespan but does not ablate the expression of certain of their characteristic phenotypic and functional features.

Alkaline Phosphatase↗

Alkaline phosphatase activity is upregulated in regenerating human periodontal cells.

The activity of alkaline phosphatase (ALP) is considered to indicate the presence of osteoblast cells and the formation of new bone. In the present study this enzyme was investigated in cells obtained from retrieved polytetrafluoroethylene membranes (M cells) of periodontal disease patients treated by guided tissue regeneration (GTR) and from the regenerated tissue underlying the membrane (RT cells). Normal periodontal ligament (PL) and gingival cells were also grown from the corresponding healthy tissues of human subjects. ALP activity was measured colourimetrically, using paranitrophenyl phosphate as the substrate, after 4 and 7 d of culture in the absence and presence of dexamethasone (DEX), a synthetic glucocorticoid which induces osteoblast differentiation. The results showed that basal levels of ALP activity were expressed by all the cells and that DEX upregulated ALP levels in the M, RT and PL cells but not in the gingival cells. Moreover, both the basal and DEX-induced ALP activities were statistically significantly higher in the RT cells than in any of the other cells. Our results suggest that both the GTR-associated and normal PL cells express osteoblast-like characteristics and, furthermore, that the RT cultures in particular contain a high proportion of osteoprogenitor cells.

Adolescent↗

Flow cytometry for assessing biocompatibility.

Flow cytometry (FCM) was examined as a possible procedure for measuring in vitro the biocompatibility of implant materials for orthopedic and dental surgery. The human osteoblast-like cell line MG63 was grown on hydroxyapatite (HA) and P2O5 glass-reinforced HA composite discs and compared with the same cells grown on polystyrene culture dishes. While morphological observation at the light and electron microscopic levels showed no major deleterious effects, FCM indicated that cell size was somewhat reduced, particularly by growth on the HA composite. Morever, this material also appeared to delay the progression of the cells from the G0/G1 into the S phase of the cell cycle. In addition to this low level of inhibition of cell growth relative to control cultures, FCM analysis also demonstrated that the glass-reinforced HA caused some down-regulation of the expression of osteocalcin and fibronectin, two antigens which play a vital part in the integrity and function of bone and soft connective tissue, respectively. These results thus show, first, that although HA and the HA composite used in these experiments were generally biocompatible, they nevertheless had certain suboptimal effects on the cells; and second, that FCM could be a highly useful procedure for effectively screening and evaluating important biological responses to implant materials.

Biocompatible Materials↗

Flow cytometry analysis of gingival and periodontal ligament cells.

Gingival and periodontal ligament (PDL) fibroblasts are the major cellular components of periodontal soft connective tissues, but the precise differences between these cells are not yet known. In the present study, we have therefore examined the phenotypic and functional features of the cells obtained from gingival and PDL biopsy samples. Spindle-shaped cells characteristic of fibroblasts were the main cell type observed in vitro, although epithelial cells were also present in primary gingival cell cultures. Flow cytometry was used to measure the size and granularity of the cultured cells, and showed that the gingival fibroblasts were smaller and less granular compared with the PDL cells. The expression of certain key extracellular matrix (ECM) proteins, fibronectin, collagen type I, and tenascin was measured by flow cytometry. Analysis of the fluorescence profiles of these cultures showed that the majority of cells expressed fibronectin and that the average fluorescence intensity of this antigen in the PDL cells was higher than that in the gingival fibroblasts. Moreover, the fibronectin-positive PDL cells apparently comprised two subpopulations which expressed fibronectin at different levels, suggesting that the cells in the PDL cultures were functionally heterogeneous. The level of collagen type I was also found to be up-regulated in the PDL compared with the gingival cells and, as with fibronectin, was expressed at two different levels by subsets of the PDL cells. In contrast, tenascin was expressed at very similar levels by both the gingival fibroblasts and PDL cells. In addition, measurement of alkaline phosphatase, a marker enzyme for mineralized tissue-forming cells, showed that the PDL cells had higher activity than the gingival fibroblasts and that the alkaline phosphatase activity in the PDL cells was far more markedly up-regulated by dexamethasone. Our findings demonstrate that, despite their similar spindle-shaped appearance, fibroblasts derived from gingival and PDL tissues appear to display distinct functional activities which are likely to play a vital part in the maintenance of tissue integrity and regenerative processes.

Adolescent↗

The evolution of clinical periodontal therapy.

Periodontal diseases are considered as old as the history of mankind, Magical, religious and herbal treatments were demonstrated in almost all of the early writings. However, methodical, carefully reasoned therapeutic approaches did not exist until the middle-ages and modern treatment with a scientific base and sophisticated instrumentation did not develop until the 18th century. Prior to the 1950s, diseases were mostly treated by root debridement and the extraction of the affected teeth. Until the 1970s, it was primarily the symptoms of periodontal diseases that were treated. The goal was radical elimination of the periodontal pocket (resective therapy). The means were gingivectomy, flap procedures and osseous surgery. The disadvantages were the massive sacrifice of periodontal tissues, lack of regeneration and clinically elongated teeth. These disadvantages, along with the realization of the importance of aetiologic agents, raised questions about the necessity of total pocket elimination, and the control of subgingival infection by a thorough scaling and root planing (nonsurgical therapy), with and without antibiotics, became a commonly used treatment during the 1980s. Comparative longitudinal studies, surgical versus nonsurgical, demonstrated that both surgical and nonsurgical therapy result in limited regeneration and healing with a long junctional epithelium. The most important aspects of today's modern concept of periodontal therapy are causal, regenerative, and specific for disease type and severity. Although the regeneration of the periodontium can be accomplished with the biological principles of guided tissue regeneration and graft materials, compared to conventional methods, the restoration of a completely normal periodontal status has not yet been achieved. We are about to reach our ultimate goals and presently, the more promising research directions for a substantial regeneration seems to lie in biological mediators. Although the future of periodontal therapy is bright, it is still of critical importance to have a preventive strategy to keep individuals healthy beforehand.

Gingival Diseases↗