Transforming growth factor-beta1 gene expression and cyclosporine A-induced gingival overgrowth: a pilot study.

AIMS: The relationship between gingival overgrowth (GO) induced by cyclosporine A (CsA) and transforming growth factor-beta1 (TGF-beta1) remains unclear. The aims of the present study were to evaluate TGF-beta1 gene expression under different immunosuppressive treatments and its association with TGF-beta1 gene functional polymorphism and GO in renal transplant recipients. MATERIAL AND METHODS: The study included 98 CsA-treated renal transplant recipients (with and without GO) and 44 tacrolimus-treated transplant patients (without GO). TGF-beta1 mRNA expression was measured using a real-time quantitative polymerase chain reaction assay. The levels were correlated with TGF-beta1 gene polymorphisms at codons 10 and 25, with different immunosuppressive treatment and GO. RESULTS: The level of TGF-beta1 gene expression was insignificantly lower in the CsA-treated group compared with the tacrolimus group, and significantly lower in the group with GO compared with patients without GO. In tacrolimus- and CsA-treated patients, but not in patients with GO, the level of TGF-beta1 gene expression was associated with functional phenotypes of TGF-beta1. The incidence, degree and extent of GO were higher in recipients with lower TGF-beta1 gene expression. CONCLUSIONS: Lower level TGF-beta1 gene expression, not functional polymorphism, in patients treated with CsA may be considered to be a risk factor for GO.

Local inflammation exacerbates cyclosporine a-induced gingival overgrowth in rats.

Gingival overgrowth (GO) is a common side effect of long-term cyclosporine A (CsA) treatment. The risk factors appraised include drug interactions with calcium channel blockers, age, cyclosporine dose, dental bacterial plaque, duration of treatment, and genetic predisposition. The relationship and mechanism between GO and local inflammation caused by dental bacterial plaque have not been clearly defined. This research was carried out to investigate the histomorphometrical alterations and serum levels of transforming growth factor beta1 (TGF-beta1) in CsA-induced GO with or without local inflammation. Thirty-four male Sprague-Dawley rats were divided into 4 groups: Group I (control); Group II (ligation); Group III (CsA); Group IV (ligation and CsA). After 9 weeks the rats were sacrificed. The morphological examination was made and the histological changes with hematoxylin and eosin (HE) staining were observed. TGF-beta1 levels in serum were detected by enzyme-linked immunosorbent assay (ELISA). We report here that obvious GO are found in Group III and Group IV after CsA treatment, especially those rats with existed gingivitis presented an aggravation of GO. TGF-beta1 levels in CsA-exposed groups were significantly higher than untreated groups, but ligation did not affect TGF-beta1 level. These findings suggest that CsA-induced GO can be exacerbated by local inflammation. TGF-beta1 may be a key factor for the development of GO.

The relationship of transforming growth factor-beta1 gene polymorphism, its plasma level, and gingival overgrowth in renal transplant recipients receiving different immunosuppressive regimens.

BACKGROUND: Cyclosporin A (CsA) induces gingival overgrowth (GO) in patients who seem to be prone to this disorder. It is still impossible to determine which patients will develop GO. Patients treated with the new immunosuppressive drug tacrolimus seem not to have GO. The aims of this study were to investigate transforming growth factor-beta1 (TGF-beta1) gene polymorphisms in renal transplant recipients treated with CsA or tacrolimus and to establish an association between these polymorphisms and TGF-beta1 plasma concentration and the incidence of GO. METHODS: The examined group consisted of 134 renal transplant recipients. Ninety-two underwent CsA treatment (50 with and 42 without GO), and 42 underwent tacrolimus treatment. Age, gender, time after transplantation, calcineurin inhibitor total dosage, number of teeth, and sulcus bleeding index were analyzed. TGF-beta1 plasma levels were estimated in 60 CsA- and 30 tacrolimus-treated patients. Two biallelic polymorphisms of the TGF-beta1 gene were studied at codon 10 (at position +869) and at codon 25 (at position +915) in patients from the examined group and in 108 healthy volunteers (the control group). RESULTS: The distribution of the high, intermediate, and low TGF-beta1 producer phenotypes was comparable in all the studied groups and in the healthy controls. The high producer phenotype was more frequent in patients with GO. TGF-beta1 levels in the CsA group showed correlation with the phenotypes. The lowest incidence of GO was observed in the 10C/C genotype, whereas the highest was observed in the 10T/C genotype. CONCLUSION: High and intermediate TGF-beta1 producer phenotypes and heterozygous genotype 10T/C might be considered risk factors for GO in patients treated with CsA.

Analysis of peripheral blood leukocytes in patients with cyclosporine A-induced gingival hyperplasia.

BACKGROUND: Gingival overgrowth is one of the major adverse effects of the immunosuppressive drug cyclosporine A (CsA). Although several studies have attempted to determine the immunological mechanisms of gingival hyperplasia (GO) due to CsA therapy, the pathogenesis remains unclear. In this study, the distribution of the peripheral blood leukocytes in a group of renal transplant patients undergoing CsA therapy was analyzed and possible correlations of periodontal and pharmacological variables to lymphocyte subpopulations, natural killer cells, and monocytes investigated. METHODS: Thirty-six patients were classified into 2 groups of 18 each according to the degree of gingival overgrowth. The periodontal evaluation included plaque index (PI), gingival index (GI), gingival overgrowth (GO), calculus index (CI), and probing depth (PD). The pharmacological variables of current doses of the therapeutic serum levels of CsA were investigated. The peripheral blood leukocytes were studied by 2-color flow cytometric analysis using anti-human CD2, CD3, CD4, CD8, CD11b, CD11c, CD16, CD19, HLA-DR, and CD3+HLA-DR+ monoclonal antibodies. RESULTS: Statistical evaluation revealed that none of the pharmacological variables varied between the 2 groups. Responders (GO >30%) had significantly higher GI, PD, and GO scores compared to nonresponders (GO < or =30%). Of the immunological parameters studied, only CD2 was higher in the responder group. None of the clinical parameters correlated to the immunological values. CONCLUSIONS: The results of this study may be useful in explaining the underlying mechanisms of drug-induced gingival overgrowth. Several previously unsuspected cells and accessory activation mechanisms for T lymphocytes could play a role in the pathogenesis.

Serum phenytoin concentration and IgG antibody titre to periodontal bacteria in patients with phenytoin-induced gingival overgrowth.

Epileptic patients taking phenytoin with gingival-overgrowth and those without gingival-overgrowth were compared for daily drug dose, plasma total phenytoin concentration, plasma free-phenytoin concentration and serum IgG antibody titre against 13 periodontal bacteria. Significantly higher daily drug dose was noted in patients with gingival overgrowth (P < 0.05) when compared with those without overgrowth. In addition, both total and free forms of plasma phenytoin concentration were significantly higher in sera of patients with gingival growth than of those without overgrowth (P < 0.01). Strong positive correlation was found between daily drug dose and serum phenytoin concentration in patients with gingival overgrowth, while weak correlation was found in patients without gingival overgrowth, suggesting a difference in drug metabolism in these two groups. However, no differences were found in serum IgG antibody titres to 13 periodontal bacteria examined between two groups. These results suggest that metabolic ability of phenytoin is one of the factors for developing gingival overgrowth, and that periodontal infection may not be a primary causative factor for gingival overgrowth but act as an additive factor which increase tissue mass for this unwanted side effect.

The effect of cyclosporine with and without nifedipine on gingival overgrowth in renal transplant patients.

PURPOSE: This investigation was performed to evaluate the effect of cyclosporine alone and in combination with nifedipine on gingival overgrowth. METHODS: One hundred and nineteen patients who had undergone renal transplantation at least 12 months previously were selected for the study. The patients were divided into 2 groups according to whether they had received cyclosporine alone (group 1, n = 98) or cyclosporine with nifedipine (group 2, n = 21). Periodontal and pharmacological characteristics were assessed for all patients. RESULTS: Marked gingival overgrowth was seen in 11 (52%) of the patients in group 2 but just 6 (6%) of those in group 1. In addition, the gingival overgrowth index was significantly greater for patients who had received both nifedipine and cyclosporine (Mann-Whitney U-test, p < 0.001). However, there were no significant differences between groups with higher and lower gingival overgrowth index in terms of age, sex, cyclosporine dose, nifedipine dose or level of cyclosporine in the serum. CONCLUSION: The combination of cyclosporine and nifedipine may increase the incidence as well as the severity of gingival overgrowth in renal transplant patients. Among patients who had received both drugs, there was a clear relationship between gingival overgrowth and duration of cyclosporine and nifedipine use.

GCF and serum myeloperoxidase and matrix metalloproteinase-13 levels in renal transplant patients.

AIM: The rationale of this study was to address whether local or systemic changes reflect proteolytic (matrix metalloproteinase-13) or oxidative (myeloperoxidase) stress in renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), in patients receiving CsA therapy and having no GO and patients receiving tacrolimus therapy. MATERIAL AND METHODS: Gingival crevicular fluid (GCF) samples were collected from sites with (GO+) and without GO (GO-) in CsA patients having GO; GO- sites in CsA patients having no GO; sites from tacrolimus, gingivitis and healthy subjects. GCF and serum myeloperoxidase (MPO) and matrix metalloproteinase-13 (MMP-13) levels were determined by ELISA. RESULTS: GO+ sites in CsA patients having GO had elevated GCF MPO levels than those of CsA patients having no GO, tacrolimus and healthy subjects (p<0.005), but comparable to those of gingivitis. GCF MPO levels were higher in GO+ compared to GO- sites in CsA patients having GO (p<0.05). Patient groups had similar, but higher GCF MMP-13 levels than healthy group. CONCLUSIONS: These results show that CsA and tacrolimus therapy have not a significant effect on GCF MPO and MMP-13 levels, and gingival inflammation seems to be the main reason for their elevations.

Increased bFGF level in the serum of patients with phenytoin-induced gingival overgrowth.

To elucidate the involvement of bFGF (basic fibroblast growth factor) in the pathogenesis of phenytoin-induced gingival overgrowth, we measured the concentration of bFGF in the serum of 36 epileptic patients taking phenytoin and in 94 normal volunteers by enzyme-linked immunosorbent assay technique. The concentration of phenytoin in serum was determined by high-performance liquid chromatography. In 34 of 36 patients taking phenytoin in this investigation, apparent gingival overgrowth was noticed. The mean concentration of bFGF was 33.9+/-18.5 pg/ml in the overgrowth group and 10.6+/-5.2 pg/ml in the volunteer group (p<0.01). The serum phenytoin level did not correlate (r=0.22, p=0.2) with the degree of gingival overgrowth but there was a significant correlation (r=0.38, p=0.023) between the degree of gingival overgrowth and the serum bFGF level. However, no correlation was observed among age, daily phenytoin dose, total phenytoin dose, duration of phenytoin therapy, serum phenytoin level, or serum bFGF level. The results suggested that enhanced serum bFGF level was implicated in the pathogenesis of phenytoin-induced gingival overgrowth.

Epidermal growth factor in saliva and serum of patients with cyclosporin-induced gingival overgrowth.

The purpose of this study was to investigate the presence of epidermal growth factor (EGF) in patients receiving cyclosporin therapy who had gingival overgrowth and to determine whether there were any differences between these patients and normal healthy controls. Seventeen patients with cyclosporin-induced gingival overgrowth and seventeen age- and sex-matched controls who were taking cyclosporin but had healthy gingiva were used for this study. Unstimulated whole saliva was collected from all individuals by expectoration. Gingival crevicular fluid (GCF) was also collected from all individuals. Blood was additionally collected from all subjects and serum was separated by keeping the samples overnight at 4 degrees C. EGF levels in all cases were measured by an ELISA assay. EGF concentrations were found to be significantly higher in the saliva of patients with cyclosporin-induced gingival overgrowth compared to the control group (401.2 +/- 31.1 pg/ml and 144.3 +/- 31.4 pg/ml, respectively), whereas the results were reversed in the serum (67.0 +/- 15.6 pg/ml and 141.6 +/- 17.7 pg/ml, respectively). EGF was not detected in the samples of GCF in either group. This study thus demonstrated an increase in EGF levels in the saliva and a decrease of EGF in the serum of patients with cyclosporin-induced gingival overgrowth.