The study of genetic predisposition on periodontitis and peri-implantitis.

Background: Peri-implant mucositis and peri-implantitis cases increase in number with the increase of implant applications. Peri-implant mucositis and peri-implantitis are defined as inflammatory diseases with inflammation and loss in soft and hard tissue, similar to the other periodontal diseases. As observed in many diseases, genetic predisposition factors also affect the progress of periodontitis and peri-implantitis. Aim: This study examines if there is any solid genetic predisposition causing periodontitis and peri-implantitis formation in Turkish patients. Patients & Methods: In order to evaluate single nucleotide polymorphism (SNP), Interleukin-8 (IL-8) and N-formyl-L-methionyl-L-leucyl-phenylalanine (fMLP), playing a role in the chemotaxis of neutrophils, and Fc Gamma Receptor IIA (FcγRIIA) and Fc Gamma Receptor IIIA (FcγRIIIA), playing a role in the antigen-antibody complexes and phagocytosis, were selected. Thirty-two Turkish non-smoking subjects, having periodontitis, thirty-three Turkish non-smoking subjects, having peri-implantitis and thirty-three Turkish non-smoking healthy subjects were selected. In total 98 adults participated in our study. Collected saliva samples from the participants were used for DNA isolation. SNPs were determined in these subgroups of the study by means of genotype-specific polymerase chain reactions. Results: When IL-8 A-251T, FcγRIIa -H131 and FcγRIIIa -V158 polymorphism were evaluated, no significant difference was found between periodontitis, peri-implantitis and healthy groups. However, this study observed that fMLP Receptor (FPR1) gene polymorphism creates a significant difference in individuals at higher risk of periodontitis or peri-implantitis. Conclusion: Results show that individuals with the G genotype have a higher risk of periodontitis, while individuals with G / C genotype have higher risk of peri-implantitis.

Apoptosis in gingival overgrowth tissues.

Variations in the balance between cell proliferation and apoptosis could contribute to the etiology of gingival overgrowth. The aim of this study was to test the hypothesis that, in fibrotic gingival lesions, fibroblast proliferation is stimulated and apoptosis is decreased. Apoptotic index, caspase 3 expression, the proliferative index, FOXO1 expression, and histological inflammation were measured in situ. Analysis of data showed that apoptosis decreased in all forms of gingival overgrowth examined (p < 0.05), and inflammation caused a small but significant increase compared with non-inflamed tissues (p < 0.05). The greatest decrease of apoptosis occurred in the most fibrotic tissues. Cell proliferation was elevated in all forms of gingival overgrowth tested, independent of inflammation (p < 0.05). To identify potential mechanisms of transcriptional regulation of apoptosis, we assessed FOXO1 and caspase 3 expression levels and found them to correlate well with diminished apoptosis. Analysis of data suggests that increased fibroblast proliferation and a simultaneous decrease in apoptosis contribute to gingival overgrowth.

Fluid dynamics of gingiva and gingival health in children with end stage renal failure.

OBJECTIVES: Impaired homeostasis and fluid balance are important physiopathological alterations in patients with chronic renal failure which may adversely affect the fluid dynamics and health status of tissues and organs. There are insufficient data about this phenomenon in periodontal tissues. The aim of this study was to evaluate the fluid dynamics of gingiva in children with end stage renal failure (ESRF), correlating this entity with gingival health in the same patient group. DESIGN: Fifteen paediatric ESRF patients undergoing peritoneal dialysis (test group) and 15 systemically healthy children (control group) who were without periodontitis participated in the study. Fluid dynamics of gingiva were assessed via the gingival crevicular fluid (GCF) volume and tissue osmotic pressure (GOP) levels in the groups. GCF volume was measured using a Periotron 8000, whereas GOP was measured using a digital osmometer. Silness and Löe Plaque index (PI) and, Löe and Silness gingival index (GI) scores were utilized to determine the gingival health status in the study population. RESULTS: There were increases in the GCF volume and GOP of the test group compared to those of the control group (p<0.01). The PI and GI scores were higher in the test group than in the control group (p<0.01). Strong and positive correlations were found between GI and GCF volume, GI and GOP and, GCF volume and GOP in both groups (p<0.01). CONCLUSIONS: Our findings suggest that the fluid dynamics of gingiva may alter in children with ESRF, and this phenomenon may consequently affect the gingival health of these patients.

Phenotype of ENAM mutations is dosage-dependent.

Five mutations in the ENAM gene have been found to cause hypoplastic amelogenesis imperfecta (AI), with phenotypes ranging from localized enamel pitting in carriers to severe hypoplastic AI. To determine the generality of ENAM mutations in hypoplastic AI, we sequenced the ENAM gene in ten Turkish families segregating autosomal hypoplastic AI. In two families, ENAM mutations were found. A novel nonsense mutation (g.12663C>A; p.S246X) was identified in one family segregating local hypoplastic AI as a dominant trait. Affected individuals in a second family segregating autosomal-recessive AI were compound heterozygotes for a novel insertion mutation (g.12946_12947insAGTCAGTACCAGTACTGTGTC) and a previously described insertion (g.13185_13186insAG) mutation. Heterozygous carriers of either insertion had a localized enamel-pitting phenotype. These findings substantiate that enamel phenotypes of ENAM mutations may be dose-dependent, with generalized hypoplastic AI segregating as a recessive trait and localized enamel pitting segregating as a dominant trait.

MMP20 active-site mutation in hypomaturation amelogenesis imperfecta.

The Amelogenesis Imperfecta (AI) are a group of clinically and genetically heterogeneous disorders that affect enamel formation. To date, mutations in 4 genes have been reported in various types of AI. Mutations in the genes encoding the 2 enamel proteases, matrix metalloproteinase 20 (MMP20) and kallikrein 4 (KLK4), have each been reported in a single family segregating autosomal-recessive hypomaturation AI. To determine the frequency of mutations in these genes, we analyzed 15 Turkish probands with autosomal-recessive hypomaturation AI for MMP20 and KLK4 gene mutations. No KLK4 mutations were found. A novel MMP20 mutation (g.16250T>A) was found in one family. This missense mutation changed the conserved active-site His226 residue of the zinc catalytic domain to Gln (p.H226Q). Zymogram analysis demonstrated that this missense mutation abolished MMP20 proteolytic activity. No MMP20 mutations were found in the remaining 14 probands, underscoring the genetic heterogeneity of hypomaturation AI.

Novel ENAM mutation responsible for autosomal recessive amelogenesis imperfecta and localised enamel defects.

The genetic basis of non-syndromic autosomal recessive forms of amelogenesis imperfecta (AI) is unknown. To evaluate five candidate genes for an aetiological role in AI. In this study 20 consanguineous families with AI were identified in whom probands suggested autosomal recessive transmission. Family members were genotyped for genetic markers spanning five candidate genes: AMBN and ENAM (4q13.3), TUFT1 (1q21), MMP20 (11q22.3-q23), and KLK4 (19q13). Genotype data were evaluated to identify homozygosity in affected individuals. Mutational analysis was by genomic sequencing. Homozygosity linkage studies were consistent for localisation of an AI locus in three families to the chromosome 4q region containing the ENAM gene. ENAM sequence analysis in families identified a 2 bp insertion mutation that introduced a premature stop codon in exon 10. All three probands were homozygous for the same g.13185_13186insAG mutation. These probands presented with a generalised hypoplastic AI phenotype and a class II openbite malocclusion. All heterozygous carriers of the g.13185_13186insAG mutation had localised hypoplastic enamel pitting defects, but none had AI or openbite. The phenotype associated with the g.13185_13186insAG ENAM mutation is dose dependent such that ARAI with openbite malocclusion segregates as a recessive trait, and enamel pitting as a dominant trait.

Evidence of a founder effect for four cathepsin C gene mutations in Papillon-Lefèvre syndrome patients.

We describe a mutation and haplotype analysis of Papillon-Lefèvre syndrome probands that provides evidence of a founder effect for four separate cathepsin C mutations. A total of 25 different cathepsin C mutations have been reported in 32 families with Papillon-Lefèvre syndrome (PLS) and associated conditions. A characteristic of these findings is the diversity of different cathepsin C mutations that have been identified. To evaluate the generality of cathepsin C mutations, PLS probands representative of five reportedly unrelated Saudi Arabian families were evaluated by mutational and haplotype analyses. Sequence analysis identified two cathepsin C gene mutations: a novel exon 7 G300D mutation was found in the proband from one family, while probands from four families shared a common R272P mutation in exon 6. The R272P mutation has been previously reported in two other non-Saudi families. The presence of the R272P mutation in probands from these four Saudi families makes this the most frequently reported cathepsin C mutation. To distinguish between the presence of a possible founder effect or a mutational hot spot for the R272P mutation, we performed haplotype analysis using six novel DNA polymorphisms that span a 165 kb interval containing the cathepsin C gene. Results of haplotype analysis for genetic polymorphisms within and flanking the cathepsin C gene are consistent with inheritance of the R272P mutation "identical by descent" from a common ancestor in these four Saudi families. Haplotype analysis of multiple PLS probands homozygous for other cathepsin C mutations (W249X, Q286X, and T153I) also supports inheritance of each of these mutations from common ancestors. These data suggest that four of the more frequently reported cathepsin C mutations have been inherited from common ancestors and provide the first direct evidence for a founder effect for cathepsin C gene mutations in PLS. Identification of these six short tandem repeat polymorphisms that span the cathepsin C gene will permit haplotype analyses to determine other founder haplotypes of cathepsin C mutations in additional PLS families.

Identification of cathepsin C mutations in ethnically diverse papillon-Lefèvre syndrome patients.

INTRODUCTION: Papillon-Lefèvre syndrome (PLS) is an autosomal recessive disorder characterised by palmoplantar keratoderma and severe, early onset periodontitis, which results from deficiency of cathepsin C activity secondary to mutations in the cathepsin C gene. To date, 13 different cathepsin C mutations have been reported in PLS patients, all of which are homozygous for a given mutation, reflecting consanguinity. AIM: To evaluate the generality of cathepsin C mutations in PLS, we studied an ethnically diverse group of 20 unrelated families. METHODS: Mutations were identified by direct automated sequencing of genomic DNA amplified for exonic regions and associated splice site junctions of the cathepsin C gene. Long range PCR was performed to determine the genomic structure of the cathepsin C gene. RESULTS: The cathepsin C gene spans over 46 kb, with six introns ranging in size from 1.6 to 22.4 kb. Eleven novel mutations and four previously reported mutations were identified in affected subjects from 14 families. Missense mutations were most common (9/15), followed by nonsense mutations (3/15), insertions (2/15), and deletions (1/15). Among these 14 probands, two were compound heterozygotes. Affected subjects with transgressions of the dermal lesions onto the knees or elbows or both had mutations in both the pro- and mature regions of the enzyme, although most were in the mature region. CONCLUSION: Mutations in the mature region of cathepsin C were more likely to be associated with the transgressions of the dermatological lesions, although the results were not statistically significant. A comprehensive list of all cathepsin C mutations described to date, representing 25 mutations from 32 families with PLS and related conditions, is also presented.

Evaluation of Local and Systemic Levels of Interleukin-17, Interleukin-23, and Myeloperoxidase in Response to Periodontal Therapy in Patients with Generalized Aggressive Periodontitis.

We aimed to investigate serum and gingival crevicular fluid levels of myeloperoxidase, interleukin-17, and interleukin-23 before and after nonsurgical periodontal therapy in generalized aggressive periodontitis patients and compare to those in healthy controls. Interleukin-17, interleukin-23, and myeloperoxidase levels were measured by enzyme-linked immunosorbent assay in gingival crevicular fluid and serum samples taken from 19 systemically healthy generalized aggressive periodontitis patients and 22 healthy controls. In addition, the levels of IL-17, IL-23, and myeloperoxidase were reassessed at 3 months after periodontal therapy in the generalized aggressive periodontitis (GAP) group. Periodontal clinical parameters were also evaluated at baseline and 3 months post-therapy. The investigated molecule levels in serum decreased significantly at 3 months as a result of the therapy (p = 0.014 for IL-17, p = 0.000 for IL-23, and p = 0.001 for myeloperoxidase (MPO)). Significant reductions were also observed in gingival crevicular fluid (GCF) IL-17, IL-23, and MPO levels at 3 months after therapy (p = 0.000 for all molecules). However, the GCF levels of IL-17, IL-23, and MPO in GAP patients were still higher than those in the controls at 3 months (p = 0.001). A significant decrease in the local and systemic levels of IL-17, IL-23, and MPO based on the therapy might indicate the role of these mediators for tissue destruction in periodontal tissues.

Sublocalization of the Papillon-Lefevre syndrome locus on 11q14-q21.

Papillon-Lefevre syndrome (PLS) is an autosomal recessive form of palmoplantar ectodermal dysplasia, characterized by palmoplantar hyperkeratosis and severe early-onset periodontitis. The presence of severe periodontitis distinguishes PLS from other palmoplantar keratodermas. As part of our efforts to study the genetic basis of periodontitis susceptibility, we performed a genome-wide search to identify major loci for PLS in 44 individuals (14 affected) from 10 consanguineous PLS families. We have identified evidence for linkage of a PLS gene on 11q14-q21. A maximum two-point logarithm of the odds (LOD) score of 8.24 was obtained for D11S1367 at a recombination fraction of theta=0.00. Multipoint analysis resulted in a LOD score of 10.45 and placed the gene for PLS within a 4-5 cM genetic interval. This genetic interval, flanked by D11S4197 and D11S931, contains more than 50 cDNAs and 200 expressed sequence tags (ESTs). This refinement of the candidate region for a PLS gene is in agreement with other recent reports of linkage for PLS to chromosome 11q14-q21 and should help in identification of the gene for PLS.

The relationship between clinical periodontal status and insulin-dependent diabetes mellitus. Results after 5 years.

The clinical periodontal status of 44 insulin-dependent diabetic children and adolescents and 20 healthy control subjects was compared for a period of approximately 5 years. Fasting blood glucose, fructosamine, and glycosylated hemoglobin (HbA1) values were determined at baseline and 5 years later. The differences in the clinical and laboratory parameters were compared during the study period. The differences between the two groups were also evaluated. The only statistically significant difference observed in the diabetic group was clinical attachment loss (CAL). The CAL was statistically significantly higher in the diabetic group compared to the controls, and a statistically significantly higher in the diabetic group compared to the controls, and a statistically significant positive correlation was observed between the duration of diabetes and CAL. Fructosamine was also correlated with the gingival index in the diabetic group while there was no correlation in the controls. It may be concluded that diabetes modifies the clinical status of the periodontal tissues and increases clinical attachment loss.

Papillon-Lefèvre syndrome. Analysis of neutrophil chemotaxis.

Papillon-lefèvre syndrome (PLS) is described as the association of palmar-plantar hyperkeratosis with precocious periodontal disease which results in exfoliation of primary and permanent dentitions. This study was planned to assess the chemotaxis of peripheral blood neutrophils in 7 patients (3 females and 4 males) with Papillon-Lefevre syndrome. The neutrophil chemotaxis was analyzed using the zymosan activated serum (ZAS) assay. Chemotaxis and spontaneous migration measurements were compared to those of the healthy control subjects. The peripheral blood neutrophil chemotaxis and spontaneous migration were depressed in all patients with Papillon-Lefèvre syndrome. The decreased chemotaxis of peripheral blood neutrophils strongly suggests that the neutrophils may act as one of the important key determinants in the pathogenesis of severe periodontal destruction in patients with PLS.

Radiologic and histomorphometric evaluation of maxillary sinus grafting with alloplastic graft materials.

BACKGROUND: Sinus lifting procedures are widely used to obtain adequate bony support for implant placement at the atrophic maxillae. The aim of this study was to compare various sinus lifting and grafting techniques and materials. METHODS: Nine maxillae were treated with delayed and 46 maxillae with immediate implant placement techniques. A total of 104 implants were inserted. Panoramic radiographs were obtained prior to, after, and 6 to 8 months after surgery. Computed tomographies were also taken before and after surgery. The height of new bone was compared. Biopsy specimens were obtained during delayed implant placement and analyzed histomorphologically. RESULTS: There were no statistically significant differences between the panoramic radiographs for delayed and immediately placed implants, or between the graft materials. We observed correlations between the panoramic radiographs and computerized tomographies. CONCLUSION: Both delayed and immediate placement of implants can be used safely for sinus lifting. There were no statistically significant differences between the various graft materials.

Clinical and immunological findings in 2 siblings with Papillon-Lefèvre syndrome.

Rapid and severe destruction of periodontal tissues in early childhood has been reported both in systemically healthy children and in children with systemic disorders. In this study, the clinical and immunological findings of two siblings in a family with Papillon-Lefèvre syndrome are presented. The peripheral blood lymphocytes were analyzed using a double colored flow cytometry and adequate monoclonal antibodies to CD2, CD3, CD4, CD5, CD8, CD11b, CD16, CD19, and HLA-DR receptors. CD11b expression was found to be higher in both siblings (35% and 37%). The elevated CD11b expression may be related to a defect in neutrophils. The expression of natural killer cells was found to be higher in one patient but the results were in normal range. The CD2+, CD3+, CD4+, CD5+, CD8+, and CD19+ lymphocytes were in normal range in both patients. We think that the depressed chemotaxis of peripheral neutrophils, and higher expression of HLA-DR and CD11b molecules in peripheral leukocytes were useful in explaining the pathogenesis of the Papillon-Lefèvre syndrome.

Fructosamine as a possible monitoring parameter in non-insulin dependent diabetes mellitus patients with periodontal disease.

Fructosamine assay is a new test used in the diagnosis and monitoring of diabetic patients. This assay may be of interest to the periodontist for, while the traditional plasma glucose value would give a general view and information about diabetic control at a certain point, the fructosamine concentration gives an indication of the plasma glucose level over a considerable period of time, such as 1 to 3 weeks. We investigated whether there was any relation between the diseased state of the periodontal tissues and plasma fructosamine and the plasma glucose values in non-insulin dependent diabetes mellitus (NIDDM) patients. We found that fructosamine correlated with the degree of gingival bleeding, however serum glucose levels had little or no correlation.

Periodontal management of Glanzmann's thrombasthenia: report of 3 cases.

Glanzmann's thrombasthenia was reported and described as a bleeding diathesis seen in children and characterized by diminished clot retraction. It is an autosomal recessive bleeding disorder. The disease is marked by frequent mucocutaneous hemorrhages either due to defective function of the platelets or lack of fibrinogen binding membrane receptor glycoproteins IIb/IIIa which are located on the surface of the platelets. Case reports on 3 siblings, a girl of 11, and 2 boys of 12 and 16 years old with Glanzmann's thrombasthenia are reviewed. The major complaint of the patients was gingival bleeding. Periodontal treatment was performed under platelet transfusion and proper oral hygiene instruction was given. The patients were followed for 6 months and no periodontal complications developed during this time. Proper periodontal care for such patients is essential both for local and systemic health.

The effect of verapamil on the prevalence and severity of cyclosporine-induced gingival overgrowth in renal allograft recipients.

Cyclosporine A (CsA) and verapamil are two agents used in renal transplantation, both of which are suspected of inducing gingival overgrowth. This study was conducted to investigate the effect of verapamil on the severity and prevalence of CsA-induced gingival overgrowth. Fifty-one (51) renal transplant recipients (total group) of whom 22 were using only CsA (Group A) and 29 of whom were prescribed CsA + verapamil (Group B) were evaluated for various periodontal and pharmacological parameters. No statistically significant differences were found in age, sex, plaque index, gingival index, calculus index, probing depth, CsA oral dose, CsA whole blood level, duration of CsA therapy, azathioprine dose, and prednisolone dose. Although the prevalence of the gingival overgrowth was more pronounced in CsA + verapamil group compared to CsA group (51.72% vs. 40.91%), the difference was not statistically significant. Similarly, the severity of gingival overgrowth, although more manifest in CsA + verapamil group than CsA patients (34.24% vs. 28.91%), was not significantly different. Gingival overgrowth scores in the main group, CsA, and CsA + verapamil groups were found to be positively correlated to periodontal probing depths (r = 0.60, r = 0.70, r = 0.52, respectively) and the gingival index (r = 0.60, r = 0.70, r = 0.54, respectively). CsA oral dose, whole blood level, and duration of CsA therapy were not found to be correlated with the gingival overgrowth in either group. Likewise, the dose of verapamil and the duration of verapamil therapy were not correlated with the gingival overgrowth in Group B. This study indicates that verapamil, when prescribed as the calcium channel blocker in renal transplant patients, has no augmenting effect on the severity and the prevalence of CsA-induced gingival overgrowth.

Clinical effects of periodontal therapy on the severity of cyclosporin A-induced gingival hyperplasia.

BACKGROUND: Gingival hyperplasia (GH) is a major side effect associated with cyclosporin A (CsA) therapy. The condition is further augmented due to the gingival inflammation. In this study, the effects of initial periodontal therapy and gingival curettage are analyzed in a group of patients with clinically significant (>30%) CsA-induced gingival hyperplasia. METHODS: The test group of 15 patients received oral hygiene instructions, supra- and subgingival scaling, polishing, and gingival curettage only oral hygiene instructions were given to 16 control subjects. Plaque index (PI), gingival index (GI), calculus index (CI), periodontal probing depth (PD), and gingival hyperplasia were recorded at baseline and repeated 8 weeks after treatment. Current doses of immunosuppressive agents, serum concentrations of CsA, and duration of CsA therapy were recorded as the pharmacological parameters. RESULTS: Statistical evaluation revealed that all clinical variables showed statistical decreases compared to baseline in the treated patients, while none of the parameters changed significantly in the control group. Initial GH scores of 53.63% in controls and 53.40% in the treated patients were 52.83% and 32.13% following treatment, respectively. A difference of 21.27% in the severity of treated GH was accompanied by a 0.56 decrease in GI scores in the test group. CONCLUSIONS: Compared to the initial observations, the results suggested that nearly 60% of the condition could be of fibrotic origin. Initial periodontal therapy and curettage resulted in the resolution of the inflammation in CsA-induced GH. Further investigation of the treated patients has shown that 7 out of 15 patients (47%) in the test group responded well and their GH scores decreased below 30% at the end of the study. The treatment in this study was effective in eliminating the necessity of more extensive surgical modes of treatment, such as gingivectomy, in 47% of cases.

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.