Granulation tissue enriched by aspirin and omega-3 fatty acids in healing experimental periodontal lesion.

AIMS: Granulation tissue (GT) and specialized pro­resolving mediators such as lipoxins and resolvins are key elements in the successful resolution of periodontitis. Aspirin­triggered lipoxins and resolvins are even more powerful than their natural analogues. Their biosynthesis can be accelerated by omega-3 fatty acids. The aim of this study was to evaluate the use of GT enriched by aspirin and omega-3 fatty acids during the surgical treatment of periodontitis in an experimental animal model (rabbit). METHODS: In each of 24 rabbits, two experimental periodontal defects were created. In total, 47 defects were treated with open-flap debridement and one of three procedures: (1) GT extracted and soaked with aspirin and omega-3 fatty acids (ASA+OMEGA3 group); (2) GT soaked with saline (PLACEBO group); or (3) GT left untreated (CONTROL group). Then, the GT was replaced in situ. Primary evaluated criteria were the probing pocket depth (PPD) and the clinical attachment level (CAL). Necropsies were harvested 2, 6, and 12 weeks after surgery. The samples were used for histological and molecular biological assessment. RESULTS: A trend of greater PPD and CAL in the ASA+OMEGA3 group was observed at 6 weeks. However, there was no significant difference between them. During the observation period, tissue levels of FGF-7, IL-1ß and TIMP-1 showed a statistically significant decrease (P<0.05). For the other variables, the ASA+OMEGA3 group was comparable with the PLACEBO and CONTROL groups. CONCLUSION: This experiment did not demonstrate the superiority of the proposed approach. However, the enriched granulation tissue did not impair healing outcomes.

Treatment outcome after neonatal cleft lip repair in 5-year-old children with unilateral cleft lip and palate.

INTRODUCTION: The aim of this study was to assess speech outcomes and dental arch relationship of 5-year-old Czech patients with unilateral cleft lip and palate (UCLP) who have undergone neonatal cleft lip repair and one-stage palatal closure. METHODS AND MATERIALS: Twenty-three patients with UCLP, born between 2009 and 2010, were included in the study. Three universal speech parameters (hypernasality, articulation and speech intelligibility) have been devised for speech recordings evaluation. Outcomes of dental arch relationship were evaluated by applying the GOSLON Yardstick and subsequently compared with the GOSLON outcome of other cleft centers. RESULTS: Moderate hypernasality was present in most cases, the mean value for articulation and speech intelligibility was 2.07 and 1.93, respectively. The Kappa values for inter-examiner agreement for all the three speech outcomes ranged from 0.786 to 0.808. Sixty-three percent of patients were scored GOSLON 1 and 2, 26% GOSLON 3, and 10% GOSLON 4. GOSLON mean score was 2.35. Interrater agreement was very good, represented by kappa value of 0.867. CONCLUSION: The treatment protocol, involving neonatal cleft lip repair and one-stage palatal repair performed up to the first year of UCLP patient's life, has shown good speech outcomes and produced very good treatment results in regard to maxillary growth, comparable with other cleft centers.

Interleukin-17A Gene Variability in Patients with Type 1 Diabetes Mellitus and Chronic Periodontitis: Its Correlation with IL-17 Levels and the Occurrence of Periodontopathic Bacteria.

Interleukin-17 contributes to the pathogenesis of type 1 diabetes mellitus (T1DM) and chronic periodontitis (CP). We analyzed IL-17A -197A/G and IL-17F +7488C/T polymorphisms in T1DM and CP and determined their associations with IL-17 production and occurrence of periopathogens. Totally 154 controls, 125 T1DM, and 244 CP patients were genotyped using 5' nuclease TaqMan(®) assays. Bacterial colonization was investigated by a DNA-microarray kit. Production of IL-17 after in vitro stimulation of mononuclear cells by mitogens and bacteria was examined by the Luminex system. Although no differences in the allele/genotype frequencies between patients with CP and T1DM + CP were found, the IL-17A -197 A allele increased the risk of T1DM (P < 0.05). Levels of HbA1c were significantly elevated in carriers of the A allele in T1DM patients (P < 0.05). Production of IL-17 by mononuclear cells of CP patients (unstimulated/stimulated by Porphyromonas gingivalis) was associated with IL-17A A allele (P < 0.05). IL-17A polymorphism increased the number of Tannerella forsythia and Treponema denticola in patients with CP and T1DM + CP, respectively (P < 0.05). IL-17A gene variability may influence control of T1DM and the "red complex" bacteria occurrence in patients with CP and T1DM + CP. Our findings demonstrated the functional relevance of the IL-17A polymorphism with higher IL-17 secretion in individuals with A allele.

Apolipoprotein E gene polymorphisms in relation to chronic periodontitis, periodontopathic bacteria, and lipid levels.

OBJECTIVE: Inflammatory periodontal diseases may be associated with common systemic conditions and, as recently described, alterations in lipid levels in the blood. The aim of this study was to determine the possible effects of apolipoprotein E (ApoE) genotypes on the lipid levels in healthy people and patients with chronic periodontitis (CP) in relation to periodontopathic bacteria. DESIGN: This case-control study comprised 469 unrelated subjects. The genomic DNA of 294 patients with CP and 175 healthy/non-periodontitis controls were genotyped, using the real-time polymerase chain reaction (RT-PCR) method, for ApoE (rs429358 and rs7412) gene polymorphisms. Subgingival bacterial colonization was investigated by the DNA microarray using a periodontal pathogen detection kit and lipid levels were measured in a subgroup of subjects (N = 275). RESULTS: There was no evidence for a significant association between ApoE gene polymorphisms and CP (P > 0.05). Patients with CP had increased levels of total cholesterol and low-density lipoprotein (LDL) compared to controls (P< 0.05); however, no significant difference was found for triglyceride and high-density lipoprotein (HDL) levels. ApoE gene variability influenced LDL levels marginally (P = 0.08) but it did not modify total cholesterol, triglyceride, and HDL levels or the occurrence of periodontal pathogens in subgingival pockets.(23) CONCLUSIONS: In the Czech population studied, ApoE genetic variations were not associated with susceptibility to CP or the presence of periodontopathic bacteria.

Haplotype analysis of interleukin-8 gene polymorphisms in chronic and aggressive periodontitis.

OBJECTIVES: Periodontitis is an inflammatory disease characterized by connective tissue loss and alveolar bone destruction. Interleukin-8 (IL8) is important in the regulation of the immune response. The aim of this study was to analyze four polymorphisms in the IL8 gene in relation to chronic (CP) and aggressive (AgP) periodontitis. METHODS: A total of 492 unrelated subjects were included in this case-control association study. Genomic DNA of 278 patients with CP, 58 patients with AgP, and 156 controls were genotyped, using the 5' nuclease TaqMan assay, for IL8 (rs4073, rs2227307, rs2227306, and rs2227532) gene polymorphisms. Subgingival bacterial colonization was investigated by the DNA-microarray detection kit in a subgroup of subjects (N = 247). RESULTS: Allele and genotype frequencies of all investigated IL8 polymorphisms were not significantly different between the subjects with CP and/or AgP and controls (P > 0.05). Nevertheless, the A(-251)/T(+396)/T(+781) and T(-251)/G(+396)/C(+781) haplotypes were significantly less frequent in patients with CP (2.0% versus 5.1%, P < 0.02, OR = 0.34, 95% CI: 0.15-0.78, resp., 2.0% versus 4.5%, P < 0.05, OR = 0.41, 95% CI: 0.18-0.97) than in controls. CONCLUSIONS: Although none of the investigated SNPs in the IL8 gene was individually associated with periodontitis, some haplotypes can be protective against CP in the Czech population.

Association of Toll-like receptor 9 haplotypes with chronic periodontitis in Czech population.

AIM: Toll-like receptors (TLRs) belong to the pattern recognition receptors family of signal molecules that recognize conserved microbial structures. The aim of this study was to analyse polymorphisms in the TLR genes and their association with chronic periodontitis (CP). MATERIAL AND METHODS: Two polymorphisms (2408G/A, i.e. Arg753Gln and -16934A/T) in TLR-2 and three variants (-1486C/T, -1237C/T and+2848A/G) in the TLR-9 genes were studied in 222 patients with CP and 259 unrelated controls. All polymorphisms were detected using the polymerase chain reaction-restriction fragment length polymorphism methods. Subgingival bacterial colonization was investigated by the VariOr Dento test. RESULTS: No significant differences were found in allele and genotype frequencies of all polymorphisms between patients and controls. Nevertheless, complex analysis revealed differences in TLR9 haplotype frequencies between both groups (p=0.001). Specifically, the haplotype T(-1486)/T(-1237)/A(2848) was significantly more frequent (9.6%versus 2.8%, p<0.000001) and the haplotype T(-1486)/T(-1237)/G (2848) of the TLR9 gene was less frequent (35.9%versus 43.3%, p=0.01) in patients than in controls. However, no significant relationships between periodontal pathogens, TLR polymorphisms and CP were found. CONCLUSIONS: In conclusion, although no significant role of the TLR2 gene in periodontitis was found, our results indicate that TLR9 haplotypes may be associated with susceptibility to CP.

Cytology of the nasal mucosa after total laryngectomy.

CONCLUSION: This study demonstrates morphological changes in the nasal mucosa as a consequence of functional changes of the nasal cavity after total laryngectomy (TLE). OBJECTIVE: Observation of the changes of the nasal mucosa in patients after TLE. PATIENTS AND METHODS: A cohort observational study contained 30 patients after TLE. These patients underwent nasal endoscopy and cytological and microbiological examination of the nasal mucosa. The control group comprised 30 patients. RESULTS: In patients who underwent TLE, morphological changes occurred in the nasal mucosa. The hyperplasia of the cells in the basal zone was the most remarkable change after TLE. The difference was statistically significant (p=0.0002) in comparison with the control group. No statistically significant difference was shown in the incidence of squamous cell metaplasia. The inflammatory changes in the nasal mucosa were found in only 10% of patients after TLE; in comparison with the control group the statistically significant decrease was proven (p=0.003). The presence of bacteria in the nasal mucosa without any signs of inflammation was demonstrated in 27% of the patients who underwent TLE. That finding was not seen in the control group. The statistically significant difference was demonstrated (p=0.0046).

The association of MMP-9 and IL-18 gene promoter polymorphisms with gingivitis in adolescents.

OBJECTIVE: Interleukin 18 (IL-18) is shown to be a proinflammatory cytokine that regulates the expression of matrix metalloproteinase 9 (MMP-9). The aim of this study was to test for differences between Czech adolescents with and without gingivitis in relation to MMP-9 and IL-18 polymorphisms. DESIGN: A total of 298 Caucasian children, aged 11-13 years, were examined to assess gingival health. DNA for genetic analysis was obtained from buccal epithelial cells, and the MMP-9 -1562C/T and IL-18 -607A/C variants were identified with PCR-RFLP. RESULTS: Gingivitis was present in 49.3% of the adolescents examined, the rest of the group was considered healthy. The IL-18 -607C and MMP-9 -1562T alleles were found in 58.9% and 8.3% of the healthy subjects, and in 62.2% and 15.0% of the patients with gingivitis, respectively. Although differences in allele frequencies were not significant for IL-18 variant, they were significant for MMP-9 polymorphism (p=0.01, p(corr) < 0.05). Furthermore, a highly significant association of the composite genotype (formed by the variants of the both genes) with gingivitis was found (p=0.004, p(corr) < 0.05). CONCLUSIONS: The -1562 T allele of MMP-9 gene could have a role in gingivitis in adolescents. In addition, interaction of the MMP-9 and IL-18 genes could be considered a risk factor for the development of gingivitis in children.

Histomorphologic study of nasal turbinates after surgical treatment: a comparison of laser surgery and radiofrequency-induced thermotherapy effects in animals.

CONCLUSIONS: From the clinical point of view, the healing process following radiofrequency-induced thermotherapy (RFITT) is faster and less painful for the patient. This corresponds with the histopathological findings, as in a case of diode laser treatment, the damage to the tissue was greater, and the regenerative and reparative processes were less prominent. OBJECTIVE: Comparison of histopathologic changes and the healing process in the turbinate tissue regarding the kind of treatment and intensity of energy used for surgery. MATERIALS AND METHODS: Pigs were chosen as laboratory animals for this study. The animals were divided into two groups. Two different techniques were used: treatment with diode laser fiber applied submucosally in contact mode and RFITT. The first tissue samples were taken on the third postoperative day; further sampling was done on the eight postoperative day. RESULTS: Histological features of the specimens taken from turbinates 8 days after the surgery included necrosis and ulceration, fibrin deposition in the mucosa, necrotizing sialometaplasia, as well as proliferation of the granulation tissue. Generally, the changes found in the turbinates treated by diode laser were more severe, with more intensive tissue damage and less prominent regenerative and reparative changes.

Association of interleukin-6 (IL-6) haplotypes with plaque-induced gingivitis in children.

OBJECTIVES: The proinflammatory cytokine interleukin-6 (IL-6) is a key regulator of the host response to microbial infection and major modulator of extracellular matrix catabolism and bone resorption. The aim of this case-control study was to test differences between children with and without gingivitis in the distribution of IL-6 alleles at positions -174, -572, and -597 and their haplotypes. MATERIAL AND METHODS: A total of 455 Caucasian children, aged 11 to 13 years, were enrolled in this study. According to gingival bleeding on probing indices, 183 were classified as healthy subjects and 272 as children with plaque-induced gingivitis. DNA for genetic analysis was obtained from buccal epithelial cells and PCR-RFLP methods were used for genotyping three selected IL-6 promoter polymorphisms. RESULTS: Complex analysis revealed significant differences in haplotype frequencies between patients and healthy subjects (p<0.01). The CGA haplotype was significantly more frequent in children with gingivitis than in healthy subjects (41.5% versus 34.1%). In subanalyses, we found that IL-6 -174C allele was more frequent in patients (44.3%) than in healthy children (36.1%, p=0.016, P(corr)<0.05). Multivariate logistic regression analysis showed that allele C remained a risk factor for gingivitis in children (p=0.03) regardless of plaque or gender. However, the proportions of the IL-6 -597 and -572 genotypes were comparable between the two groups. CONCLUSIONS. Our results indicate that the three promoter polymorphisms in the IL-6 gene act in a cooperative fashion and suggest that IL-6 haplotypes could play a role in the pathogenesis of gingivitis in Caucasian children.

IMRT with the use of simultaneous integrated boost in treatment of head and neck cancer: acute toxicity evaluation.

Acute toxicity has been evaluated in head and neck cancer patients treated with intensity-modulated radiotherapy using simultaneous integrated boost (SIB-IMRT). The basis of the treatment protocol is an irradiation in 30 fractions with a total dose: 66 Gy to the region of macroscopic tumor, 60 Gy to the region of high-risk subclinical disease and 54 Gy to the region of low-risk subclinical disease. Between December 2003 and September 2005, 38 patients with carcinoma of different locations in the head and neck region were irradiated. Five patients underwent concurrent chemotherapy (weekly cisplatin). Acute toxicity was evaluated according to Radiation Therapy Oncology Group toxicity scale for skin, mucous membrane, salivary glands, pharynx and esophagus and larynx. All 38 patients completed the therapy without urgency of interruption due to acute toxicity of radiotherapy. No patient experienced grade 4 toxicity. More severe toxicity was observed in patients with concurrent chemotherapy. The results confirm that the irradiation according to our SIB-IMRT protocol is a therapy with acceptable toxicity and there is a space for radiobiological enhancement of this regimen by concurrent chemotherapy, e.g. weekly cisplatin.

Pediatric tumors of the parapharyngeal space. Three case reports and a literature review.

Three cases of pediatric primary parapharyngeal space tumors, namely a Burkitt's lymphoma, lipoblastoma and a ganglioneuroma are described here, along with a literature review. These disorders are very rare, accounting for less than 20% of all parapharyngeal neoplasms. In comparison to adults, there are differences in pathomorphologic spectrum of pediatric parapharyngeal tumors. The authors describe the prevalence of malignant tumors, mainly soft tissues sarcomas. The salivary gland tumors and paragangliomas which dominate in adult populations, are extremely rare in pediatric populations where the most common benign tumor to be encountered is that of neurogenic origin. It follows that these features modify the diagnostic and therapeutic approach.

Neurofibromatosis 2: two case reports.

Our study presents two cases of neurofibromatosis 2 (NF2) that have been diagnosed at the Ear, Nose, and Throat Department of Hradec Králové (Czech Republic). The first case involved a young man with a history of sudden hearing loss accompanied by tinnitus on the left side. The diagnosis of NF2 was made, and an operation for left acoustic neuroma was performed. Looking toward the future, the acoustic neuroma on the right side should be resolved as well. The second case concerned a woman (the mother of our patient 1) examined at the same Ear, Nose, and Throat Department in 1980, after 4 years of gait instability and progressive loss of hearing and tinnitus on the right side. Computed tomography scan detected a bilateral expansion in the pontocerebellar angles, and a large tumor on the right side was removed. The patient is deaf and has facial palsy without progression of symptomatology during long-term follow-up. These two cases document the rare but serious hereditary disease of NF2. Its most frequent first presentation is acoustic neuroma; further, benign tumors of the nervous system and juvenile cortical cataract also are often detected. The variability of number, location, and biological behavior of tumors associated with NF2 require an individual patient treatment approach, long-term follow-up, and insertion of appropriate hearing aids. Important also is a genetic examination to exclude pathological NF2 genes in the first-degree relatives of the affected individuals.