CD14 and TNFα single nucleotide polymorphisms are candidates for genetic biomarkers of peri-implantitis.

OBJECTIVES: This study aims to investigate whether CD14-159 C/T and TNFα -308 A/G single nucleotide polymorphisms are associated with peri-implantitis and to evaluate their effects on bone resorption by correlating with local levels of receptor activator nuclear factor kappa-B ligand (RANKL) and osteoprotegerin (OPG). MATERIAL AND METHODS: Study population included 369 Southeastern Europe Caucasians (180 with peri-implantitis and 189 with healthy peri-implant tissues). Genotyping was performed using polymerase chain reaction from the periphery blood samples, while RANKL and OPG were evaluated in peri-implant crevicular fluid specimens using enzyme-linked immunosorbent assay. RESULTS: Analysis of CD14-159 C/T polymorphism showed that genotype of CC nucleic acid combination was associated with peri-implantitis demonstrating a fivefold increased risk in these carriers. Furthermore, for TNFα -308 A/G polymorphisms, it was evidenced that AG genotype was associated with peri-implantitis and a fivefold increased risk in these carriers. Peri-implantitis patients with CC genotype at CD14-159 exhibited significantly higher concentrations of RANKL and relative ratio RANKL/OPG when compared to patients with CT genotype, while concentration of biomarkers between different genotypes at TNFα -308 remained insignificant. CONCLUSION: Within the limitations of the study, we can conclude that CD14-159 C/T and TNFα -308 A/G polymorphisms are associated with peri-implantitis and may present biomarkers for peri-implantitis. CLINICAL RELEVANCE: Investigated genetic markers might serve as precious parameters in clinical practice in course of treatment planning and prognosis, since preventive and treatment approach could be positively shifted and adjusted depending on present genotype.

Estimation of bone loss biomarkers as a diagnostic tool for peri-implantitis.

BACKGROUND: The aims of this study are to estimate the profile of bone loss biomarkers in peri-implant tissues and to identify potential prognostic biomarkers of peri-implantitis. METHODS: Peri-implant crevicular fluid samples collected from 164 participants (52 patients with peri-implantitis, 54 with mucositis, and 58 with healthy peri-implant tissues) were analyzed using enzyme-linked immunosorbent assays to evaluate concentrations of the receptor activator of nuclear factor-κB (RANK), soluble RANK ligand (sRANKL), osteoprotegerin (OPG), cathepsin-K, and sclerostin. RESULTS: Concentrations of RANK, sRANKL, OPG, and sclerostin were significantly increased in patients with peri-implantitis compared with patients with healthy peri-implant tissues. Comparisons between peri-implantitis and mucositis demonstrated significantly higher values of sclerostin in peri-implantitis samples. Comparisons between mucositis and healthy peri-implant tissues showed significantly increased levels of RANK and cathepsin-K in mucositis. CONCLUSION: These results are suggestive of a role of sRANKL, OPG, and sclerostin as prognostic biomarkers in peri-implantitis.

Impact of dental implant insertion method on the peri-implant bone tissue--an experimental study.

BACKGROUND/AIM: The function of dental implants depends on their stability in bone tissue over extended period of time, i.e. on osseointegration. The process through which osseointegration is achieved depends on several factors, surgical insertion method being one of them. The aim of this study was to histopathologically compare the impact of the surgical method of implant insertion on the peri-implant bone tissue. METHODS: The experiment was performed on 9 dogs. Eight weeks following the extraction of lower premolars implants were inserted using the one-stage method on the right mandibular side and two-stage method on the left side. Three months after implantation the animals were sacrificed. Three distinct regions of bone tissue were histopathologically analyzed, the results were scored and compared. RESULTS: In the specimens of one-stage implants increased amount of collagen fibers was found in 5 specimens where tissue necrosis was also observed. Only moderate osteoblastic activity was found in 3 sections. The analysis of bone-to-implant contact region revealed statistically significantly better results regarding the amount of collagen tissue fibers for the implants inserted in the two-stage method (Wa = 59 < 66.5, alpha = 0.05), but necrosis was found in all specimens, and no osteoblastic activity. Histopathological analysis of bone-implant interface of one-stage implants revealed increased amount of collagen fibers in all specimens, moderate osteoblastic activity and neovascularization in 2 specimens. No inflammation was observed. The analysis of two-stage implants revealed a marked increase of collagen fibers in 5 specimens, inflammation and bone necrosis were found in only one specimen. There were no statistically significant differences between the two methods regarding bone-implant interface region. Histopathological analysis of bone tissue adjacent to the one-stage implant revealed moderate increase of collagen tissue in only 1 specimen, moderate increase of osteoblasts and osteocytes in 3 specimens. No necrotic tissue was found. The analyzed specimens of bone adjacent to two-stage implants revealed a moderate increase in the number of osteocytes in 3 and a marked increase in 6 specimens respectively. This difference was statistically significant (Wb = 106.5 > 105, alpha = 0.05). No necrosis and osteoblastic activity were observed. CONCLUSION: Better results were achieved by the two-stage method in bone-to-implant contact region regarding the amount of collagen tissue, while the results were identical regarding the osteoblastic activity and bone tissue necrosis. There was no difference between the methods in the bone-implant interface region. In the bone tissue adjacent to the implant the results were identical regarding the amount of collagen tissue, osteoblastic reaction and bone tissue necrosis, while better results were achieved by the two-stage method regarding the number of osteocytes.

The impact of thickness of resorbable membrane of human origin on the ossification of bone defects: a pathohistologic study.

BACKGROUND/AIM: A wide range of resorbable and non-resorbable membranes have been investigated over the last two decades. The barrier membrane protects the defect from ingrowth of soft tissue cells and allows bone progenitor cells to develop bone within a blood clot that is formed beneath the barrier membrane. The membranes are applied to reconstruct small bony defect prior to implantation, to cover dehiscences and fenestrations around dental implants. The aim of this study was to evaluate the influence of human resorbable demineralized membrane (RHDM) thickness on bone regeneration. METHODS: The experiment, approved by Ethical Committee, was performed on 6 dogs and conducted into three phases. Bone defects were created in all the 6 dogs on the left side of the mandible, 8 weeks after extraction of second, third and fourth premolars. One defect was covered with RHDM 100 micro thick, one with RHDM 200 micro thick, and the third defect left empty (control defect). The histopathological analysis was done 2, 4 and 6 months after the surgery. In the third phase samples of bone tissue were taken and subjected to histopathological analysis. RESULTS: In all the 6 dogs the defects treated with RHDM 200 micro thick showed higher level of bone regeneration in comparison with the defect treated with RHDM 100 micro thick and especially with empty defect. CONCLUSION: Our results demonstrated that the thicker membrane showed the least soft tissue ingrowths and promoted better bone formation at 6 months compared with a thinner one.

[Impact of dental implant insertion method on the periimplant epithelial seal--experimental study on dogs].

BACKGROUND/AIM: There is a controversial opinion among implantologists on the method of dental implantation that provides more favourable response of soft and bone tissues. The aim of this study was to pathohistologicaly compare the influences of one- and two-phase implant surgical methods on the periimplant epithelial tissues. METHODS: The experiment was performed on 10 dogs. Eight weeks following tooth extractions implants were inserted using one phase method on the right side of the mandible, and two-phase method on the left one. Three months after implantation the animals were sacrificed. Contact regions of epithelial and subepithelial tissues and implants were pathohistologicaly examined, the elements of analysis were scored and compared. RESULTS: Epithelial tissue was not found in the two samples of one-phase implants, while in the remainder of samples the structure of basal membrane was preserved, massive inflammatory infiltrate was found in one, and partial necrosis was found in three samples. Epithelial tissue was not found in the three samples of two-phase implants, in three samples the structure of basal membrane was intact, while in three remained samples the membrane could not be detected; moderate inflammatory infiltrate was found in four samples and massive inflammatory infiltrate in both two remained samples; tissue necrosis, which was observed in the half of the samples, was complete. In subepithelial gingival tissues around one-phase implants the number of blood vessels was increased in three samples, accompanied by the thickening and dilatation of vascular walls, proliferation of blood elements, altered vascular walls and inflammatory cell infiltrate was found in four samples. CONCLUSION: On the base of the analized characteristics of epithelial and subepithelial tissues in contact with dental implants, one-phase method of implantation showed a more favorable tissue response.

[Association between clinical parameters and the presence of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis in patients with progressive periodontal lesions].

BACKGROUND/AIM: Periodontitis is a chronic inflammatory disease of periodontal tissues with consequential is bone loss as a result of host immunological reactions caused by periopathogens. The aim of the study was to investigate if there is a correlation between clinical parameters and the presence of two most aggressive periopathogens (Aggregatibacter actinomycetemcomitans--Aa and Porphyromonas gingivalis--Pg) in patients with progressive periodontal lesions. METHODS: A total of 34 systemic healthy people, 23 to 70 years old, were included in the study. The patients were clinically and radiologically examined, and after that, the representative pocket with greatest pocket depth was chosen and the sample was collected from that place. The measured clinic parameters were: gingival index, index of gingival bleeding, pocket depth and plaque indices. The multiplex Polymerase Chain Reaction (PCR) method was used for detection of periopathogens. After obtaining results, appropriate statistical tests were used to correlate the clinical and microbiological results. RESULTS: Aa and Pg were detected in the same percentage of samples. Aa and Pg were detected in 35.29% samples alone, and in 29.41% both were detected. The values of measured clinical parameters did not show a statistical significance between the groups. In analysis of correlations among clinical parameters inside the groups, a statistical significance was found only between gingival and plaque index in the group with Aa. CONCLUSION: Clinical course of periodontitis in the developed stage does not differ in relation to the presence of different periopathogens as the major inductors of immunologically guided destructive processes.

[Mobility of B.C.T. dental implants inserted by one- and two-phase surgical method--an experimental study].

BACKGROUND/AIM: Achievement of the osseointegration of dental implants is of crucial importance for their long-term survival. One of the factors that influence the osseointegration is a surgical method of implantation. The outcome of osseointegration can be evaluated on the basis of implant mobility in bone. The aim of this study was to investigate and compare the mobility of B.C.T. dental titanium implants inserted to experimental animals using an one- and two-phase method. METHODS: The investigation was performed using a split-mouth design on nine dogs, male german shephards, average age of 3.5 years and average weight of 32 kg. Extractions of the third and fourth lower premolars were performed under intravenous (i.v.) anaesthesia with 5% ketamine chloride. Eight weeks after the extractions, the implants, diameter of 4.5 mm each, with four threads 13.7 mm long, were inserted. Eighteen implants were inserted one side of the jaw using a one-phase method, and another 18 implants were inserted contralaterally using a two-phase method. Three months after the implantation, the implant mobility was evaluated. Three measurements were performed with a Periotest device, and average values were calculated. The implant mobility was classified according to the Periotest scale in four groups of Periotest values (PTV) and compared. A total of 36 implants were inserted in 9 experimental dogs. The PTV ranged from -7.666 to + 50. RESULTS: According to the Periotest scale, 14 one-phase implants (78%) were classified into the 0 group of PTV, and 4 one-phase implants (22%) in the 3rd group. Thirteen two-phase implants (72%) were classified in the 0 group, and 5 implants in the 3rd group of Periotest scale. The difference in the average values of PTV between the two methods was 0.879 which was not statistically significant. CONCLUSION: This study showed that the method of implant insertion had no influence on the implant mobility, i.e. satisfactory osseointegration could be achieved by both methods. Further clinical parameters, as well as pathohistological and histomorphometric ones, have to be evaluated in order to assess better outcome of a particular method.

[Local application of concentrated trombocytes and the level of osseointegration of dental titanium implants: a histomorphometrical analysis].

BACKGROUND/AIM: In order to achieve better, and, thus an increased bone-implant interface, growth factors have been used over the past few years. All growth factors considered have fundamental role in the growth and development of cells and tissues. Concentrated trombocytes from platelet-rich plasma (CT-PRP) are fraction of the blood. Thrombocytes contain a number of growth factors namely PDGF, TGF-beta, IGF, VEGF and many others, which contribute to the achievement of the increased bone-implant interface, the increased stability of implants and the faster functional loading of implants. The aim of this study was to establish the effect of CT-PRP on bone-implant interface. METHODS: This experimental study included six dogs in which 24 BCT implants were inserted (4 implants per dog). On the left side of the lower jaw 2 implants were placed with CT-PRP, while on the right side the implants were placed without CT-PRP. The animals were sacrificed after 42, 70, and 98 days. The specimens were examined histomorphometrically, and analyzed 42, 70 and 98 days after the implant insertion. The contacts bone to implant in 16 zones for each analyzed implant were measured according to the established protocol. RESULTS: Results obtained with histomorphometrical analysis imply the increased bone-to-implant contact by use CT-PRP. The difference of the bone-to-implant contacts between these two groups of inserted implants has been particularly pronounced at six weeks after the implant insertion. CONCLUSION: According to the obtained results in the measurement of the level of osseointegration of the inserted implants, it should be advisable to use the CT-PRP method because it provides the higher level of osseointegration.