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Presence of Trichomonas Tenax and Entamoeba Gingivalis in Peri-Implantitis Lesions

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Presence of Trichomonas Tenax and Entamoeba Gingivalis in Peri-Implantitis Lesions IMPLANTOLOGY Presence of Trichomonas tenax and Entamoeba gingivalis in peri-implantitis lesions Osman Fatih Arpağ, PhD/Özlem Makbule Kaya, PhD Objective: The aim was to investigate the presence of Entam- Results: Although there was no presence of parasite around oeba gingivalis and Trichomonas tenaxx in peri-implantitis le- the healthy implants, two parasites were detected in peri- sions. Method and materials: A total of 141 individuals were implantitis lesions. Out of 101 lesions, 31 (30.7%) showed included in this study, of which 40 had clinically healthy im- E gingivalis, and 34 (33.6%) presented with T tenax. There was a plants (group H); the remaining were associated with peri-im- statistically significant difference between the presence of plantitis (group P). Gingival crevicular fluid was collected using E gingivalis and demographic data including gender, education absorbent paper, followed by a dental plaque sample from the status, frequency of dental visits, and brushing frequency. Pres- peri-implant sulcus/pocket using a titanium curette. The sam- ence of T tenaxx in lesions was correlated with frequency of den- ples were transferred into an Eppendorf tube. Each specimen tal visits (P < .05). It was observed that E gingivalis and T tenax was divided into two parts. One part was examined under a were mostly detected in the mandible (P = .004 and .014, re- light microscope at a 10 × and 40 × magnification to detect spectively) in comparison with the maxilla. Conclusion: This parasites. The other part was spread on a microscope slide, study showed that peri-implantitis lesions were involved with stained with Giemsa stain, and examined under a microscope E gingivalis and T tenax, in contrast to the healthy areas. at 100 × magnification. Pearson chi-square test was used in the (Quintessence Int 2020;51:212–218; doi: 10.3290/j.qi.a43948) statistical analysis of data, with a significance level of P < .05. Key words: dental implants, direct microscopy, Giemsa staining, microbiology, parasite, peri-implantitis In recent years, dental implants have been widely used in den- some of the common types of bacteria found in peri-implantitis tistry to replace missing teeth. Accumulation of microbial bio- lesions.2-4 film around the dental implant causes a breakdown of the Periodontal lesions also contain viruses, and fungal and implant-supporting tissues such as alveolar bone and gingiva. protozoa species.5 However, the effect of parasites as a part of This condition, which is called peri-implantitis, clinically pres- the pathogenesis of periodontium is still debated. Previous ents as gingival bleeding on probing and alveolar bone loss, studies have reported that two protozoans, Trichomonas tenax eventually leading to implant loss.1 Several microorganisms are and Entamoeba gingivalis, are among the types of parasites involved in the pathogenesis of peri-implantitis lesions. Some that are found in the dental plaque biofilm of patients with studies have reported that microbial plaque samples around periodontal disease.6,7 T tenax is an anaerobic motile-flagel- the healthy implant contain bacilli and cocci, and the samples lated protozoan that is classified in the same genus as Tricho- obtained from peri-implantitis lesions contain numerous fusi- monas vaginalis. T tenax plays a role in gland infections, pleuro- form bacteria, spirochetes, and motile rods. Porphyromonas pulmonary infections, sinusitis, and tonsillitis, which occur gingivalis, Prevotella intermedia, Prevotella nigrescens, Tannerella outside the oral cavity. T tenax can be transmitted through forsythia, Treponema denticola, and Fusobacterium nucleatum saliva, droplet spray, kissing, drinking water, and contaminated play a role in the progression of periodontal diseases, and are dishes. The prevalence of T tenax is correlated with demo- 212 QUINTESSENCE INTERNATIONAL | volume 51 • number 3 • March 2020 Arpağ/Kaya Table 1 Comparison of demographic data in the groups Parameter Variable Group H Group P P value* Gender, n (%) Male 14 (35.0%) 52 (51.5%) .077 Female 26 (65.0%) 49 (48.5%) Total 40 (100.0%) 101 (100.0%) Age, y (mean) Male 57.14 53.86 .594 Female 53.16 56.53 Overall 55.15 55.19 Education status, n (%) University 9 (22.5%) 16 (15.8%) .115 High school 13 (32.5%) 18 (17.8%) Primary school 10 (25.0%) 42 (41.6%) Illiterate 8 (20.0%) 25 (24.8%) Frequency of dental visit, n (%) Regular 11 (27.5%) 45 (44.6%) .062 Irregular 29 (72.5%) 56 (55.4%) Brushing frequency (daily), n (%) Twice or more 4 (10.0%) 9 (8.9%) .867 Once 24 (60.0%) 57 (56.4%) Sporadic 12 (30.0%) 35 (34.7%) n, count of individuals in same row; Group H, healthy patients; Group P, patients with peri-implantitis. *P value for significance level is < .05. graphic and systemic diseases such as diabetes and Down syn- dental implants (group H) were enrolled in the study. When a drome.8 E gingivalis is an opportunistic pathogen; it may cause patient had more than one implant, only one implant, consis- periodontal diseases in immunocompromised patients.9 More- tent with the inclusion criteria, was evaluated for that patient. over, E gingivalis has been identified in patients with advanced Patients who had used any antibiotic, antimicrobial, or anti- periodontitis, and it was detected in the dental plaque samples protozoal drugs in the previous 3 months and who had under- of disease-free individuals. Some studies have reported that gone any periodontal treatment within the last 6 months were the presence of E gingivalis in the oral cavity impairs gingival excluded. Systemically compromised patients, pregnant pa- health over time.10,11 tients, and smokers were also excluded. As previously noted, peri-implantitis is an infectious condi- The following criteria were used for inclusion: tion caused by several periodontopathic bacteria. The microbi- ■ individuals who were 18 years of age or older ota of peri-implantitis lesions was examined in previous stud- ■ for both groups, having implants loaded with a prosthetic ies.2,3 In the literature, no study investigating the presence of restoration at least 1 year before the study began parasites in peri-implantitis cases was found. Thus, the aim of ■ for group H, having implants without any peri-implant the present study was to investigate the presence of E gingivalis mucositis or peri-implantitis lesion and T tenax in peri-implantitis lesions. ■ for group P, having implants with a probing depth > 5 mm, gingival bleeding on probing, and radiographically detect- able bone loss distance > 2 mm between the implant collar Method and materials and the alveolar crest. Patient selection and study groups Sample collection Before the study began, all the ethical requirements were ful- filled (ethical approval number: 17.01.2019/08). Informed con- In the patients in both groups, the implant area was isolated sent was obtained from all the participants. In total, 101 patients with rolled cotton, and saliva was aspirated using suction. First, with peri-implantitis (group P) and 40 individuals with healthy gingival crevicular fluid was collected with an absorbent paper QUINTESSENCE INTERNATIONAL | volume 51 • number 3 • March 2020 213 IMPLANTOLOGY Table 2 Number of lesions with and without parasite in the groups Table 3 Distribution of species detected in combination or separately Group H (n = 40) Group P (n = 101) Species Count of lesions n (%) Species (+) (−) (+) (−) Entamoeba gingivalis (alone) 7 (17.1%) 0 (0.0%) 40 31 70 Entamoeba gingivalis Trichomonas tenaxx (alone) 10 (24.4%) (100.0%) (30.7%) (69.3%) Both species (together) 24 (58.5%) 0 (0.0%) 40 34 67 Trichomonas tenax (100.0%) (33.6%) (66.4%) Number of all cases detected 41 (100.0%) n, number of cases; +, presence of parasite; −, absence of parasite. n, count of lesions in which parasites were present together or alone. point that was inserted into the gingival sulcus for 10 seconds. study groups. Differences were considered to be statistically A dental plaque sample was then taken from the same site significant when P < .05. using a titanium curette.12 In group P, the deepest site of the pocket around the implant was selected for the samples. The Results samples were transferred into an Eppendorf tube filled with 0.9% NaCl solution (0.5 mL). Each sample was delivered to the No problems occurred when collecting and transferring the parasitology laboratory for examination, as quickly as possible. samples. Thus, a total of 144 individuals were evaluated for the presence of parasites. Table 1 presents the patients’ demographic data, including Microscopic determination their gender, age, education status, frequency of dental visits, The dilution sample drawn with a pipette was incubated at and brushing frequency. The distribution of the variables in the 37°C for 20 minutes. Two different methods were used to detect two groups was not statistically significant (P > .05). the parasites. In the first method, one droplet of the sample The prevalence of E gingivalis and T tenax in the two groups taken from the incubated dilution was placed on a microscope is presented in Table 2. From the incidence rate, it is clear that slide and closed with a glass cover. The specimen was exam- both parasites were associated with the peri-implantitis lesions. ined under a light microscope at 10 × magnification and 40 × E gingivalis and/or T tenax were found in almost one-third of magnification. In the second method, the specimen on a micro- the peri-implantitis cases. Forty-one out of 101 peri-implantitis scope slide was stained with a Giemsa stain for 45 minutes. lesions involved at least one of the parasites. The presence of Then, the slide was gently washed under clean water and air- both parasites in the same lesion was observed in 24 patients dried. After immersion oil was dropped onto the slide, the (58.5%) with peri-implantitis (Table 3).
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