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SUMMARYPeriodontitis is an infectious disease concerning

supporting tissues of the teeth. The primary etiological agentfor disease development and progression is the subgingivalbiofilm, but recently it is known that host factors may modifythe pathological process or may affect the severity and /orextent. The increasing levels of some specific pathogenicsubgingival bacteria such as Aggregatibacteractinomycetemcomitans, Porphyromonas gingivalis,Treponema denticola, Tannerella forsythia, Fusobacteriumnucleatum, Prevotella intermedia and others can result inperiodontal destruction and possibly correlate with diseaseseverity (3, 4, 5). Data from controlled studies show highprevalence of P. gingivalis, T. forsythia and Tr. denticolawhich represent the red complex (coexistence of these threespecies) in patients with moderate and severe chronicperiodontitis (3, 5, 9). Parallel investigation of probing depth(PD) and clinical attachment level (CAL) with themicrobiological testing may give a confirmation of relationbetween subgingival pathogenic bacteria and severity ofperiodontitis (1, 2, 7).

Key words: periodontal pathogens, chronicperiodontitis, clinical periodontal examination.

INTRODUCTION:Chronic periodontitis is one of the most significant

diseases that cause teeth loss. The key role of severalanaerobic bacteria in subgingival microbiota has beendiscussed in the literature (pathogenic potential of the redand the orange complexes) (4, 5, 9, 10). There is a need forfurther studies on the correlation of clinical parameters PDand CAL and the levels of key periodontal pathogenssubgingivally.

AIM:To evaluate the presence and the levels of main

periodontal pathogens in the periodontal pockets in patientswith chronic periodontitis in relation with clinicalmeasurements.

CLINICAL AND MICROBIOLOGICAL DATA INPATIENTS WITH CHRONIC PERIODONTITIS

Christina Popova1, Velichka Dosseva-Panova1, Vladimir E. Panov2,1) Department of Periodontology, Faculty of Dental Medicine, MedicalUniversity - Sofia,2) Department of Conservative dental treatment and Pediatric dental medicine,Faculty of Dental Medicine, Medical University - Varna, Bulgaria.

Journal of IMAB - Annual Proceeding (Scientific Papers) 2013, vol. 19, issue 4ISSN: 1312-773X (Online)

MATERIALS AND METHODS:In this study 20 patients with chronic periodontal

disease were evaluated based on their clinical parameters(gingival bleeding, pocket depth, clinical attachment level,recession, furcation involvement, mobility, hygiene status)and microbiological analysis. Microbial evaluation includedthe detection of three periodontopathogenic bacteria:Aggregatibacter actinomycetemcomitans, Porphyromonasgingivalis, Treponema denticola, by means of real-timepolymerase chain reaction (MIP Pharma GmbH – PET test).

RESULTS:In our study T. denticola and P. gingivalis were

established in great percent of subgingival simples (P.gingivalis - in 95,8 %; T. denticola – in 98,2%). Thesepathogens were closely associated with moderate and deepperiodontal sites but they also present in lower levels inshallow periodontal pockets (table 1 and diagram 1.). Noone of investigated periodontal sites demonstrates thepresence of A. actinomycetemcomitas.

Diagram 2 presents the results of the evaluation ofgingival bleeding on probing. The data showed high PBIaverage values in all investigated patients except one whodemonstrates average value of 0.5.

Table 1. Levels of periodontal pathogens in differentperiodontal pockets.

MOPeriodontal P. gingivalis T. denticolapocketsshallow 7,4x104 5,9x104

moderate 3,4x105 1,6x105

deep 5,3x105 7,4x105

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Diagram 1. Levels of periodontalpathogens in different periodontalpockets in graphic.

Diagram 2. Evaluation of gingivalbleeding.

Diagram 3. Clinical parameters.

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Diagram 3 allows direct comparison of three clinicalmeasurements: periodontal pocket depth, gingival bleedingand loss of attachment. There may be simultaneously viewedevaluated clinical parameters in each patient. It is evidentthe correlation between the loss of attachment and the pocketdepth and gingival bleeding in the investigated patients.

Diagram 4 and Diagram 5 present available amountsof P. gingivalis and respectively T. denticola depending tothe measured PPD. In this investigation periodontal pocketswere differentiated as shallow (<4mm), medium (4-6mm)and deep (>6mm). Periodontitis we observed were moderateand severe according to received average values of CAL.In regard with this we found a clear distinction in the levelsof the detected pathogens, that correlate to the increasingdepth of the periodontal pockets (average values) as wellas to changes in the CAL (average values) and the disease’sseverity.

DISCUSSION:The similar analyses in specialized literature are

focused on the detection of the presence of A.actinomycetemcomitas, T. denticola and P. gingivalis assignificant pathogens implicated in periodontal lesions (1,3, 5, 8, 10, and 11). Recently various authors suggest thatthese main periodontal pathogens present in supragingivalplaque as well as subgingivally in periodontitis subjects (6).

In current investigations T. denticola and P. gingivalis weredetected in subgingival plaque samples from activeperiodontal sites in high levels. Our microbiological dataincluded expression of these bacteria as counts x10n levelsand % sites colonized – prevalence. In our study greatproportions of T. denticola and P. gingivalis were observedin samples of periodontal sites in diseased individuals. Theamount of the investigated bacteria is low in the shallowpockets and they levels increase with the increasing pocketdepth. Possibly these pathogens could exist in smallproportion in shallow periodontal sites and proliferate dueto inflammatory process, and multiplicate in favorable forthem environment with enough nutrients and impairedrelations between the epithelial cells. This may to someextent explain higher levels of detected periodontalpathogens in deeper sampling sites.

Our study was limited in evaluation of only threepathogen bacteria. Our results show any presence of A.actinomycetemcomitas in all plaque samples in investigatedpatients. Based on literature data for microbial etiology ofperiodontal diseases this microorganism seems to be relatedwith aggressive and refractory periodontitis more than thechronic periodontitis. Like other investigators we attempt totrace levels of detected periodontal pathogens with severityof periodontal destruction, measured with the loss of both -attachment and bone.

Diagram 4. Distribution of P.gingivalis.

Diagram 5. Distribution of T.denticola.

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Address for correspondence:Velichka Dosseva-Panova,Department of Periodontology, Faculty of Dental Medicine, Medical University– Sofia;1, Georgi Sofiiski str., 1431 Sofia, BulgariaE-mail: vdosseva@yahoo.com,

1. Boyanova L, Setchanova L,Gergova G, Kostyanev T, Yordanov D,Popova Chr, et al. Microbiologicaldiagnosis of the severe chronicperiodontitis. J of IMAB. 2009; 15(2):89-94. [CrossRef]

2. Dosseva V, Chr. Popova.Microbial monitoring in periodontaldiseases. Review. Dental review. 2008,1:47-56. [in Bulgarian]

3. Colombo APV, Boches SK,Cotton SL, Goodson JM, Kent R,Haffajee AD, et al. Comparisons ofsubgingival microbial profiles ofrefractory periodontitis, severeperiodontitis and periodontal healthusing the human oral microbeidentification microarray. JPeriodontol. 2009; 80(9):1421-1432.[PMC] [CrossRef]

4. Dahlen G. Role of suspectedperiodontopathogens in microbiolo-gical monitoring of periodontitis. AdvDent Res. 1993 Aug;7(2):163-174.[PubMed]

REFERENCES:5. Holt SC, Emersole JL.

Porphyromonas gingivalis, Treponemadenticola and Tannerella forsythia: a“red complex” a prototipe polybacterialpathogenic consorcium in periodontitis.Periodontol 2000. 2005 Jun;38(1):72-122. [PubMed] [CrossRef]

6. Ximenez-Fyvie LA, Haffajee AD,Socransky SS. Comparison of themicrobiota of supra- and subgingivalplaque in health and periodontitis. JClin Periodontol. 2000 Sep;27(9):648-657. [Pubmed] [CrossRef]

7. Listgarten MA, Loomer PM.Microbial identification in themanagement of periodontal diseases: Asystematic review. Ann Periodontol.2003 Dec;8(1):182-192. [PubMed]

8. Sanz M, Lau L, Herrera D,Morillo JM, Silva A. Methods ofdetection of Actinobacillus actino-mycetemcomitans, Porphyromonasgingivalis and Tannerella forsythensisin periodontal microbiology, withspecial enphasis on advanced

molecular techniques: a review. J ClinPeriodontol. 2004; 31:1034-1047.

9. Socransky SS, Haffajee AD,Cugini MA, Smith C, Kent RL Jr.Microbial complexes in subgingivalplaque. J Clin Periodontol. 1998 Feb;25(2):134-144. [PubMed] [CrossRef]

10. Yang HW, Huang YF, ChouMY. Occurrence of Porphyromonasgingivalis and Tannerella fosythensis inperiodontally diseased and healthysubjects. J Periodontol. 2004 Aug;75(8):1077-1083. [PubMed][CrossRef]

11. Yano-Higuchi K, Takamatsu N,He T, Umeda M, Ishikawa I.Prevalence of Bacteroides forsythus,Porphyromonas gingivalis andActinobacillus actinomycetemcomitansinsubgingival microflora of Japanesepatients with adult and rapidlyprogressive periodontitis. J ClinPeriodontol. 2000 Aug;27(8):597-602.[PubMed] [CrossRef]

Conclusion:The results of this investigation confirm the strong

relation between the levels of T. denticola and P. gingivalisand their presence together in progressive periodontallesions, evaluated with clinical measurements. Diminutionand/or elimination the proportion of pathogen species belongto red and orange complexes as well as the total bacterialnumber subgingivally could be main goal of infection

Acknowledgement:This presentation is a part of a project Grant, financed by Medical university- Sofia, scientific investigation, contract

1/17. 07. 2012

control in periodontal treatment.The results obtained in the present study are

preliminary. The severest limitation of this study being thelow number of subjects included. Further studies are plannedto increase the number of patients and to obtain statisticallyreliable results.