IMMUNO-BIOCHEMICAL AND CLINICAL RESEARCHES ON THE EVOLUTION OF ANTIOXIDANTS LEVEL IN THE ETIOPATHOGENY OF PERIODONTAL PATHOLOGIES

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1 Periodontology IMMUNO-BIOCHEMICAL AND CLINICAL RESEARCHES ON THE EVOLUTION OF ANTIOXIDANTS LEVEL IN THE ETIOPATHOGENY OF PERIODONTAL PATHOLOGIES Oana Potârnichie¹, Sonia N\nescu 2, Liliana P\s\rin¹, Ioana Rudnic¹, Liliana Foia 3, Silvia Mâr]u 4 1. Assist Prof. PhD, Dept Periodontology, Faculty of Medical Dentistry Gr. T. Popa U.M.Ph., Ia[i 2. PhD Student, Dept Periodontology, Faculty of Medical Dentistry Gr. T. Popa U.M.Ph., Ia[i 3. Prof. PhD, Dept of Biochemistry, Faculty of Medical Dentistry Gr. T. Popa U.M.Ph., Ia[i 4. Prof. PhD, Dept Periodontology, Faculty of Medical Dentistry Gr. T. Popa U.M.Ph., Ia[i Corresponding author: Silvia Martu: parodontologie1@yahoo.com Abstract Introduction: A comprehensive, recent analysis on the topic reached the conclusion that oxidative stress is the main cause provoking destruction of the periodontal tissue resulting from the host-microbe interaction. Scope of the study: To investigate the correlation between periodontal health condition and the seric levels of some antioxidants. Materials and method: A representative group of 58 patients, with ages between 30 and 70 years, were examined, between , as to their periodontal condition, after which peripheric blood was taken over and retinol, -tocopherol, -tocopherol, -caroten, -caroten, -criptoxantine, zeaxantine, luteine and licopen were extracted from the serum. The values of the antioxidants were measured by high-performance chromatographic liquid technique. Results: The levels of and -caroten, -criptoxantine and zeaxantine were significantly lower in the patients with minimum periodonthopaty debut (p<0.001), as well as in those with increased intensity of periodontopathy debut. -caroten and -criptoxantine were the only antioxidants associated with a higher risk of severe periodontic pathologies. Conclusions: The low levels registered for some carotinoides, especially -caroten and -criptoxantine, are directly proportional with a higher prevalence of periodonthy installation. Keywords: oxidative stress, antioxidants, -caroten and -criptoxantine, periodontal risk INTRODUCTION Periodontopathy is an inflammatory disease, representing the response of the periodontal tissues to the action of the lipopolysaccharide of a Gramm-negative-type anaerobic bacteria. A comprehensive, recent analysis on the topic reached the conclusion that oxidative stress is the main cause provoking destruction of the periodontal tissue resulting from the host-microbe interaction. [1]. Tissue destruction results from a disturb caused by chronic inflammation, which is the consequence of phagocytes mobilization and activation. [2] The production, in excess, of cytokines, proteinases and of the reactive species (RS) contribute to chronicization of the inflammatory lesion [3,4]. SCOPE OF THE STUDY The scope of the study was to investigate the correlation between periodontal health condition and the seric levels of some antioxidants, among which carotinoides, retinol and vitamin E[4], on a group of patients selected in the Clinics of Periodontology Ia[i. MATERIALS AND METHOD A representative froup of 58 patients, with ages between 30 and 70 years, selected among those having addressed the Periodontology Clinics of the Gr.T.Popa U.M.Ph. of Ia[i, were examined and treated in Each subject included din the study, having at least 6 teeth, filled in a questionnaire and was evaluated as to his/her periodontal condition. In a subsequent stage, peripheric blood was taken over, in the Clinics of Geriatry at Parhon Hospital, the samples obtained being analyzed in the laboratories of the Faculty of Chemistry Gh. Asachi Technical University. The seric levels of soluble lipids have been determined by chromatographic techniques. 52 volume 2 issue 1 January / March 2012

2 AIMMUNO-BIOCHEMICAL AND CLINICAL RESEARCHES ON THE EVOLUTION OF ANTIOXIDANTS LEVEL IN THE ETIOPATHOGENY OF PERIODONTAL PATHOLOGIES The criterion on the basis of which these parameters were included in the present investigation was the correlation between a valid measurement of the antioxidants and a clinical periodontal examination of the patients with at least 6 teeth. Out of the 58 patients forming the group, 2.7% had lees than 6 teeth while, in 7.4% of them, no suitable measurement of antioxidants level was possible, because the blood sample had not been reliable or, in less than 1% of the cases, the device indicated erroneous results. Each of the subjects having met the imposed conditions filled in a questionnaire, providing data on their demografic and socio-economic conditions, as well as on their smoke habits. In parallels, measurements of height and weight were performed. The scope and nature of the study were explained in detail to the participants, and informed consent was obtained from their part. During clinical periodontal examination, each tooth the third molar included as well as the mezial, distal and palatine/lingual sides were measured. The clinical attachment levels were registered with periodontal Michigan probes. The parameters analyzed included: index of plaque (PI), gingival index (GI) and the loss of attachment (ILA). Data of each examination were computerized and updated in a data base. Along the whole study, the mezial and vestibular sites of each tooth were constantly registered, as selected for evaluation purposes. The frequency of attachment loss distribution on various surfaces, at different teeth, was evaluated in time. Samples of venous blood were collected after 12 hr, centrifuged within 4 hr, then freezed at a temperature of 80 C. [5]. The antioxidants should remain stable, if considering samples storing at 80 C, which should not affect the antioxidants levels determined in subsequent analyses [6]. Retinol, -tocopherol, -tocopherol, -caroten, -caroten, -criptoxantine, zeaxantine, luteine and lycopene were extracted from serum and measured by high performance liquid chromatography [7]. Analysis was standardized with seric samples of known concentrations, provided by the Faculty of Chemistry of the Gh. Asachi Technical University of Iasi. The index of bodily weight (IMC) was calculated according to the weight/stature² formula (kg/m²). The IMC values calculated between were classified according to WHO regulations (2000): normal weight IMC<25 kg/ m², supra-ponderal =25 <30kg/m² and obese =30kg/m². The smokers were divided into former smokers, active smokers and non-smokers. Diabetes was classified according to antecedents. The socio-economic status was established as high, average and low, according to the responses given to standard questions (job, place of residence, income etc.) The data corresponding to the antioxidants level were not normally distributed, so that, prior to analysis, they had been transformed into logarithms. The results were summarized by the geometrical method. The antioxidants levels of the patients were compared between those suffering and not suffering from periodontal pathologies, by means of independent samples, such as the t-test, while the ratio of geometrical measurements was employed for summarizing the comparisons made. Multivariable analysis was based on logistic regression, for obtaining an equilibrium of forces adjustable to the possible external variables. RESULTS AND DISCUSSION The present investigation was based on clinical and paraclinical examinations of the 58 patients forming the group, each having at least 6 teeth (which is a compulsory condition for being included in the experimental group), evaluated clinically, gingivo-periodontally, and subjected to measurements of the antioxidant levels (table 1). The average age of the participants was of 44.2 years, with limits ranging between 48.8 and Only 7% of them were older than 70 years. Age constituted an important criterion for excluding the hypothesis that the patients might suffer from aggressive. Most of the patients were smokers; among them, 44% are ex-smokers and 17% active smokers. International Journal of Medical Dentistry 53

3 Oana Potârnichie, Sonia N\nescu, Liliana P\s\rin, Ioana Rudnic, Liliana Foia, Silvia Mâr]u The mean IMC value was of 27.4kg/m², 21% of the men being considered as obese (IMC> 30kg/m²). A quarter of the patients (25%) suffered from with reduced intensity debut (superficial or moderate ), while a much lower ratio (8%) showed obvious signs of severe periodontopathy (severe ) (table 1). The and -caroten, -criptoxantine and zeaxantine levels were significantly lower in the patients with minimum debut of periodontopathy (p<0.001). Lower values of these carotinoides were also recorded in patients with increased intensity of. No significant differences of the luteine, licopen, - and -tocopherol or retinol levels in correlation with the were registered. In the wholly corected patterns, some inversion was recorded in the relation between the number of carotinoides ( and -caroten, criptoxantine) and the periodontopathy with minimum debut. -caroten and -criptoxantine were the only antioxidants associated with a higher risk of severe perodontitis. The additional ratio ajusted for the with high intensity debut, between the fifth lowest value and the fifth highest value of -caroten was of 4.02 (p=0.003). The ratios of the geometrical measurements show that the - and -caroten, -criptoxantine and zeaxantine levels have significantly low values (p<0.001) in the males with low intensity debut of the periodontal disease. Low values of these carotinoides were also recorded in the patients with high intensity debut of. No major differences in the levels of luteine, licopen, - and - tocopherol, or retinol were registered, versus the severity degree of the perodontitis but significantly different according or not with the presence of periodontal pathology (fig. 1-3). Figure 1. -caroten level in patients with or without periodontal pathology Table 1 Percent distribution of the evaluated cases and elements Participants Age(years), average (SD) Nr. teeth, average IMC, average Diabetes, n(%) Smoking, n(%) Never Former smokers Active smokers Material situation, n(%) High Average Low Nr=58 Incipient Nr=20 Average Nr=18 Severe Nr= (2.9) 64.6 (2.9) 64.1 (2.9) 64.5 (2.9) Grave, but not severe Nr= (2.9) 19.9 (5.6) 18.1 (5.6) 20.5 (5.4)* 15.3 (5.0) 20.3 (5.4)* 27.4 (3.5) 9 (5.5) 27.9 (3.8) 28 (8.8) 27.2 (3.3)* 41 (4.4)* 28.4 (4.1) 11 (11.5) 27.3 (3.4)* 58 (5.0)* 22 (37.9) 25 (43.1) 6 (30.0) 9 (45.0) 8 (44.4)* 8 (44.4) 6 (42.9)* 6 (42.9) 11 (18.9) 5 (25.0) 2 (11.2) 2 (14.2) 25 (44.8) 12 (24.1) 21 (31.1) 8 (40.0) 4 (20.0) 8 (40.0) 8 (44.5) 4 (22.2) 6 (33.3) 1 (16.7) 3 (50) 7 (50.0) 3 (21.4) 4 (28.6) *significant at p< volume 2 issue 1 January / March 2012

4 AIMMUNO-BIOCHEMICAL AND CLINICAL RESEARCHES ON THE EVOLUTION OF ANTIOXIDANTS LEVEL IN THE ETIOPATHOGENY OF PERIODONTAL PATHOLOGIES -caroten and -criptoxantine were the only antioxidants associated with a higher risk of severe. No obvious interaction could be established between smoking and the antioxidants, in any of the logistic patterns. Figure 2. -criptoxantine level in patients with or without periodontal diseases Figure 6. Antioxidants levels in patients with incipient superifical parodontopathy DISCUSSION Figure 3. -caroten level in patients with or without periodontal diseases Figure 4. -caroten, -criptoxantine, luteine, licopen levels in patients without periodontal pathology but risking a severe form of the disease Figure 5. -caroten, -criptoxantine, luteine, licopen levels in patients with severe periodontal pathology The major achievement of the present study was to evidence the fact that the low levels of carotinoides discovered in the subjects under analysis are associated with a higher risk of debut of superficial. The correlation between carotinoides and with low intensity debut (superficial) exceeded the adjustments made for the external variables that might affect the periodontal status. When was evaluated on two planes, as proposed in literature, [8,9] it was only -caroten and -criptoxantine that were especially associated with severe. The most important observation of the present research was the correlation established between -criptoxantine and. At bone level, -criptoxantine plays an anabolic part, an aspect not evidenced in the other carotinoides, such as luteine or licopene. [9] A close correlation also existed between - caroten and with a minimum intensity debut, as well as between -caroten and the minimum and severe, a possible explanation involving the effects of these carotinoides or, alternatively, the role they play in the modulation of the immune response. International Journal of Medical Dentistry 55

5 Oana Potârnichie, Sonia N\nescu, Liliana P\s\rin, Ioana Rudnic, Liliana Foia, Silvia Mâr]u A longitudinal study developed along 2 years showed that the persons with low - and - caroten levels (the total of carotinoides having IL-6, as an index of systemic inflammation) had a precarious health state. The reduced prevalence of severe, identified in only 8% of the subjects, results from an insufficient capacity of discovering the factors associated to the etiopathogeny of the periodontal pathology. In spite of this, the association between -criptoxantine and -caroten, on one side, and the with a severe debut is possible. Generally, the patients having registered the fifth lowest level of antioxidants were exposed to different risk factors, comparatively with those recording high levels. According to the present study, the patients having recorded lowest antioxidants levels had a higher smoking prevalence; heavy smokers register the highest values of IMC and the lowest values of SES, all being known as severe risk factors for [10,11]. The conclusion of the study is that low carotenoide levels may cause superficial. A significant association was also put into evidence between the low -caroten and - criptoxantine levels, on one had, and severe, on the other. CONCLUSIONS The present study permits the conclusion that low levels of some carotinoides, especially - caroten and -criptoxantine, are directly proportional to an increased prevalence of in such a homogeneous group of patients, with ages between approximately 40 and 70 years. The low carotinoide levels may reflect an unsuitable life and alimentary style, which does not assure periodontal equilibrium, the main causes being unfavourable socio-economic conditions, smoke habits, obesity and poor food. Last but not least, some low carotinoide levels (especially - caroten and - criptoxantine) might have benefic effects in the treatment of, which is a challege for some additional investigations. References 1. Borrell L.N., Papapanou P.N.: [2005] Analytical epidemiology of. Journal of Clinical Periodontology, 32: Chapple I.L.C., Matthews J.B.: [2007] The role of reactive oxygen and antioxidant species in periodontal tissue destruction. Periodontology 2000, 43: Akalin F.A., Baltacioglu E., Alver A., Karabulut E., [2007], Lipid peroxidation levels and total oxidant status in serum, saliva and gingival crevicular fluid in patients with chronic. Journal of Clinical Periodontology, 34: Nanescu Sonia, Ciomaga Georgeta, Martu Silvia, Racovita Stefania, Foia Liliana. [2010], Antioxidant effects of some vitamins-implications at oral level. Proceedings of the int. Congress of the Romanian Dental Association for Education, The Publishing House of the Romanian Academy, Erlinger T.P., Gualla E., Miller E.R., Stolzenberg- Solomon R., Appel L., [2001], Relationship between systemic markers of inflammation and serum betacarotene levels. Archives of Internal Medicine 161: Brock G.R., Butterworth C.J., Matthews J.B., Chapple I.L.C., [2004], Local and systemic total antioxidant capacity in and health. Journal of Clinical Periodontology 31: Tsai C.C., Chen H.S., Chen S.L., Ho Y.P., Ho K.Y., Wu Y.M., Hung C.C., [2005], Lipid peroxidation: a possible role in the induction and progression of chronic. Journal of Periodontal Research, 40: Chapple I.L.C., Brock G., Eftimiadi C., Matthews J.B., [2002], Glutathione in gingival crevicular fluid and its relation to local antioxidant capacity in periodontal health and disease. Journal of Clinical Pathology-Molecular Pathology, 55: Chapple I.L.C., Milward M.R., Dietrich T., [2007], The prevalence of inflammatory is negatively associated with serum antioxidant concentrations. Journal of Nutrition, 137: Yamaguchi M., Uchiyama S., [2004], Betacryptoxanthin stimulates bone formation and inhibits bone resorption in tissue culture în vitro. Molec. and Cel.Biochem., 258: Chew, B.P. & Park, J.S. [2004] Carotenoid action on the immune system. Journal of Nutrition 134: 257S 261S. 56 volume 2 issue 1 January / March 2012