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Comparison of Etoricoxib and Indomethacin for the Treatment of Experimental Periodontitis in Rats

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Comparison of Etoricoxib and Indomethacin for the Treatment of Experimental Periodontitis in Rats Brazilian Journal of Medical and Biological Research (2007) 40: 117-125 Etoricoxib in experimental periodontitis 117 ISSN 0100-879X Comparison of etoricoxib and indomethacin for the treatment of experimental periodontitis in rats M.C.F. Azoubel1, 1Departamento de Fisiologia e Farmacologia, A.M.A. Menezes1, 2Departamento de Morfologia, Faculdade de Medicina, D. Bezerra1, R.B. Oriá2, Universidade Federal do Ceará, Fortaleza, CE, Brasil R.A. Ribeiro1 and G.A.C. Brito2 Abstract Correspondence We investigated the effect of etoricoxib, a selective cyclooxygenase- Key words G.A.C. Brito 2 inhibitor, and indomethacin, a non-selective cyclooxygenase inhib- • Alveolar bone loss Departamento de Fisiologia itor, on experimental periodontitis, and compared their gastrointesti- • Inflammation e Farmacologia nal side effects. A ligature was placed around the second upper left • Periodontitis Faculdade de Medicina, UFC molars of female Wistar rats (160 to 200 g). Animals (6 per group) • Cyclooxygenase inhibitors Rua Coronel Nunes de Melo, 1127 • were treated daily with oral doses of 3 or 9 mg/kg etoricoxib, 5 mg/kg Etoricoxib 60430-270 Fortaleza, CE • Indomethacin Brasil indomethacin, or 0.2 mL saline, starting 5 days after the induction of Fax: +55-85-3366-8333 periodontitis, when bone resorption was detected, until the sacrifice E-mail: [email protected] on the 11th day. The weight and survival rate were monitored. Alveolar bone loss (ABL) was measured as the sum of distances Research supported by CNPq and between the cusp tips and the alveolar bone. The gastric mucosa was Fundação Cearense de Pesquisa e examined macroscopically and the periodontium and gastric and Cultura (FUNCAP). intestinal mucosa were examined by histopathology. The ongoing ABL was significantly inhibited (P < 0.05) by 3 and 9 mg/kg etoricoxib and by indomethacin: control = 4.08 ± 0.47 mm; etoricoxib (3 mg/kg) Received April 17, 2006 = 1.89 ± 0.26 mm; etoricoxib (9 mg/kg) = 1.02 ± 0.14 mm; indometh- Accepted October 27, 2006 acin = 0.64 ± 0.15 mm. Histopathology of periodontium showed that etoricoxib and indomethacin reduced inflammatory cell infiltration, ABL, and cementum and collagen fiber destruction. Macroscopic and histopathological analysis of gastric and intestinal mucosa demon- strated that etoricoxib induces less damage than indomethacin. Ani- mals that received indomethacin presented weight loss starting on the 7th day, and higher mortality rate (58.3%) compared to etoricoxib (0%). Treatment with etoricoxib, even starting when ABL is detected, reduces inflammation and cementum and bone resorption, with fewer gastrointestinal side effects. Introduction Although bacteria are considered to be the primary etiologic agents of periodontal dis- Periodontitis is a chronic inflammatory ease (3,4), the pathogenesis of periodontitis disease characterized by destruction of the involves host-bacterial interaction followed tooth-supporting connective tissue and ce- by release of inflammatory mediators such mentum, bone resorption, leukocyte infiltra- as eicosanoids and cytokines (5). Arachi- tion, and periodontal pocket formation (1,2). donic acid metabolites, mainly prostaglan- Braz J Med Biol Res 40(1) 2007 118 M.C.F. Azoubel et al. dins of the E series (PGE2), seem to be preventive use of NSAIDs can modify the critical mediators in the progression of peri- progression of experimental periodontitis in odontal disease. There is a large body of rats (17). A recent study has documented a evidence suggesting an association of PGE2 preventive benefit of etoricoxib (18). How- levels within the periodontal tissues and ever, the mode of treatment with etoricoxib, within the crevicular fluid with the expres- a selective COX-2 inhibitor, has been poorly sion of clinical manifestations and bone de- studied in periodontal disease models. struction due to periodontal disease (6,7). In our experimental model, we have used PGs are synthesized from cell membrane a previously unexplored delayed initiation phospholipids by action of cyclooxygenase of treatment with etoricoxib 5 days after (COX) enzymes (6,7). Three isoforms of COX experimentally induced periodontal disease have been recognized thus far (8). COX-1, the in order to mimic the conditions of normal constitutive isoform, is found in almost all dentistry practice. Therefore, the aim of the cells and has a role in normal physiological present study was to evaluate the effect of functions such as gastric cytoprotection and treatment with etoricoxib, a selective COX- vascular and renal homeostasis. In contrast, 2 inhibitor, compared with indomethacin, a COX-2 is expressed during the inflammatory non-selective COX inhibitor, on the pro- reaction and mediates cell differentiation, mi- gression of experimental periodontal dis- togenesis and specialized signal transduction ease, and to correlate the outcome with the (7,9,10). A third isoform of cyclooxygenase, potential gastric and intestinal mucosa side named COX-3, has been recently described as effects of these drugs. a variant of COX-1 which is most abundant in cerebral cortex and heart (8). Material and Methods It has been suggested that the therapeutic activities of nonsteroidal anti-inflammatory Animals drugs (NSAIDs) are derived from COX-2 blockade, whereas unwanted side effects such Female Wistar rats (160-200 g) from our as gastrointestinal ulceration and bleeding, own animal facilities were housed in tem- renal damage and platelet dysfunction are perature-controlled rooms and received wa- due to inhibition of COX-1 (11,12). Thus, a ter and food ad libitum. All experiments selective blockade of the COX-2 isoform were conducted in accordance with local should lead to the inhibition of inflammation guidelines on the welfare of experimental and pain without causing the COX-1-de- animals and with the approval of the Ethics pendent side effects in gastric tissue and Committee of Animal Research of the Fede- kidney (13,14). However, it is important to ral University of Ceará. emphasize that COX-2 inhibitors are con- traindicated for patients with or at high risk Induction of inflammatory periodontal disease of ischemic heart disease or stroke (15). Etoricoxib is believed to provide gas- A sterilized nylon (000) thread ligature trointestinal safety compared with nonselec- was placed around the cervix of the second tive NSAIDs and is rapidly and completely left upper molar of rats anesthetized with absorbed following oral administration, with 10% chloral hydrate (400 mg/kg, intraperi- a rapid onset of action (16). For this reason, toneally) (19). The ligature was knotted on we evaluated the oral administration of etori- the buccal side of the tooth, resulting in a coxib in the present study using a model of subgingival position palatally and in supra- periodontitis. gingival position buccally. The contralateral Various studies have reported that the right side was used as the unligated control Braz J Med Biol Res 40(1) 2007 Etoricoxib in experimental periodontitis 119 (20). Animals were weighed and survival obtained by taking the sum of the recordings rate was monitored daily. from the buccal tooth surface and subtract- ing the value of the right maxilla (unligated Drug treatments control) from the left, in millimeters. Al- though it is claimed that laboratory rats may Animals subjected to experimental peri- harbor ongoing baseline periodontitis (23), odontitis (see above) were divided into equal in the present study we used the contralateral groups of 6 animals each to receive an oral right maxilla as the unligated control in or- dose of 5 mg/kg indomethacin or 3 or 9 mg/ der to avoid the influence of a baseline peri- kg etoricoxib, po, starting 5 days after the odontitis on our results. induction of periodontitis and daily until sacrifice on the 11th day. Etoricoxib (Merck Histopathological analysis Sharp and Dohme, Withehouse Station, NJ, USA) was diluted in 2% DMSO (w/v) and The excised maxillae were fixed in 10% sterile saline solution. Indomethacin (Merck (v/v) buffered formalin and demineralized Sharp and Dohme) was diluted in 5% so- in a 7% (v/v) nitric acid, followed by dehy- dium bicarbonate (w/v) in sterile saline solu- dration and paraffin embedding. The speci- tion. The indomethacin (5 mg/kg) and etori- mens were stained either with hematoxylin- coxib (3 or 9 mg/kg) doses were chosen as eosin or Mallory trichrome which stain blue described elsewhere (21). the collagen fibers of alveolar bone, cemen- The results of these groups were com- tum, and periodontal ligament. Six-microme- pared to those for animals subjected to peri- ter thick sections obtained from 6 animals in odontitis that received 0.1 mL/40 g saline, each experimental group, corresponding to po (experimental periodontal disease (EPD) the area between the first and second molars group; N = 12) and also to those for a group where the ligature had been placed, were of 6 rats that received no manipulation (na- examined by light microscopy. A 0 to 3 scale ive group). The same procedure was re- was used for analysis: 0, absence of or only peated in order to obtain maxillas for histo- mild cell infiltration (the inflammatory cell pathology. infiltration is sparse and restricted to the region of the marginal gingiva), preserved Measurement of alveolar bone loss alveolar process and cementum; 1, moderate cell infiltration (the inflammatory cell infil- The animals were sacrificed on the 11th tration is present over the entire inserted day after the induction of periodontitis with gingiva), minor alveolar process resorption an overdose of ether. The maxillae were and intact cementum; 2, severe cell infiltra- excised and fixed in 10% neutral formalin. tion (the inflammatory cell infiltration is Both maxillary halves were then defleshed present in both the gingivae and the peri- and stained with 1% aqueous methylene blue odontal ligament), extensive degradation of in order to differentiate bone from teeth. The the alveolar process, and partial destruction bone loss was analyzed using a stereoscope of cementum; 3, severe cell infiltration, total loupe (4X magnification) to measure the destruction of the alveolar process and se- distance between the cusp tip and the alveo- vere destruction of the cementum (24).
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