Full Proceeding Paper EFFECT OF LIPOPOLYSACCHARIDE INDUCTION OF PORPHYROMONAS GINGIVALIS ON OSTEOCLAST AND OSTEOBLAST CELL NUMBER OF WISTAR RATS’ (RATTUS NORVEGICUS) ALVEOLAR BONE TANTIN ERMAWATI 1 , DESY FUTRI INTAN GANDINI ABDI NAGARI 2 , DEPI PRAHARANI 3 , DESI SANDRA SARI 3 1 Department of Biomedical Science, Faculty of Dentistry, University of Jember, Jember, Indonesia, 2 Faculty of Dentistry, University of Jember, Jember, Indonesia, 3 Department of Periodontology, Faculty of Dentistry, University of Jember, Jember, Indonesia Email: tantin.ermawati@gmail.com Received: 20 Jan 2019, Revised and Accepted: 25 May 2019 ABSTRACT Objective: This experimental laboratory research too serves the effect of LPS P. gingivalis induction on the osteoclast and osteoblast cell number of Wistar rats’ alveolar bone. Methods: in vivo laboratory, experimental research was conducted using posttest only control group design. The samples were 20 male Wistar rats divided into 4 groups. Groups I and II were groups injected using LPS P. gingivalis for 6 w and were decapited on the day 3 and day 7, groups III and IV were the control groups (not injected using LPS P. gingivalis) and decapited on the day 3 and day 7. Subsequently, conducting tissue preparation, staining using haematoxilin eosin, and calculating the number of osteoclasts and osteoblasts cells using a microscope (Optilab) with 400x magnification. The results of osteoblast and osteoclast cell calculation were analyzed using ANOVA and LSD one-way test. Results: Induction of LPS P. gingivalis affected the number of osteoclasts and osteoblasts cell number of Wistar rats’ alveolar bone. Conclusion: Induction of LPS P. gingivalis in Wistar rats (Rattusnorvegicus) increases osteoclast cell number and decreases osteoblast cell number. Keywords: Lipopolysaccharide, Osteoblast, Osteoclasts, Periodontitis, Porphyromonas gingivalis © 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4.0/) DOI: http://dx.doi.org/10.22159/ijap.2019.v11s4.35294 INTRODUCTION Riset Kesehatan Dasar (Riskesdas) described that prevalence of dental and oral disease increase, included periodontal diseases, from 23.2% in 2007 to 25.9% in 2013 [1]. According to the oral health services program in 2012, periodontal diseases was the second of oral diseases in Indonesia, particularly periodontitis [2, 3]. Periodontitis is inflammation disease in periodontal tissue or tooth-supporting tissue which there is loss attachment between tooth, root surface, cementum and alveolar bone [4]. Periodontitis is usually related with presence or enhancement specific pathogen bacteria, such as Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia and Bacteroides forsytus [5]. Porphyromonas gingivalis (P. gingivalis) is the dominant of oral microorganism which is able to greatly colonize in the oral cavity [6]. These bacteria are able to express several virulence factors, as well as fimbriae; lipopolysaccharide (LPS); proteinase; organic metabolite, such as butyric acid; and enzymes, such as arginine, gingipain, Collagenase, gelatinase and hyaluronidase [7]. LPS is Gram negative-endotoxin that is one of the pathogenic factors and identified as the main cause of sepsis [8]. LPS of P. gingivalis has the ability to activate host response and disturb alveolar bone remodeling. LPS stimulates macrophage producing pro- inflammatory cytokines, such as interleukin-1α (IL-1α), tumor necrosis factor-α (TNF-α) and prostaglandin E2 (PGE2), which cause alveolar bone resorption [9]. Alveolar bone resorption is a dynamic process resulting in an imbalance of osteoclast and osteoblast number, whereas the number of osteoclasts are higher than osteoblast. LPS is played a role in bone remodeling disturbance started as LPS penetrates into periradicular tissue and causes inflammation and bone resorption [9, 10]. Alveolar bone resorption studies were performed using rats as the animal model because the genome was as similar or homolog as human [11]. Likewise, in the periodontal tissue of molar area, the anatomy and histology resemble with the human. With that reasons, this recent study used rats as the animal model of the study [11, 12]. The objective of this study was to know the effect of the lipopolysaccharide of P. gingivalis to the number of osteoclast and osteoblast in alveolar bone of Wistar rats (Rattusnorvegicus). MATERIALS AND METHODS Material Ethical clearance This study was experimental laboratories (in vivo study) with the posttest only control group design. All of the procedures were approved by Research Ethic Commission of Dentistry Faculty of Gadjah Mada University, Yogyakarta No. 0748/KKEP/FKG-UGM/EC/2016. Model periodontitis Animal models (rats) were acclimatized about a week and divided into 4 groups (I=rats were injected LPS of P. gingivalis for 6 w and euthanized on the 3 rd day; II= rats were injected LPS of P. gingivalis for 6 w and euthanized on the 7 th day; III= rats were a control group without LPS of P. gingivalis injection and euthanized on the 3 rd day; IV rats were a control group without LPS of P. gingivalis injection and euthanized on the 7 th day). Every group was consisted five rats. 10 µl of 0.5 mg/ml LPS of P. gingivalis was injected in the gingival sulcus of the proximal area of the mandible first and the second molar using 30G tuberculin syringe. The injections were three times a week for 6 w [13]. After that, on the 3 rd and 7 th day the rats were euthanized using the overdose of ether per inhalation. Micro-computed tomography (Micro-CT)-based investigation The sample was positioned on a horizontally rotating holder of the Micro-CT scanning device (Bruker Micro-CT SkyScan 1173, High Energy Micro-CT, FMIPA ITB). The specimens were scanned using a source X- Ray voltage of 40 kV. The current of the source is 130 mA with exposure time of 500 ms and using a 1.0-mm aluminium filter. The sample was scanned through a 180° rotation in a rotation step of 0.2 °. During image acquisition, the 10 frames were averaged; and the scanning process took about 2 h per sample. By using the camera binning of 1×1, the produced projection images have a dimension of 2240×2240 [14]. Hematoxylin and eosin staining Then, the mandibles were removed and transversally (buccal-lingual direction) sectioned in the first and second molar area. The International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 11, Special Issue 4, 2019