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Volume 1 | Number 2 |August 2010

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Asia Pacific Dental Students Journal

Asia Pacific Dental Students Journal August 2010 Vol. 1 No.2

Editorial Editors-in-chief: Melissa Yap Stanley Kamadjaja

Manusript of Papers presented at Scientific Research Competition, 37th Asia Pacific Dental Students Association Congress, Japan. Poster Presentation 3. PP1: THE AGE-DEPENDENT OSTEOGENIC DIFFERENTIATIONOF ADIPOSEDERIVED STEM CELLS Tsai, Meng-Wen, Fu-Hung Tseng, Hen-Yu Liu

16. PP2: ORAL HEALTH STATUS AND ORAL HEALTH RELATED TO QUALITY OF LIFE OF DENTAL STUDENTS IN UNIVERSITI SAINS ISLAM MALAYSIA Nusaibah Mutmainnah Bt Mohamad Azmi, Mursyidatun Najihah Mohamad Nasar, Zulkarnain Sinor

28. PP3: THE EFFECT OF SOY MILK ON MANDIBLE BONE DENSITY IN RAT MODEL STUDY Devi Gunawan, Diana S. Djohan, Tamara Gladysia E, William Adi Santoso, Lee Yan Ying

Publisher Asia Pacific Dental Students Association (APDSA) Executive Committee APDSA 2009/2010 President: Rumi Sato Secretary: Jun Ai Chong Treasurer: Karen Voon Editor: Stanley Kamadjaja International Liaison Officer: Jack Chao Ji SRC Coordinator: Melissa Yap President Elect: Peerapat Kaweewongprasert Country Representatives: Korea: Koo Seung Hwan Taiwan: Meng-Hsuan Tu Singapore: Syazwan Lim Cambodia: Sok Chenh Chhean

39. PP4: TEST OF INHIBITORY POWER OF ESSENTIAL OIL EXTRACT NUTMEG SEED (MYRISTICA FRAGRANS) ON GROWTH OF STAPHYLOCOCCUS AUREUS IN ANGULAR CHEILITIS Novita Eka Lestari, Aisyah Bella Azzanjani, Ita Purnama Alwi

53. PP5 : THE ANTI-INFLAMMATORY EFFECT OF CAHEW FRUIT EXTRACT ON ARTIFICIAL EDEMA IN WISTAR RAT Tiar Rennyka, Euis Mila Savista

62. PP6: SYNTHESIZE AND CHARACTERIZE THE YTTRIUM STABILIZED ZIRCONIA (YSZ) AS DENTAL RESTORATION MATERIAL Arifialda. A

71. PP7: THE SIZES OF MAXILLARY ANTERIOR TEETH IN ASIAN AND ITS CORRELATION WITH SELECTIVE CRANIOFACIAL ANTHROPOMETRIC MEASUREMENTS Renette Gan Siok Lynn, Karthiravan Purmal, Ngeow Wei Cheong, Yeoh Oon Take

88. PP8: INJURY TO THE ORAL CAVITY EXPERIENCED BY BRASS WIND INSTRUMENT PLAYERS OF BANDUNG MARCHING BAND UNIT Randita Diany Yordian, Sayed Mohamad Ridhwan, Puput Nurani

102. PP9:BACTERICIDAL AND CYTOTOXIC EFFECTS OF Erythrina fusca LEAVES AQUADEST EXTRACT Timotius Andi Kadrianto, Nadya Saputri Halim, Melinia

113. PP10: DENTAL SANTRI SCHOOL PROGRAM AS A SOLUTION FOR INTEGRATED DENTAL HEALTH EDUCATION IN ISLAMIC BOARDING SCHOOL Aditya Mukti Setyaji, Renna Maulana Yunus, Ira Willyanti, Dhea Adittya

124. PP11: PREVALENCE OF PERIODONTITIS IN DENTAL STUDENTS IN UNIVERSITY TECHNOLOGY MARA

Azwin Assilah bte Kamaruddin, Aiman Nadiah Ahmad Tajuddin, Farah Hidayah Mohd The Asia Pacific Dental Students Journal is Fazli, Siti Sarah Nor Rizan, Maziahtul Zawani Munshi, Fouad Hussain M.H Al-Bayaty

the official scientific journal for the Asia 134. PP12: ORAL HEALTH STATUS OF CHINESE ELDERLY PEOPLE WITH Pacific Dental Students Association DEMENTIA (APDSA). Tam Hoy Suet Ailsa, Chan Yau Chuen, Cheung Wing Pan, Ho Tek Ka, Lau Chon Kit, Mak Copyright: All rights reserved. No part of Ka Man, Ng Alice, Woo Cheuk Hang Timothy this publication may be produced, stored in a retrieval system or transmitted, in any form 165. PP13: THE EFFECT OF ETHANOLIC EXTRACT OF PINEAPPLE-STEM ( ANANAS COMOSUS (L.) MERR) ON INCREASING APOPTOSIS OF A HUMAN or by any means (electronic, mechanical, photocopying, recording or otherwise, ORAL TONGUE CANCER CELL ( SP – C1 ) IN VITRO Agnes Bhakti Pratiwi, Muhammad Isa, Wisda Septiana Chandra Devi, Supriatno without the written permission of the publisher. 174. PP14: THE HEMOSTATIC EFFECT OF ARTEMISIA VULGARIS EXTRACT IN TRAUMATIC BLEEDING ON MUS MUSCULUS

© 2010 Asia Pacific Dental Students Association

Grace Angelina Samuel, Melisa Budipramana, Dian Lupita Sari, Astari Puteri, Anisha Giantini, Jessica Theresia

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PP1: THE AGE-DEPENDENT OSTEOGENIC DIFFERENTIATIONOF ADIPOSE-DERIVED STEM CELLS Tsai, Meng-Wen ,Fu-Hung Tseng, Hen-Yu Liu Objectives: Adipose tissue is an ideal source of stem cells since we can obtain the adiposederived stem cells(ADSCs) from the tissue in a large quantity through simple surgery. In vivo and in vitro studies indicate that a sub-population of ADSCs has potential to differentiate into multiple cell types, including osteoblasts. Since the differentiation capacity of stem cells is much affected by age, in this study, we want to compare the the growth kinetics and differentiation potential on ADSCs in different ages. Methods: ADSCs were isolated from adipose tissue of female SAMP8-1M and SAMP8-10M mice. First, we used flow cytometry to examine the surface marker expression of CD45, CD34, Sca-1, CD44 and CD105 to characterize the isolated cells. Furthermore, we compared the cell number and the gene expression in PCR analysis between the older(ADSC-10M) and the younger(ADSC-1M) after induction in osteogenic medium. Finally, we use Alizarin red staining to show the osteogenic potential.Results: In cell number comparison, ADSC-1M grew faster than ADSC-10M. In PCR analysis, ADSC-1M showed greater expression of osteoblast marker genes Runx2 and OPN than ADSC-10M .In the meanwhile, the Alizarin red staining showed the same resalt. Conclusions: Accordint to in vitro study, we can tell that the younger ADSCs have better osteogenic capacity.

INTRODUCTION Osteoporosis is a disorder characterized by compromised bone quality which predisposes increased risks for fractures particularly in the hips, spine, and limbs, resulted from insignificant trauma1. It is estimated over 200 million people worldwide have osteoporosis 2. The prevalence of osteoporosis is continuing to escalate with the increasingly elderly population. The major complication of osteoporosis is an increase in fragility fractures leading to morbidity, mortality, and decreased quality of life. Hence osteoporosis is also known as ”the silence killer”. The majority of current therapeutic protocols are focused on preventing excessive bone loss in osteoporotic patients. For instance, bisphosphonates have been shown effective as antiresorptive agents and approved by the FDA3. These agents exert their actions by inhibition of farnesyl diphosphate synthase, a key enzyme of the mevaloneate pathway, leading to osteoclast apoptosis 4. However, these therapies are not focused on the major factor in the pathogenesis of senile osteoporosis: the loss of functional osteoblasts during aging5-8. In our study, we hypothesized that osteoblast dysfunction and the 3

Asia Pacific Dental Students Journal

loss of osteoblasts leading to osteoporosis could potentially be prevented by administration of stem cells that have been pre-differentiated to the osteoprogenitor phenotype. Bone marrow is a common source of multipotent stem cells 9-11; however, there might be a better choice. In previous studies indicates that a sub-population of adipose-derived stem cells(ADSCs) has potential to differentiate into multiple types of cells, including osteocytes, adipocytes and chondrocytes12. In the meanwhile, the proliferation rate of ADSCs is higher than bone marrow stem cells(BMSCs), and the osteogenesis potential of ADSCs is better as well 13. Since there are more and more people undergoing liposuction surgery, surplus adipose tissue has become common medical waste. Therefore, adipose tissue is an ideal source of multipotent stem cells because of easy obtainment. Since the differentiation capacity of stem cells is much affected by age 14and ADSCs have better ability in osteogenesis than in adipocytogenesis, in this study, we compare the growth kinetics and osteogenic potential on ADSCs in different ages. Thus, whether age affects the osteogenesis of ADSCs will likely shed lights on the mechanism of osteoporosis and the development of its therapeutic tools. METHOD 1. Outline

First, we obtained ADSCs from adipose tissue of SAMP8-1M and SAMP8-10M mice. Since the differentiation and self-renewal ability are significant characteristics of stem cells, we compared the cell number/colony forming and the gene expression in PCR analysis between the older(ADSC-10M) and the younger(ADSC-1M) after inducing the cells in osteogenic medium. Finally, we use Alizarin red 4

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staining to observe the osteogenic potential. 2. ADSCs extraction

To obtain ADSCs, we have to prepare digest medium first. Standard digest medium contains collagenase type IV 5mg/5c.c., hyaluornic acid 1.7mg/5c.c. and α-MEM(Minimum Essential Medium Alpha Medium with 10% Fetal Bovine Serum and 1% PSA) 5cc filtrated through 0.22um filter. Then we sacrificed the SAMP8 1M and 10M mice, and cut off the adipose tissue around the abdominal region. Put the tissue in digest medium, cut it into small pieces, and shaking for 1 hour in 37 ℃ incubator. Next, we centrifuged it of 1500 RPM for 10 minutes, seeding the cells on the dish. 3. ADSCs cell culture It usually takes 3~5 days to achieve 90% confluency in primary culture of ADSCs. We used 0.25% trypsin-EDTA to "digest" the proteins that facilitate adhesion to the container and between cells, and we could then seed the cells on a new dish. This process is done to permit subculture of the cells to a new container, observation for experimentation, or reduction of the degree of confluency in the dish. 4. Cytochemistry Staining

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Asia Pacific Dental Students Journal

1.0 Alizarin Red S Staining Alizarin Red ,may be used to identify calcium in tissue sections. Calcium forms an Alizarin Red S-calcium complex in a chelation process, in which the end product would be red. First we used 10% formaldehyde to fix the cells and applied 2% Alizarin Red staining for 15minutes. For quantification, we added 10% cetylpyridnium chloride with 8mM Na 2HPO4 and 1.5mM KH2PO4 to separate out the red color. 2.0 Oil Red O Staining Oil Red O is a lysochrome (fat-soluble dye) diazo dye used for staining of neutral triglycerides and lipids. First we used 10% formaldehyde to fix the cells. Then we mixed Oil Red O 0.4g and isopropanol 80ml, diluting the stock with ddH2O. Finally we added the working solution to stain the cell for 15 minutes so we could observe the results. 5. Osteogenic Medium Preparation To prpare osteogenic medium, we had to prepare α-MEM with 10% FBS and 1% PSA first. Then we added 0.1uM dexamethasone, β-glycerol phosphate and 50uM ascrobate into α-MEM. 6. mRNA Extraction After we trypsinized the ADSCs, we centrifuged the cells over 1500 RPM for 5 minutes. Use 1ml TRIzol to lysis the cells. Then we added 0.2ml chloroform and shook it for 15 seconds. 3 minutes later, we centrifuged it over 4℃, 12000 RPM for 15 minutes. After centrifuged, the solution would be 6

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separated into 3 layers. The upper layer contained RNA and should be removed into a new eppendorf. Then we added Isopropanol of the same volume into the eppendorf, mixing well. We put the eppendorf in -20℃ for 15 minutes, and centrifuged over 12000 RPM, 4℃ for 15 minutes to get the white pellet. After removing the upper solution, we added 1 ml 75% ETOH. Then, we centrifuged it again and removed the upper solution. Finally, we added 20 ul DEPC-ddH2O and mixed well. 7. Reverse transcription(RT) The mRNA which we extracted would turn into complementary DNA (cDNA) through reverse transcription. RT materials comprised SuperscripTM III, Oligo dT primer, 10mM dNTP mix, 10X RT buffer, 25mM MgCl2, 0.1M DTT, RNase inhibitor and 4 ug mRNA. The total volume was 20 ul and the reaction took place in PCR machine. 8. Polymerase Chain Reaction(PCR) After RT, we did the PCR to enlarge the DNA fragment of our choosing. We added cDNA into ddH2O, 2.5mM dNTP, 25mM MgCl2, upstream/downstream primer and Taq polymerase. The PCR reaction would take place in PCR machine and go through 35 cycles of denaturation, annealing and extension. 9. Agarose gel preparation We added 0.5g Agarose powder into 50ml TAE buffer, heating with a microwave oven for 4 minutes. After mixed well, the gel was then poured into a mold. Finally, we inserted a comb and the gel could be used after cooled. RESULTS

I.

The Cell Culture of ADSCs

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Asia Pacific Dental Students Journal

Fig.1 As Fig.1 shows above, ADSCs-1M were spindle-like and posses a more compact morphology. On the other hand, ADSCs-10M were comparably larger and were more spread out. II.

Define the isolation cells on stem cell characterization 12. The Self-renewal ability of ADSCs 1.1.

Colony forming Unit

Fig.2 We seeded ADSCs-1M and ADSCs-10M in two dishes. 14 days later, we used crystal violet staining to assist in the observation of the number of cell colony. As Fig.2 shows above, the younger 8

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ADSCs formed about 150 colonies. In comparison, the older ADSCs formed only about 40 colonies under the same condition. This result empirically demonstrates that the self-renewal ability gradually gets worse as aging progresses. 1.2.

Cell Proliferation

Fig.3 We counted the cell number on the 1 st,3rd,5th,7thand 9th day and drew the growth curves of ADSCs-1M and ADSCs-10M. As Fig.3 shows above, the younger cells grew faster and the doubling time was about 5 hours less than the older ones. 2. The Differentiation Potential of ADSCs

Fig.4

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Asia Pacific Dental Students Journal

Mesenchymal stem cells have the potential to differentiate into multiple types of cells, including osteocytes and adipocytes. We seeded the ADSCs-1M and ADSCs-10M in the induction medium to compare their differentiation potential. As Fig.4 shows above, both 1M and 10M cells showed the tendency towards osteogenesis and adipogenesis. III.

Semi-quantitative PCR Analysis

Fig.5

Fig.6

*P19 years was chosen. Other inclusion criteria were dentate which defined as having at least one natural root or tooth in the mouth (MOH, 2004). As for exclusion criteria, we exclude those who have been certified by medical doctor of having systemic diseases. Pregnancy can play a role in periodontal disease development; hence, pregnant mothers were also excluded. Patients on long term medication were also excluded to avoid influence on saliva quality and also development of periodontal disease and dental caries. Patients who had undergone invasive periodontal surgical procedure were also excluded as this may affect the measurement for periodontal status. Sample size calculation Since all dental students available during this study period were taken, therefore no sample size was calculated. Data collection procedure This study started with a session of giving self-administered GOHAI questionnaire to the respondents. This procedure was followed by oral examination of the respondents to determine oral health status, and was carried out in the dental treatment room. Oral health examinations were carried out on a dental chair using mouth mirrors, probe number 9, CPI probe number 621 and under attached dental light. The data were recorded into data form by a trained dental assistant. 18

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Research tools Questionnaire In this study, we used 12 item GOHAI questionnaires. Subjects were asked about three dimensions of oral health related quality of life which are physical function including eating, speech and swallowing. Secondly about psychosocial function including worry or concern about oral health, dissatisfaction with appearance, self-consciousness about oral health and avoidance of social contacts because of oral problems and thirdly is about pain or discomfort including the use of medication to relieve pain or discomfort from the mouth. In addition, the questionnaire also included socio-demographic characteristics such as age, sex, medical problem and orthodontic appliance. Clinical examination Caries measurement The diagnosis of dental caries was based on the WHO criteria (WHO, 1998) using DMFT index. Dental caries is defined as cavitated lesion found on visual observation. The examinations involve inspection of the occlusal, facial, distal, lingual and mesial surfaces. A visual examination is carried out using a mirror and is aided by an air syringe to dry a tooth surface. An explorer was used in caries diagnosis as a tool to remove plaque and debris and check the surface characteristics of suspected carious lesions. This explorer was handled with light pressure. The score was taken as the dependent or outcome variable for the study where the score was range from 0 – 28 score. The exclusively visual criteria of caries recorded as D, missing of tooth from arch recorded as M, and tooth surfaces fill with any restorative material recorded as F. The total accumulated score of decayed, missing and filled was taken as the dependent variable for analysis. Periodontal disease measurement Basic periodontal examination (BPE) index was used in this study. The standardized periodontal probe with light pressure was used to examine the tissue for bleeding, plaque retentive factors and pocket depth. The score was pen down according to standardized code, as follow: Code 0

No bleeding or pocketing detected

1

Bleeding on probing - no pocketing > 3.5mm 19

Asia Pacific Dental Students Journal

2

Plaque retentive factors present - no pocketing > 3.5mm

3

Pockets > 3.5mm but 5.5 mm in depth

Statistical analysis Data was entered and analyzed using Statistical Package for Social Sciences (SPSS, version 17.0) software. Descriptive statistic was presented in form of means and SD and for categorical variables frequency and percentage. To test the relationship between variables, Pearson coefficient correlation was used. Level of significant was set at 0.05. RESULT Table 1: Descriptive statistics variables understudy (n=62)* Characteristics

Mean (SD)

Freq (%)

Sex Male Female Overall Male Female

Age

18 (29.0) 44 (71.0) 19.7 (0.85) 19.4 (0.85) 19.7 (0.84)

Past Medical History No Yes

57(91.9) 5 (8.1)

No Yes

60 (96.8) 2 (3.2)

Healthy BOD Calculus Deep Pocket

5 (8.1) 12 (19.4) 44 (71.0) 1 (1.6)

Good Fair Poor Overall Male Female

56 (90.3) 5 (8.1) 1 (1.6)

Wearing Orthodontic appliance BPE score

Oral Hygiene

DMF Score

3.7 (3.42) 2.1 (2.07) 4.4 (3.42)

*

all respondent are Malay 20

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Table 2: Prevalence of Periodontal Disease and dental Caries among Respondents (n=62) _______________________________________ Variable freq (%)_______________ Dental caries Yes 33 (53.2) No 29 (46.8) Periodontal disease Yes No

17 (27.4) 45 (72.6)_______________

Table 3: Oral Health Quality of Life Score (n=62) _________________________________________________ Variable mean (SD)__________ Total GOHAI Score (12 item) 11.1 (5.50) GOHAI item 1 0.9 (1.11) GOHAI item 2 0.8 (0.70) GOHAI item 3 0.4 (0.64) GOHAI item 4 0.4 (0.71) GOHAI item 5 0.6 (0.88) GOHAI item 6 0.4 (0.72) GOHAI item 7 1.3 (1.07) GOHAI item 8 0.9 (0.91) GOHAI item 9 1.9 (0.98) GOHAI item 10 1.2 (1.06) GOHAI item 11 0.7 (0.83) GOHAI item 12 1.5 (0.84)__________ Table 4: Relationship between oral health status and Oral Health related Quality of Life score (GOHAI Score) (n=62) ________________________________________________________ Variables b1 coeff.(95% CI) p-value___ DMF

0.73 (0.35, 1.10)

0.05). The percentage inhibition of inflammation will be higher when the AUC showed a low value (Table II). Extract of cashew at concentration 2.5% and 7.5% inhibited the inflammation by 61.3% and 64.78% (Figure II). Extract of cashew at concentration 5% also exhibited the inflammation inhibition although less than 50%. Table II. The average AUC of edema volume after 7 hours and the percentage inhibition of inflammation for each group

Treatment Control Extract of cashew Extract of cashew Extract of cashew Diclofenac sodium

Dose/

AUC1-7

% Inhibition of

Concentration 10 ml/kg WB 2.5% 5% 7.5% 40 mg/kg WB

( Mean ± SD) Score 0,602 ± 0,123 0,233 ± 0,036* 0,358 ± 0,099* 0,212 ± 0,102* 0,208 ± 0,058*

Inflammation 61,3 40,53 64,78 65,45

Values are mean ± SD; n = 3; significantly difference from control group *p < 0.01 One-way ANOVA followed by LSD tests

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Asia Pacific Dental Students Journal

Figure II. Anti-inflammatory effect of the extract of cashew fruits on carrageenaninduced rat paw edema in rats DISCUSSION Carrageenan-induced paw edema is a suitable experimental animal model for evaluating an antiedematous effect. Edema developed following injection of carrageenan serves as an index of acute inflammatory changes, was and can be determined from differences in the paw volume measured immediately after carrageenan injection and then every hour for 7 hours.8 Carrageenan is the phlogistic agent of choice for testing antiinflammatory drugs as it is not known to be antigenic and is devoid of apparent systemic effects.9 In the carrageenan-induced paw edema test the development of edema (inflammatory response) is a biphasic event with a maintence phase in between (2-3 hours): initial non phagocytic exudative inflammatory phase lasting up to 2 hours and a delayed phagocytic inflammatory phase from 3-5 hours. The initial phase is primarily mediated by histamine, serotonin and increase in prostaglandin synthesis in the surroundings of the damaged tissue while the late phase is mediated by leukotrienes, mobilized phagocytic cells, polymorphonuclear cells, monocytes, macrophages, prostaglandins produced by tissue macrophages, oxygen free radicals, nitric oxide, proteolytic enzymes and platelet activating factor.10 In this study, the extracts did not 58

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show a significant effect in edema formation at the early phase but showed significant effect at the later phases after 3 hours. The inhibitory activity shown by the extract of cashew (2.5% and 7.5%) over a period of 7 hr in carrageenan-induced paw inflammation was quite similar to that observed in the group treated with diclofenac sodium. This anti-inflammatory effect of the extract observed might be due to the presence of tannin and flavonoid in the plant. According to Jeffers (2006)11, the role of tannin as anti-inflammatory is to inhibit the production of prostaglandin E2 (PGE2), whereas flavonoids inhibit the lipoxygenase and cyclooxygenase pathways of arachidonate metabolism.12 Another anti-inflammatory property of flavonoid was also demonstrated by its ability to inhibit neutrophils degranulation. This is a direct way to diminish the release arachidonic by neutrophils and other immune cells.13 Therefore, it can be assumed that the inhibitory effect of the extract of cashew on carrageenan-induced inflammation could be due to the inhibition of the enzyme cyclooxygenase, leading to the inhibition of prostaglandin synthesis by tannin and flavonoids. Cyclooxygenase (COX) is an enzyme needed for the conversion of arachidonic acid into prostaglandin. This enzyme has two isoforms of COX-I and COX-II. COX-I represents a key enzyme in many cells and tissues, whereas COX-II was induced by inflammatory stimuli so that the enzyme is not found in normal cells.14 Extract of cashew at concentration 2.5% and 7.5% exhibited the inhibitory activity more than 50%, whereas at concentration 5% less than 50%. The difference in the concentration of the extract is suspected in affecting the effectiveness of each antiinflammatory compounds that also indirectly affect the anti-inflammatory effect of cashew extract. The highest percentage inhibition of inflammation was exhibited by cashew extract at concentration 7.5%. This might be due to the higher concentration of an extract, the higher active substances contained therein, thus more anti-inflammatory substances and resulted better anti-inflammatory effect. In contrast, the inhibitory activity extract of cashew at concentration 2.5% was better compared to concentration 5%. This might be because of substance or compound that only produce a response at low 59

Asia Pacific Dental Students Journal

concentrations, so that if the concentration is increased, the compound will produce a constant response or opposite response (aggravate inflammation). Variance response mechanism of the body (biological variation) experimental animals also suspected of affecting the result. According to Katzung (1997),15 the relationship between drug dose (concentration of the extract) and the clinical response can be quite complex, because the response of each individual/animal to a drug can vary widely. These responses can be caused by genetic differences in drug metabolism or immunological mechanisms. ACKNOWLEDGEMENTS The authors would like to thank Dr. drg. Juni Handajani, M. Kes and drg. Heni Susilowati, M. Kes, Ph.D from department of oral biology of Gadjah Mada University for their advice and input to this study. CONCLUSION This study concluded that extract of cashew at concentration 2.5% and 7.5% has anti-inflammatory properties on carrageenan-induced rat paw edema model. The presence of one or more phytochemical constituents present in the extract further study is warranted, for isolation of the constituents responsible for the activity and also to explore the exact mechanism of action of the activity. REFERENCES 1.

Olajide OA, Aderogba MA, Adedapo ADA, Makinde JM. Effects of Anacardium occidentale stem bark extract on in vivo inflammatory models. J Ethno pharmacol 2004; 95, 139-142

2.

Tedong L, Dzeufiet PDD, Dimo T, Asongalem EA, Sokeng SN, Flejou J, Callard P, Kamtchouing P. Acute And Subchronic Toxicity Of Anacardium

Occidentale Linn (Anacardiaceae) Leaves Hexane Extract In Mice, Afr. J.

Traditional 2007; 4 (2): 140-147. Available at: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2816447/pdf/AJT0402-0140.pdf. Accessed Aug 2, 2010

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Pereira JV, Sampaio FC, Pereira M, Melo AFM, Higino JS, Carvalho A. In vitro Antimicrobial Activity of an Extract from Anacardium occidentale Linn. On Streptococcus mitis, Streptococcus mutans and Streptococcus sanguis. Odontolgia. Clin. Cientif 2006; 5 (2): 137-141

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Raharni, Mutiatikum D, Saroni. Uji Antiiflamsi

Ekstrak Buah Semu Jambu Mede

(Anacardium occidentale L) (Abstr.), Available at:http://www.litbang.depkes.go.id/risbinkes/Buku %20laporan%20penelitian%201997-2006/26-uji_antiiflamsi_ekstrak_buah_sem.htm. Accessed May 6, 2009. 5.

Syawalia ZA, Fadlilah R, Rennyka T, Werdiningsih NE, Ningsih JR. Pemberian Topikal Ekstrak Buah Jambu mete (Anacardium occidentale L) Konsentrasi 2% pada Epitel mukosa Rongga Mulut Wanita Penderita Recurrent Apthous Stomatitis, Laporan Penelitian Program Kreativitas Mahasiswa, Universitas Gadjah Mada, Yogyakarta, 2009. [Indonesian]

6.

Mansjoer S. Efek Antiradang Minyak Atsiri Temu Putih (Curcuma Zedoria Rosc.) Terhadap Udem Buatan Pada Tikus Putih Betina Galur Wistar. Majalah Farmasi Indonesia 1997; 8: 35-41. [Indonesian]

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Hapsari HD, Handajani J, Tandelilin RTC. Efektivitas Ekstrak Etanol Buah Mengkudu sebagai Bahan Antiinflamasi pada Tikus Wistar. Majalah Ilmiah Kedokteran Gigi 2006; 21(2): 60-68. [Indonesian]

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Buadonpri W, Wichitnithad W, Rojsitthisak P, and Towiwat P. Synthetic Curcumin Inhibits Carrageenan-Induced Paw Edema in Rats. J Health Res 2009; 23(1): 11-16.

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Sudjarwo SA. The Potency of Piperine as Antiinflammatory and Analgesic, Folia Medica Indonesiana 2005; 41(3): 190194.

10. Ratnasooriya WD and Fernando TSP. Anti-inflammatory Activity of Sri Lankan Black Tea (Camellia sinensis L.) in rats, Phcog Res [serial online] 2009; 1:11-20. Available at: http://www.phcogres.com/text.asp?2009/1/1/11/58142. Accessed Aug 2, 2010. 11.

Jeffers MD. Tannins As Anti-Innflammatory Agents. Thesis. Miami University Oxford: Ohio, 2006.

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Ebadi M. Pharmacodynamic Basis of Herbal Medicine. CRC Press LLC: Florida, 2002: 393-395.

13. Nijveldt RJ, Nood EV, Hoorn DECV, Boelens PG, Norren KV, Leeuwen PAMV. Flavonoids: a review of probable mechanisms of action and potential applications [online]. Available at: http://www.39kf.com/cooperate/qk/AmericanSociety-for Nutrition/017404/2008-12-28-550119.shtml. Accessed March 5, 2010. 14.

Cheng Z, Nolan AM, and Mc Kellar QA. Measurement of Cyclooxygenase Inhibition in vivo: A Study of Two NonSteroidal Anti-Inflammatory Drugs in Sheep. Inflammation 1998; 22(4): 353-366.

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Katzung BG. Farmakologi Dasar dan Klinik. 6th ed. EGC: Jakarta, 1997: 564-565.

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PP6: SYNTHESIZE AND CHARACTERIZE THE YTTRIUM STABILIZED ZIRCONIA (YSZ) AS DENTAL RESTORATION MATERIAL Arifialda. A Objectives:To get the nanoparticle size from Yttrium Stabilized Zirconia (YSZ) on sol-gel method as new bioceramics in dental restoration material.Methods:The precursor concentration (Zirconium Chloride/ZrCl 4) that used in this research was 0.05 M and 0.01 M which allow the retention of tetragonal structure and efficiently arrest crack propagation. 3% and 8% of Yttrium Nitrate/ Y(NO3) added as stabilizer despite use pulp acasia as template. This research involved the using of Scanning Electron Microscope (S.E.M) type JSM – 6360 to characterize the visual grain size (particle size) also the morphology of particle and X-ray Diffraction (XRD) to characterize the cristal structure (structure phase) of Zirconia. Results:There are differences on visual grain size between samples with precursor concentration 0.05 M and 0.01 M, which grain 0,05 M has smaller particle size (14.2 nanometer) than 0,1 M (17.7 nanometer). The crystal structure that shaped are tetragonal and monoclinic. The colour result is white opaque.Conclusion:It could be concluded from S.E.M result table that the particle size depends on precursor concentration. As small as precursor concentration, the particle size becomes smaller. The YSZ has greatly stable tetragonal structure because of small particle size which is able to reduct the free energy where can be forming. The good characteristic of YSZ is shape memory ceramics where can be used to apply in posterior teeth with good mechanical characteristic.

INTRODUCTION As a current Indonesian mineral resource, zircon is very potential to be a basic material of synthesis of partially stabilized zirconia (PSZ) that constitutes an important component in advanced ceramics. In manufacture industry, we can find zirconia extensively as a basic material in tile production and refractory material to be a layer of melting furnace, kiln furnace, nozzles, crucible, as a component of sensory ceramics and SOFC, healthy application (especially as heads for hydroxyapatite), as well as jewelries. The variousness of the application of zirconia (ZrO2) is absolutely related to its specific

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characteristics and its ability; for instance, it has a very high refractoriness, namely, about 2750° C for pure zirconia, its easiness to phase transform to result in mechanical characteristics intended, its good ionic conductance as well as its easiness to be stabilized by the other metal oxide to modify the physical, mechanical and chemical characteristics. The existence of ZrSiO4 in Indonesia has been recognized since many years ago. It can be found in water logged area Bangka-Belitung as alluvial sediment a long with tin sand and other mineral resources. Furthermore, zircon is also existed along the watershed of rural region in Central Kalimantan along with alluvial sediment of gold. Until the beginning of year 2000, zircon from Bangka-Belitung was still considered as residue of ore that did not have much attention but had given the good result, until now the use of zirconia with high quality still depends on import products, where as the existence of zircon (ZrSiO4) as a main source of zirconia has a big reserve potency. Zircon sand (ZrSiO4) that is existed in an abundant amount in South Kalimantan presently has not been used optimally as we can see in the mineral map distribution below.

This big potency of natural resource has not been used optimally to produce the products that have value added and high utility and therefore have a certain up sell. Because the abundance of zircon sand in Indonesia, it is important to develop the

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function of zirconia in order to increase the value added of Indonesian mineral resources both in bioceramics and especially in dental material. This condition is appropriate with the research of the success of zirconia applications in orthopedic or implant that encourages the practioners of dentistry to explore the possibility of the use of material fulfill aesthetic aspects as support material (alloy substitute) or, in other words, if it is viewed from its mechanical aspect, it has resemblance to metal alloy. Nowadays, the need of the use of zirconia is significantly increasing along with global era that emphasizes the aesthetic aspects in one side and has superiority in mechanical aspects in another side. Traditionally, alumina has been used as implant or as addition into dentifrice. Nevertheless, the experts of dentistry prefer to develop zirconia compared to alumina because zirconia is stronger than alumina and the other bioceramics besides it has a good resistance to fracture. Zirconia substitutes gold or stainless steel that has traditionally been used in dentistry as a tooth mantle because it is more translucent and transparent to x-ray. In recent years, zirconia has been used in cosmetics dentistry for the purpose of aesthetic application. In addition, it is extensively selected as the application of dental material. There are many researches to renew zirconia since the invention of transformation toughening capabilities of zirconia in the middle of 1970. Zirconia is one of bioceramics that has been introduced intensively in prosthetics to be used as the material of crown, bridge or fixed partial denture because of its characteristic that is inert to body liquid and aesthetic. Bioceramics as dental material must fulfill the following requirements (1) has compression strength (2) wear and corrosion resistance (3) can be polished (4) bioactive or inert, appropriate with the indication (5) fatigue resistance (6) easy to sterilize (7) easy to fabrication, and (8) shape memory where the material is able to return to the initial form after getting the pressure. From biomaterial aspect zirconia constitutes a selected material because it fulfills the requirements of protheses, namely, it must be bioactive, bionert and biocompatible where it is able to adopt and integrate well with minimal adverse reaction. There are some types of zirconia, that is; Tetragonal Zirconia Polycrystals (TZP), Fully Stabilized Zirconia (FSZ) and Partially Stabilized Zirconia (PSZ) that constitutes a 64

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type of zirconia used as dental material. As dental material, zirconia has good characteristics with physical, mechanical, chemical and biological characteristics. From the literature aspect, zirconia described as bioceramics material partially stabilized. Accordingly, it needs the addition of single oxide in order to be absolutely stable in room temperature by adding Yttrium Oxide (Y2O3), Calcium Oxide (CaO) or Magnesium Oxide (MgO). The selected oxide in this research is Yttrium Oxide (Y 2O3), that is able to retain the tetragonal structure and retain it, in order not to be monoclinic structure. In the field of dental material, Yttrium Stabilized Zirconia (YSZ) is very potential to develop, because it is stable both chemically and biologically besides it is very strong mechanically with the toughness that reaches 1000 Mpa. METHODS The method of synthesis of Yttrium stabilized zirconia (YS2) that will be used in this research is the method of sol gel with the purpose of obtaining nonparticles of YSZ. It is important to process this material to be nanoparticles because it is smaller size of the particle, the more active it is, then the genuine characteristics will be emerged, namely, electrical, magnetical and mechanical characteristics. In addition, the process will be more efficient because as smaller as particles, the process of temperature used can be reduced. The method of sol gel is a process of producing the advanced ceramics constituing bottom-up process that involves the formation of colloid (sol) and the formation of polymer chains (gelation) from the sol in the medium (gel). In the process of producing zirconia, precursor used is zirconium chloride (ZrCl 4). In this research, the process of gelation will be substituted by the pulp of paper as template. Process of Producing Process of producing zirconia involves two steps, as follow: 8.0 Hydrolysis The reaction of cutting metal tie by water that, then, forms a compound of hydroxide 65

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metal. Si(OR)4 + H2O  HO-Si(OR)3 + ROH 9.0 Condensation The reaction polymerization hydroxide metal to form the bigger particle or molecule. (OR)3Si-OH + HO-Si(OR)3 (OR)3Si-O-Si(OR)3 + H2O or (OR)3Si-OH + HO-Si(OR)3 (OR)3Si-O-Si(OR)3 + ROH The Processing Methods of The Yttrium Stabilized Zirconia Grain AQUA BIDESTILATA

Precursor ZrCl4

Mixing and Aging for 2 days

Adding and mixing with pulp acasia

Stirring continuing methods

Y(NO3)3.6H2O

Heating on temperatures 90-100oC (1 days)

Heating on temperatures 1652oF/900oC in order to get nanoparticles YSZ form 66

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Get specimen with 3 point bending methods

Characterizing (XRD, SEM)

Bending test The precursor concentration (Zirconium Chloride/ ZrCl4) that used in this research was 0.05 M and 0.01 M which allow the retention of tetragonal structure and efficiently arrest crack propagation. 3% and 8% of Yttrium Nitrate/ Y(NO 3) added as stabilizer despite use pulp acasia as template. This research involved the using of Scanning Electron Microscope (S.E.M) type JSM – 6360 to characterize the visual grain size (particle size) also the

morphology of particle and X-ray Diffraction (XRD) to

characterize the cristal structure (structure phase) of Zirconia.

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Picture 1 SEM Result 20000X ZrCl4 0,1 M (17,7 nanometer)

Picture 2 XRD Result of ZrCl4 0,1 M

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Picture 3 SEM Result 20000X ZrCl4 0,05 M (14,2 nanometer)

Picture 4 XRD Result of ZrCl4 0,05 M

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RESULTS There are differences on visual grain size between samples with precursor concentration 0.05 M and 0.1 M, which grain 0,05 M has smaller particle size (14.2 nanometer) than 0,1 M (17.7 nanometer). The crystal structure that shaped are tetragonal and monoclinic. The colour result is white opaque. CONCLUSION It could be concluded from S.E.M result table that the particle size depends on precursor concentration. As small as precursor concentration, the particle size becomes smaller. The YSZ has greatly stable tetragonal structure because of small particle size which is able to reduct the free energy where can be forming. The good characteristic of YSZ is shape memory ceramics where can be used to apply in posterior teeth with good mechanical characteristic. REFERENCE 1.

Park, J.B., Bronzino, J.D. 2003. Biomaterials: Principles and Applications. New York: CRC Press. 38, 41-42

2. Black, J., Hastings, G. 1998. Handbook of Biomaterial Properties. London: Chapmann & Hall. 340, 343.

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PP7: THE SIZES OF MAXILLARY ANTERIOR TEETH IN ASIAN AND ITS CORRELATION WITH SELECTIVE CRANIOFACIAL ANTHROPOMETRIC MEASUREMENTS Renette Gan Siok Lynn, Karthiravan Purmal, Ngeow Wei Cheong, Yeoh Oon Take Selecting appropriate anterior teeth size becomes difficult when there are no preextraction records of the missing natural teeth. Objective: This study determines the sizes of maxillary anterior teeth in Asian, and investigates if selective craniofacial anthropometric measurements could be used to predict anterior teeth sizes. Material & Methods: This study was conducted on a group of Asian subjects (mean age: 22.3 + 1.7 years; range 19-25 years) with Class I occlusion, where direct measurements were made of the anterior maxillary teeth on stone casts. Selective craniofacial anthropometric measurements were correlated to the sizes of their anterior teeth. Results: The mesiodistal diameter of the maxillary central incisor, lateral incisor and canine were 8.67 + 0.64 mm, 7.00 + 0.65 mm and 7.91 + 0.71 mm respectively. The anterior arch width, as represented by the inter-canine cusp tip distance was 34.87 + 2.19 mm. Sex differences in tooth sizes were seen in all anterior teeth. However, there were no racial differences between 3 major races (Malay, Chinese and Indian) recruited. The intercanine distance was similar to that reported for Singaporean Chinese, and this was larger than Caucasian’s. The height, and facial, mandibular and mouth widths of the subjects were significantly greater in males. The mandibular width showed significant correlation with all anterior teeth concerned. Conclusion: The maxillary anterior teeth of men were greater than those of women in the Asian population studied. The lateral incisor and canine of Asian males were larger than that reported for other races. The anterior arch width was similar to that reported for Singaporean Chinese.

INTRODUCTION Ideal selection and arrangement of anterior teeth in prosthodontics can be challenging in the absence of pre-extraction records. In this aspect, the mesio distal width of the anterior teeth is harder to determine compared to its height (also called tooth length). 1-3 The height of the anterior teeth can usually be determined according to the length of lip at smiling and rest position.4

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Although several authors have attempted to use biometric guide as an aid in the positioning of the anterior teeth, there is still a lack of agreement in relating successive width of the anterior teeth.3

5 6

This can be attributed to the fact that tooth size and

antropometric measure are race and gender specific.7 Table 1 provides a summary of the size of the central incisor in various races. Lombardi8 indicated that there is a recurring ratio in the width of successive teeth from the central incisor. Lombardi also mentioned that central incisor should be larger than the lateral incisor to dominate the composition and bring order and coherence. Levin 9 and others10

11

expanded this concept to come up with proportions that was deemed

aesthetically pleasing in the arrangement of teeth. They used the Fibonacci Number of 1.618 (simplified in literatures as ‘the golden proportions’) as used in the ancient Greek architecture and adapted it to suit the use in dentistry. If the same ratio between the width of the central incisor and lateral incisor is repeated between the lateral incisor and the amount of cuspid shown, and between the cuspid and bicuspid, each tooth size will be different but related because of the repetition of the same ratio. This is bringing order by organizing the elements according to a principle. 8 However there also has been conflicting reports that pleasing arrangement of teeth does not necessarily follow the golden proportions.12 13 For example Ward

12

believed that when the golden proportion is

used, the lateral incisor appears too narrow, and the resulting canine is not prevalent enough. Preston14 reported that the golden proportion was found in the relationship between the maxillary central and lateral incisors in only 17% of the casts of patients he studied when viewed from the frontal. Therefore the purpose of this study was to determine the width of maxillary anterior teeth and maxillary intercanine width of adult Asians residing in Malaysia, and investigate if selected craniofacial antropometric measurents could be used to predict the sizes of anterior teeth. The existence of the golden proportion in the maxillary anterior teeth were also explored. 72

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Author

Size of central incisor

Intercanine width

Year

Sample Size

Race

Zakiah et al

2010

22 (male) 38 (female)

46 Malay 14 Chinese

8.50 ± 0.50

NA

Woodhead C.M

1977

284

Caucasian

9.00±0.40

NA

1972

60 (male) 60 (female)

40 Caucasian

Lavelle, C.L.B

40 Negroid 40 Mongoloids

Mack P.J

Keng S.B and Fong K.W.C

1981

1996

200

64 (male)

8.79 ± 0.45

NA

9.33±0.59

NA

8.67±0.42

NA

100 Nigerians

9.9 ± 0.74

37.32 ±3

100 British

8.8 ± 0.66

34.68±2.49

Chinese

8.85±0.59

35.74±2.17

Findings The widths of the anterior teeth can be predicted by a combination of the interpupillary width, inner canthal distance, and inter alar distance.

The moulds available for artificial teeth were narrower then the width of incisor measured

Tooth size is highly correlated in Negroids as compared to Caucasians. Tooth size may be a factor in malocclusions.

Maxillary Arch dimensions and central incisor widths of patients of Nigerian birth are in general greater than those of British origin.

42.8 percent of the studied sample had maxillary central mesiodistal diameters greater than 9 mm.

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Table 1: The size of the central incisor in various studies. MATERIAL AND METHODS Sample Selection criteria The subjects consisted of convenient samples of students of the University of Malaya and another nearby learning institution. They were recruited on a voluntary basis. The subjects were generally healthy and exhibited no craniofacial abnormalities. Subjects of mixed parentage were excluded from this study. Ninety four students volunteered for the project, but 4 were excluded because of their prior orthodontic treatment. In addition, damages to study cast resulted in exclusion of another 6 subjects. Therefore only 84 subjects were recruited into this study. The inclusion criteria were: 1. Healthy state of gingival and periodontium 2. Full complement of teeth which is caries free from second molar to second molar in both arches 3. No supernumerary teeth present 4. Normal molar and canine relationship (Angle Class I relationship) 5. Normal overjet (< 3 mm) and overbite (0.05). Non significant differences were found in total number of upper teeth between males(± 4.100) and females (± 3.671).however, significant differences between male(± 4.233) and female (± 3.686) were found in total number of lower teeth (p0.05) compared to females with a mean alveolar bone loss of 0.296mm (p>0.05). There is a significant difference between males and females (p>0.05). These findings were in consistent with other studies conducted by several researchers (Albandar et al. 1999, Kelly et al. 2000, Krustrup & Erik Petersen 2006, Bourgeois et al. 2007, Suominen-Taipale et al. 2008). The fact that women had substantially less documented periodontal disease might be due to differences in periodontal risk factors, sociocultural determinants, or differences in dental and general health behaviour. Smoking patterns, for example, were different across genders, favoring males (ever smokers: 59.5% versus 52.5% in adults and 61.4% versus 35.9% in seniors).( Paulander et al.2004). Our results revealed that the percentage of alveolar bone loss of 0.1 mm to 1 mm was 91.9 % and alveolar bone loss of 1.1 mm to 2 mm was 7.6 %. While the percentage of alveolar bone loss for 2.1 mm to 4 mm is 0.4%. This suggests that despite the high prevalence of periodontitis in the dental students, the amount of alveolar bone loss is still minimal. Future management programs should be scheduled to prevent progression of bone loss among dental students.. 131

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CONCLUSION High prevalence of periodontitis was found among all dental students. Males showed higher prevalence than females. Future management programs should be scheduled to prevent progression of bone loss among dental students. REFERENCES 1.

Albandar, J. M. (2007) Periodontal disease surveillance. Journal of Periodontology 78, 1179–1181.

2.

Albandar, J. M., Brunelle, J. A. & Kingman, A. (1999) Destructive periodontal disease in adults 30 years of age and older in the United States, 1988–1994. Journal of Periodontology 70, 13–29. Albandar, J. M. & Rams.

3.

Carranza FA, Takei HH, Newman MG. Clinical periodontology. 9th ed. Massachusetts: W.B. Saunders CO; 2002. p. 35469, 491-2.

4.

Dummer PHM, Jenkins SM, Newcombe RG, Adday M, Kingdon A. An assessment of approximal bone height in the posterior segments of 15 to 16-year old children using bitewing radiographs. J Oral Rehabil 1995, 22: 249-5

5.

Eliasson S, Lavestedt S, Ljungheimer C. Radiographic study of alveolar bone height related to tooth and root length. Comm Dentistry Epidemiology 19986; 14:169-171

6.

Hugoson A, Norderyd O. Has the prevalence of periodontitis changed during the last 30 years? Journal of Clinical Periodontology 2008;35: 338-345

7.

Krustrup, U. & Erik Petersen, P. (2006) Periodontal conditions in 35–44 and 65–74-yearold adults in Denmark. Acta Odontologica Scandinavica 64, 65–73.

8.

Marshall-Day CD, Stephens RG, Quigley LE JNR. Periodontal disease: Prevalence and incidence. J Periodontal 1955; 26:185

9.

Mol A. Imaging methods in Periodontology. Periodontol 2000 2004; 34: 34-8

10.

Lindhe J, Karring T. The anatomy of the periodontium in Lindhe. (Ed). Journal of Clinical Periodontology and Implant Dentistry 4th ed. Munksgaard, Copenhagen 2003; 19-69

11.

Paulander J, Wennstrom JL, Axelsson P, Lindhe J. Some risk factors for periodontal bone loss in 50-year-old individual. A 10 year cohort study. Journal of Clinical Periodontology 2004: 7: 489-6

12.

Mehdizadeh M, Amintavakoli M, Allahverdi M. The Effect of X–Ray Vertical Angulation on Radiographic Assessment of Alveolar Bone Loss. Dental Research Journal 2005. 2;2

13.

Papapanou, P.N., Wennstrom, J.L. & Grondahl, K. (1989). A 10 year retrospective study of periodontal disease progression. Journal of Clinical Periodontology. 16; 403 - 411

14.

Reynolds MA. Gender Differences in Destructive Periodontal Disease: A Systemic Review. Journal of Clinical Periodontology 2010; 0:1-18

15.

Sood M, Kumar A, Kumar N. Evaluation of periodontal disease in dental students. Contemp Clin Dent 2010;1:14-6

16.

Theilade .j An evaluation of reliability of radiographs in the measurement of bone loss in periodontal disease. J periodontal 1960 ; 31 : 143 – 53

17. Wolf B, Bethlenfalvy E, Hassfeld S, Staehle HJ, Eickholz P. Reliability of assessing interproximal bone loss by digital radiography: intrabony defects. 132

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Journal of Clinical Periodontology 2001. 28;9:869-87

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PP12: ORAL HEALTH STATUS OF CHINESE ELDERLY PEOPLE WITH DEMENTIA Tam Hoy Suet Ailsa, Chan Yau Chuen, Cheung Wing Pan, Ho Tek Ka, Lau Chon Kit, Mak Ka Man, Ng Alice, Woo Cheuk Hang Timothy Aims: To compare oral hygiene habits and oral health status of Chinese elderly people with and without dementia and to promote the oral health to elderly people with dementia. Methods: Chinese elderly people with dementia attending day-care centers were invited for this study. Age and gender matched people without dementia were recruited as control. All participants and their care-takers were informed of the study purposes and procedures, and consent was sought. The study comprised of questionnaire survey, saliva collection and clinical examination. Oral hygiene habits, use of dental aids, dental service utilization were explored in the questionnaire survey. Unstimulated salivary flow rate was measured. Caries experience, oral hygiene status, periodontal status, were measured by DMFT index, Visible Plaque Index (VPI) and Community Periodontal Index (CPI). Results: Fifty-nine Chinese elderly people with dementia (47 female) were recruited, and their mean age was 80±7. Compare with the control, there were more people with dementia who received assistance on tooth-brushing (31% vs. 5%; p