RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BANGALORE ANNEXURE-II PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

Name of the candidate and DR. APURVA J. GUJARATHI address A-12, TRISHNA SOC,GILBERT HILL ROAD, ANDHERI WEST, MUMBAI-400058

Name of the institution

MARATHA MANDALS NATHAJIRAO G. HALGEKAR INSTITUTE OF DENTAL SCIENCES & RESEARCH CENTRE, BELGAUM-590010

MASTER OF DENTAL SURGERY 3 Course of the study and subject

(PEDODONTICS AND PREVENTIVE DENTISTRY)

Date of admission

31/05/2011

Title of the topic

EVALUATION

OF

REMINERALIZING

POTENTIAL OF COMMERCIALLY AVAILABLE CHILD FORMULA DENTIFRICES: AN IN VITRO STUDY.

Brief resume of intended work 6.1. Need for the study The effectiveness of fluoridated dentifrices in caries prevention has been accepted for nearly four decades and researchers now speculate that the use of such toothpastes may be contributing significantly to the large decline in dental caries.1 The most common source of topical fluoride for majority of children is dentifrice. It is generally accepted that the use of fluoride containing dentifrices, while simultaneously providing the benefits associated with improved oral hygiene has also been one of the most important factors in decline of dental caries. Many studies have shown that early carious lesions can be remineralized, with fluoride being one of the most significant agents for promoting remineralization.2 Fluorides are added to community water supplies and are available in the form of dietary supplements, topical applications, mouthwashes, and dentifrice. Today sodium fluoride (NaF) and sodium monofluorophosphate are two fluoride agents most widely used in tooth pastes.1 There are only a few studies conducted to evaluate remineralizing potential of fluoride dentifrices on primary teeth. Consequently the effectiveness of low fluoride dentifrice on primary teeth remains unclear.2 Thus the aim of this in vitro study is to evaluate the remineralizing potential of low fluoride child formula dentifrice on primary teeth and secondary aim is to visualize structural changes occurring within the artificial carious lesion by low fluoride concentrated dentifrice with the help of scanning electron microscopy.3

6.2. Review of literature Nelson DG, Coote GE, Shariati M, Featherstone JD (1992)4 used pH cycling model to evaluate fluoride uptake and lesion progression for NaF based dentifrice, NaF based mouth rinse and a monofluorophosphate based dentifrice. Their observation showed that NaF dentifrice and mouth rinse showed considerably higher uptake of fluoride in the lesion than Monofluorophosphate dentifrice. However the mineral content profile for all specimens was not significantly different.

Itthagarun A, Wei SH, Wefel JS (1999)5 compared the calcium fluoride-like reaction products formed, both on the surface and in the sub surface caries like lesions of enamel, from different commercial dentifrices manufactured locally from developed and developing countries. Surface and subsurface of the lesions, before and after treatment, were examined using scanning electron microscopy. Varying degrees of fine globular patterns of calcium fluoride were observed on the enamel surfaces of five out of eight groups and in subsurface of four groups and supports that when compared to multinational dentifrices some Chinese and Indian dentifrices locally failed to induce the typical morphological appearance of globules as seen with fluoride dentifrices.

Itthagarun A, Wei SH, Wefel JS (2000)6 evaluated and compared remineralization effects of dentifrices manufactured locally in some developing countries stated that control group showed an increase in lesion depth of 70 % with non fluoridated control and also suggest that, when compared to multinational dentifrices, Chinese and Indian dentifrices manufactured locally failed to show equivalent healing efficacy even though they claimed to contain varying levels of fluoride.

Thaveesangpanich P, Itthagarun A, King NM, Wefel JS (2005)7 conducted an in vitro study to evaluate the effect of child formula tooth pastes on enamel caries using two in vitro pH-cycling models with use of polarized light microscopy and micro radiography. Model 1 (group A,B,C) without added fluoride and pH cycling for 7 days and Model 2 (group D,E,F)

fluoridated and pH-cycling for 10 days. Group A and D exposed to non fluoridated tooth paste, group B and E to half pea sized tooth paste and group C and F to pea sized portion of 500 ppm F toothpaste . They found that both 10 days and 7 days model were suitable for studying caries lesion progression in primary teeth and pea sized portion of 500 ppm F toothpaste slowed down the demineralization progression better than half pea sized portion and non fluoridated tooth paste.

Ekambaram M, Itthagarun A, King NM (2011)

conducted an in vitro study to evaluate

remineralizing potential of child formula dentifrice. Primary teeth were placed in demineralizing solution for 96 hour to produce artificial carious lesions 100m deep, and then cut longitudinally into 50 sections 100-150m thick and randomly assigned to five groups. Group A-500 ppm AmF, Group B-500 ppm MFP, Group C-500 ppm MFP and xylitol, Group D-500 ppm NaF and Group E- non fluoridated dentifrice. Group D sections exhibited significant decrease in lesion depth where as Group E showed a significant increase in depth (p<0.05, paired t-test). Decrease lesion progression was observed in Group A, B and C. It was concluded that 500 ppm NaF dentifrice demonstrated remineralization of carious lesions by virtue of significant decrease in lesion depth; whereas dentifrices that contained AmF, MFP and MFP with xylitol decelerated the progression of demineralization. 6.3. Objectives of the study 1. To evaluate remineralizing potential of Colgate (anti tooth decay), Colgate- Palmolive Co India 2. To evaluate remineralizing potential of Cheeriogel, Dr Reddys Laboratories LTD, India 3. To compare remineralizing potentials of Colgate (anti tooth decay) and Cheeriogel.

Materials and Methods 7.1. Source The study will be conducted at the department of Pedodontics and Preventive Dentistry and Department of Oral Pathology, Maratha Mandals, Nathajirao G. Halgekar Institute of Dental Sciences & Research Centre, Belgaum-590010. Thirty sound extracted or naturally exfoliated primary anterior teeth will be collected from department of pedodontics Maratha Mandals Nathajirao G. Halgekar Institute of Dental sciences & research centre, 10 samples with approximately 100 m artificial carious lesion will be prepared using 7 days pH cycling for each of three commercially available tooth paste, (two fluoride containing tooth pastes and one non fluoridated toothpaste) will be used for this study.

7.2. Method of collection of data 30 extracted or naturally exfoliated anterior teeth will be studied. Teeth will be stored in sterile distilled water. 10 samples with approximately 100 m artificial carious lesions will be prepared for each of three commercially available tooth pastes. Following three toothpaste will be used for the study-

Test material used for the studySr no 1 Colgate tooth (anti Sodium fluoride decay 500 ppm Trade name Content ppm fluoride Manufacturer name ColgatePalmolive Co India Sodium monofluorophosphate 458 ppm Dr Reddys Laboratories LTD, India 3 Vicco Non fluoridated tooth NIL paste Vicco Laboratories, India

toothpaste) 2 Cheeriogel gel

Dentifrice slurry preparation- Dentifrice slurry will be prepared in 3:1 ratio of deionised water to dentifrice. To achieve this 17 gm of dentifrice will be dispensed from the respective tube and then transferred into three tubes to witch 51 ml of deionised water will be added and to stir with stirring rod until well mixed.

Demineralizing and remineralizing solution preparationDemineralizing solution2.2 mM calcium chloride 2.2 mM sodium hypophosphate 0.05 M acetic acid 1 M potassium hydroxide is added to adjust pH to 4.5 Remineralizing solution1.5 mM calcium chloride 0.9 mM sodium hypophosphate 0.15 M potassium chloride with pH of 7.

Lesion formationThirty sound extracted teeth will be cleaned to remove soft tissue debris and will be inspected for cracks, hypoplasia and white spot lesions. Teeth will be coated with acid resistant nail varnish. Leaving 1mm narrow window on sound intact buccal or lingual surface. It will be kept in demineralizing solution for 96 hour to produce 100 m lesions. Approx 100-150 m longitudinal enamel section will be prepared by using hard tissue microtome. 10 sections will be randomly assigned to each of the three experimental groups. Polarized light microscopy and scanning electron microscopy will be used to record the depth and mineral content of lesion before and after treatment with the different dentifrice.

pH cycling model - Specimens will be placed in pH cycling system on an orbital shaker for 7 days. Each cycle will involve 3 hour demineralization twice daily and 2 hour of remineralization in between 1 min treatment with dentifrice slurry before 1st demineralizing and before and after 2nd demineralizing solution. Freshly prepared solution for each cycle will used. After 7 days nail varnish will be removed with acetone.

Evaluation techniquePolarized light microscopy- (Olympus research microscope) will be used for evaluating the lesion depth pretreatment and post treatment by photo-micrographs taken before and after the experiment at the same magnification. Scanning electron microscopy-(JSM Model 6380) will be used to visualize structural changes occurring within the artificial carious lesion.

Statistical analysis- Analysis of variance test will be done to compare the pre treatment and post treatment lesion depth. 7.3. Does the study require any investigation or interventions to be conducted on patients or other NO humans or animals? If so, please describe briefly. 7.4. Has ethical clearance been obtained from your institution in case of 7.3? YES

References 1) Stookey GK, DePaola PF, Featherstone JD, Fejerskov O, Mller IJ et al. A critical review of the relative anticaries efficacy of sodium fluoride and sodium monofluorophosphate dentifrices. Caries research 1992;27:337-360. 2) Ekambaram M, Itthagarun A, King NM. Comparison of the remineralizing potential of child formula dentifrices. IJPD 2011;21:132-140. 3) Hubbard Mj. Correlated light and scanning electron microscopy of artificial carious lesions. J Dent Res 1982; 61:14-19. 4) Nelson DG, Coote GE, Shariati M, Featherstone JD. High resolution fluoride profiles of artificial in vitro lesion treated with fluoride dentifrices and mouth rinses during pH cycling conditions. Caries Research 1992; 26:254-262. 5) Itthagarun A, Wei SH, Wefel JS. Morphology of initial lesions of enamel treated with different commercial dentifrices using a pH-cycling model: Scanning electron microscopy observations. International Dental Journal 1999;49:352-360. 6) Itthagarun A, Wei SH, Wefel JS. The effects of different commercial dentifrices on enamel lesion progression: an in vitro pH-cycling study. International Dental Journal 2000; 50:2128. 7) Thaveesangpanich P, Itthagarun A, King NM, Wefel JS. The effects of child formula toothpastes on enamel caries using two in vitro pH-cycling models. International Dental Journal 2005; 55:217-223.

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Signature of candidate Remarks of the guide Name and designation of(in block letter) 11.1. Guide DR. SANTOSH SHOLAPURMATH , M D S READER, DEPT OF PEDODONTICS AND PREVENTIVE

DENTISTRY MARATHA MANDALS N.G.H.I.D.S. & R.C. BELGAUM-590010 11.2. Signature 11.3 Co-guide
DR. PRAVEENKUMAR S. MANDROLI, M D S PROFESSOR, DEPT. OF PEDODONTICS AND PREVENTIVE DENTISTRY MARATHA MANDALS N.G.H.I.D.S. & R.C. BELGAUM-590010

11.4. Signature 11.5. Head of the department

DR. PRAVEENKUMAR S. MANDROLI, M D S PROFESSOR, DEPT. OF PEDODONTICS AND PREVENTIVE DENTISTRY MARATHA MANDALS N.G.H.I.D.S. & R.C. BELGAUM-590010

11.6. Signature 12 12.1. Remarks of the Principal 12.2. Signature