FULVIC ACID JC.pptxhhhhhhhhhhhhhhhhhhhhh

ggggggggggggggggggggggggggggggggggggggggggggggggggggg

1. Effect of 5% Carbohydrate Derived-Fulvic Acid on Smear Layer Removal and Root Dentin Microhardness – An In Vitro study Devadurai Ravindar Arun ,  Venkatappan Sujatha , Sekar Mahalaxmi Presented by: DR. PRIYANKA IPPAR PG-2ND YEAR GUIDED BY: DR. RANA K. VARGHESE, PROFESSOR AND HOD. DR. MALWIKA SISODIYA, READER. DR. NAVEEN KUMAR GUPTA, READER. DR. DR. ANITA CHANDRAKAR, SENIOR LECT. DR. CHANDRABHAN GENDLY, SENIOR LECT.

2. Content INTRODUCTION MATERIALS AND METHODS IRRIGATION PROTOCOL SEM EVALUATION VICKERS MICROHARDNESS TESTING STATISTICAL ANALYSIS RESULTS DISCUSSION CROSS REFERENCE

3. INTRODUCTION Root canal preparation comprises combined action of endodontic instruments and irrigating solutions for thorough disinfection The mechanical instrumentation of the canal creates a granular,amorphous smear layer (smear layer) that contains both organic and inorganic substances which cover the canal wall and occlude the openings of the dentinal tubules. The presence of SL favours the adhesion and colonization of microorganisms and limits effective disinfection by preventing the irrigants and intracanal medicaments from penetrating into the dentinal tubules

4. The action of sodium hypochlorite (NaOCl), the most routinely used irrigant is limited to the removal of the organic component of SL when used alone, hence necessitating its association with chelating agents such as ethylenediaminetetraacetic acid (EDTA) as a final irrigant, for removal of SL by acting on its inorganic component as well. However, use of EDTA has a strong demineralizing effect on root dentin. This decreases the microhardness and fracture resistance of root dentin. The intra and peritubular erosion when as_x0002_sociated with NaOCl leave the naked collagen fibrils exposed which are prone to bacterial adherence. Further EDTA has a cytotoxic effect when extruded beyond the periapical tissues and is considered a weak antibacterial agent.

5. There is a constant search for natural biocompatible irrigants that can minimize the undesirable changes in dentin microstructure and low toxic and antibacterial efficency. Humic substances are natural degradation products of organic matter formed from decomposed plant and animal residues comprising humins, humic acid and fulvic acid. Fulvic acid (FA), is a major constituent of “humic” substances that is water soluble under all pH conditions. FA biological chelator is lower in molecular size and weight and has lower colour intensity than humic acid it is reported to have chelating, anti-inflammatory, antioxidant, antibacterial, antiviral and antifungal properties. there are very few article publish that compare the eof EDTA and CD-FA and there efficacy on dentine microhardness.

6. MATERIALS AND METHODS A total of 56 human mandibular single canal premolars with mature roots Teeth with severely curved roots standardised using Schneider technique, calcifiedcanals,morphological defects, caries and fracture were excluded. The collected teeth were cleaned and stored in 0.1% thymol solution at 37°C to maintain hydration . CHD-FA (Mineralife Nutraceuticals, USA) used in this study was diluted to 5% concentration using distilled water.a/c pilot study various concentration of FA use for removal of SL but only 5% CHA-FA is effictive in removal of SL. The teeth were decoronated to obtain a standardized root length of 12 mm using a high-speed diamond disc. The canal patency was established using 10 size K-file passing through the apical foramen. Working length was determined to be 1mm less than the length of size 15 K-file just exiting the foramen.

7. the sample is randomly assigned into 3 differnt group G roup1- NaOCl /EDTA G roup2-NaOCl/CHD-FA G roup3-distilled water The root canals were prepared with Pro Taper Universal rotary instruments using crown_x0002_down technique.The root apices were covered with sticky wax to prevent outflow of the irrigants . The irrigation procedure was done using 30-gauge side vented needle placed 1 mm short of working length.After the final rinse with distilled water to remove any resid_x0002_ual irrigating solution, the canals were dried using size 30 paper points. Two longitudinal grooves were made on the outer surface of the roots using a diamond discat slow speed under water coolant without perforating the canal to facilitate splitting of the roots into two equal halves (n=112). The teeth were split with the aid of a surgical chisel and a mallet.

8. one half being subjected to Scanning Electron Microscopic (SEM) analysis for evaluation of smear layer and the other half subjected to Vickers Microhardness Test_x0002_ing (VMT) for evaluation of microhardness. SEM evaluation- One half of the samples were delineated into coronal, middle and apical thirds using an indelible marker, mounted on a metallic stub and viewed under SEM at 2000x magnification. The images were scored according to the criteria given by Takeda et al. score1-surface completely covered by smear layer score2- partially covered with smear layer & few dentinal tubules where visible. score3-, surface with little smear layer and more tubules visible. score4-smear layer-free surface.

9. 17% EDTA (group 1) CHA-FA(group 2)

10. distilled water ((group 3)

11. Vickers microhardness testing - The remaining root samples were horizontally mounted in autopolymerizing acrylic resin with the canal lumen exposed. Three indentations were made on the apical third (4 mm from apex) of root dentin using Vickers diamond indenter under 300 g load and a dwell time of 20 sec. Statistical analysis - all the groups were compared using non-para_x0002_metric Kruskal-Wallis test with significance level fixed at 0.05. For pairwise comparison, post-hoc test was done. RESULTS - The pH of 5% CHD-FA solution measured was 2.7. Both 17% EDTA and 5% CHD-FA exhibited a score of 4 in the coronal and middle thirds. apical third, 5% CHD-FA showed complete removal having a score of 4, while 17% EDTA exhibited moderate smear layer removal having a score of 3

12. apical third, 5% CHD-FA showed complete removal having a score of 4, while 17% EDTA exhibited moderate smear layer. VHN values Group 3 showed the highest mean VHN, followed by groups 2 and 1.

13. DISCUSSION- The chelating agents improve the chemo-mechanical de_x0002_bridement of the root canal by removing the smear layer Chelators have also been found beneficial in negotiating constricted and calcified canals by softening the dentin, how_x0002_ever the degree of softening and demineralization induced by these agents can have an adverse effect on dentin leading to reduction in microhardness The SEM results indicated that irrigation protocol using 5mL of both group 1 (NaOCl/EDTA) and group 2 (NaOCl/CHD-FA) for 1 min was significantly more effective in the removal of smear layer from coronal and middle third of instrumented root canal surfaces when compared to group3 (Distilled water) It was observed that 17% EDTA was significantly less effective in removing the smear layer when compared to 5% CHD-FA in the apical third.

14. This reduced efficacy of EDTA can be ascribed to its increased surface tension and the higher molecular size. Conversely, be_x0002_cause of the relatively small size of fulvic acid (FA) molecules CHD-FA can readily enter the apical portion of the canal. In fulvic acid, chelation occurs at salicylate-like bidentate sites for copper (Cu2+) a biva_x0002_lent ion similar to calcium (Ca2+), by the formation of Ca-fulvic acid complex with hydroxyapatite with the chelate occurring at 1:1 ratio. Further, both strong acidic COOH and weak COOH are involved in the formation of complexes and the sta_x0002_bility of such complexes is pH dependent . For fulvic acid, the number of released protons decreases as pH increases and complexation of calcium to fulvic acid involves mainly strong acidic groups. Formation of Ca-fulvic complex is not possible at acidic pH probably due to higher concentration of H+ compared to that of Ca2+ for dissociated COO–.

15. This initial lower pH of CHD_x0002_FA may be the reason for lesser reduction in microhardness for CHD-FA when compared to EDTA (pH-7.8). It is also believed that configuration changes in FA occurs at basic pH and allows stability of the complexes formed between fulvic acid and bivalent cations, contributing to the significantly better smear layer removal. CHD-FA, a novel antimicrobial agent was shown to possess antifungal high activity against oral bacterial biofilms (A. actinomycetemcomitans, P. gingivalis, F. nucleatum, S. mitis), with a MIC of 0.5%. CHD-FA a naturally derived chelator with supplementary antimicrobial, antioxidant and anti-inflamma_x0002_tory properties can be suggested as a promising final irrigant.

16. CONCLUSION 5% CHD-FA was equivalent to 17% EDTA in smear layer removal at coronal and middle third of the root whereas in apical third CHD-FA removed smear layer more effectively with significantly lesser reduction in microhardness than EDTA.

17. CROSS REFERENCES 1)Botes ME,et al in 2018 Carbohydrate-derived fulvic acid wellness drink: its tolerability, safety and effect on disease markers in pre-ART HIV-1 positive subjects concluded that there was no side effect of CHA-FA drink. 2)Rath PPet al in 2020 The effects of sequential and continuous chelation on dentin.Sequential chelation (NaOCl/EDTA) exposed thick collagen bundles on the surface, while the continuous chelation (NaOCl/HEDP) protocol resulted in a thin, frail layer of exposed collagen on the surface.

18. REFRENCES . Pashley DH, Michelich V, Kehl T. Dentin permeability: effects of smear layer removal.  J Prosthet Dent.  1981;46(5):531–7. [ PubMed ] [ Google Scholar ] 2. Rath PP, Yiu CKY, Matinlinna JP, Kishen A, Neelakantan P. The effects of sequential and continuous chelation on dentin.  Dent Mater.  2020;36(12):1655–65. [ PubMed ] [ Google Scholar ] 3. Calt S, Serper A. Time-dependent effects of EDTA on dentin structures.  J Endod .  2002;28(1):17–9. [ PubMed ] [ Google Scholar ] 4.  Niu W, Yoshioka T, Kobayashi C, Suda H. A scanning electron microscopic study of dentinal erosion by final irrigation with EDTA and NaOCl solutions.  Int Endod J.  2002;35(11):934–9. [ PubMed ] [ Google Scholar ] 5.  Karkehabadi H, Yousefifakhr H, Zadsirjan S. Cytotoxicity of endodontic ırrigants on human periodontal ligament cells.  Iran Endod J.  2018;13(3):390–4. [ PMC free article ] [ PubMed ] [ Google Scholar ] 6. Silva PV, Guedes DF, Nakadi FV, Pécora JD, Cruz-Filho AM. Chitosan: a new solution for removal of smear layer after root canal instrumentation.  Int Endod J.  2013;46(4):332–8. [ PubMed ] [ Google Scholar ] 7. Kirchhoff AL, Viapiana R, Miranda CE, Sousa Neto MD, Cruz Filho AM. Comparison of the apple vinegar with other chelating solutions on smear layer and calcium ions removal from the root canal.  Indian J Dent Res.  2014;25(3):370–4. [ PubMed ] [ Google Scholar ]

19. 8. Botes ME, Gilada IS, Snyman JR, Labuschagne JP. Carbohydrate-derived fulvic acid wellness drink: its tolerability, safety and effect on disease markers in pre-ART HIV-1 positive subjects.  South African Family Practice.  2018;60(3):91–6. [ Google Scholar ] 9. Takeda FH, Harashima T, Kimura Y, Matsumoto K. Efficacy of Er:YAG laser irradiation in removing debris and smear layer on root canal walls.  J Endod .  1998;24(8):548–51. [ PubMed ] [ Google Scholar ] 10.  McComb D, Smith DC. A preliminary scanning electron microscopic study of root canals after endodontic procedures.  J Endod .  1975;1(7):238–42. [ PubMed ] [ Google Scholar ] 11.  Baldasso FER, Roleto L, Silva VDD, Morgental RD, Kopper PMP. Effect of final irrigation protocols on microhardness reduction and erosion of root canal dentin.  Braz Oral Res.  2017;31:e40. [ PubMed ] [ Google Scholar ] 12.  Hülsmann M, Heckendorff M, Lennon A. Chelating agents in root canal treatment: mode of action and indications for their use.  Int Endod J.  2003;36(12):810–30. [ PubMed ] [ Google Scholar ] 13.  Hennequin M, Pajot J, Avignant D. Effects of different pH values of citric acid solutions on the calcium and phosphorus contents of human root dentin.  J Endod .  1994;20(11):551–4. [ PubMed ] [ Google Scholar ] 14. Zehnder M. Root canal irrigants .  J Endod .  2006;32(5):389–98. [ PubMed ] [ Google Scholar ]

20. THANK YOU