FIELD OF INVENTION
The present invention relates to the development of gel formulation with extracts of Cocos nucifera for medical and dental applications. It specifically relates to the development of gel formulation with ethanolic extracts of husk of Cocos nucifera against the Enterococcus faecalis, Prevotella intermedia and Porphyromonas gingivalis and as pulpotomy agent, healing gel, for ulcer and extracted wound, as a local drug delivery after conventional scaling and root planning. The invention also pertains to the development of process for preparation of gel formulation with ethanolic extracts of Cocos nucifera for medical and dental applications.
BACKGROUND OF THE INVENTION
Nature is the source of all true Knowledge. She has her own logic, her own laws, she has no effect without cause nor invention without necessity.
-Leonardo Da Vinci
Despite modern advances in prevention of dental caries, many teeth are lost prematurely in children. Primary teeth are a guide for the eruption of permanent dentition and are an important contributing factor for development of jaws, chewing process and growth of the child. Premature loss of teeth can produce problems in the eruption of succedaneous teeth, phonetic disturbances and harmful oral habits like tongue thrusting (Fabris AS, Nakano V, Avila-Campos MJ. Bacteriological analysis of necrotic pulp and fistulae in primary teeth. J Appl Oral Sci. 2014; 22(2): 118-24). One of the primary objectives of Pediatric dentistry is to preserve the health and integrity of primary teeth until they naturally exfoliate. When the infectious and inflammatory process is advanced, it is not possible to carry conservative pulpal treatment rendering pulpectomy as the ideal treatment to eliminate micro-organisms

through chemo-mechanical disinfection of the root canal system (Valdez-Gonzalez C, Mendez- Gonzalez V, Torre-Delgadillo G, Flores-Reyes H, Gaitan- fonseca C, Pozos-Guillen AJ. Effectiveness of oxidative potential water as an irrigant in pulpectomized primary teeth. J Clin Pediatr Dent 2012; 37(1): 31-35). The peculiar morphology of the deciduous dentition with apical delta and significantly abundant lateral and accessory canals makes the infectious process difficult to ward off, especially in necrotic canals, abscesses and sinus tract infections (Jolly M, Singh N, Rathore M, Tandon S, Banerjee M. Propolis and commonly used intracanal irrigants: Comparative evaluation of antimicrobial potential. J Cin Pediatr Dent 2013; 37(3): 243-249). Belanger (Belanger GK. Pulp therapy for the primary dentition. In: Pinkham JR, ed. Pediatric Dentistry. Philadelphia, PA, USA: Saunders; 1988) mentioned that an intact primary tooth adequately disinfected and restored natural tooth acts as a best maintainer of space than any artificial appliance. Due to these reasons much importance is given to chemical cleaning and disinfection of canals as opposed to thorough mechanical shaping of the canals. The pathway of progression of the infection and the ecology of infecting microorganisms are different in the deciduous dentition as compared to the permanent dentition (Ruviere DB, Leonardo MR, da Silva LAD, Ito IY, Nelson- Filho P. Assessment of the Microbiota in Root Canals of Human Primary Teeth by Checkerboard DNA - DNA Hybridization. Journal of Dentistry for Children 2007; 74(2): 118-23). Hence, for complete elimination of infection irrigation is an important step for the success of pulpectomy. Studies have demonstrated that despite the use of antimicrobial agents, microorganisms such as Enterococcus faecalis, Prevotella intermedia and Porphyromonas gingivalis still persists in the root canal of the primary teeth. Therefore, an ideal irrigating solution should provide mechanical flushing, be

bactericidal, and dissolve the remnants of the organic tissue without damaging the periodontium. It must be biocompatible and nontoxic both locally and systemically (Zehnder M. Root canal irrigants. J of Endod May 2006; 32(5): 389-98). Sodium hypochlorite (NaOCl) is one of the most popular irrigating solutions. It has antibacterial activity and the ability to dissolve necrotic tissue remnants. But it is cytotoxic and has potential risk of injuring the permanent tooth germ.2 Even though it has been considered as a gold standard and there is significant documentation in literature for its use is unpleasant taste, foul smell, high toxicity, allergic potential, bleaching effect and inability to remove the smear layer has led to the need to find other alternatives. Sodium hypochlorite (NaOCl) is an effective antibacterial agent but is harmful when extrudes outside the root canal and causes damage when it contacts the periradicular tissues (Hulsmann M, Hahn W. Complications during root canal irrigation- literature review and case reports. Int Endod Journal 2000; 33: 186-93). Chlorhexidine gluconate (CHX) has been suggested as an effective irrigating solution for pulpectomy treatment of necrotic primary teeth. It was developed in the late 1940s in the research laboratories of Imperial chemical Industries Ltd. (Macclesfield, England). Aqueous solutions of 0.1% to 0.2% have been used for plaque removal purpose, while 2% is the concentration of root canal irrigants usually found in the endodontic literature. It was reported that 2% chlorhexidine (CHX) showed a greater reduction of intracanal bacterial loading and suggested that this irrigating solution is an alternative for pulpectomy treatment of necrotic primary teeth. But it may be cautioned here that chlorhexidine is unable to dissolve necrotic tissue remnants and it is less effective against gram negative bacteria which predominate the primary endodontic infections. Also, the constant increase in antibiotic resistance and side effects caused by the synthetic agents, has led to search for other natural alternatives

(Prabhakar P, Senthilkumar M, Priya MS, Mahalakshmi K, Sehgal K, Sukumaran VG. Evaluation of antimicrobial efficacy of herbal alternatives (triphala and green tea polyphenols), MTAD, and 5% Sodium hypochlorite against Enterococcus faecalis biofilm formed on tooth substrate: An in vitro study. J of Endod Jan 2010; 36(1):83-6).
The World Health Organization (WHO) estimates that 80 percent of the world's population presently uses herbal medicine for some aspect of primary health care (Mandroli PK and Bhat K. An in-vitro evaluation of antibacterial activity of curcumin against common endodontic bacteria. J of Appl Pharmaceutical Sci Oct 2013; 3(10): 106-8). The use of plants as medicines predates written human history. Over the past decade, interest in drugs derived from medicinal plants has markedly increased. Phytomedicine has been used in dentistry as anti-inflammatory, antibiotic, analgesic, sedative and also as endodontic irrigant. Herbal preparations can be derived from the root, leaves, seeds, stem, and flowers (Sinha DJ, Sinha AA. Natural medicaments in dentistry. Ayu. 2014;35(2): 113-118). Taking into consideration the ineffectiveness, potential side-effects and safety concerns of synthetic drugs, the herbal alternatives for endodontic usage might prove to be advantageous. Amongst the many herbal products that are being used in dentistry, the husk of Cocos nucifera is one such herb that has not been explored yet as a irrigating solution. Also, very few studies have been carried out in Indian scenario utilizing this herb as an endodontic irrigant.
Cocos nucifera (coconut) belongs to Arecaceae family has been used since ancient times as a traditional remedy. The beneficial medicinal effects of Cocos nucifera including the antibacterial activity result from the secondary products present in the plant, although it is usually not attributed to a single compound but a combination of

the metabolites (Parekh J, Jadeja D, Chanda S. Efficacy of aqueous and methanol extracts of some medicinal plants for potential antibacterial activity. Turk J Biol 2005; 29:203-10). One of the mechanisms suggested is hydrophobic activities, which enable them to partition the lipids of the bacterial cell membrane and mitochondria, disturbing the cell structures and rendering them more permeable. This result in extensive leakage from bacterial cells or the exit of critical molecules and ions will lead to death (Sikkema J, De Bont AM, Poolman BM. Interaction of cyclic hydrocarbons with biological membranes. J Biol Chem 1994; 269:8022-8). Tannins present in the plant extracts also have an astringent effect on the mucous membrane, and they form a layer over enamel, thus providing protection against dental caries (Prashanth GM, Chandu GN. The effect of mango and neem extract on four organisms causing dental caries: Streptococcus mutans, Streptococcus salivarius, Streptococcus mitis, Streptococcus sanguis: An in vitro study. Indian J Dent Res 2007; 18:148-51). Phytochemical screening of C. nucifera conducted by Alviano et al. has reported that this plant material is rich in alkaloids, flavonoids, catechin and epicatechin together with condensed tannins, which confers on its potent antimicrobial properties (Alviano WS, Alviano DS, Diniz CG, Antoniolli AR, Alviano CS, Farias LM, et al. In vitro antioxidant potential of medicinal plant extracts and their activities against oral bacteria based on Brazilian folk medicine. Arch Oral Biol 2008; 53: 545-52).
The substances that have been used during chemomechanical preparation are usually in liquid form. Some authors have suggested the use of a viscous irrigant that might have better lubricant action and enhancement of the antimicrobial property. Nevertheless the viscous bases used in these irrigants are little soluble in water, icavingU'esiiluesoju, UieNdtimirial walls that damag'e; the'hnal obturation of the root

canal system. Therefore the Cocos nucifera husk extract gel is used in the present study which can be used as gel for chemomechanical preparation in endodontics procedures. It can also be used as a pulpotomy agent, as a gel for for ulcer, mucositis, healing of extraction wound, as a local drug delivery after scaling and root planning. It has shown other properties like analgesic, antifungal, antiviral, antioxidant, anti neoplastic, anti parasitic, anti leishmania and anti malarial.
Since inception Periodontal treatment has focused on the reduction of bacterial infection by mechanical removal of infectious agents but despite spectacular advances in medical sciences treatment of periodontal diseases has changed very little in its approach over the years. A large evidence based studies now exist to demonstrate the efficacy of non surgical periodontal therapy.
Scaling and root planing (SRP) remains the 'gold standard" treatment for periodontal diseases against which other treatments are compared. However, after non surgical therapy several deep periodontal pockets may persist and in such cases the treatment consists of surgical procedures. Moreover, antibiotics and antiseptics have been successfully used to treat moderate to severe periodontal disease (Rams TE, Slots J. Local delivery of antimicrobial agents-in the periodontal pocket. Periodontology 2000. 1996 Feb; 10: 139-159)
It is found that the local application of antimicrobials that are effective against periodontal pathogens can reduce periodontal pocket depths (Okuda K, Wolff L, Oliver R, Osborn J, Stoltenberg J, Bereuter J, et al. Minocycline slow-release formulation effect on subgingival bacteria. J Periodontol 1992;63:73-9)

Controlled release antimicrobial delivery systems have also been tested as mono therapies independent of SRP (Knoll-Kohler E. Metronidazole dental gel as an alternative to scaling and root planing in the treatment of localized adult periodontitis. Eur J Oral Sci 1999;107:415-21)or in combination with mechanical debridement as adjunctive therapies (Vinholis AH, Figueiredo LC, Marcantonio Junior E, Marcantonio RA, Salvador SL, Goissis G. Subgingival utilization of a 1% chlorhexidine collagen gel for the treatment of periodontal pockets. A clinical and microbiological study. Braz Dent J 2001;12:209-13)
Treatment of periodontal disease is routinely based on mechanical debridement of the tooth surface and appropriate meticulous maintenance of oral hygiene. It alone may fail to eliminate the periodontal pathogens because of their location within the gingival and dental tissues which are inaccessible to periodontal instrumentation. As an adjunctive approach, systemic or local administrations of antibiotics are used because of the complex microbial etiology of periodontal disease.
Since the systemic antibiotics therapy has various disadvantages like hypersensitivity reaction, organ toxicity, development of resistant bacteria and also requires higher dosage to attain required GCF concentration at the target site this led to use of local drug delivery system.( Amitha R, Thomas Biju, SathishM. Comparative evaluation of sub-gingival application of chlorhexidine varnish and chlorhexidine gel as an adjunct to full mouth scaling and root planing in the treatment of moderate to deep periodontal pockets- A clinical study. Journal of Indian Dental Association2011; 5(3): 369-372).

As an antiseptic, chlorhexidine has been used effectively for over 30 years in the treatment of periodontal disease. It shows a broad spectrum of topical antimicrobial activity, safety, effectiveness, substantivity and lack of toxicity (Lo"e H, Schiott CR. The effect of mouthrinses and topical application of chlorhexidine on the development of dental plaque and gingivitis in man. J Periodontal Res 1970;5:79-83). However, after prolonged use it may lead to undesirable side-effect such as the pigmentation of teeth and oral tissues (Eriksen HM, Nordbo H, Kantanen H, Ellingsen JE. Chemical plaque control and extrinsic tooth discoloration. A review of possible mechanisms. J Clin Periodontol 1985;12:345-50)
To attain and maintain periodontal health bacterial plaque must be eliminated. Scaling and root planing are considered the gold standard for achieving this goal however, in pockets >5mm and furcations, debridement often fails to remove plaque and calculus. To reduce surgical treatment needs at these pockets, chemo mechanical treatment concepts have been developed using systemic antibiotics or locally applied antimicrobial agents.
Chlorhexidine is a highly effective anti microbial agent. A study by Loe and Schiott showed that chlorhexidine is effective as a mouth rinse against supra gingival plaque in prevention and treatment of gingivitis.
A degradable subgingivally placed local drug delivery system containing 2.5 mg chlorhexidine gluconate was developed by Steinberg et al and other investigators such as Soskolne et al. and Heasman et al tested 2.5 mg chlorhexidine gluconate in association with scaling and root planing and concluded that this mode of treatment ...wnHality wns hr.ttr.r. thnn .fiKT> nlnnc (Pnrincirti C, Pnnlnntonio . M, Cordelia C,

D'Ercole S, Serra E, Piccolomini R.Clinical and microbiological effects of subgingival administration of two actice gels on persistent pockets of chronic periodontitis patients. J Clin Periodontol 2004; 31:273-281).
Local delivery devices are systems designed to deliver agents locally into periodontal pocket to retain therapeutic levels for a longer duration of time. The periodic use of local drug delivery systems along with periodontal therapy help in reducing probing depths, stabilizing attachment levels and minimizing bleeding so that it would allow better control of the disease.
Devices that have been developed include fibers, chips & injectible systems. Different drugs used for local delivery are tetracyclines including doxycycline and minocycline, metronidazole and chlorhexidine.
Patients with mild to moderate chronic periodontitis thus can be treated by non-surgical therapy along with local drug delivery to halt the periodontal disease and limit the need of surgical intervention in the future.
India has a rich source of medicinal plants that are widely distributed. Known in English as Coconut and botanically called as Cocos nucifera, Coconut, belongs to the family Arecaceae. (Lima EB, Sousa CN, Meneses LN, Ximenes NC, Junior S, Vasconcelos GS, Lima NB, Patrocinio MC, Macedo D, Vasconcelos SM. Cocos nucifera (L.)(Arecaceae): A phytochemical and pharmacological review. Brazilian Journal of Medical and Biological Research. 2015 Nov;48(ll):953-64). The plant is originally from Southeast Asia (Malaysia, Indonesia, and the Philippines) and the islands between the Indian and Pacific Oceans. The plant is an arborescent . . monpcotyledonous tree of around 25 m.in height.(giant rnrnnnt) with a densp canopy

The root of the coconut system is fasciculated. The stem is an unbranched type, and at its apex, a tuft of leaves protects a single apical bud. The pinnate leaves are feather-shaped, having a petiole, rachis and leaflets. Under favorable environmental conditions, the giant adult coconut emits 12-14 inflorescence spikes per year, while the adult dwarf coconut can emit 18 spikes in the same period. The axillary inflorescence has globular clusters of female flowers. The plant is monoecious (male and female reproductive organs on the same plant) (EEM Steps. Morphology of the coconut. The coconut tree culture in Brazil. 2nd edn. Brasilia: Embrapa - Information Production Service; 1998,). The coconut fruit comprises an outer epicarp, a mesocarp, and an inner endocarp. The epicarp, which is the outer skin of the fruit, and the mesocarp, which is heavy, fibrous, and tanned when dry, have many industrial uses. The endocarp is the hard dark core. Inside is a solid white albumen of varied thickness, depending on the age of the fruit, and with an oily pulp consistency and a liquid albumen called coconut water that is thick, sweet, and slightly acidic (EEM Steps. Morphology of the coconut. The coconut tree culture in Brazil. 2nd edn. Brasilia: Embrapa - Information Production Service; 1998, Andrade AM, Passos PRA, Marques LGC, Oliveira LB, Vidaurre GB, Roch JDS. Pyrolysis of cocoa residues (Cocos nucifera Linn) and charcoal analysis. Rev Arvore 2004; 28: 707-714). All parts of the fruit of the coconut tree can be used. Both the green coconut water and solid albumen ripe fruits are used industrially and in home cooking in many ways (Rosa M de F, Santos FJ de S, Montenegro AAT, Abreu FAP, Correia D, Araujo FBS, et al. Characterization of the powder of the green coconut shell used as an agricultural substrate. Fortaleza: Embrapa Tropical Agroindustry, Technical Communication, 54; 2001). Additionally, several parts of the fruit and plant have been used by people in different countries for the treatment of various pathological

conditions. The husk fiber of Cocos nucifera Linn. (Palmae) is rich in catechin and epicatechin together with condensed tannins, which confers to its aqueous extract a potent antioxidant characteristic. Antibacterial, antiviral, antileishmanial, anti-lymphoproliferative and anti-neoplasic activities were also observed. (Alviano WS, Alviano DS, Diniz CG, Antoniolli AR, Alviano CS, Farias LM, et al. In vitro antioxidant potential of medicinal plant extracts and their activities against oral bacteria based on Brazilian folk medicine. Arch Oral Biol 2008; 53: 545-52). Phytochemical studies of the coconut fiber (mesocarp) ethanolic extract revealed that the presence of phenols, tannins, leucoanthocyanidins, flavonoids, triterpenes, steroids and alkaloids ((Matos FJA. Introduction to experimental physics. 2nd edn. Fortaleza: University Press; 1997.),), while a butanol extract recovered triterpenes, saponins, and condensed tannins (Costa CT, Bevilaqua CM, Morais SM, Camurca-Vasconcelos AL, Maciel MV, Braga RR, et al. Anthelmintic activity of Cocos nucifera L. on intestinal nematodes of mice. Res Vet Sci 2010; 88: 101-103.). The beneficial medicinal effects of plant materials including the antibacterial activity typically result from the secondary products present in the plant, although it is usually not attributed to a single compound but a combination of the metabolites [Parekh J, Jadeja D, Chanda S. Efficacy of Aqueous and Methanol Extracts of Some Medicinal Plants for Potential Antibacterial Activity. Turk J Biol 2005;29:203-10]. The exact mechanism by which the polyphenols of the plant materials contribute to anti-bacterial activity is not known. One of the mechanisms suggested is hydrophobic activities, which enable them to partition the lipids of the bacterial cell membrane and mitochondria, disturbing the cell structures and rendering them more permeable. This result in extensive leakage from bacterial cells or the exit of critical molecules and ions will lead to death. [Sikkema J, De Bont AM, Poolman BM. Interaction of cyclic

hydrocarbons with biological membranes. J Biol Chem 1994;269:8022-8]. Tannins present in the plant extracts also have an astringent effect on the mucous membrane, and they form a layer over enamel, thus providing protection against dental caries.f Prashanth GM, Chandu GN. The effect of mango and neem extract on four organisms ' causing dental caries: Streptococcus mutans, Streptococcus salivarius, Streptococcus mitis, Streptococcus sanguis: An in vitro study. Indian J Dent Res 2007;18:148-51] Zablotowics et al., reported that saponins have the ability to cause leakage of proteins from bacterial cells. Cocos nucifera extract revealed the presence of this compound and thus might have also contributed to the leakage of protoplasmic materials. (Zablotowicz RM, Hoagland RE, Wagner SC. Effect of saponins on the growth and activity of rhizosphere bacteria. Adv Exp Med Biol. 1996;405:83-95).
Table 1: Details of the Cocos nucifera plant and its parts along with their applications (Lima EB, Sousa CN, Meneses LN, Ximenes NC, Junior S, Vasconcelos GS, Lima KB, Patrocinio MC, Macedo D, Vasconcelos SM. Cocos nucifera (L.)(Arecaceae): A phytochemical and pharmacological review. Brazilian Journal of Medical and Biological Research. 2015 Nov;48(l l):953-64).

A study was conducted to evaluate antimicrobials and antiviral activities of polyphenols from Cocos nucifera Linn. (Palmae) husk fiber extract. Cocos nucifera Linn. (Palmae) is fully ripe, the husk turns brown, dry and very fibrous with a high

content of pentoses, cellulose and lignin. The husk fiber decoction has been used in north eastern Brazil for treatment of diarrhoea and arthritis. The alcoholic extract of ripe dried coconut shell has antifungal activity against Microsprum canis, M. gypseum, M. audouins, T. mentagrophytes, T. rubrum and its due to its high content of phenolic compounds. The results presented can be considered promising with antibacterial and antiviral activity which could be due to the presence of catechins and epicatechins on the coconut husk fiber extract. Catechins were capable of inhibiting tumour cell lines. Researchers are increasingly giving attention to folk medicine, looking for new leads to developing better drugs against microbial infections. The antibacterial activity of C. nucifera suggests that it is useful for topical application in wound healing (Esquenazi D, Wigg MD, Miranda MM, Rodrigues HM, Tostes JB, Rozental S, da Silva AJ, Alviano CS. Antimicrobial and antiviral activities of polyphenolics from Cocos nucifera Linn(Palmae) husk fiber extract. Research in microbiology. 2002 Dec l;153(l0):647-52).
A study was carried out to evaluate the antioxidant and antimicrobial properties of methanolic extract from Cocos nucifera mesocarp. C. nucifera L (family Arecaceae) commonly known as coconut, is an important crop in tropical countries. The fruit wall consists of three layers exocarp, mesocarp and endocarp. Due to extensive cross linking between phenolics, lignin and polysaccharides, the mesocarp becomes hard and fibrous. As a traditional medicine in Brazil, Husks of C. nucifera have been used for the treatment of diarrhoea and arthritis. It was identified that the caffeic acid derivatives present in the methanolic extract of the young, mature and old coconut mesocarp showed antioxidant property. Also, caffeoyl derivatives like chlorogenic acid (CGA) are strong antioxidants which inhibit the in vitro oxidation of Low
i _ .-..__ i- _:J_ /T r\r \ A „*.;««:—nu:n1 n/*fiwitw r»f motUannMn f*vtn\rt QVIHWC Qtrnncr

activity against S. aureus and an inhibition zone was formed against Bacillus sublilis. However, it shows no response,against E. coli and Pseudomonas aeruginosa. This study will definitely open up a scope for future utilization of these agro wastes for therapeutic purpose (Chakraborty M, Mitra A. The antioxidant and antimicrobial properties of the methanolic extract from Cocos nucifera mesocarp. Food Chemistry. 2008 Apr l;107(3):994-9).
A study evaluated the antibiofilm activity of coconut {Cocos nucifera linn.) husk fiber extract. The fibers are found between the husk and the fibrous outer shell (mesocarp) of a coconut. The individual fiber cells are narrow, shallow with thick walls made of cellulose. The aqueous extract of the coconut husk has anti-microbial activity. The results showed that the coconut husk extract was capable of inhibiting the growth of biofilm forming bacteria like Pseudomonas sp. Alteromonae sp. and Gallionelle sp. The coconut husk extract showed inhibitory activity on the growth of all the bacterial strains tested. The adhesion of bacterial cells to glass slides submerged in coconut husk extract medium was assessed using Pseudomonas sp. as target organisms. Coconut husk is similar to hard wood in chemical composition composed mainly of lignin and cellulose. It is a source of chemical compounds mainly phenolic compounds. During the retting process (soaking of husks in water) organic substances such as pectin, tannins and phenols may be liberated into the water. Phenolic compounds exhibit properties like anti-allergic, anti-atherogenic, anti-inflammatory, antimicrobial and vasodilatory effects. Coconut husk extract also affected the hydrophobicity of the biofilm forming bacterial strains. The physio-chemical theory for bacterial adhesion implies that a decrease in bacterial cell surface hydrophobicity may result in increased adhesion rate to hydrophilic surfaces, so, coconut husk extract . nnuM ho .used ng.an inhibitory compound to control biofilm development on surfaces

submerged in aquatic environments (Viju N, Satheesh S, Vincent SG. Antibiofilm activity of coconut (Cocos nucifera Linn.) husk fibre extract. Saudi journal of biological sciences. 2013 Jan 31;20(1):85-91).
The aqueous crude extract of husk fibre from Cocos nucifera is used in Brazilian folk medicine to treat arthritis and diarrhoea. Recently it has shown that it has anti¬bacterial, antiviral, antioxidant, antineoplastic and anti-inflammatory actions. In this study, anti-inflammatory action of aqueous extract of husk fibre from C. nucifera was assessed in animal models. The antioxidant study was evaluated by 2, 2-diphenyl-l-picryl-hydrazyl-hydrate photometric array. The extract was also screened for C. albicans, E. coli, S. aureus and methicillin resistant S. aureus by agar diffusion method. The minimum inhibitory concentration and minimum bactericidal concentration were determined by the broth microdilution assay. The extract showed anti-inflammatory action by reducing cell migration, protein extravasation and TNF-a production. Minimum inhibitory concentration and bacterial concentration found were 1.024(ig/ml and showed against S. aureus. They concluded that extract exhibits anti-inflammatory activity and mixture of extract constituents with methicillin can lead to the development of new anti-biotic against Multi resistant Staphylococcus aureus infection (Silva RR, e Silva DO, Fontes HR, Alviano CS, Fernandes PD, Alviano DS. Anti-inflammatory, antioxidant, and antimicrobial activities of Cocos nucifera var. typica. BMC complementary and alternative medicine. 2013 May 16;13(1):107).
A study was designed to find out anti-bacterial properties of aqueous extract of coconut husk against cariogenic bacteria. Herbal medicines are known to mankind and medicinal plants have been used in treating diseases. In coastal India husk of coconut is used as herbal chewing sticks instead of regular toothbrushes for brushing ^ 'thleirLteeui. The1 aa.u'eous^extract'oT husk of cbc6nut'was prepared"1 and subjected to
■1 ~l

antimicrobial evaluation against S. mutans, S. salivarius, S. mitis and L. acidophilus. The results showed a concentration dependent activity with zone of inhibition ranging from 4.44 to 15.33mm. The antibacterial activity depends on hydrophobic action, which enables them to partition the lipids of the bacterial cell membrane and mitochondria, disturbing the cell structure and rendering them more permeable. These results in extensive leakage from bacterial cells or the exit of critical molecules and ions leads to death. This study confirmed the beneficial effects of coconut husk with antimicrobial effect on cariogenic bacteria. As alcohol is a better solvent capable of extracting most active components further studies are recommended using alcoholic extracts (Cyriac MB, Pai V, Shantaram M, Jose M. Antimicrobial properties of coconut husk aqueous extract on cariogenic bacteria. Archives of Medicine and Health Sciences. 2013 Jul 1;1(2):126).
A study was carried out to find out antimicrobial properties of C. nucifera husk against oral microflora. The natural products derived from medicinal plants are of biologically active compounds. The coconut (C. nucifera) belongs to the Arecaceae family, is native to the coastal areas of South East Asia. When fibrous coconut husk is used for cleaning the teeth, it will mechanically remove the colonized microorganisms from the tooth surface. The present study was designed to find out antimicrobial action against oral pathogens. Alcoholic extract of the husk of C. nucifera was prepared and antimicrobial properties were found using agar well diffusion method. The action was more significant on cariogenic organisms and Candida as compared to periodontogenic. However, effect was lesser than chlorhexidine. The phytochemical screening of C. nucifera shows that it is rich in alkaloids, flavonoids, catechins and epicatechins along with tannins gives antimicrobial properties. Catechin is also
Jaitiduirarjd-cpiuui^hiiLJDhlbltod S. mutans =

by inhibiting glucoryltransferase [GTF]. C. nucifera has a significant inhibitory action against common oral pathogens so it can contribute to oral health to a great extent for control of oral diseases (Jose M, Cyriac MB, Pai V, Varghese I, Shantaram M. Antimicrobial properties of Cocos nucifera (coconut) husk: An extrapolation to oral health. Journal of natural science, biology, and medicine. 2014 Jul;5(2):359).
A study was designed to find out probable bactericidal mechanisms of husk extract of C. nucifera and fractions on bacteria isolates. As resistance to antibiotics is more common in clinical dentistry which is multi drug resistant (MDR) and these infractions are difficult to manage. So, in such cases medicinal plants are of great importance because of innate phytochemicals. One of the great advantages is availability and low side effects. Powdered husk of C. nucifera was cold extracted using mixture of methanol and distilled water. The minimum inhibitory concentration of ethyl acetate fraction was found to be more active than chloroform, n-butanol fractions. The MIC of extract ranged between 0.39 and 12.50mg/ml and those fractions between 0.16 to 0.5 mg/ml. A total of 27.8% organisms killed at IX minimum inhibitory concentration after 15minutes and 95% at 120 minutes. Leakage of potassium ions in these fractions show that it inhibits or kills the susceptible isolates. It increases as the fraction concentration increases which in turn led to the loss of nucleotides and damage to the cell membrane. The ability of C. nucifera extract to kill gram positive and gram-negative bacteria proved can be effectively used in multi-drug resistant infections (Akinpelu DA, Alayande KA, Aiyegoro OA, Akinpelu OF, Okoh AI. Probable mechanisms of biocidal action of Cocos nucifera Husk extract and fractions on bacteria isolates. BMC complementary and alternative medicine. 2015 Apr 14; 15(1):116).

Though there are reports in the literature to testify the efficacy of natural various alternatives for medical and dental applications but none of the reports on gel formulation with ethanolic extracts of husk of Cocos nucifera for various dental and medical applications. Thus, the present inventors have developed the effective gel formulation with ethanolic extracts of husk of Cocos nucifera and compared and evaluated the antibacterial effectiveness of ethanolic extracts of husk of Cocos nucifera and against the Enterococcus faecalis, Prevotella intermedia and Porphyromonas gingivalis. Also, the developed the gel formulation of Cocos nucifera as pulpotomy agent, healing gel, for ulcer and extracted wound, as a local drug delivery after conventional scaling and root planning. Cocos nucifera extract gel is very useful as an adjunct to conventional scaling and root planning.
OBJECTIVES OF THE INVENTION
The primary object of the present invention is the development of gel formulation with extracts of Cocos nucifera.
Object of the present invention is the development of gel formulation with ethanolic extracts of husk of Cocos nucifera.
Another object of the present invention is the development of gel formulation with ethanolic extracts of husk of Cocos nucifera against the Enterococcus faecalis, Prevotella intermedia and Porphyromonas gingivalis.
Another object of the present invention is the development of gel formulation with ethanolic extracts of husk of Cocos nucifera as pulpotomy agent, healing gel, for ulcer and extracted wound, as a local drug delivery after conventional scaling and root planning.

Another object of the present invention is the development of process for preparation of gel formulation with ethanolic extracts of Cocos nucifera.
A further object of the invention is to develop the gel formulation with naturally existing agent such as Cocos nucifera, which do not produce any undesirable by-products.
A further object of the invention is to develop the gel formulation with ethanolic extracts of husk of Cocos nucifera, which are safe and practical to use with little technical expertise.
It is a further object of the present invention to develop the gel formulation with ethanolic extracts of husk of Cocos nucifera having a long shelf life.
A further object of the invention is to develop the gel formulation with ethanolic extracts of husk of Cocos nucifera, which are inexpensive and cost effective as compared to the existing formulations.
STATEMENT OF THE INVENTION
Gel formulation of Cocos nucifera for medical and dental applications comprising the effective concentration of extract of Cocos nucifera complexed with p- cyclodextrin and other excipients. The extract is the ethanolic extract of Cocos nucifera from the husk. The ratio of the complexed ethanolic extract of husk of Cocos nucifera and p-cyclodextrin is 1:2. The effective concentration of the complex of ethanolic extract of husk of Cocos nucifera and p- cyclodextrin is 1 to 20% (w/w) and preferably 2% (w/w). The excipients are selected from Carbopol 934 (CRB), Triethanolamine, Sodium Saccharin.

Process for preparation of Gel formulation of Cocos nucifera for medical and dental applications comprising:
a) preparation of ethanolic extract of husk of Cocos nucifera;
b) ethanolic extract of step (a) is complexed with (3- cyclodextrin; and
c) complex formed at step (b) at effective concentration is then mixed with excipients to form the uniform gel formulation of Cocos nucifera.
The concentration of the complex of ethanolic extract of husk of Cocos nucifera and (3- cyclodextrin is 1 to 20% (w/w) and preferably 2% (w/w) in the process. The excipients are selected from Carbopol 934 (CRB), Triethanolamine, Sodium Saccharin in the process for gel formulation of Cocos nucifera.
BRIEF DESCRIPTION OF THE FIGURES/DRAWINGS
Figure 1. Flow chart detailing the action of the extracts of Cocos nucifera (Ahmad A, Kaleem M, Ahmed Z, Shafiq H. Therapeutic potential of flavonoids and their mechanism of action against microbial and viral infections—A review. Food Research International. 2015 Nov 30;77:22l-35.)
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to the development of gel formulation with extracts of
Cocos nucifera. It specifically relates to the development of gel formulation with
ethanolic extracts of husk of Cocos nucifera against the Enterococcus faecalis,
Prevotella intermedia and Porphyromonas gingivalis and as pulpotomy agent, healing
gel, for ulcer and extracted wound, as a local drug delivery after conventional scaling
and root planning. The invention also pertains to the development of process for

preparation of gel formulation with ethanolic extracts of Cocos nucifera against the Enterococcus faecalis, Prevotella intermedia and Porphyromonas gingivalis. The study was conducted in the Department of Pedodontics and Preventive Dentistry at KLE University's KLE VK Institute of Dental Sciences, Belagavi and KLE University's Dr. Prabhakar Kore Basic Science Research Center, Belagavi. Outpatients with the inclusion criteria reporting to the Department of Pedodontics & Preventive Dentistry and Department of Oral and Maxillofacial surgery for the purpose of extraction of primary and permanent teeth at the KLE University's VK Institute of Dental Sciences, Belagavi were selected for the study. Ethical clearance for the study was obtained from the Institutional review board of the college.
SOURCE OF DATA:
The study was conducted at Department of Pedodontics and Preventive Dentistry, KLE University's KLE V.K. Institute of Dental Sciences, Belagavi.
The active component of any natural product has to be extracted using a solvent in which the compounds of interest are dissolvable. The most commonly used solvents for extraction of the components
WORKING EXAMPLES:
PREPARATION OF EXTRACT OF COCOS NUCIFERA:
The extract was prepared in the Department of Pharmaceutics, College of Pharmacy, KLE University, Belagavi the same is disclosed and claimed in our Co-pending application which is incorporated here for reference only.
Preparation of ethanolic extract from husk of Cocos nucifera:

The husks of Cocos nucifera used in the study was obtained and verified from the KLE University's Shri. BMK Ayurveda Mahavidyalaya, Belagavi and Regional Medical Research Center (RMRC) branch of Indian Council of Medical Research (ICMR), Belagavi.
The husk was dried in hot-air oven at 45°C until a constant weight of the sample is reached. This was ground into fine powder using a husk grinder available in the local market. Exactly 1.5kg of the powdered sample was extracted with ethanol in ratio of 3:2 (v/v) for four days. The supernatant collected was filtered into clean sterile dried conical flask. The filtrate was concentrated in vacuo and lyophilized. The yield obtained was obtained was dark brown in colour and weighed 110.5grn.
According to the preliminary study carried out with the four extracts obtained, it was found that ethanolic extract of husk of Cocos nucifera showed best results for Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) against Porphyromonas. gingivalis, Prevotella intermedia and Enterococcus faecalis as disclosed in our co-pending application which is incorporated here for reference only.
Method of Preparation of Cocos nucifera 2% gel (w/w)
Weighed quantity of (3- cyclodextrin is complexed with Cocos nucifera ethanolic extract by wet kneading method in pestle and mortar for ten minutes. The kneaded mass was triturated in a beaker with addition of 5 ml water. Weighed quantity of Carbopol (CRB) 934 powder was added slowly to the above solution with constant stirring on magnetic stirrer. Stirring was continued for 3 hours to get homogenous solution. The solution was kept for overnight for uniform dispersion. Sodium
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saccharin was added and stirred for half an hour. The final solution pH was adjusted to 6.8 to 7 using Trithanolamine to get a uniform gel.

The Cocos nucifera husk extract gel developed in the present invention which can be used as gel for various medical and dental applications:
1) 2% Cocos nucifera husk extract gel can be used for chemo-mechanical preparation
in endodontics procedures.
2) 2% Cocos nucifera husk extract gel can be used as a pulpotomy agent.
3) 2% Cocos nucifera husk extract gel can be used as a gel for ulcer, mucositis.
4) 2% Cocos nucifera husk extract gel can be used for healing of extraction wound
after extraction of teeth.
5) 2% Cocos nucifera husk extract gel can be used as a local drug delivery after
scaling and root planning.

The developed invention gel formulation has shown other properties like analgesic, anti-inflammatory, antifungal, antiviral, antioxidant, anti-neoplastic, anti-parasitic, anti leishmania and antimalaria.

We Claim,
1. Gel formulation of Cocos nucifera for medical and dental applications comprising the effective concentration of extract of Cocos nucifera complexed with p-cyclodextrin and other excipients.
2. Gel formulation of Cocos nucifera as claimed in claim 1 wherein the extract is the
ethanolic extract of Cocos nucifera.
3. Gel formulation of Cocos nucifera as claimed in claim 1 or 2 wherein the extract
is the ethanolic extract of husk of Cocos nucifera.
4. Gel formulation of Cocos nucifera as claimed in claim 1 wherein the ratio of the
complexed ethanolic extract of husk of Cocos nucifera and p- cyclodextrin is 1:2.
5. Gel formulation of Cocos nucifera as claimed in claim 1 wherein the effective concentration of the complex of ethanolic extract of husk of Cocos nucifera and p-cyclodextrin is 1 to 20% (w/w).
6. Gel formulation of Cocos nucifera as claimed in claim 5 wherein the effective concentration of the complex of ethanolic extract of husk of Cocos nucifera and p-cyclodextrin is preferably 2% (w/w).
7. Gel formulation of Cocos nucifera as claimed in claim 1 wherein the excipients are selected from Carbopol 934 (CRB), Triethanolamine, Sodium Saccharin.

8. Process for preparation of Gel formulation of Cocas nucifera for medical and dental applications comprising:
a) preparation of ethanolic extract of husk of Cocos nucifera;
b) ethanolic extract of step (a) is complexed with p- cyclodextrin; and
c) complex formed at step (b) at effective concentartion is then mixed with excipients to form the uniform gel formulation of Cocos nucifera.

9. Process as claimed in claim 8 wherein the concentration of the complex of ethanolic extract of husk of Cocos nucifera and p. cyclodextrin is 1 to 20% (w/w) and preferably 2% (w/w).
10. Process as claimed in claim 8 wherein the excipients are selected from Carbopol 934 (CRB), Triethanolamine, Sodium Saccharin.