“EVALUATION OF LEAF EXTRACTS OF “XIMENIA AMERICANA” LINN.FOR WOUND HEALING ACTIVITY ON ALBINO RATS”
SYNOPSIS FOR REGISTRATION
Of
M.PHARM DISSERTATION
Submitted toRAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,
KARNATAKA
By
PRIYANKA SINGH
Ist M.PHARMACY
Under the Guidance of
Dr.B.Rajkapoor
Department Of Pharmacology
DAYANANDA SAGAR COLLEGE OF PHARMACY
2009
ANNEXURE - II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
1. / Name of the Candidateand Address /
Priyanka Singh
HIG.-16 ,Prem Nagar, Near Trivatinath Temple,BDA Colony, Dharmkanta, Barriely (U.P.)
2. / Name of the Institution / Dayananda Sagar College of Pharmacy,
Kumaraswamy Layout,
Bangalore – 560 078
3. / Course of Study and
Subject / M. Pharmacy -Pharmacology
4. / Date of Admission / 22/05/2009
5. Title of the Topic:
“Evaluation Of Leaf Extracts Of Ximenia Americana LINN. For Wound Healing Activity on Albino Rats”.
6.0 / Brief resume of the intended work:
6.1 – Need of the study:
A wound can be defined as a break in the continuity of the soft tissues like skin, mucous membranes, tissue surface etc. caused by physical, chemical or biological insult. It can also called as traumatic lesion .It can be classified as-
1. External Wound
2. Internal Wound
* An External wound is one with a varying degree of damage to the tissue including skin e.g. incised wounds, lacerated wounds, punctured wounds, penetrating wounds.
* An Internal wound damages the underlying tissue to varying degree having the skin intact. e.g. contusions, brusies and hematomas 1.
Wound infections are most common in developing countries because of poor hygienic conditions .staphylococcus aureus, streptococcus pyrogens, corynebacterium spp. pseudomonas aeruginosa and E. coil 2.
· Acute wound healing occurs in 4 stages namely haemostasis, inflammation, proliferation and remodeling. The majority of such investigations take the form of in vitro assays based on cell culture models of the various phases of healing.
· In chronic wound the efficacy of many advanced therapeutic agents has been less then what had been predicted from in vitro studies, e.g. in bedridden and diabetic patient 3 .
The use of medicine of plant origin has been in practice for well over 5000 years. The plants or chemical entities derived from plants need to be identified and formulated for treatment and management of wounds4.
In this direction a number of herbal products are been investigated at present so “Ximenia Americana plant in different models induced in albino rats”5.
6.2 –About the plant :
An ethno pharmacological survey of six medicinal plants “Ximenia Americana” Linn. Family- Olacaceae is a variable shrubby tree up to 5m high. It is found in Senegal to west Cameroon and is also widely distributed in tropical Africa, America and Asia. In India it is found in Tamilnadu, Madurrai district.6
In Mali (South Africa) wounds are treated traditionally with Ximenia leaves or bark or by applying powder. It is used in dysmenorrhoea, ailments, febrifugal schistosomiasis7, 8.
With Annona chrysophylla it is used in the treatment of sleeping sickness and in insect sting9. It has anti-trypanosomal activity10 itself. Widely used in gum trouble11. It shows antiviral12, anticancer13 and antimicrobial14 activity. Roots are used in fever, guinea worm infection, leprosy, jaundice, and stomatitis etc.
Plant contains volatile oils15 in leaves. The major constituents are benzaldehyde (63.5%), hydroxybenzyl cyanide (13.00%), and isophorone (3.5%). In plant saponins, cyanogenetic glycosides, flavonoides and tannins are present16. It is a edible plant which is used in skin care and eye care due to presence of ximenyic acid.
Based on above chemical constituents the Ximenia americana which is belongs to Olacaceae family used in wound healing due to presence of tannins and flavonoides.
6.3 – Review of the literature:
1. Sabah Handoo (2006) reported Survey of plants used for wound healing in animals
2. Muthusamy Senthil Kumar, Ramasamy Sripriya, Harinarayan vijaya Raghavan and Praveen Kumar Sehgal (2006), reported wound healing potential of Cassia fistula on infected albino rat model.Cassia fistula treated rats showed better wound closure, inprooved tissue regeneration at the wound site.
3. P.Krishnan (2006) reported scientific study of herbal wound healing therapies. The investigation of wound healing herbal therapies should be directed toward clinical trials. For the purpose of the joint demonstration of efficacy and elucidation of drug metabolism.
4. Qidwai W, Waheed S, Ayubs, Syed, 1-A (2009) reported Indian journal for the practicing doctors.They found widespread use of honey for a variety of medical condition.
5. P.K.Verma and N.K.Pankaj (2008) reported medicinal plants and their role in wound healing. Plants and their extracts have immense potential for management and treatment of wounds. The natural agents induce healing and regeneration of the lost tissue by multiple mechanism.
6. Tom Erik Gronhaug, Silje Glaeserud, Mona Skogsrud, Ngolo Ballo, Sekau Bah, Drissa Diallo and Berit Smestad Paulsen (2008) reported Ethnopharmacological survey of six medicinal plants from Mali, West- Africa. More than sixty medical indication are reported for the use of these plants in traditional medicine. The most frequently used plants part is leaves and stem bark.
7. Diallo D, Sogn C, Samake FB, Paulsen BS, Michaelson TE Keita (2002) reported wound healing plants in Mali the Bamako region. According to the literature the plant is well known for its medicinal properties and wounds are treated with a decoction of leaves or a combination of a wash with a decoction of bark.
8. Inngjerdingen K, Nergard Cecilie S, Diallo D, Mounkoro Pakuy P, Paulsen Berits (2004) An Ethnopharmacological survey of plants used for wound healing in Dogonland, Mali, West-Africa. According to the literature the aim to identify medicinal plants used in the treatment of wounds. Plants used as first aids, in the washing of wounds, extraction of pus, as coagulants as well as for infected wounds.
9. Burkil HM (Ed) (1997) reported the useful plants of west tropical Africa. The root bark is put into febrifugal medicine and feed in boiled water with crushed bark. The plant is used in schistosomiasis and with Anona in sleeping sickness.
10. Maikai V.A., Nok J.A., Adaudi A.O and Klawa (2008) reported an in-vitro antitrypanosomal activity of aqueous and methanolic crude extracts of stem bark of Ximenia americana on Trypanosoma congolense.
11. S Ganesan (2008) reported traditional oral care medicinal plants survey of Tamilnadu. An oral care medicinal plants survey was conducted in different district of Tamilnadu during the period of 2004-08.Most of plant used to relief tooth ache, tooth brush and common dental diseases.
12. Asresk, Bucar F, Kartnig T, Witvrovw M, Pannecouque C, De Clercq E (2001) reported an antiviral activity against HIV-1 and HIV-2 of ethnobotanically selected Ehiopian medicinal plant.
13.Voss C, Eyol E, Frank M, Vorder Lieth, Berger MR (2006) reported identification and characterization of ripoximin a new type-ц ribosome-inactivating protein with antineoplastic activity from Ximinea Americana.
14. Omer MEFA, Elinima EI (2003) reported an antimicrobial activity of Ximenia Americana. According to the literature extract of the bark, leaves, root and stem of Ximenia americana were tested for their antimicrobial and antifungal activity.
15. Jean Philippe Mevy, Jean Marie Bessiere, Stephane Greff, Gerard Zombre and Josette viano (2006) reported the composition of volatile oil in leaves of Ximenia Americana. Volatile oil of leaves was analysed by GC.MS. Major constituent identified were Benzaldehyde (63.5%) , Hydroxybenzylcyanide(13%) and Isophorone (3.5%). The occurance of Hydroxybenzylcyanide as well as Benzaldehyde was discussed in this paper.
16. D.S.Ogunleye and S.F. Ibitoye (2003) reported an antimicrobial activity and chemical constituent of Ximenia Americana. The leaves extract was active against the test organism E. coli. , Pseudomonas aeruginosa and Candida albicans. Tannins, Flavonoides, Alkaloides, Saponins, Quinones, Starch and Glycoside are present.
6.4 – Objective of the study:
The main objective of the study was to evaluate wound healing and phytochemical investigation of leaf extracts of Ximenia americana
The work consists –
A. Collection and authentification of leaves of Ximenia americana Linn.
B. Extraction of dried leaves with ethanol in a soxhlet apparatus:
The leaves will be shade dried and powdered mechanically. Powdered materials were subjected to successive extraction with petroleum ether, chloroform, ethanol and distilled water in the increasing order of polarity in a soxhlet apparatus.
C. Preliminary phytochemical screening of crude extracts:
The crude extract will be subjected to preliminary phytochemical screening for the presence of carbohydrates, flavonoids, tannins and cyanogenetic glycosides.
D. Toxicity studies:
E. Evaluation of wound healing activity by following methods:
1. Excision Method: In this method rats are inflicted with excision wounds under light anesthesia. A circular wound of about 500 mm2 made on depilated ethanol sterilized dorsal thoracic of rats. The animals are divided into 6 groups of 6 animals each. The group 1 will consider as control and group 2 served as reference standard. Group 3rd, 4th, 5th and 6th animals are received plant extract. The wounds are trace on mm2 graph paper on 4th, 8th , 12th and 16th day and there after on alternative days until healing will completed.
2. Incision Method: In this method two para vertebral straight incision of 6 cm. each will be made through the entire thickness of the skin on either side of the vertebral column with the help of surgical blade anaesthetized animals. The animals are divided into 6 groups of 6 animals each. The group 1 will consider as control and group 2 served as reference standard. Group 3rd, 4th, 5th and 6th animals are received plant extract. Removal of sutures will be done on 8th post wounding day and tensile strength of the healed wound will measured on 10th post wound day.
3. Dead Space Methods: In this method animals are divided into 5 groups of 6 rats in each group. The groups 1 will be considered as control. The animals of group 2nd ,3rd ,4th and 5th received plant extracts respectively. Under light anesthesia dead space wound are created by subcutaneous implantation of sterilizes cylindrical grass piths, one on either side of the dorsal para vertebral surface of rat.
F. Histological studies: Histopathological study evaluates the effect of the extract on collagen formation.
G. Statistical analysis: The data will subjected to one-way ANOVA followed by Tukey’s multiple comparision test.
Materials and methods:
7.1-Source of data:
Journal of Ethno pharmacology, Indian journal of forestry, International journal of pharmacology, Indian journal of experimental biology, Complementary and alternative medicine, Phytotherapy research, Fitoterapia, Indian journal of physiology and pharmacology
Standard books:
Indian medicinal plants by Kirtikar KR, Basu BD
Indian Materia Medica
Experimental pharmacology by M.N.Ghosh
Pharmacographica Indica
Medicinal plants research
7.2-Method of collection of the data:
The plant will be collected from Andhra Pradesh, India. Authenticated by Dr. K.Madhavachetti, Assistant Professor and botanist from Sri Venkateshwara University Tirupathi. All drugs, chemicals and solvents used in the study would be procured from reputed manufacturers like Merck, BDK.
7.3:Does the study require any investigations or interventions to be conducted on
Patients or other humans or animals? If so, Please describe briefly.
Yes, the study requires investigation on albino rats.
7.4: Has ethical clearance been obtained from your Institution in case of 7.3?
Yes, Approved by CPCSEA.
8.0 / List Of Reference:
1. Handoo Sabah, Survey of plants used for wound healing in animals. Indian Vet J 2006;1(1)
2.Kumar Senthil Muthusamy , Sripriya Rmaswamy, Raghavan Vijaya Harinarayan and Sehgal Praveen Kumar, wound healing potential of Cassia fistula on infected albino rat model. J.Surgical Res 2006; 131:283-289.
3. Krishnan P., scientific study of herbal wound healing therapies. Current Anaesthesia and critical care 2006; 17:21-27.
4. Qidwai W, Waheed S, Ayubs, Syed, 1-A,Medicinal Use Of honey: Result of a survey of patient visiting family doctors at a teaching hospital, Indian journal for the practicing doctors.2009;5:6.
5. Verma P.K. and Pankaj N.K., medicinal plants and their role in wound healing. Vet J 2008;3(1).
6. Gronhaug Erik Tom , Glaeserud Silje, Skogsrud Mona, Ballo Ngolo, Bah Sekau, Diallo Drissa and Paulsen Smestad Berit, Ethno pharmacological survey of six medicinal plants from Mali, West- Africa. J Ethnobiol and Ethno med 2008; 4:26.
7. D Diallo, C Sogn, FB Samake, BS Paulsen, Keita TE Michaelson, wound healing plants in Mali the Bamako region. Pharmaceutical Biol 2002; 40:117-128.
8. K Inngjerdingen, S Cecilie Nergard, D Diallo, P Pakuy Mounkoro, Berits Paulsen, An Ethnopharmacological survey of plants used for wound healing in Dogonland, Mali, West-Africa. J Ethno Biol and Etnomed 2004; 92:233-234.
9. HM Burkil (Ed), The useful plants of west tropical Africa. Royal Botanic Gardens1997; 4(2).
10. V.A Maikai., J.A.Nok, A.O Adaudi and Klawa, An in-vitro antitrypanosomal activity of aqueous and methanolic crude extracts of stem bark of Ximenia americana on Trypanosoma congolense. J Med Plants Res 2008;2(3):055-058.
11. Ganesan S, Traditional oral care medicinal plants survey of Tamilnadu. Natural Product Radiance 2008; 7(2):166-172.
12. Asresk, F Bucar, T Kartnig, M Witvrovw, C Pannecouque, E Clercq De ,An antiviral activity against HIV-1 and HIV-2 of ethnobotanically selected Ehiopian medicinal plant. Phytother Res 2001; 15:62-69.
13.C Voss, E Eyol, M Frank, Lieth Vorder, MR Berger , Identification and characterization of ripoximin a new type-ц ribosome-inactivating protein with antineoplastic activity from Ximinea Americana. J Federation of American societies for Exp Biol 2006; 20:1194-1196.
14 MEFA. Omer, EI Elinima, An antimicrobial activity of Ximenia Americana. Fitoter 2003;74(1-2):122-126
15. Mevy Philippe Jean, Bessiere Marie Jean, Greff Stephane, Zombre Gerard and viano Josette, The composition of volatile oil in leaves of Ximenia Americana. Biochem Systematic and Ecology 2006;34(7):549-553.
16. Ogunleye D.S. and Ibitoye S.F. An antimicrobial activity and chemical constituent of Ximenia Americana.Trop J Pharm Res 2003; 2(2):239-241.
17. P. Umachigi Sanjay, Wound healing activity of medicinal plants formulated in pluronic F 127 in rats.Pharmainfo.net 2009;15:36.
18. Kumar. B, Vijaykumar. M, Govindarajan. R, Pushpangadan. P, Ethno pharmacological approaches to wound healing exploring medicinal plants of India. J Ethnopharmacol 2007; 114:103-113.
19. Reddy Suresh J, Rao Rajeswara. P, Reddy Mada S, Wound healing effects of Heliotropium indicum, Plumbago zeylanicum and Acalypha Indica in rats. J Ethnopharmacol 2002; 79:249-251.
20. Jalalpure Sunil S, Agrawal Nitin, Patil MB, Chimkode. R, Tripathi Ashish, Antimicrobial and wound healing activity of Alternanthera sessilis. Intrnational J Green Pharmacy 2008; 2(3):141-144.