Effect of Wound Healing Potential of Plumeria obtusa (Champa) Spray

 

Abhishek R. Bura*

Dr. Vithalrao Vikhe Patil Foundation’s College of Pharmacy, Vilad Ghat, Ahmednagar- 414001, Maharashtra, India.

*Corresponding Author E-mail: abhishekbura007@gmail.com

 

ABSTRACT:

Plumeria obtusa is commonly known as Champa (Hindi), White Frangipani (Australia) belonging to family Apocynaceae. The aim of this project is to study the wound healing activity of ethanolic extract of Plumeria obtusa leaves spray. The wound healing activity was studied by the in-vivo method. In Vivo method by excision model the % wound closure rate on the epidermal skin of white Wistar albino rats was studied and was compared with standard Hansaplast spray. In this animals were divided into 4 groups of 5 each. Group 1 was left untreated and considered as control. Group 2 was treated with the standard Hansaplast spray. Group 3 was treated with 5% test Plumeria obtusa spray and group 4 was treated with 10% test Plumeria obtusa spray. In the incision wound model, the tensile strength or skin breaking strength was measured and studied. The tensile strength was measured on the 10th post-wounding day and compared to the control group. The studies carried out in these both experimental models shows that 10% Plumeria obtusa extract spray shows better and efficient wound healing activity than 5% Plumeria obtusa extract spray. The futures expect is to isolate and study the activity of specific chemical constituent responsible for wound healing activity from the Plumeria obtusa leaf extract.

 

KEYWORDS: Champa, Plumeria obtusa, Wound contraction rate, Tensile strength, Extract.

 

 


INTRODUCTION:

Wound healing is the process by which skin or other body tissue repairs itself after trauma. Wound healing is muchorganized and highly precise process characterized by four distinct phases. I.e. Haemostasis, Inflammation, Proliferation, and Maturation orRemodelling.2 The repair process needs the coordination of various cells, growth factors and cytokines12. Plumeria belongs to family Apocynaceae and has a wide family of about 300 Genera with more than 1400 Species.

acutifolia and Plumeria alba.3

 

This plant is mainly grown for its ornamental purpose and fragrant flowers. The commonly known species having medicinal importance are Plumeria Rubra, Plumeria obtusa, Plumeria Plumeriaobtusa is commonly known as Champa (Hindi), Champagne (Konkani), Arali (Tamil), Kathgolop (Bengali) in India and White Frangipani (Australia), Melia (Hawaii), Araliya (Sri Lanka), Temple Tree (United Kingdom), Hong Ji Dan Hua (China). Plumeria obtusa is native to the Bahamas and the Greater Antilles in Central America. They are Prolific in Hawaii16.

 

Plumeria obtusa is used in traditional medicine as healers to diabetes mellitus. Leaves are used as purgative, treat a headache, wounds. Roots are used in the treatment of skin and liver diseases, leprosy, tumours, ulcers, etc.4 the latex of bark known to be purgative and diuretic property. Flowers are used for the ornamental purpose, perfumes; etc.1the extract of leaves contains the following phytochemicals flavonoids, tannins, glycosides, cardiac glycosides, alkaloids, saponins, terpenoids, phenols, etc present.5 Aerosols are a new drug delivery system in wound healing which minimizes the efforts and discomforts caused to the person and hence increases their acceptance and patient compliance. Sprays are easy to use and cause lesser contamination and discomfort as that of dressing16. The aim of this research is to study the wound healing property of Plumeria obtusa leaves extract spray.

 

MATERIAL AND METHODS:

Plant Material Collection:

Fresh leaves of plant Plumeria obtusa were collected in the month of September 2016. The leaves were authenticated from the Radhabai Kale Mahila Mahavidyalaya Ahmednagar. The leaves were washed with water and shade dried at room temperature. The dried leaves were then ground in local grinder machine. The powder was then subjected for further process.

 

Preparation of Extract:

The extraction process was carried out by the maceration technique. About 100gm of ground leaf powder was soaked in 600ml of ethanol for not less than 7 days at room temperature. The mixture was shaken after every 5 hours during this period. After 7 days the powder mixture was filtered using Whatman filter paper grade 1 and extract was evaporated with rotary vacuum evaporator at 60oC. The percentage yield of the extract was calculated and is showed in table no 1. Phytochemical tests were carried out for the extract and showed flavonoids, glycosides, cardiac glycosides, phenols, tannins, terpenoids present.16

 

Preparation of spray:

The Plumeria obtusa spray was prepared in two formulations- 1st formulation contains Plumeria obtusa 10% extract as an active ingredient and 2nd formulation contains Plumeria obtusa 5% extract as an active ingredient. The solvent used in the preparation of both sprays is ethanol. Both formulations also contain propylparaben added as preservative quantity sufficient and Rose oil used as a perfume. The formulation was then filled into a suitable container and caped.6,10,11

 

Experimental Animals:

Swiss albino Wistar rats of either sex weighing about (200-250gm) were used for animal studies for wound healing. The experimental protocol and procedure were approved by the Committee for the purpose of control and supervision of experiments on animals (CPCSEA) by Ref no. Dr.V.V.P.F's COP/IAEC/Avish/2016/1. Animals were divided into four groups of five each. The animals were housed in proper polypropylene cages under controlled conditions of temperature (20-250C) and were given normal food and diet.

 

In Vivo models:

Excision Wound Model:

The animal was anesthetized by using ketamine hydrochloride inj. (10mg/100g BW-IP). Half an hour prior to administration of ketamine inj. hairs of animals were removed about 2X2 cm square from the back of rat which has been untouched before. The wound was then made on the shaved back of rat about 350mm square and 2mm deep 21. The wound area was cleaned with cotton soaked in water12, 13 the animals were closely observed for any bacterial infection and if observed they were separated.1The animals were divided into four groups of five each. Group 1 was considered as Control. Group 2 was treated with Standard drug Hansaplast spray. Group 3 was treated with Plumeria obtusa spray containing 5% extract and group 4 was treated with Plumeria obtusa spray containing 10% extract. The Plumeria obtusa spray and Standard drug i.e. Hansaplast spray was applied daily on the wound area till complete healing was observed. The wound area was measured on the days 3, 7, 11, 16 and 1718. The percentage of wound contraction/closure rate was calculated by the formula-

% of Wound closure= (Initial wound size – Specific day wound size)/ Initial wound size.

 

Incision Wound Model:

The rats were anesthetized with ketamine hydrochloride inj. (10mg/100g BW-IP). The hairs on its back were removed by electric clipper. Paravertebral incision of about 5cm long was made on the skin of its back leaving about 1.5 cm2 from either side of the back of the rat. The wound was then cleaned with cotton soaked in normal saline. After hemostasis wound was closed by giving intermittent sutures about 1cm apart with help of surgical thread and curved needle no 16. The spray was applied twice daily on the wound area for about 8 days. The sutures were removed on the 9th post wounding day. The skin breaking strength or the tensile strength was estimated on the 10th postwounding day by the method described by ‘Ehrlich and Hunt'7, 8.

 

Formula- Tensile strength=

Total breaking load/ Cross sectional area.

 

RESULT:

In both models and experimental studies, the wound healing property of the leaf extract of Plumeria Obtusa was estimated for its potency. Table No. 1 shows the wound contraction rate and the period required by different groups of experimental animals for complete epithelisation of the wound by the excision wound model. The wound contraction/closure rate was measured on the 3rd, 7th, 11th and 16th day. The same is also shown in the photos. According to the results, complete wound healing was observed on day 21 in the control group and day 19 by application of 5% Plumeria obtusa extract spray. The complete wound healing was observed on day 17 by 10% Plumeria obtusa extract spray and the results were compared to the standard i.e. Hansaplast spray which showed complete wound healing on day 16.


 

 

 

 

 

0-day

3rd day

7th-day

 

 

11th-day

16th day

Figure 1- Effect of 10% Plumeria obtusa extract spray on the excision wound model in rats measured on 0, 3rd, 7th, 11th and 16th day.

 

 

 

 

Table no 1- Effect of Plumeria obtusa sprays on wound contraction/closure rates and epithelization period in excision wound model.

Groups

% of Wound healing

Period of Epithelization

(Days)

3rd day

7th day

11th day

16th day

Control

14.87±2.04

31.62±1.89

55.16±1.21

77.28±1.37

21

P. obtusa 5% Spray

23.38±2.24*

41.61±1.92*

62.57±1.34**

81.19±1.62**

19

P. obtusa 10% Spray

28.42±2.17*

47.6±2.06**

69.91±1.57*

93.57±1.46*

17

Hansaplast Spray

30.23±1.91*

51.05±2.13**

76.66±1.23*

99.33±1.74**

16

Values are expressed as mean ± SE, (n=5 animals), where * P= <0.05 and ** P= <0.01 when compared to control.

 

 

Fig 2- Wound contraction or closure rate at 3, 7, 11 and 16 days of different groups of animals treated by the Plumeria obtusa test 5%, 10% spray and standard Hansaplast spray. Values are mean of ± SE, (n=5 animals), where * P= <0.05 and ** P= <0.01 when compared to control.


The wound healing effect of Plumeria obtusa sprays in incision wound model is shown in table no 2. The tensile strength of the skin was measured on the 10th post wounding day. The skin breaking strength was found to be 385gm by application of 5% Plumeria obtusa extract spray and 403gm by the application of 10% Plumeria obtusa extract. The 10% Plumeria obtusa extract spray showed significant skin breaking strength as compared to the results shown by the control group.

 

Table no 2- Effect of Plumeria obtusa sprays on the tensile strength and epithelization period in incision wound model.

Group

Epithelization period (days)

Tensile strength (g)

Control

17.12

330±2.74

P. obtusa 5% Spray

14.68

385±3.8

P. obtusa 10% Spray

12.34

403±3.54*

Std

11.2

435±2.14*

Values are expressed as mean ± SE, (n=5 animals), where * P= <0.01 when compared to control.

 


 

Fig 3- Tensile strength of different groups of animals treated by the Plumeria obtusa test 5%, 10% spray and standard Hansaplast spray. Values are mean of ± SE, (n=5 animals), where * P= <0.01 when compared to control.

 

 

Table no 3- Isolation of crude extract from the leaves of Plumeria obtusa by maceration process.

A weight of powder (gm)

A solvent used for extraction (ml)

An extract obtained (gm)

% yield of crude extract

P. obtusa leaves 100 gm

Ethanol- 600 ml

13.46

13.46%

 

 


DISCUSSION:

Wound occurs when the integrity of any tissue is compromised and wound healing is a general repair response or process of the body immediately after the disruption of the skin integrity. Wound healing is a physiological process and generally does not require much help in treating but still wounds cause distress and are susceptible to infection or may lead information on chronic wounds. Whereas some diseases like diabetes, ischemia, local infection, malnutrition, and aging also causes a delay in wound healing process. Hence the use of further wound healing modulators or agents which facilitate the healing process is indicated.19, 20, 21

 

In our present study, we evaluated the wound healing activity of ethanolic extract of leaves of Plumeria obtusa and also compared the wound healing potency of 5% and 10% Plumeria obtusa extract spray by in vivo studies in both excision and incision models. The results obtained were compared with the standard marketed Hansaplast spray. Our aim was to formulate an herbal wound healing spray and evaluate its activity for checking the potency of the formulation.14, 24

 

The preliminary phytochemical analysis of Plumeria obtusa shows the presence of tannins, flavonoids, saponins, and terpenoids present. These phytochemicals act by different mechanisms as tannins, saponins facilitate wound healing by an antimicrobial and astringent activity whereas flavonoids possess potent antioxidant and free radical scavenging activity and promote wound healing activity. Plumeria obtusa leaf extract also possesses antibacterial activity and this also, in turn, may facilitate the wound healing process.22, 23

 

ACKNOWLEDGMENT:

The authors are thankful to the Dr. Vithalrao Vikhe Patil Foundation's College of Pharmacy, Vilad Ghat, Ahmednagar, for providing all the necessary facilities, chemicals and support to carry out this research work, also Prof. Nilesh Mhaske Sir for their Guidance.

 

REFERENCES:

1.         N. Ali, M Junaid, D. Ahmad, M. U.Rahman, G. Katzenmeier, Antibacterial and Antifungal activity of solvent extracts from Plumeria obtusa Linn. Trop. Biomedicine. 2014; 31: 607-615.

2.         All Things Frangipani, 2016. Frangipani Species. http://www.allthingsfrangipani.com/frangipani-species.html/. 2016

3.         M. Choudhary, V. Kumar, S. Singh, Phytochemical and Pharmacological activity of Genus Plumeria: An updated review. Int. J. of Biomed. and Advance Res. 2014; 5: 266-271.

4.         S. Demirci, A. Dogan, Y. Demirci, F. Sachin, In vitro wound healing activity of methanol extract of Verbascum specious. Int. J. of Applied Res. in Nat. Prod. 7: 37-44.

5.         Devprakash, R. Tembare, S. Gaurav, S.G.P. Kumar, T.T. Mani, An review of Phytochemical constituents and Pharmacological activity of Plumeria species. Int. J. of Curr. Pharmaceutical Res. 2012; 4: 1-6.

6.         H.P. Ehrlich and T.K. Hunt, Effects of cortisone and Vitamin A on wound healing. Ann Surg. 1968;16(7):324-328

7.         Febriyenti, A.M. Noor, S.B. Bai, Physical evaluations of Haruan spray for wound dressing and wound healing. Int. J. of Drug Delivery.2011; 3: 115-124.

8.         R.J. Goldman, Many ways to measure a wound: an overview of techniques. Adv. Skin Wound Care. 2002; 15:236.

9.         C.W.T. Jacqueline, M.L. Kit, L.L. Cheuk, H.T. Ming, F.K. Hin, P.L. Ching, H.K. Khun, C.L. Ping, P.F. Kwok, The in vivo and in vitro diabetic wound healing effects of a 2-herb formula and its mechanisms of action. J. of Ethnopharmacol. 2011;134: 831-838.

10.       R.K. Jha, N. Garud, R.K. Nema, Excision and Incision Wound Healing Activity of Flower Head Alcoholic extract of Sphaeranthus indicus Linn. In Albino rats. Global J. of Pharmacol.2009;3: 32-37.

11.       M. Karami, D. Yazdani, M.R.J. Nadoushan, S. Geravand, In vivo wound healing activity of herbal ointment in a rat. Res. J of Pharmacognosy. 2016; 3: 9-16.

12.       D. Kumarasamyraja and M. Swamivelmamickam, Evaluation of In vivo and In vitro wound healing activity of aqueous extract of Acalypha indica. Int. Res. J. of Pharm. 2015; 6: 57-61.

13.       L. Laila, Febriyenti, S.M. Salhimi, S. Baie, Wound healing effect of Haruan (Channa striatus) spray. Int. Wound J. 2011: 1-8.

14.       K.H. Lee, Studies on a mechanism of action of salicylates II, Retardation of wound healing by aspirin. J. Pharm. Sci. 1968;57: 1042-1043.

15.       T.K. Lim, Edible Medicinal and Non-Medicinal Plants. Springs, Vol. 7- Flowers, 2014: 87-93.

16.       P. Mallefet and A.C. Dweck, Mechanisms involved in wound healing- Focusing on Injuries. The Biomedical Scientist. 2008: 609-614.

17.       A.F. Mechesso, A. Tadese, R. Tesfaye, W. Tamiru, T. Eguale, Experimental evaluation of wound healing activity of Croton macrostachyus in a rat. African J. of Pharm. And Pharmacol.2016; 10: 832-838.

18.       J.J. Morton and M.H. Malone, Evaluation of vulnerary activity by an open wound procedure in rats. Arch. Int. Pharmacodyn. Ther.1972; 19(6):117-126.

19.       S. Muhammad, A. Nasim, R. Naheed, A. Muhammad Shafiq, J. Abdul, Isolation and characterization of Secondary metabolites from Plumeria obtusa. J. of Asian Nat. Prod. Res.2011; 13: 1122-1127..

20.       S. Murthy, M.K. Gautam, S. Goel, V. Purohit, H. Sharma, R.K. Goel, Evaluation of In Vivo Wound Healing activity of Bacopa monniera on Different Wound model in rats. Biomed Res. Int.2013:1-9.

21.       A. Nasir, D. Ahmad, J. Bakht, S. Shah, F. Ullah, M. Rehman, Antimicrobial activity of leaves extracted samples from medicinally important Plumeria obtusa. J. of Med. Plants Res. 2013;7: 1121-1128.

22.       S. Nayak, P. Nalabothu, S. Sandiford, Evaluation of wound healing activity of Allamanda cathartica. L. and Laurus nobilis. L. extract on rats. BMC Complementary and Alternative Med. 2006;6: 12.

23.       K.S. Priya, A. Gnanamani, N. Radhakrishnan, M. Babu, the Healing potential of Datura alba on burn wound in albino rats. J of Ethnopharmacol.2002; 83: 193-9.

24.       K. Roy, H. Shivakumar, Wound Healing Potential of Leaf extract of Ficus religiosa on Wistar albino strain rats. Int. J. of Pharm Tech Res..2009;1:506-508.

25.       P.A. Sanders, 1974. Propellants and Solvents, in the Science and Technology of Aerosol Packaging, J.J. Sciarra and L. Stoller, Editors. John Wiley and Sons, New York. 1974: 97-149.

26.       S. Sasidharan, S. Logeswaran, L.Y Latha, Wound Healing Activity of Flaeis guineensis Leaf extract ointment. Int. J. of Mol. Sci.2012;13: 336-347.

27.       P.R. Shinde, P.S. Patil, U.A. Bairagi, Phytopharmacological Review of Plumeria Species, Scholars Academic J. of Pharm. 2014;3:217-227.

28.       A. Shukla, A.M. Rasik, G.K. Jain, R. Shankar, D.K. Kulshrestha, B.N. Dhawan, In vitro and in vivo Wound healing activity of asiaticoside isolated from Centella asiatica. J. of Ethnopharmacol, 1999; 65: 1–11.

29.       B.S. Siddique, Firdous, S, Begum, Two triterpenoids from the leaves of Plumeria obtusa. Phytochemistry, 1999; 52: 1111-1115.

30.       B.S. Siddiqui, I. Firdous, M. Rasheed, S. Begum, Chemical Constituents of leaves and stem bark of Plumeria obtusa. Phytochemistry, 2004; 65: 2077-2084.

31.       H. Soni and A.K. Singhai, A recent update of botanicals for wound healing activity. Int. Res. J. of Pharm, 2012; 3: 1-7.

32.       M. Subalakshmi, A. Saranya, M.U. Maheshwari, A. Jarina, S. Kavimani, R. Murali, An overview of the current Methodologies used fort for the evaluation of drugs having wound healing activity. Int. J. of Exp. Pharmacol, 2014; 4: 127-131.

 

 

 

 

 

 

Received on 24.07.2018       Accepted on 30.08.2018     

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Res. 2018; 8(4): 231-235.

DOI: 10.5958/2231-5691.2018.00039.4