The role of the skin in the body is very significant, especially in protecting the deeper tissues of the body and it acts as an effective barrier from foreign substances from the environment. It also regulates the body temperature through sweating (Zhao, Liang, Clarke, Jackson, & Xue, 2016).
The wound is a physical injury that may result in the disruption, damage, breaking or opening of the skin that is caused by chemical, thermal, microbial or immunological inflammation of the skin tissues. It categorizes as an open wound, where the blood escapes from the body and causes bleeding that is visible and the second classification is the closed wound where the blood escapes inside the body (Nagar, et al., 2016; Sharma, Jeyabalan, Singh, & Semwal, 2013).
According to the Wound Healing Society, the cause of the disturbance in the normal anatomy and functions of the skin is due to the wound that results in opening or breaking of the tissues (Alam, Singh, & Singh, 2011). According to Thomas and Harding 2002, the leading cause of morbidity in the United Kingdom is the wound, where it affects more than 1% of the population with a treatment that cost at least £1 million per year (Ezealisiji, Omotosho, Udoh, & Agbo, 2014).
When wounds have failed to repair within its normal range of development of the damaged skin tissue within 4-8 weeks are known to be chronic wounds. If the damaged skin is not treated, it may become a threat to an individual because of the susceptibility of it to acquire infections that may cause severe skin disease. This type of wound that failed to improve within its normal stage of healing, it may occur continually. This result is considered as a healthcare problem of the society, causing over 4 million American are affected by an annual treatment cost of 4 billion dollars (Izadi & Ganchi, 2005). According to Roig et al., 2017, damage skin can affect the quality of life of an individual that can last for a long time spending years in treating damage skin (Roig, Fabrellas, Rubio, Wilson, & Kate, 2017).
Wound healing is a complicated process that involves the restoration of the structural and functional integrity of the skin tissue. When the body is suffering from an injury, it stimulates the inflammatory and immune response, and the process of healing starts immediately. The repairing of the skin tissue occurred in two major activities of regeneration and reparation and divided into four phases. The hemostasis phase where the immediate response happens by stopping the flow of blood. The inflammation phase where the clotting of proteins occurs to prevent the infection of the wound. The proliferation phase is the rapid reproduction of the cells to repair the defects and lastly, the remodeling phase where the development of the usual epithelium and maturation of the tissue will change and will alter its structure (Young & McNaught, 2011; Marieb, 2014). The delayed and prolonged healing process of the wound is due to several factors such as improper diet, infection at the wound site, and tissue perfusion to the wound area, drugs (Sharma, Jeyabalan, Singh, & Semwal, 2013). If the injured tissue heals inappropriately, it may develop into either chronic wound such as the venous ulcer or pathological scarring such as keloid scar (Thakur, Jain, Pathak, & Sandhu, 2011).
Allium cepa from the family of Liliaceae is an aromatic vegetable that is being used worldwide and is commonly known as the onion. It can be cultivated worldwide with different varieties. The bulb of the white onion (Allium cepa) are allowed to grow under the ground, and it bears small flowers that are white or purple. The green stems and leaves can grow and reach up to 3ft in height. It contains chemical substances that produce pungent odor, and at the same time, it irritates the eyes once the white onion (Allium cepa) is chopped. This plant has a high value because of its therapeutic properties that show the positive result in controlling type 2 diabetes mellitus, in wound healing and anti-scar. Its extract can be used as an anticancer where it suppresses the growth of various tumor cells. And as anti-inflammatory because of the flavonoid that is present (Upadhyay, 2016; Kumar, Bhowmik, Chiranjib., Biswajit., & Tiwari, 2010).
According to the study of Shenoy et al. 2009, they used the alcoholic extract of the onion (Allium cepa) in rats for wound healing. As the result of their experiment, in the 18th day, the treated group of rats showed broad growth of granulation, it also started along its surface, and it indicates the complete healing of the wound. The phytochemical screening of their study exhibit the presence of flavonoid that is one of the major constituents of wound healing (Shenoy, Patil, Kumar, & Patil, 2009).
Flavonoids are natural compounds that are present in plants, and it is thought to have high antioxidant properties and high biological properties that help in promoting the health of human and lessen the risk to acquire diseases. It has been used by humans for about 4 million years because of its therapeutic potential that can be easily found in different medicinal plants. The presence of flavonoids in food produces color, taste, and protection of vitamins and enzymes. Many studies suggested that flavonoids have a protective effect against many infections and degenerative diseases. This compound possesses biological activities such as antioxidant activity, antibacterial activity, anti-inflammatory activity and anti-cancer activity (Kumar & Pandey, 2013). Numerous studies were conducted related to the wound healing properties of some plants such as papaya, and turmeric (Aruna, Sravanthi, Sri, Priya, & Rao, 2015).
From the ideas mentioned above, the researcher aims to assess and confirm the wound healing potential of the traditional plant white onion bulb. The ethanolic extract of white onion bulb was investigated to evaluate the rate of wound healing enclosure to the wounded rats. And the purpose of this research is to prove whether the flavonoid present in white onion bulb is potent or not in wound healing and to formulate an ointment from the semi-purified extract of white onion bulb that has a lesser unwanted side effect.
Wound healing is a challenging process of repairing the damaged skin and other soft skin tissues of the body. It is a systemic process of renewing cells that generate reconstitution, and restoration of the injured skin tissue (Niulugal, Perumal, Ugander., & Chitor, 2014). This process is categorized into four phases of wound healing: Coagulation and hemostasis, it is the immediate process of platelet aggregation and clot formation to reduce or avoid blood loss. Inflammation phase, it establishes the barrier to protect the wound from microorganisms. Proliferative is the process by which rapid growth of new parts and cells of the injured skin tissue occurs. Remodeling, the last stage of wound healing where it is responsible for the formation of new collagen and in developing tensile strength of new cells and tissues of the skin. It may last for up to 1 to 2 years of repairing the damaged skin tissue or sometimes for an extended period (Velnar, Bailey, & Smrkolj, 2009).
The ointment is a semisolid preparation which contains medicaments and is usually applied to the skin. The ideal characteristics should be chemically and physically stable, the active ingredient should be finely divided and distributed uniformly, and it should be free from grittiness. The advantages of ointments are, it will avoid the first pass metabolism of the drug, it is convenient to use for unconscious patients, and they are easy to handle, and they are more chemically stable. The selection of the base to be used is very important in formulating this type of dosage form because it controls the performance of the ointment on the skin (Usha & Ashish, 2015).
The plant Allium cepa or most commonly known as the onion that belongs to the family of Liliaceae originated from Egypt and is being cultivated in different countries. The word onion is described as “one large pearl” that is derived from the Latin word “unio”, and also called by the Chinese as “jewel among vegetables.” It has been cultivated for more than five thousand years and is still using in alternative herbal medication and as well as a flavoring agent or a vegetable that is cooked or can be eaten raw. It also became important in the daily diet of many people by using it in their vegetable salad, pickle sauce, and others. This plant can grow in the dry season that is required to ripen the bulb of the onion. The best soil that can be used in cultivating should be dry and sandy loams that contain organic substances. The seedlings should be planted first in the nursery before it will be transferred in the field. And the harvesting time depends on the geographical condition (Srestha, 2004).
Testing the effectiveness of formulated ointment for wound healing
b) 6.6% Formulated white onion bulb ointment
Formulated ointment for wound healing
Solcoseryl® Statistical analysis
This research study aims to test and evaluate the effectiveness of white onion, Allium cepa (Fam. Liliaceae) ointment. It sought to answer the following question:
1. Is there any significant difference between the different concentration of 3.3% and 6.6% white onion bulb ointment?
2. Is there any significant difference between the formulated 6.6% white onion bulb ointment and the Solcoseryl® ointment?
1. There are no differences between the different concentrations of 3.3% and 6.6% white onion bulb ointment.
2. The effect of the 6.6% white onion bulb ointment is the same as the effect of the Sorcoseryl® ointment used as positive control in wound healing.
The study regarding the wound healing activity of the white onion, Allium cepa ointment (Fam. Liliaceae) signifies the number of uses that will be significant to the following:
Pharmaceutical Industries. It can be prepared as a pharmaceutical drug product that can be used in wound healing since onion is being cultivated worldwide and is common in the Philippines.
Researchers. The outcome of this study can help and will serve as a reference in conducting further studies about the pharmacological effect of the plant.
Families. It provides additional knowledge about the natural medications with use of alternative herbal medicines mainly found in kitchen wherein it helps to save money by purchasing other products.
Citizens. The result of this study will provide alternative, efficacious, safe, natural, and economical medication to help improve the healing process of wounds especially to those persons who cannot afford to buy expensive products.
Students. The given data from this study will provide meaningful information and reference inside the class discussion about alternative uses of medicinal plants and its affordability and accessibility in the market.
This research was conducted from September 2017 until next year April 2018 in Urdaneta City University, San Vicente West Urdaneta City Pangasinan. This research is concerned primarily with the semi-purification extraction of white onion, bulb and in making wound healing ointment that is made from white onion bulb extract. Due to its proven wound healing property, the focus of this research is to formulate and to compare the wound healing effectivity to commercial Solcoseryl®.
This chapter presents a discussion regarding the research design of the study, data gathering tools, duration and locale of the study, subjects of the articles and sampling scheme. The procedures that will be applied to answer the given questions presented in chapter 1, and also it explains the proper incision of the wound in albino rats and the right handling.
The experimental method was used in this research and study to get the ethanolic extract of the bulb of white onion and the difference between the two concentrations from it. The two different amount of active ingredients were used to evaluate the capacity of the wound healing property of the ethanolic extract of the bulb of white onion bulb in albino rats.
In table 1, twenty (20) healthy albino rats of either sex that weighs between 150- 200 grams were used as test animals. They are assigned to each group with the following treatments that consist of four (4) replicates. The first group received Solcoseryl® ointment (AP1). The second group received 3.3% ointment of white onion bulb ointment (AP2). The third group received 6.6% ointment of white onion bulb ointment (AP3). And the fourth group received no treatment (AP4). Percentage of wound incision size will be computed to determine the significant difference of the positive control Solcoseryl® to the different concentration of the ointment of the white onion.
Experimental procedures such as the extraction process, preparation of different concentration of ethanolic extract of white onion bulb, and assessment of wound healing property was handled from September 2017 to April 2018. The extraction process was conducted at Virgen Milagrosa University Foundation, San Carlos City, Pangasinan. And the wound healing test was performed at the Pharmaceutical Laboratory of College of Pharmacy at Urdaneta City University, San Vicente West, Urdaneta City Pangasinan.
This study used a total number of twenty (20) healthy albino rats of either sex weighing 150- 200 grams. They are divided into four groupings with five rats in each group. The twenty (20) healthy albino rats were acclimatized to the standard laboratory for seven days and fed them with pellet diet and tap water. The ethical clearance through animal permit was accomplished from Bureau of Animal Industry at San Juan, San Carlos City, Pangasinan before using the animals in the experimental procedure.
A. Collection of Plant Material
The white onion bulb was harvested from San Jose, Nueva Ecija and were identified by the botanist from the Department of Agriculture, Urdaneta City Pangasinan. The collected white onion were transported to Virgen Milagrosa University Foundation, San Carlos City, Pangasinan.
Preparation of the Ethanolic Extract of White Onion (Allium cepa) bulb
The collected white onion bulb was washed first to remove dirt and unwanted substances. After cleaning the white onion bulb, it was chopped into small. The fresh white onion bulb was macerated in 95% ethyl alcohol for 72 hours. The white onion bulb undergoes the semi- purification process to obtain the semi-purified extract. Then the semi-purified extract was used in the preparation of the different concentrations which are 3.3% and 6.6%. And it was used in formulating ointment for wound healing.
B. Phytochemical Screening
Screening for Flavonoids
1. Sodium Hydroxide Test
A 0.2 ml of diluted sodium hydroxide was added to 0.2 ml of the extract and shaken gently.
Positive Result: Golden yellow precipitate obtained
(Itodo, Oyero, Umeh, Ben, & Etubi, 2014)
2. Ferric Chloride Test
A 0.2 ml of 10% ferric chloride is added to the extract. The mixture is shaken together.
Positive Result: Woody brownish
(Itodo, Oyero, Umeh, Ben, & Etubi, 2014)
3. Lead Acetate Test
A 0.2 ml of the sample extract is added to 0.2 ml of 10% of lead acetate. It was gently shaken to avoid emulsion.
Positive Result: Dirty brownish precipitate
(Itodo, Oyero, Umeh, Ben, & Etubi, 2014)
C. Preparation of the different concentrations of the ointment of white onion bulb
1. 3.3% ointment of white onion bulb
The stearyl alcohol and white petrolatum was melted on a 70oC steam bath. One gram of the active ingredient was added, and the remaining ingredients were dissolved in the water and heated to 70oC. The oleaginous phase was added slowly to the aqueous phase with constant stirring. Then remove it from heat and stir the mixture until it congeals.
2. 6.6% ointment of white onion bulb
The stearyl alcohol and white petrolatum were melted on a 70oC steam bath. Two grams of the active ingredient were added, and the remaining constituent was dissolved in the water and heated to 70oC. The oleaginous phase was added slowly to the aqueous phase with constant stirring. Then remove it from heat and stir the mixture until it congeals.
D. Animal Handling
Make sure that the animals are aware in the presence of the handler by placing the gloved hand inside the cage and allowing the rats to become used to it. The approach should not be quick so that the rats will not be frightened. Handling the rats by holding the base of the tail to move the rat from one place to another. The rat is grasped carefully and firmly by placing the hand over its back with the thumb placed over its shoulder. The index finger is placed on the head, and the remaining three fingers are closed together, pulling a fold of skin.
E. Incision of Wound in Albino Rats
The wound site was prepared and identified by the veterinarian. Followed by the incision of the skin. The hair of the albino rat was shaved on the dorsal thoracic region. The wound size will be 4 cm long. The albino rats were treated once daily from 1st up to the 7th day. The wound size was measured every other day to see the improvement of the excision site of the wound.
This chapter presents the analysis and interpretation of the data about the stated problems of the study such as: (1) Is there any significant difference between the different concentrations an of (a) 3.3% and (b) 6.6%, white onion bulb ointment? (2) Is there any significant difference between the formulated 6.6% white onion bulb ointment and the Solcoseryl® ointment?
The Effect of Solcoseryl Ointment in Wound Healing as shown by the Differences of wound sizes
The table 2 shows that the wound healing activity observed in albino rats after givig the Solcoseryl® ointment as the positive control. From the 1st rat to the last rat, the table shows that after the first day and third day of applying the Socoseryl® ointment, the effect of it was slightly effective. And on the fifth day of applying the Solcoseryl® ointment, it shows that it was effective and on its seventh day, it was already highly effective and the wound was not fully contracted.
The Effect of 3.3% of White Onion Ointment as shown by the Differences of wound sizes
3.3% of white onion ointment
The table 3 that on the first day and third day of application the effect was slightly effective and on the fifth day of, it became effective and on the last day of application, it shows that the 3.3% of white onion ointment is highly effective and the wound was completely healed.
Shows the Effect of 6.6 % of White Onion Ointment as shown by the Differences of wound sizes
6.6 % of white onion ointment
The table 4 shows that the wound healing activity of the 6.6% of white onion observed in the test animals (Albino rats), from the first day up to the third day of application, the effect of the ointment was moderately effective. On the fifth day and seventh day of application of ointment, it shows that the effect was highly effective.
Significant differences between the 3.3% white onion ointment, 6.6% white ointment and positive control (Solcoseryl® Ointment)
?- level= 0.01
The table 5 shown if there are no significant differences between the 3.3% white onion (Allium cepa) ointment, 6.6% white onion ointment and positive control (Solcoseryl® Ointment) employed on the albino rats. The first row and second showed that there is no significant difference between the 3.3% white onion ointment, 6.6% white onion ointment because the t-computed significance value of 0.999 is lesser than the t-critical value of 2.896 at the degree of freedom of 8.
Second row and the third row showed that there is no significant difference between 6.6% white onion ointment and positive control (Solcoseryl® Ointment) because the t-computed significance value of 2.028 is less than the t-critical value of 2.896 at the degree of freedom of 8.
This chapter presents the summary of the study, the findings of the study, the conclusion based from the data gathered and recommendation endorsed by the researchers regarding the wound healing property of the ethanolic extract of the onion bulb.
This research study aims to test the effectiveness of the white onion in wound healing activity. The T-test is used as a statistical tool to determine the difference between the two different concentrations of white onion bulb.
The bulb of onion was collected from Nueva Ecija. The collected bulb of onion was cleaned, and extracted with 95% ethanol. It was macerated for 72 hours. The extract that was collected was evaporated under the water bath until a syrupy liquid is obtained. And the extract that was obtained was cooled for the formulation of different concentrations needed for the study.
The study used four (4) replicates which include: Group 1, that was treated with Solcoceryl® ointment; Group 2 was treated with 3.3% ointment; Group 3 was treated with 6.6% ointment of onion ethanolic extract; and the Group 4 received no treatment as the negative control. The percentage of wound incision size was computed using two different concentrations for seven days. The wound closure on each Albino rats was measured in two- day interval until the wound is completely healed.
Given the problems stated in this study, the following are the findings:
1. There is no significant difference between the different concentrations prepared of white onion bulb ointment and the positive control, Solcoceryl® in the healing of the wound of the wounded albino rats.
2. The ethanolic extract of the white onion bulb contains flavonoid which is responsible for the healing of the wound of the albino rats.
Based on the summary of the findings, the following conclusions arrived:
1. The white onion bulb has a wound healing property since it was able to contract the wound of the wounded albino rats.
2. A different concentration of the wound healing ointment can be prepared from the ethanolic extract of white onion bulb.
3. The higher the concentration of the ethanolic extract of the onion bulb, the faster the wound to be healed.
1. To formulate the white onion bulb in other suitable dosage form that has wound healing property.
2. To conduct similar study using other wound models such as excision and burn model
3. To conduct study on the toxic effect of white onion bulb.
4. To conduct similar study by using another plant part of onion.