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INTRODUCTION
Nowadays, we have more advanced equipment, stuffs, technologies and activities
compared to the past days. Unfortunately, more access of these also means people are more
prone to injuries, usually wounds. A wound is an injury that causes damage to ones body. It is a
general term that refers to harm caused by accidents, falls, hits, weapons, and more. These
injuries range from minor to life-threatening. Wounds are injuries that break the skin or other
body tissues. They include cuts, scrapes, scratches, and punctured skin. They often happen
because of an accident, but surgery, sutures, and stitches also cause wounds. If ever a person
acquired a wound, there should be an act of first-aid taken for they can cause infections or
threaten ones life if leaved untreated for a time. Usually, wounds heal themselves overtime. But
with wound applicants applied, they will have faster healing time since they promote new skin
production. As the skin cells increase the faster the wound can heal. Wound dressings, usually a
gauze or a piece of cloth, are used to coat and cover cuts, prevents infection, disallow wound
exposure, and cover wound away from contacts contaminations. However, they cant promote
blood clotting and accelerate the wound healing process.
If we get wounded, our skin cells in the part of our body that is wounded dies. Skin cells
can increase by applying collagen to our skin. We can get collagen from skin applicants like
lotions. Technology, nowadays provide us many applicants in order to increase our skin cells but
surely many of them are ineffective.
In the meanwhile, Bali-bali (with a scienfic name of Euphorbia tirucalli in the modern Plant
Kingdom) is Balinese traditional medical treatment is known as "Usada". The term comes from a
Sanskrit word "Ausadhi", meaning plants that are used for medicinal purposes since in ancient
times as a tradition. There are more than 491 species of medicinal plant in Bali. These Balinese

medicinal plants hold valuable potential for global medicinal plant biodiversity. A worrying trend
of the extinction of many of these Balinese medicinal plants has become a concern at the Eka
Karya Bali Botanic Garden.
It is rumored by the elderly and native people, here in the Philippines, from the
countryside that Bali-bali plant (Also known as Pobreng Kahoy in the locals) has ability to heal
the wounds efficiently, have anti-infection and blood clotting properties. Since that the Bali-bali
plant can be commonly found in our country, more of our fellow countrymen use it in treating
wounds. They usually squeeze the stems to obtain its extract and apply it to the wound directly,
making it as an alternative applicant instead of commercial medicines even having no idea or
information if the Bali-bali plant can really be effective in healing wounds and safe to be applied
in the open cuts of the skin.
In this matter, the researchers are motivated to determine if the Bali-bali plant extract is
effective to be an applicant for treating the wounds and can promote new skin cell production;
and determine if there are significant differences between the Bali-bali plant extract and the
commercial wound applicant in healing processes and parameters.

MATERIALS AND METHODS

A. Materials and Equipment

The study required the use of the following materials and equipment: One (1) kilograms
of Bali-bali plant stalks, shears, factory plastic bags, commercial wound applicant (PovidoneIodine, Betadine), synthetic gauze pads (Band-Aid branded), bandage tapes, cottons, scalpel,
cages (with suitable environment setting for the mice), white mice, surgical gloves, Petri dishes,
surgical scissors, glass jars, fine strainers, droplets, razor, marker, plastics sheets, plastic block
containers, plaster, laboratory juicer and microscope.

B. Research design
The study used the posttest only design in terms of the healing time parameter and the
skin cells production effect parameter. Three set-ups was laid out in the study (Set up A:
treatment with the use of synthetic gauze pad with Bali-bali plant extract as an applicant, Set up
B: treatment with the use of synthetic gauze pad with commercial wound applicant, and Set up
C: Untreated) and was replicated five (5) times. Each mouse represented a replicate.

C. General Procedure
C.1 Collection of Bali-bali plant stalks
One kilogram (1) of Bali-bali plant stalks were collected in Pahanocoy Village, Villa
Lourdes, Bacolod City. With the use of freshly sharpened shears, the stalks were cut in its main
stalk two (2) inches away from its roots. The collected stalks were thoroughly washed and was

kept and sealed with the use of the factory plastic bags for safe transporting. The stalks were
transported at one of the researchers residence.

C.1.2 Extraction of Bali-bali Plant Extract

The collected Bali-bali plant stalks were squeezed with the use of juicer to acquire its
extract. The collected extract was strained to the fine strainer to remove the unwanted small parts
of stalks and solidified juice, and then it will be strained again to the finer strainer to ensure no
solid materials was in the solution. The strained liquid was kept and covered with the use of
plastic sheets and glass lids in sterilized and dry glass jars provided by the researcher to prevent
storage contamination. Wearing of surgical gloves was observed in the entire procedure.

C.1.3 Preparation of Synthetic Gauze Pads

The gauze pads were used in the study should be freshly unpacked from its factory
package and cover, and surgical gloves should be worn when unpacking the pads to prevent
contamination. These pads were cut into smaller pieces, about 2 inches square in measurement.
Five (5) pieces of gauze pads were applied by the varying types of applicants based where set up
was used. With the use of droplets, each of five (5) gauze pads were applied by three (0.3) mL of
Bali-bali plant extract for the Set up A. Another five (5) of gauze pads were applied by three
(0.3) mL of commercial wound applicant for the positive control or Set up B. And the other five
(5) gauze pads were left unapplied for the negative control or Set up C. These gauze pads were
kept and covered in plastic containers after the application processes.

C.1.4 Preparation of White Mice

Twenty (20) healthy white mice were bought from Sta. Fe Petshop, Bacolod City. Fifteen
(15) mice were randomly picked from the group and were put in their respective glass tanks
while the other five (5) will serve as reserves. The mice were randomly distributed to three set
ups, five (5) for each set up (Set up A: treatment with the use of synthetic gauze pad with Balibali plant extract as an applicant, Set up B: treatment with the use of synthetic gauze pad with
commercial wound applicant, and Set up C: Untreated). The mice undergo through
acclimatization for two (2) weeks before the start of the experiment. They were fed with
recommended rodent pellet of the veterinary hospital and distilled water under strict hygienic
conditions.

C.1.5. Casting of the Wound on White Mice

The mice samples were brought to the Provincial Veterinary Office near the Provincial
Capitol Lagoon, Bacolod City. The researchers were assisted by a licensed veterinarian. All of
the Fifteen (15) mice were carefully transferred from their glass tank the operating table. With
the use of razor, four (4) centimeter square of fur in back part of the mice was removed. Using
the marker, the researchers mark the shaved part at the back of the mice to serve as a marking
template of the wound to be casted. The location of the wound area was intentionally chosen to
prevent the mice to reach the wound by itself. The area was cleaned first by the 70% Isopropyl
alcohol. Then with the use of scalpel, two (2) centimeter of wound length was cast on the skin of
the mice with almost one (1) centimeter depth. Wearing of surgical gloves was observed in the
entire procedure.

C.1.6 Application of Gauze Pads on the White mice

After the wound casting procedure, the gauze pads were applied immediately on the
wounds of the mice. The Set up A (treatment using the gauze pads with Bali-bali plant extract)
wound samples were assisted by bandage tape to ensure that it will not loosen to the body of the
mice. Similar procedure was done for the Set up B (treatment using the gauze pads with
commercial wound applicant). The Set up C (untreated) samples were left untreated with no use
of any medicine but was covered by the fresh gauze pads to avoid direct contact to the wound
and infection. After application, the Set ups was left at one of the researchers residence for five
(5) days. The entire procedure was done daily at the same time of the day.

C.1.7 Determining the New Skin Production Effect on the White Mice
After three (3) days, where the wounds are freshly contracted, the mice samples were
brought to the Villanueva Laboratory, Villanueva, Negros Occidental. With the supervision of a
licensed biologist, the researchers get a two (2) by one (1) wound contraction from each mouse
using a small surgical scissors. Then, the samples obtained were given to the personnel of the
laboratory to count the new skin cells in the contraction under the microscope. It took three (3)
days for the data to be acquired. After obtaining the new skin cell count data of each set ups, the
data was analyzed and compared using the statistical tool of the study.

C.1.8 Data Gathering and Statistical Analysis Tool

The mice were observed every day and recorded the time used by the applicant to heal
the wound (determined as the wound area has been replaced by a new skin and there is no sign of

contraction) by each set ups. The data of the new skin count was analyzed immediately after the
laboratory acquired them.
The data obtained from the study were subjected to the following descriptive and
inferential statistical treatments using the Microsoft Excel Tool Pak 2010.With the use of
following statistical analysis tool, the researchers was successfully analyze and compare the data
from each set ups.
1. Mean was

used to determine the average scores of the results of the

treatments in this study; and

2. ANOVA was used to determine the difference between two or more means, set
at 0.05 level of significance.
After the researchers have acquired the final data with the used of mentioned statistical
tools, the data were compared from each other depending on their set-ups.

ACKNOWLEDGEMENT

The researchers would like to express their utmost gratitude and appreciation to the
following that exerted efforts and shared their knowledge to make this research study successful:
To the parents of the researchers, for their never ending support and for the
encouragement in times of hardships and problems.
To the research advisers, Mrs. Villagracia and Mr. Gorre, for their knowledge and giving
encouragement and advices.
To Mr. Renante J. Decena, DVM, Ph.D, Head Laboratory, Provincial Veterinary Office
for assisting in casting of the mice and allowing the researchers to conduct in their laboratory.
To Mr. John A. Calumpiano, Station Manager, National Plant Quarantine Services
Division for certifying our Bali-bali (Euphorbia tirucalli) plant.
Most of all, to the Almighty God for the strength and wisdom provided to the researchers.
-The researchers

BIBLIOGRAPHY
Abdou, E. S., Nagy, K. S. A., & Elsabee, M.Z. (2008). Extraction and characterization of Balibali plant from local sources. Bioresource Technology: 99 13591367.
Agapito, Y., Canaco, M., Cruz, K., et. al. (2013). Determination of the Wound Healing Property
of Indian Rubber Tree (Ficus elastic) Leaf Extract in Male Sprague- dawley Rats on
Circular Excision and linear Incision Model. Omics Group Conferences. 1 (1). p. 1.
Black, J., Matassarin-Jacobs, E. and Silver, F. (1997). Wound Healing Physiology. Retrieved
January 15, 2014 from http://woundhealing.in/wound-healing-physiology.html.
Dai T, Tanaka M, Huang YY, Hambin R (2011) Euphorbia preparations for wounds and burns:
antimicrobial and wound healing effects. Expert Rev Anti Infect Ther 9(7): 85779.
Dutta P, Dutta J, Tripathi VS (2004) Euphorbia tirucalli: chemistry, properties and
applications. J Sci Industrial Res 63: 2031.
Helander I, Nurmiaho-Lassila E, Ahvenainen R, Rhoades J, Roller S. (2001). Euphorbia milii
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Hertling, D. (2005). Wound Healing. Columbia University Medical Center.
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Je, J., & Kim, S. (2006). Potential Eurphorbia tirucalli on bacteria-killing by disrupting the outer
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APPENDIX

Figure 1. The juicer used in extracting

the Bali-bali plant.

Figure 3. The Bali-bali plant was put in

bali plant.
the extracting machine.

Figure 5. The materials used in casting of

bali Plant
wounds on the mouse.
Extract)

Figure 2. The Bali-bali to be used.

Figure 4. The Extract of Bali-

Figure 6. Treatment A (Bali-

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Figure 7. Treatment B (commercial wound applicant)

(Untreated)
(Untreated)

Figure 8. Treatment C

Figure 9. The mouse used in casting wounds.

Figure 10. Injecting the
Lidocaine to the mouse

Figure 11. Disinfecting the mouse

mouse

Figure 12. Shaving the

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Figure 13. Casting wound in the mouse

Casted mice

Figure 14. The

Figure 15. Preparing the application of gauze Figure 16. The application of
the gauze to the
applied to the mouse
mouse