The Effectiveness of Ethanol Extract in Muli Banana Peels (Musa Acuminata) to Heal Cut Wounds in Mice (Mus Musculus L.)

*Correspondence Address: eriolamaulidya97@gmail.com Muli banana peels contain flavonoids, saponins, and tannins that foster the process of wound healing. The purpose of this study was to determine the effect of ethanol extract in muli banana peels (Musa acuminata) on cut wounds healing process in mice (Mus musculus). The study design was the completely randomized design by using 24 male mice which were divided into 6 groups, namely (K+, K-, P1, P2, P3, and P4). Each treatment consisted of 4 mice. All groups of mice were treated with a 1 cm cut wound on the back. The K+ group as the positive control group were applied with povidone-iodine, the Kgroup as the negative control group were applied with distilled water, the P1 group was applied with a concentration of 10% extract, the P2 group with 20% extract, the P3 group with 30% extract, and the P4 group with 40% extract for 14 days. On the 15th day, histological slides were made by taking 1 sample from each group. The results showed that the ethanol extract of muli banana peels played a role in the cut wound healing process.


INTRODUCTION
The skin is the largest organ because it encloses the entire outer surface of the body (Muthia, Titin, & Neneng, 2019). The thicker skin is located on the back, shoulders, soles of the feet, palms, and buttocks (Putrinda, Budi, & Rudy, 2014). The skin protects parts of the body from physical and mechanical disturbances that occurred outside of the body (Hadi, 2006;Nuzantry & Widayati, 2015). The disturbances from outside of the body are traction, harmful chemicals, pressure, friction, fungal infections, radiation, and UV exposure (Maulidya, 2019). Body injuries can be caused by explosions, electric shock, animal bites, changes in temperature, chemicals, and trauma by sharp or blunt objects (Darwis, Hardiningsih, & Nurlidar, 2013;Ningsih & Paturusi, 2015).
Based on research by (Yunanda & Rinanda, 2016), onion extract is often used to heal the banana plants' wounds, such as leaves, flowers, pseudo-stems, tubers, and banana peels. Another research by (Onyenekwe, Okereke, & Owolewa, 2013), reveals that the ointments of Ambon banana stem extract accelerate the wound healing process. Bananas contain secondary metabolites in the form of alkaloids, glycosides, terpenoids, saponins, flavonoids, and tannins (Budianto & Basuki, 2018). Based on the statement above, it is necessary to research to test the Muli banana peels extract (Musa acuminata Colla) on the cut wound healing process on the back of mice.

METHODS
This research was conducted at the Zoology Laboratory of the Faculty of Mathematics and Natural Sciences of the University of Lampung, the Laboratory of Analytical and Instrumentation Chemistry of the Mathematics and Natural Sciences Faculty of the University of Lampung, and Pathology Laboratory Regional Veterinary Investigation and Testing Center (BPPV) Bandar Lampung in February to April 2019. The tools used in this study included the maintenance tools for mice (cages, feeds, and drinking containers), calipers, analytical balance, scissors and shavers, scalpels, permanent markers, beaker cups, Erlenmeyer flask, measuring cups, stirring rods, pipettes drop, aluminum foil, plastic wrap, extraction equipment (blenders, ovens, filter paper, Buchner funnels, and rotary evaporators), and cameras for documentation.
The materials used in this research included 24 male mice (Mus musculus), Muli banana skin (Musa acuminata), 96% ethanol, povidone-iodine, 2% lidocaine, chloroform, vegetable pellets to feed the mice, and tap water for the mice to drink. The first analysis of the obtained data the homogeneity test and then followed by the analysis of variance or ANOVA using the SPSS and Minitab programs.

RESULTS AND DISCUSSION
The length of the wound closure was measured using calipers. The following is the figure of the average percentage of the length of wound closure from day 1 to day 7.  Figure 1, the average percentage of cut wound closure at day 1 was 0% because the wounds had just formed and the healing process had not occurred yet. After 7 days of treatment, the highest wound closure percentage was found in P4 with the average percentage of wound closure length reaching 73.25%. Consecutively, the second-highest wound closure percentage was obtained by P2 with the average percentage of 72% and then followed by P1 with the average percentage of wound closure length of 71.50%, K+ with the average percentage of wound closure length of 67.25%, P3 with the average percentage of wound closure length of 49.50%, and K-with the average percentage of wound closure length of 48.50%.
Wounds on mice smeared with 40% concentration of Muli banana peels ethanol extract had a faster recovery rate than the positive control (K+) smeared with povidone-iodine and negative control (K-) smeared with distilled water. ANOVA test results showed that there were no significant differences between treatments ( > 0.05). Based on data, the average wound healing time in mice for 14 days can be seen in Figure 2.  Preparing the test materials. Muli banana peels were cut into small pieces and then washed clean to remove the dirt that sticks under running water.
Making the cut wounds on the mice's back. Before the back of the mice was cut, the hair had been shaved clean using a razor or scissors.
Administering the extracts. The cut wounds in the K+ group (positive control) were smeared with povide iodine and the cut wounds in the K-group (normal control) were smeared with distilled water.
Making histology slides. One mouse was taken from each group of mice to make histological slides.
Macroscopic and microscopic observations. The observations covered the wound healing time, length measurements measured using calipers, hyperemic features, granulation, and crusting. properties. Excessive antioxidant activity in the wound healing process can inhibit the wound healing process (Kurahashi & Fujii, 2015).
Each experimental group experienced a healing process. The positive control group was smeared with povidone-iodine. Povidone-iodine is a commonly used drug for wound healing because it is an antiseptic so that it helps to accelerate wound healing (Bigliardi et al., 2017). However, the negative control group also experienced the wound healing process even in a longer period because the body has the natural ability to protect and restore itself from injury (Govindam, Kuchi, Balekari, & Rani, 2011). On the 5 th -day, the wound on the back of the mice had begun to shrink proven by the formation of granulation and crusts marked by the formation of dried exudate or scab in each group. The observation on the 7 th -day of the wound on the P4 group showed a sign of total wound healing with the absence of the attached crust as can be seen in Figure   5. Macroscopic observations were made by describing and scoring the results of the histology of mice's skin based on the description table. A 40x magnification microscope was utilized to see differences in results epithelialization, the formation of new blood vessels, and the number of inflammatory cells. Based on the results of the crosssectional histology of the skin of mice, the histological score of the skin can be made which can be seen in the table below.  Table 1 shows that normal epitalization occurred in the treatment group that was given Muli banana peels ethanol extract of 10%, 20%, and 40%. The epithelium is a protective tissue that protects the tissue underneath from external disturbances in the form of mechanical or microbial (Honari, Andersen, & Maibach, 2017). There were more than 2 new blood vessels found in the histology of the skin of the treatment group that was given the Muli banana peels ethanol extract of 20% and 40%. When the tissue is regenerating blood vessels, its function is to carry oxygen and nutrients to help heal wounds (Reinke & Sorg, 2012). There were 6-10 inflammatory cells per microscope field of view found in the skin histology of the treatment group that was given Muli banana peels ethanol extract of 40%. Inflammatory cells show an inflammatory process that can function to avoid tissue from contamination caused by microorganisms (Panche, Diwan, & Chandra, 2016).
Active compounds contained in banana peels such as flavonoids act as bacterial growth inhibition. Besides, the flavonoids and the antioxidant activity can increase epitalization (Thakur, Jain, Pathak, & Sandhu, 2011). Saponins have antifungal properties, enhance the immune system, reduce the occurrence of blood clots and stimulate the formation of new epithelial cells, thereby, accelerating the process of reepitalization during the wound healing process (Prasetyo, Wientarsih, & Priosoeryanto, 2010). Tannins serve as  antimicrobial, antidiarrheal, and anthelmintic (Tiwari, Kumar, Kaur, Kaur, & Kaur, 2011).
Based on research by (Yunanda & Rinanda, 2016), metabolite compounds are good for wound healing by accelerating the formation of granulation tissue, contraction of the wound, fibroblast maturation, and reducing hyperemia around the wound. Muli banana plant (Musa acuminate Colla) type AAA genome, is a plant that grows in Indonesia because it is much loved by people from various backgrounds. Banana Muli is often served as a dessert in various events because it is sweet and small.
Besides, other parts of the banana plant such as leaves, flowers, pseudostems, tubers, and banana peels are often used for treatment (Onyenekwe et al., 2013). The Ambon banana stem extract ointment at a concentration of 20% can increase the number of infiltrations of inflammatory cells by four, improve neocapillary shape, accelerate the formation of fibroblasts, thus accelerating the wound healing process. Based on the statement above, it is necessary to research testing the extract of Muli banana peels (Musa acuminata Colla) on the wound healing process on the back of mice.

CONCLUSIONS AND SUGGESTIONS
Based on the results of the research, it can be concluded that the Muli banana (Musa acuminata) peels ethanol extract with a concentration of 40% was the best compared to positive and negative controls. Ethanol extract of Muli banana peels (Musa acuminata) has effects on the wound healing process, including the level of epitalization, the number of new blood vessels, and the number of inflammatory cells.
It is suggested for further researchers to conduct further research on the ethanol extract of Muli banana peels. This research can be used as a reference for subsequent research.