GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Indonesian Scholars’ Alliance Open Access Research Article Effectiveness of Gedong Gincu Mango Leaf Extract Gel (Mangifera indica L. var. Gedong gincu) on Cut Wound Healing in Male White Rats (Rattus norvegicus) of the Wistar Strain Azzahro Maharani Loren1,* , Ignatius Hapsoro Wirandoko1,# , Sri Marfuati2 1Departement of Basic Medical Sciences, Faculty of Medicine, Universitas Swadaya Gunung Jati, Indonesia, 2Departement of Pharmacology, Faculty of Medicine, Universitas Swadaya Gunung Jati, Indonesia *Corresponding author’s e-mail: azzahromaharani04@gmail.com* , ignatiushapsorowirandoko@gmail.com# DOI: 10.35898/ghmj-741041 ABSTRACT Background: The Indonesia Basic Health Research data show the prevalence of wounds including incision/scratches/stab wounds reaches 20.1%. Natural herbal products play an important role in the wound healing process functioning as anti-inflammatories, antioxidants, antimicrobials, and collagen formation. Mango leaves are often considered waste even though they have many health benefits with the high content of coactive compounds, such as alkaloids, flavonoids, tannins, and saponins which are good for cut wound healing. The use of gel is more stable and controlled release compared to other topical preparations. Gedong Gincu mangoes originating from West Java have unique properties and they are marketed widely in some areas including Cirebon. Aims: This study aims to identify the most effective concentration of Gedong Gincu mango leaf extract on cut wound healing in male white rats (Rattus norvegicus) of the Wistar Strain. Methods: This study used a post-test-only control group design. The extract gel used three levels of concentration, namely 40%, 50%, and 60%. The positive control used 10% povidone iodine while the negative control used a gel base. The gel was applied to the cut wound twice a day for 14 days. Results: The average length of the cut wound was 0.916 cm, 0.912 cm, and 1.050 cm for concentration levels of 40%, 50%, and 60%. For the positive and negative controls, it reached 0.996 cm and 0.702 cm respectively. This indicates no significant difference in the length of the cut wound with a p-value of 0.121 (p>0.05). The wound healing process was effective at a concentration level of 60% as indicated by the disappearance of erythema and edema. Conclusion: Mango leaf extract gel (Mangifera indica L.var. Gedong Gincu) is effective in healing cut wounds with the disappearance of erythema and edema in male white rats (Rattus norvegicus) Wistar strain with a concentration of 60%. Keywords: Natural products, Mangifera indica, Wound healing, Povidone iodine. Received: 25 September 2024 Reviewed: 19 October 2024 Revised: 30 November 2024 Accepted: 15 December 2024 © Yayasan Aliansi Cendekiawan Indonesia Thailand (Indonesian Scholars’ Alliance). This is an open-access following Creative Commons License Deed - Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) mailto:azzahromaharani04@gmail.com* mailto:ignatiushapsorowirandoko@gmail.com https://doi.org/10.35898/ghmj-741041 https://orcid.org/0009-0008-6145-845X https://orcid.org/0009-0004-6024-5605 https://orcid.org/0009-0006-1320-4848 325 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. 1. Introduction Wounds are damage or loss of part of the tissue (Sjamsuhidajat et al., 2017). The Indonesia Basic Health Research data show that in Indonesia, the prevalence of wounds including incisions/cuts/stabs reaches 20.1% (RISKESDAS, 2018). This result ranks third after abrasions/bruises/contusions and sprains. This value is based on national data, while the percentage result in West Java reaches 21.4% (RISKESDAS, 2018). Wound healing methods are varied including the use of modern products. Natural products made from herbs provide anti-inflammatories, antioxidants, antimicrobials, and collagen formation for the wound healing process with few side effects (El-Sherbeni S. A. et al., 2023; Trinh X. et al., 2022; Criollo-Mendoza M. S. et al., 2023). Gedong gincu mangoes are exotic and have unique characteristics. These fruits are from West Java and spread across Majalengka, Indramayu, and Sumedang Districts (Ariningsih E. et al., 2021; Bangun RHB. et al., 2020; Umam K. et al., 2021; Sumantri K. et al., 2021). They have been widely distributed and marketed to various regions including Cirebon District (Awaliyah F. et al., 2020). Mango leaves are often considered waste but they have many health benefits as they contain many bioactive compounds (Kumar M. et al., 2021). Previous studies revealed that mango leaves contain alkaloids, flavonoids, tannins, and saponins. Those compounds play a role in the wound- healing process (Suhatri et al., 2022). Previous studies focusing on the use of gedong gincu mango peel extract gel for cut wound healing showed that mango leaves contain alkaloids, flavonoids, tannins, saponins, phenols, triterpenoids, and steroids. These metabolite compounds have a mechanism of action on the assessment of wound length, erythema, edema, and wound healing time. In this study, the concentration that provides the best wound healing effect was 5% and 10% (Fathunnisa, et al., 2024). The topical preparation formulation in the form of mango leaf extract gel with concentrations of 40%, 50%, and 60% according to previous studies with cassava leaf extract on wound healing in New Zealand white male rabbits with a good formulation with organoleptic tests, homogeneity tests, pH tests, and gel preparation spread tests, as well as final experiments for the effect on the wound healing process. The concentration of 60% provided the best wound-healing effects (Megawati S. et al., 2020). The use of topical preparations in the management of skin diseases is common. Gel formulations applied in the management of skin diseases are a safe and effective choice. Gels are more stable with more controlled release than other topical preparations. As a topical drug, gels have good absorption (Umam K. et al., 2021). Based on the explanation above, this study tries to experiment with making Gedong Gincu mango leaf extract. Gedong gincu mangoes originating from West Java have unique properties as wound healers with various secondary metabolites. The determination of the sampling concentration refers to previous studies and produced significant results with different gel extracts. Therefore, this study aims to identify the use of Gedong Gincu mango leaf extract as a secondary metabolite in cut wound healing. 2. Methods Study design/ Research procedures This experimental laboratory study used a post-test-only control group design. This study aims to determine the effectiveness of Gedong gincu mango leaf extract gel (Mangifera indica L. var. gedong gincu) on cut wound healing in male white rats (Rattus norvegicus) of the Wistar strain. The researcher collected data on the length of the cut wound and the presence of erythema and edema for 14 days. The inclusion criteria were male white rats of the Wistar strain aged 2-3 months with a body weight of 150-250 grams and an cut wound on the back. The exclusion criteria were male white rats of the Wistar strain who got sick during treatment. The male white rats of the Wistar strain that died were dropped out. This research was conducted in the research laboratory of the Faculty of Medicine, Swadaya Gunung Jati University. Preparation of mango gedong gincu leaf extract The preparation of mango leaf extracts used the maceration method. The mango leaves were obtained from plantations in the Sedong area, Cirebon District. The sun-dried mango leaves were blended and sieved. The results Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 326 were added with 70% ethanol with a ratio of solute and solvent of 1:10. Thus, 1 kg of simplicia powder was added with 70% ethanol. The results were macerated for 3x24 hours and stirred twice with a rotary evaporator at a temperature of 50oC. The results were weighted and put into the oven to produce a thick extract. Phytochemistry Test Of Mango Gedong Gincu Extract Gel For the alkaloid test, 0.5 grams of mango leaf extract extract was added with 1 mL of 2 N HCl and 9 mL of hot distilled water, then filtered. The first tube was added with bouchardat reagent with a positive result if getting black- brown sediment. Dragendorff reagent was added to the second tube with a positive result if getting a white fog sediment (Ifmaily I. et al., 2023). Hager reagent was added to the third tube with a positive result when getting a yellow sediment. Mayer reagent, methanol, and bouchardat reagent were added to the fourth tube and the result was considered positive if getting brownish-black sediment. For the flavonoid test and phenolic test, 0.5 grams of mango leaf extract were added with 2 mL of methanol. They were then filtered and divided into two different tubes. The first tube was added with 10% NaOH and considered to have positive results if getting red hydroquinone phenol (Ifmaily I. et al., 2023; Rusnedy R. et al., 2023). Concentrated H2SO4 was added to the second tube with a positive indication when getting a red flavonoid compound. For the tannin test, 1 gram of mango leaf extract was added with 10 mL of distilled water and then filtered. The results were added with FeCl3 with a positive result if getting a dark blue or greenish black color. For the saponin test, 0.5 grams of mango leaf extract in the test tube was added with 10 mL of distilled water, and then shaken. It was left for 2 minutes and added with 1 drop of HCl 2 N. The results will be positive if getting foams for± 10 minutes (Ifmaily I. et al., 2023). For the triterpenoid test, 0.5 grams of mango leaf extract was added with Bouchardat reagent. If it gets a red or purple color, it is declared positive. For the steroid test, 0.5 grams of mango leaf extract was added with ether, 3 drops of acetic acid, and 1 drop of concentrated H2SO4. If getting a green color, it is declared positive. Preparation of gel mango gedong gincu leaf extract Table 1 shows the formulation of mango leaf extract gel covering Na-CMC and hot distilled water which are homogenized, glycerin, propylene glycol, and methylparaben which are homogenized. Mango leaf extracts are added with that formulation with a certain concentration. After getting the gel, it was kept away from sunlight for one night (Hasanudin MN. et al., 2023). Then, the gel was tested using organoleptic tests, pH tests, homogeneity tests, and spreadability tests (Budi S. et al., 2020; Nurlatifah N. et al; 2022). Tabel 1. Gedong gincu mango leaf extract gel formulation Ingredient Formulations group (grams) K1 (40%) K2 (50%) K3 (60%) KN Mango gedong gincu leaf extract 20 25 30 - Na-CMC 0,5 0,5 0,5 0,5 Metyl paraben 0,25 0,25 0,25 0,25 Glyserin 5 5 5 5 Propylene glycol 2,5 2,5 2,5 2,5 Aquadest ad 50 50 50 50 Evaluation of mango gedong gincu leaf extract gel preparation results 1. Organoleptic test The results of the organoleptic test for gedong gincu mango leaf extract covered the physical form, smell, and color of the gel preparation (Budi S. et al., 2020). 327 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. 2. pH test The pH test aims to assess the acidity level of the gedong gincu mango leaf extract gel preparation. The test used a pH meter (Budi S. et al., 2020). A good gel preparation has a value of 4.5-6.5 (Nurlatifah N. et al; 2022). 3. Homogeneity test The homogeneity test aims to assess the distribution of the gedong gincu mango leaf extract gel content evenly (Budi S. et al., 2020). This test was conducted by applying the gel to clear glass and looking at whether coarse grains appear or not. The result of the test is considered good if all groups based on extract concentration do not contain coarse grains (Nurlatifah N. et al; 2022). 4. Spreadability test This test aims to assess the spreadability of the gedong gincu mango leaf extract gel preparation. A value of 5-7 cm is considered good. This test was carried out by dropping 0.5 grams of gel on an object glass and then covering the top with another object glass. The results were in the form of spreadability value (Nurlatifah N. et al; 2022). Making cut wounds on male white rats The researcher used white male rats of the Wistar strain with cut treatment. Before getting the cut, the rats were given 2% lidocaine subcutaneously and got their back fur shaved. The cut area was cleaned with alcohol cotton. The cut was made using a scalpel with a length of ± 2 cm and a depth of ± 0.3 cm. The cut wound healing was based on 3 criteria, namely the length of the cut, erythema stopped, and edema stopped. The Gedong gincu mango leaf extract gel was applied twice a day for 14 days. Measurements In this research, the independent variable was Gedong gincu mango leaf extract gel (Mangifera indica L. var. gedong gincu). Then, the dependent variable was the cut wound healing in male white rats of the Wistar strain. The sample size was based on the Federer formula and the determination of the sample used a simple random sampling technique. Statistical analysis The cut wound healing was seen from 3 criteria, namely the length of the cut, erythema elliminated, and edema disappeared. The treatment was performed by applying mango leaf extract gel twice a day for 14 days. Statistical analysis used SPSS (Statistical Package for the Social Sciences). The data obtained were analyzed using the following steps. The length of the cut was assessed first with a normality test using the Shapiro-Wilk test because the sample was less than 50, namely only 25 male white rats. Second, the homogeneity test used the Levene Test. Third, the comparison test used the One-Way Annova test when the requirements for normality and homogeneity are met or (p (Sig.) > 0.05). Then the assessment of erythema and edema was based on direct observation to determine whether the erythema and edema have disappeared or not. Ethical clearance This study has received approval from the Ethics Commission of the Faculty of Medicine, Swadaya Gunung Jati University, Cirebon Number 8/EC/FKUGJ/IV/2024, dated April 8, 2024. The mango plants used in this study have been tested at the UNNES Biology Laboratory with the number 54/UN.37/SHP/Lab. Plant Taxonomy/III /2024. Male white rats of the Wistar strain have an animal identity certificate number 09.141/RTP/VI/2024 which comes from Pahlevi's house rats. Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 328 3. Results The researcher obtained 311 grams of thick mango leaf extract. The characteristics of this mango leaf extract gel were thick with a blackish-brown color and had a distinctive mango leaf odor and a bitter taste. Table 2 shows that the Gedong gincu mango leaf extract (Mangifera indica L. var. Gedong gincu) contains secondary metabolite compounds such as phenolics, flavonoids, tannins, saponins, triterpenoids and steroids. Tabel 2. Results phytochemistry test Phytochemicals Reagent Results Depiction Alkaloids Bouchardat - Photo 1 Dragendroff - Photo 2 Hager - Photo 3 Mayer+methanol+bauchardat - Photo 4 Fenolics NaOH 10% ++ Photo 5 Flavonoids H2SO4 concentration + Photo 6 Tannins FeCl3 +++ Photo 7 Saponins HCl 2 N +++ Photo 8 Triterpenoids Bouchardat + Photo 9 Steroids Eter+3 drops anhydrous+1 drop of H2SO4 concentration + Photo 10 Description: 1. Negative results = (-) 2. Positive results: a. Weak = (+) b. Strong = (++) c. Very strong = (+++) d. Extremely strong = (++++) Table 3 shows that the physical form, odor, and color of gel are in accordance with the extracts used. The physical form of the gel should be semi-solid, have a mango leaf extract odor, and a blackish brown color. The results of the organoleptic test showed that the mango leaf extract gel preparation is as expected. Tabel 3. Results organoleptic test Groups Results Physical form Odor Colour 40% concentration Semi-solid/viscous Mango leaf extract Brown-black 50% concentration Semi-solid/viscous Mango leaf extract Brown-black 60% concentration Semi-solid/viscous Mango leaf extract Brown-black Gel base Semi-solid/viscous Don’t have odor Pellucid Table 4 shows that the concentrations of 40%, 50%, and 60% have good standard normal pH values. The negative control, namely the gel base preparation, did not show a normal pH value. So, concentrations of 40%, 50%, and 60% show good pH results. Tabel 4. pH test (Photo 11) Groups Normal value Results 40% concentration 4,5-6,5 6 50% concentration 4,5-6,5 6 60% conecntartion 4,5-6,5 6 Gel base 4,5-6,5 7 329 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. Photo 1. Bouchardat alkaloids (-) Photo 2. Dragendroff alkaloids (-) Photo 3. Hager alkaloids (-) Photo 4. Mayer alkaloids (-) Photo 5. Phenolics (++) Photo 6. Flavonoids (+) Photo 7. Tannins (+++) Photo 8. Saponins (+++) Photo 9. Triterpenoids (+) Photo 10. Steroids (+) Photo 11. Results of pH test of mango leaf extract gel Table 5 shows that the spreadability at concentrations of 40%, 60%, and the negative control obtained good spreadability. The 50% concentration had poor spreadability as indicated by a value of lower than 5-7 cm. Tabel 5. Spread test Groups Normal value Results Depiction 40% concentration 5-7 cm 5,1 cm Photo 12 50% concentration 5-7 cm 4,9 cm Photo 13 60% concentration 5-7 cm 5,2 cm Photo 14 Gel base 5-7 cm 5,3 cm Photo 15 Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 330 Photo 12. 40% consentation Photo 13. 50% concentration Photo 14. 60% concentration Photo 15. Gel base Table 6 shows that the gel preparations with concentrations of 40%, 50%, 60% and negative control obtained good results that the gel was homogeneous and did not clot. Tabel 6. Homogeneous test Groups Results Depiction 40% concentration Homogeneous, don’t have lumps Photo 16 50% consentration Homogeneous, don’t have lumps Photo 17 60% concentration Homogeneous, don’t have lumps Photo 18 Gel base Homogeneous, don’t have lumps Photo 19 Photo 16. 40% concentation Photo 17. 50% concentration Photo 18. 60% Concentration Photo 19. Gel base 331 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. Table 7 shows that the average length of the cut wound in each treatment group did not differ significantly. Tabel 7. Measurement results of the length of the cut wound Concentration Average length of cut wound (cm) 1 2 3 4 5 40% 1,81 1,83 2,29 1,73 1,63 50% 1,67 1,48 1,86 1,41 2,03 60% 1,71 1,60 1,98 1,68 1,98 (+) 1,46 1,23 1,67 1,50 1,99 (-) 1,68 1,52 1,90 1,93 1,81 Table 8 shows that 3 rats did not experience healing of erythema. In the group with a concentration of 60% and positive control, all rats experienced healing of erythema. Tabel 8. Results of erythema observation Group Rat Erythema disappeared The day the erythema disappeared 40% concentration 1 Yes Day 14 2 No - 3 Yes Day 11 4 Yes Day 11 5 Yes Day 11 50% concentration 1 Yes Day 13 2 Yes Day 12 3 No - 4 Yes Day 12 5 Yes Day 14 60% concentration 1 Yes Day 12 2 Yes Day 12 3 Yes Day 11 4 Yes Day 11 5 Yes Day 13 Positive control (+) 1 Yes Day 7 2 Yes Day 13 3 Yes Day 11 4 Yes Day 9 5 Yes Day 13 Negative control (-) 1 Yes Day 11 2 No - 3 Yes Day 11 4 Yes Day 12 5 Yes Day 14 Table 9 shows that only group 3 with a concentration of 60% experienced edema healing. At a concentration of 40% and positive control, edema did not disappear in one rat. At a concentration of 50%, two rats whose edema did not disappear. In the negative control with a gel base, edema was difficult to assess in one rat because it was covered by scabs. Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 332 Tabel 9. Results of edema observation Group Rat Edema disappeared The day the edema disappeared 40% concentration 1 Yes Day 8 2 No - 3 Yes Day 13 4 Yes Day 11 5 Yes Day 10 50% concentration 1 No - 2 Yes Day 14 3 Yes Day 11 4 Yes Day 12 5 No - 60% concentration 1 Yes Day 11 2 Yes Day 13 3 Yes Day 10 4 Yes Day 13 5 Yes Day 10 Positive control (+) 1 Yes Day 6 2 No - 3 Yes Day 11 4 Yes Day 10 5 Yes Day 12 Negative control (-) 1 Yes Day 7 2 It’s hard to judge - 3 Yes Day 9 4 Yes Day 11 5 Yes Day 14 Table 10 shows that this research involved 25 sample rats. So, the normality test used was the Shapiro-Wilk Test. Data are considered normally distributed if the significance value is higher than 0.05 (p>0.05). The data are the result of dividing Day 14 by Day 1. In group 1 with a concentration of 40%, the Sig. value was 0.611. In group 2 with a concentration of 50%, the Sig. value was 0.232. In group 3 with a concentration of 60%, the Sig value was 0.555. In group 4 with the administration of 10% povidone-iodine as a positive control, the Sig value was 0.994. Group 5 with gel base as a negative control obtained a Sig. Value of 0.206. So, the results of the normality test presented in table 10 show that the data are normally distributed because the p-value is higher than 0.05. Tabel 10. Normality test for the cut wound length Groups Amount Sig. Description 40% concentration 5 0,611 Normal 50% concentration 5 0,232 Normal 60% concentration 5 0,555 Normal Positive control (+) 5 0,994 Normal Negative control (-) 5 0,206 Normal *Shapiro-Wilk test 333 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. Table 11 shows the results of the homogeneity test using the Levene test. The data are homogeneous with a significance value of 0.232 meaning that the p-value is higher than 0.05. Tabel 11. Results of homogeneous cut wounds Variable Sig. Description Length of cut wound 0,232 Homogeneous *Levene test Table 12 shows that the data are normally distributed so it is continued with a one-way ANOVA test to see the effect of treatment on the length of the cut wound based on the group in the sample of male white rats of the Wistar strain. A p-value lower than 0.05 (p <0.05) is considered significant. Tabel 12. Results of One-way Annova Test Kelompok Mean±SD Sig. 40% concentration 0,916±0,171 50% consentration 0,912±0,107 60% concentration 1,050±0,251 0,121 Positive control (+) 0,996±0,309 Negative control (-) 0,702±0,101 *One Way Annova Test The treatment group 1 with a concentration of 40% obtained a mean value of 0.916 (Photo 20). The treatment group 2 with a concentration of 50% obtained a mean value of 0.912 (Photo 21). The treatment group 3 with a concentration of 60% obtained a mean value is 1.050 (Photo 22). The treatment group 4 with 10% povidone iodine as a positive control obtained a mean value of 0.996 (Photo 23). The treatment group 5 with the administration of gel base as a negative control obtained a mean value of 0.702 (Photo 24). The analysis using One-Way ANOVA obtained a p-value of 0.121 which is lower than 0.05. Thus, there is no significant difference in all treatment groups for cut wounds. Photo 20. 40% concentration Photo 21. 50% concentration Photo 22. 60% concentration Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 334 4. Discussion Mechanism of secondary metabolite compounds in wound healing process Wound healing processes cover 3 important stages, namely the inflammation, proliferation, and remodeling phases. The inflammation phase is characterized by the appearance of erythema and edema (Sjamsuhidajat, et al., 2017). Previous studies examining cut wounds with gedong gincu mango leaf extract showed that mango leaves contain flavonoids, tannins, phenols, triterpenoids, steroids, alkaloids, and saponins. The optimum concentrations were 5% and 10% which provided the best wound healing effect (Fathunnisa, et al., 2024). Gedong gincu mango leaves have various secondary metabolite compounds that can help the wound healing process. Phytochemical tests show that mango leaves contain phenolic compounds, flavonoids, tannins, saponins, triterpenoids, and steroids. However, they do not contain alkaloids. Generally, plants that have close taxonomic classifications will have secondary metabolite compounds that are not much different (Julianto TS, et al., 2019). Secondary metabolites are the result of biosynthesis metabolism that passes through 3 pathways and the results of this metabolism can be influenced by some factors (Anggraito YU, et al., 2018). Secondary metabolite compounds in plants depend on plant growth conditions, such as season, year, and environment. Plant growth is influenced by genetic factors in the biosynthesis metabolism pathway for the formation of secondary metabolite compounds (Li Y, et al., 2020). This contradicts the results of a previous study that obtained positive alkaloid results (Suhatri, et al., 2022). Flavonoids are compounds with a chemical formula of C6C3C6 whose fifteen-carbon basic framework consists of two benzene rings (A and B) and is connected by a heterocyclic pyran ring (C) (Julianto TS, 2019). Flavonoids are secondary metabolite compounds that work by inhibiting the migration of leukocyte cells that appear after getting wound, encouraging the formation of apoptosis, and maintaining cell survival (Anisa N, et al., 2019; Al-Khayri JM, et al., 2022). In this study, erythema and edema did not disappear in the concentrations of 40% and 50%. The inflammatory phase is influenced by other secondary metabolic compounds, namely phenolics and tannins. In the wound healing process, phenolics work by counteracting free radicals to prevent tissue damage and inhibiting the formation of enzymes that work in the inflammatory phase, namely the cyclooxygenase enzyme (Anisa N, et al., 2019). Tannins are natural compounds contained in various plants (Das AK, et al., 2020). The molecular weight of tannins ranges from 500-3000 (gallic acid ester) and higher than 20,000 (proanthocyanidins) (Julianto TS, 2019; Anisa N, et al., 2019). Tannins function to prevent free radicals, reduce edema, and prevent pus in wounds (Suhatri, et al., 2022; Anggraito YU, et al., 2018). In the negative control with a gel base, rats discharged pus from their wounds indicating that the mango leaf extracts prevent the formation of pus because the negative control was not provided with the mango leaf extract. Tannins act as anti-inflammatories and antibacterials by reducing mucosal permeability and strengthening mucosal bonds causing difficulties for bacteria to penetrate wounds and inhibit bacterial proliferation. Therefore, the cut wound heals faster without being worsened by a bacterial infection (Mustika S.Y., et al., 2020). Photo 23. Positive control (+) Photo 24. Negative control (-) 335 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. Saponins have properties like soap and detergent because they contain a fat-soluble group (sugar) in one molecule (Das AK, et al., 2020). Shaking saponin can produce foam on the surface of the water because saponin has the property of reducing the tension on the surface of the water (Mustikasari SY, et al., 2020). Saponins can strengthen the edges of open wounds to close faster. Saponin plays a role in the formation of collagen and tissue epithelialization in the proliferation phase. Besides, saponin encourages the formation of type 1 collagen which helps wound healing process (Sjamsuhidajat, et al., 2017; Mustikasari SY, et al., 2020; Anggraeni PP, et al., 2023). Most of the rats in each treatment group closed their wounds on day 4. Gedong gincu mango leaves contain other secondary metabolite compounds, namely steroids. Ki et al (2011) revealed that steroids play a role in the epithelialization mechanism, where the epithelium will be formed faster enabling faster closure of epithelialization (Dewi A.A., et al., 2020). Based on the phytochemical tests, Gedong gincu mango leaves contain secondary metabolite compounds, namely triterpenoids. Triterpenoids inhibit ROS (Reactive oxygen species) interference in the wound-healing process. Thus, the presence of triterpenoids can reduce interference with the healing process. Besides, triterpenoids act as compounds that increase antioxidant activity (Vitale S, et al., 2022). This compound helps the wound healing process. Cut wound healing process in different concentration levels Treatment groups with 40% concentration levels show that the average length of the cut wound is not significantly different as the cut line does not disappear until the last day of the treatment. Erythema and edema disappeared and almost all rats experienced wound healing. However, the 2nd rat did not experience healing indicated by the presence of erythema, edema, and abscesses. A total of 4 treatment rats experienced healing until the last day of treatment. Negative alkaloids in Gedong gincu mango leaves affect the wound healing process. The 2nd rat did not experience the wound-healing process. The treatment groups with 50% concentration levels show a difference in the average length of the cut wound. Erythema and edema did not completely disappear in all rats. Only 2 rats in this group experienced the wound-healing process. This shows that this concentration level is not as good as the 40% concentration level. Based on the spreadability test of the mango leaf extract, the 50% concentration showed poor spreadability, namely less than 5 cm. The group with 60% concentration level shows a difference in the average length of the cut wound. In all rats, erythema and edema disappeared with wound healing until the last day of treatment. In this group, the day of wound healing did not differ much among rats. This shows that the 60% concentration is good for the wound- healing process. The positive control used 10% povidone iodine. Previous studies revealed that the use of 10% povidone- iodine does not have an anti-inflammatory and antiseptic effect and can interfere with collagen synthesis because it can attack healthy cells (Mustikasari SY, et al., 2020). In this group, erythema disappeared in all treated rats, but the edema did not disappear in the 2nd rat. The negative control uses a gel base to ensure whether the mango extract plays a role in the wound healing process. Based on the result of the study, the negative control showed wound healing in four rats only. However, in the second rat, there is no healing process, erythema does not disappear, and edema is difficult to assess due to being covered by scars. This means that the gel base gel does not contain mango leaf extract so it does not contain secondary metabolite compounds. Tannin affects the discharge of pus. The gel base does not contain this compound, so pus comes out of the wound. Previous studies with the same concentration of 40%, 50%, and 60% with topical extract gel preparations from cassava leaves. This study assessed the percentage of cut wound length on wound healing. The highest concentration of 60% provided the best effect in accelerating the wound healing process (Megawati S. et al., 2020). The same concentration of 40%, 50%, and 60% in this study showed insignificant results on the length of the cut wound. However, for other wound healing criteria, namely erythema and edema, a concentration of 60% is the best concentration for eliminating erythema and edema. Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 336 Another study with concentrations of 5%, 10%, and 15% of gedong gincu mango extract showed the best cut wound healing at the concentrations of 5% and 10%. The best healing of erythema and edema was at a concentration of 5% and 10% respectively on the same day (Fathunnisa. et al., 2024). In this research, based on the length of the cut wound, the wound healing criteria were not significant in all treatment groups. However, at a concentration of 40%, 4 out of 5 rats experienced healing until the end of the study. At a concentration of 50%, 3 out of 5 rats experienced healing. At a concentration of 60%, all rats experienced a healing process for both erythema and edema. In the positive control group, the use of 10% povidone-iodine indicated that all rats experienced healing with the disappeared erythema. However, only 4 rats experienced healing with the disappeared edema with no influence on the length of the cut wound between the positive control and other treatment groups. Moreover, in this study, the best concentration to eliminate erythema and edema in all rats was 60%. The group with 50% concentration has good quality in terms of shape, smell, color, and pH with poor spreadability. The 60% concentration was able to eliminate erythema and edema. The 60% concentration was the best concentration in eliminating erythema and edema until the last day of treatment in five rats compared to the other groups. Results of Statistical Testing and Wound Healing The significance of the length of the cut wound was tested using one-way ANOVA test. It obtained a p-value of 0.121 which is higher than 0.05. The statistical test showed that the length of the cut wound was not significant among all treatment groups. The intervention group with 60% concentration level experienced healing with erythema and edema disappearing in all treated rats. 5. Conclusion Mango leaf extract (Mangifera indica L.var. Gedong Gincu) contains secondary metabolite compounds such as flavonoids, phenolics, tannins, saponins, triterpenoids, and steroids. This extract has good quality in terms of shape, smell, color, pH, and poor spreadability at a concentration of 50%. Mango leaf extract gel has no effect on the length of the cut wound in male white rats of the Wistar strain. The group with a concentration of 60% can make erythema and edema disappear in male white rats of the Wistar strain. Mango leaf extract gel (Mangifera indica L.var. Gedong gincu) managed to help heal cut wounds in male white rats (Rattus norvegicus) of the Wistar strain. 6. Limitations This study has two limitations. First, the levels of active substances in mango leaf extract that affect the wound healing process have not been tested. Therefore, the extent to which the active substance influences the wound- healing process cannot be identified. Second, the levels of toxicity and toxic substances contained in Gedong Gincu mangoes are not known yet. Gedong gincu mango leaves have not been tested. Thus, which substances that have side effects on wound healing cannot be identified. 7. Clinical and Future Studies Further studies are expected to prepare extracts with different treatments, and leaf extract gels with dependent variables of burns or cuts. Further studies can examine different topical preparations, creams, ointments or gel fractions, not only on the length of the wound, but also the width and area of the wound to assess its effectiveness. . Conflict of Interest The authors declare no conflict of interest for the results. 337 GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 Loren, et. al. References Sjamsuhidajat, R., Zunilda S. Bustami, Leo, Arya Rendy Kristandyo, Aryandhito Widhi Nugroho, Bisono, Theddeus O H Prasetyono, Reno Budiman, Ignatius Riwanto, Paul Tahalele. 2017. Buku Ajar Ilmu Bedah Masalah, Pertimbangan Klinis Bedah, Dan Metode Pembedahan. 4th ed. Jakarta: EGC. Badan Penelitian dan Pengembangan Kesehatan RI. 2018. "Laporan Riskesdas 2018 Nasional.Pdf." Lembaga Penerbit Balitbangkes: 156. El-Sherbeni, S. A., and W. A. Negm. 2023. "The Wound Healing Effect of Botanicals and Pure Natural Substances Used in in Vivo Models." Inflammopharmacology 31(2): 755-72. https://doi.org/10.1007/s10787-023- 01157-5 Trinh, Xuan Tung et al. 2022. "A Comprehensive Review of Natural Compounds for Wound Healing: Targeting Bioactivity Perspective." International Journal of Molecular Sciences 23(17). https://doi.org/10.3390/ijms23179573 Marilyn S. Criollo-Mendoza et al. 2023. "Wound Healing Properties of Natural Products: Mechanisms of Action." Molecules 28: 598. https://doi.org/10.3390/molecules28020598 Ariningsih, Ening et al. 2021. "Kinerja Agribisnis Mangga Gedong Gincu Dan Potensinya Sebagai Produk Ekspor Pertanian Unggulan." Forum penelitian Agro Ekonomi 39(1): 49. https://doi.org/10.21082/fae.v39n1.2021.49-71 Bangun, Rita Herawaty Br. 2020. "Potensi Komoditi Mangga Gedong Gincu Dalam Pengembangan Wilayah Dan Perekonomian Kabupaten Majalengka." Jurnal Sosial Ekonomi Pertanian 16(3): 213-25. Megawati, Sefi, Nur'aini Nur'aini, and Dewi Kurniasih. 2020. "Uji Efektivitas Gel Ekstrak Etanol 96% Daun Singkong (Manihot Esculenta Crantz.) Pada Penyembuhan Luka Sayat Kelinci Jantan Galur New Zealand White." Jurnal Farmagazine 7(1): 1. https://doi.org/10.47653/farm.v7i1.159 Umam, Khoirul, Rokhani Hasbullah, and Mohamad Solahudin. 2021. "Karakteristik Dan Kelayakan Finansial Usahatani Mangga Gedong Gincu Di Kabupaten Indramayu." Jurnal Keteknikan Pertanian 9(3): 119-26. https://doi.org/10.19028/jtep.09.3.119-126 Sumantri, Kosasih. 2021. "Strategi Pemasaran Mangga Gedong Gincu Kabupaten Sumedang." Agrivet : Jurnal Ilmu- Ilmu Pertanian dan Peternakan (Journal of Agricultural Sciences and Veteriner) 9(2): 200-205. https://doi.org/10.31949/agrivet.v9i2.1749 Awaliyah, Fitri, and Bobby Rachmat Saefudin. 2020. "Efisisensi Pemasaran Komoditas Mangga Gedong Gincu Di Kabupaten Cirebon." Paradigma Agribisnis 3(1). https://doi.org/10.33603/.v3i1.3543 Fathunnisa F, Marfuati S, Fitriani H. 2024. "Effectiveness of Mango Gedong Gincu Peel Extract Gel ( Mangifera Indica L .) on Cut Wounds in Male Wistar White Rats ( Rattus Norvegicus )." GHMJ (Global Health Management Journal) 7(3): 1-9. https://doi.org/10.35898/ghmj-731060 Kumar, Manoj et al. 2021. "Mango (Mangifera Indica l.) Leaves: Nutritional Composition, Phytochemical Profile, and Health-Promoting Bioactivities." Antioxidants 10(2): 1-23. https://doi.org/10.3390/antiox10020299 Suhatri, Fidiyani, Dwi, Nia Azzahra, and Muslim Suardi. 2022. "Effectiveness of Mango Leaf Extract (Mangifera Indica L) on Healing of Second Level Cut Wound in Male White Mice (Mus Musculus)." JURNAL KESEHATAN TERAPAN 9: 66-71. https://doi.org/10.54816/jk.v9i2.520 Ifmaily, Ifmaily, Ulfa Anggraini, and Putri Rizki Fitriani. 2023. "Pengaruh Salep Ekstrak Biji Buah Mangga Arumanis (Mangifera Indica L ) Terhadap Penyembuhan Luka Eksisi Secara In Vivo." Jurnal Ventilator: Jurnal riset ilmu kesehatan dan Keperawatan 1(4): 362-77. https://doi.org/10.59680/ventilator.v1i4.830 Rusnedy, Rahmayati, Mira Febrina, and Cindy Patika Sari. 2023. "Uji Aktivitas Wound Healing Ekstrak Etanol Buah Averrhoa BilimbiL. (Belimbing Wuluh) Pada Mencit Putih Jantan (Mus Musculus)." Pharmacon: Jurnal Farmasi Indonesia 20(1): 50-60. https://doi.org/10.23917/pharmacon.v20i1.22677 https://doi.org/10.1007/s10787-023-01157-5 https://doi.org/10.1007/s10787-023-01157-5 https://doi.org/10.3390/ijms23179573 https://doi.org/10.3390/molecules28020598 https://doi.org/10.21082/fae.v39n1.2021.49-71 https://doi.org/10.47653/farm.v7i1.159 https://doi.org/10.19028/jtep.09.3.119-126 https://doi.org/10.31949/agrivet.v9i2.1749 https://doi.org/10.33603/.v3i1.3543 https://doi.org/10.35898/ghmj-731060 https://doi.org/10.3390/antiox10020299 https://doi.org/10.54816/jk.v9i2.520 https://doi.org/10.59680/ventilator.v1i4.830 https://doi.org/10.23917/pharmacon.v20i1.22677 Loren, et. al. GHMJ (Global Health Management Journal) 2024, Vol. 7, No. 4 338 Muhammad Nurul Hasanudin, Muhammad Alfian, Moch. Fathul Mujib. 2023. "Uji Aktivitas Gel Ekstrak Etanol 70% Daun Kitolod (Isotoma Longiflora (L.) C Presl) Terhadap Penyembuhan Luka Sayat Pada Mencit (Mus Musculus)." Sains Indonesia: Jurnal Ilmiah Nusantara 1: 118-28. Budi, Setia, and Mila Rahmawati. 2020. "Pengembangan Formula Gel Ekstrak Pegagan (Centella Asiatica (L.) Urb ) Sebagai Antijerawat." Jurnal Farmasi Dan Ilmu Kefarmasian Indonesia 6(2): 51. https://doi.org/10.20473/jfiki.v6i22019.51-55 Nurlatifah, Nurlatifah et al. 2022. "Formulasi Sediaan Gel Dari Ekstrak Etanol Herba Pegagan (Centella Asiatica (L) Urb)." Journal of Pharmaceutical and Health Research 3(1): 10-14. https://doi.org/10.47065/jharma.v3i1.1366 Julianto, Tatang Shabur. 2019. Fitokimia Tinjauan Metabolit Sekunder Dan Skrining Fitokimia. Yogyakarta: Universitas Islam Indonesia. Anggraito, Yustinus Ulung et al. 2018. Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Negeri Semarang Metabolit Sekunder Dari Tanaman : Aplikasi Dan Produksi. Li, Yanqun et al. 2020. "The Effect of Developmental and Environmental Factors on Secondary Metabolites in Medicinal Plants." Plant Physiology and Biochemistry 148(January): 80-89. https://doi.org/10.1016/j.plaphy.2020.01.006 Anisa, Nurul et al. 2019. "Efektifitas Anti Inflamasi Daun Mangga (Mangifera Indica) Terhadap Luka Bakar Derajat Dua The Effectiveness of Anti Inflation Mangoes Leaves (Mangifera Indica) Against Burns Degrees Two." Jurnal B-Dent VIII(1): 1-7. https://doi.org/10.35580/sainsmat81101182019 Al-Khayri, Jameel M. et al. 2022. "Flavonoids as Potential Anti-Inflammatory Molecules: A Review." Molecules 27(9). https://doi.org/10.3390/molecules27092901 Das, Atanu Kumar et al. 2020. "Review on Tannins: Extraction Processes, Applications and Possibilities." South African Journal of Botany 135(October): 58-70. https://doi.org/10.1016/j.sajb.2020.08.008 Mustikasari, Susi Yanuari, Ignatius Hapsoro Wirandoko, and Ika Komala. 2020. "Efektifitas Ekstrak Daun Afrika (Vernonia Amygdalina) Terhadap Ketebalan Epitelisasi Pada Luka Insisi Mencit." Jurnal Kedokteran & Kesehatan 6(3): 12-18. Anggraeni Putri, Puspa, Moralita Chatri, and Linda Advinda. 2023. "Characteristics of Saponin Secondary Metabolite Compounds in Plants Karakteristik Saponin Senyawa Metabolit Sekunder Pada Tumbuhan." Serambi Biologi 8(2): 251-58. Dewi, Ayu Utami, and Imam Adi Wicaksono. 2020. "Review Artikel: Tanaman Herbal Yang Memiliki Aktivitas Penyembuhan Luka." Jurnal Farmaka Universitas Padjadjaran 18(2): 191-207. Vitale, Stefania et al. 2022. "Phytochemistry and Biological Activity of Medicinal Plants in Wound." Molecules 27(11): 1-30. https://doi.org/10.3390/molecules27113566 Cite this article as: Loren, A. M., Wirandoko, I. H., & Marfuati, S. (2024). Effectiveness of Gedong Gincu Mango Leaf Extract Gel (Mangifera indica L. var. Gedong gincu) on Cut Wound Healing in Male White Rats (Rattus norvegicus) of the Wistar Strain. GHMJ (Global Health Management Journal), 7(4), 324–338. https://doi.org/10.35898/ghmj-741041 https://doi.org/10.20473/jfiki.v6i22019.51-55 https://doi.org/10.47065/jharma.v3i1.1366 https://doi.org/10.1016/j.plaphy.2020.01.006 https://doi.org/10.35580/sainsmat81101182019 https://doi.org/10.3390/molecules27092901 https://doi.org/10.1016/j.sajb.2020.08.008 https://doi.org/10.3390/molecules27113566 https://doi.org/10.35898/ghmj-741041