THE ENHANCEMENT OF COLLAGEN SYNTHESIS PROCESS ON DIABETIC WOUND BY MERREMIA MAMMOSA (LOUR.) EXTRACT FRACTION
Objective: This research aimed to evaluate the effect of fractionation of Merremia mammosa Lour. (Mm (Lour.)) extract on diabetic wound healing by observing the collagen synthesis process and to search the most potent fraction.
Methods: Wistar rats were divided into five groups (n=5), i.e., K-(negative control), K+(positive control), K1 (ethyl acetate fraction), K2 (water fraction), and K3 (n-hexane fraction). The Mm (Lour.) was extracted with ethanol 70%, then fractionated by using three solvents which have different polarity. The rats were adapted in 7 d, then induced into diabetic by streptozotocin dose 40 mg/kg body weight. The wound was made by Morton excision method. Treatment was given every two days and a skin biopsy was done on day 11. Analysis of collagen density was done by photomicrograph of histopathology preparations in Masson’s trichome stained by using trinocular microscope with 400x magnification in 6 fields of view, then processed by imageJ software and analyzed by appropriate statistic tool.
Results: The results of this research showed that fractionation of Mm (Lour.) extract significantly enhanced diabetic wound healing based on macroscopic (percentage of wound healing) and collagen density with p-value<0.05 when compared with negative control, especially the water fraction (p=0.000). The follow-up post hoc analysis showed that there was no significant (p=0.989) or there was no meaningful difference in the group of water fraction when compared to positive control.
Conclusion: Water fraction is the extract fraction of Mm (Lour.) which has the most significant influence on diabetic wound healing showed by enhancement of collagen synthesis.
2. Ministry of Health of the Republic of Indonesia. Situation and analysis of diabetes. Data and information center of Ministry of Health of the Republic of Indonesia, Indonesia; 2014.
3. Chin LCH, Boulton AJM. The diabetic foot: epidemiology, risk factor, and standards of care in general surgery. Springer; 2009. p. 1867-76.
4. Nurlitasari N. Wound healing effect of perorally administration of combination sirih merah (Piper cf. fragile, Benth) leaves and pegagan (Centella asiatica, (L.) Urb) herbs infusions in the diabetic rat. Unpublished script. Universitas Indonesia, Depok, Indonesia; 2015.
5. Hidayat FK, Elfiah U, Sofiana KD. Comparison of the number of macrophage in full thickness wound incision between Merremia mammosa extract treatment and NaCl in male wistar rats. J Agromed Med Sci 2015;1:9–13.
6. Sofiana KD, Elfiah U, Ulfa EU. The effect of Merremia mammosa Lour. on wound healing of hyperglycemic male wistar rats. Unpublished articles. Fakultas Kedokteran Universitas Jember, Jember, Indonesia; 2015. Available from: http://repository. unej.ac.id [Last accessed on 20 Aug 2018].
7. Julianto IGP. Effect of Merremia mammosa (lour) on wound healing process and blood sugar levels in hyperglycemic male wistar rats. Unpublished script. Fakultas Kedokteran Universitas Jember, Jember, Indonesia; 2015.
8. Mukhriani T. Extraction, separation of compounds and identification of active compounds. J Kesehatan 2014;7:361-7.
9. Novriansyah R. The difference of collagen density around wistar mice wound incision dressing with conventional gauze and occlusive hydrocolloid for 2 and 14 d. Unpublished thesis. Universitas Diponegoro, Semarang, Indonesia; 2008.
10. Dewi AS. Antioxidant test of ethyl acetate fraction and water fraction of green tea ethanol extract through hydroxyl radical examination with deoxyribose method. Unpublished script. Universitas Sanata Dharma, Yogyakarta, Indonesia; 2007.
11. Sugara TH, Irawadi TT, Suprapto HH, Hanafi M. Anti-bacteria activity of ethyl acetate fraction of bandotan leaves (Agerantum conyzoides L). Jurnal Ilmiah Ibnu Sina 2016;1:88-96.
12. Damasceno DC, Netto AO, Iessi IL, Gallego FQ, Corvino SB, Dallaqua B, et al. Streptozotocin-induced diabetes models: pathophysiological mechanisms and fetal outcomes. Hindawi Publishing Corporation. BioMed Res Int 2014;1-11. http://dx.doi.org/10.1155/2014/819065
13. Morton JJ, Malone MH. Evaluation of vulnerary activity by an open wound procedure in rats. Arch Int Pharmacodyn Ther 1972;196:117-26.
14. Wathoni N, Hasanah AN, Mohammed AFA, Pratiwi ED, Mahmudah R. Accelerated wound healing ability of sacran hydrogel film by keratinocyte growth factor in alloxan-induced diabetic mice. Int J Appl Pharm 2018;10:57-61.
15. Badr G. Camel whey protein enhances diabetic wound healing in a streptozotocin-induced diabetic mouse model: the critical role of ?-defensin-1,-2 and-3. Lipids Health Disease 2013;12:46.
16. Chen Y, Yu Q, Xu C. A convenient method for quantifying collagen fibers in atherosclerotic lesions by imagej software. Int J Clin Exp Med 2017;10:14904-10.
17. Kirubandanan S, Bharathi R, Renganathan S. Histological and biochemical evaluation of wound regeneration potential of Terminalia chebula fruits. Asian J Pharm Clin Res 2016;9:228-33.
18. Amaliya S, Soemantri B, Utami YW. Effects of pegagan leaves (Centellaasiatica) extract in accelerating contaminated wound on Rattus novergicus wistar strain. Jurnal Ilmu Keperawatan 2013;1:19-25.
19. Waspadji S. Kaki Diabetes. Editor Setiati S, Alwi I, Sudoyo AW, MSK, B Setiyohadi, Syam AF. Buku Ajar Ilmu Penyakit Dalam. (Book of Internal Medicine). Edisi Empat. Jilid 2. Jakarta: Interna Publishing; 2015.
20. Prahesti NR, Suzery M, Cahyono B. The antioxidant activities, phenolic total, and cytotoxicity of extract and fractions of Aloe Vera Linn). Jurnal Sains Dan Matematika 2015;23:50-4.
21. Rahayu S, Kurniasih N, Amalia V. Ekstraksi dan identifikasi senyawa flavonoid dari limbah kulit bawang merah sebagai antioksidan alami. Al Kimiya 2015;2:1-8.
22. Pratiwi L, Fudholi A, Martien R, Pramono S. Ethanol extract, ethyl acetate extract, ethyl acetate fraction, and n-heksan fraction mangosteen peels (Garcinia mangostana L.) as the source of bioactive substance free-radical scavengers. J Pharm Sci Clin Res 2016;1:71–82.
23. Faisal AP, Azhari A. Identification of secondary metabolites and antioxidant activity of mojo leaves (Aegle Marmelos L.) against DPPH (1,1 Difenil-2-pikrilhidrazil) free radicals. Mahakam Med Laboratory Technol J 2017;11:27-36.
24. Pasilala FB, Daniel C, Saleh C. Toxicity test (brine shrimp lethality test) and antioxidant activity of sintrong leaves (Crassocephalum Crepidioides) 2,2-diphenyl-1-picrylhidrazil (DPPH) method. Jurnal Kimia Mulawarman 2016;14:13-4.
25. Tanaya V, Retnowati R, Suratmo S. Fraksi semi polar dari daun mangga kasturi (Semi polar fractions of Mangifera casturi Kosterm). Kimia Student J 2015;1:778-84.