The Enhancement of Collagen Synthesis Process on Diabetic Wound by Merremia Mammosa (Lour.) Extract Fraction

Abstract

Objective: This research aimed to evaluate the effect of fractionation of Merremia mammosa Lour. (Mm (Lour.)) extract on diabetic wound healing by observing 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 days, then induced into diabetic by streptozotocin dose 40mg/kg body weight. The wound was made by Morton excision method. Treatment was given every two days and 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.Result: 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.

Gel formulations of Merremia mammosa (Lour.) accelerated wound healing of the wound in diabetic rats

“… 26 The transition from the inflammatory phase to the proliferative phase was hampered. This was relevant to the our previous studies 11 , 12 , 13 of diabetic wound, which showed that wound healing in the negative control took longer than in the positive control and treatment groups, as assessed via several wound parameters. This was also similar with the result of Ackermann et al.…”

“…This result was consistent with our previous study on Mm(Lour) extract fractions. 11 , 12 , 13 Therefore, the findings in this study allowed the presumption that the gel formulation of Mm (Lour.) water fraction possibly restored the delayed process of diabetic wound healing.…”

“…Connective tissues were observed at day 10 after excision using histopathological examination with modified Masson’s Trichrome (MT) staining specific to collagen, which stained collagen fibers a bright blue color, 13 , 24 and measuring hydroxyproline levels. Observation of collagen density was carried out with 400x magnification at 6 fields of view, and then, pictures were taken using Olympus DP21 series microscope.…”

“…The percentage of collagen density was measured using imageJ software according to a previous study. 13 Hydroxyproline level measurement followed the General Hydroxyproline Assay Kit ® protocol. A standard curve to measure concentration of hydroxyproline was made by measuring standard concentration 0; 7,5; 15; 30; 60; 120; and 240 ng/mL at 450 nm wavelength absorbance.…”

“…showed that each could accelerate wound closure in diabetic wound healing and increase the density of collagen in diabetic wounds. 11 , 12 , 13 The effective dose of a water extract fraction of Mm ( Lour .) was 50 mg/625 mm 2 wounded area.…”