Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
53359/mfi.
Development of Pineapple Skin Extract Hydrogel Patch Applications (Ananas comosus L.
) as Wound Dressing in Diabetic Ulcers in Mice (Mus musculu.
Putik Titian Citra Hening.
Anisah Putri Yuana Sari.
Marshanda Nikita.
Fajrina Rizqi Riyadi.
Ahmad Maulana Al Fatah.
Ihsanti Dwi Rahayu1*) Pharmacy Study Program.
Department of Pharmacy.
Faculty of Medicine.
University of Lampung.
Lampung.
Indonesia.
Medical Education Study Program.
Faculty of Medicine.
University of Lampung.
Lampung.
Indonesia.
Applied Biology Study Program.
Faculty of Mathematics and Natural Sciences.
University of Lampung.
Lampung.
Indonesia Email : ihsanti.
rahayu@fk.
Abstract Diabetes mellitus is a chronic metabolic disorder caused by high blood glucose levels.
One common complication in diabetes mellitus sufferers is diabetic ulcers.
Research has shown that pineapple skin contains flavonoid, an active compound with anti-inflammatory and antimicrobial properties, which can accelerate wound healing.
This study aims to determine the effect of applying a hydrogel patch containing pineapple peel extract (Ananas comosus L.
) as a wound dressing on male white mice (Mus musculu.
with diabetic ulcers.
The study involved several stages, including pineapple peel extraction, hydrogel patch preparation, antibacterial activity testing, and application of the hydrogel patch on test animals.
The research used 20 male white mice of the Deutschland Denken Yoken (DDY) strain divided into 5 groups.
The negative control group received a hydrogel patch without extract, while the positive control group received Hydrocolloid Dressing.
Treatment groups I.
II, and i were given hydrogel patches with 10%, 20%, and 30% pineapple peel extract, respectively.
The results showed that the hydrogel patch with pineapple peel extract accelerated wound healing in diabetic ulcer models.
The antibacterial activity test indicated antimicrobial activity against Staphylococcus aureus, commonly found in diabetic ulcers.
The hydrogel patch application reduced inflammation and improved skin tissue structure in test animals, demonstrating the potential of pineapple peel extract hydrogel patches as an effective alternative wound dressing for diabetic ulcers.
Keywords: diabetic ulcer, pineapple peel extract, hydrogel patch, wound dressing INTRODUCTION The World Health Organization (WHO) stated that in 2021 there will be more than 537 million adults living with diabetes According to the International Diabetes Federation (IDF), the number of diabetes patients in Indonesia increased rapidly by 167% over the past ten years, as of Diabetes mellitus often leads to complications such as diabetic ulcers.
Currently, the incidence of diabetic ulcers in Indonesia is quite high, with an annual prevalence of 5-7.
Therefore, the Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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management of diabetic ulcers requires a comprehensive approach, including good glycemic control and proper wound care using appropriate wound dressings.
On the other hand, the Central Statistics Agency (BPS) stated that in 2022 Lampung Province will be the largest pineapple producer in Indonesia with total production reaching 861,706 tons.
However, in general, the majority of Lampung people only consume pineapple flesh, while pineapple tubers and skin are often considered waste.
Pineapple skin has secondary metabolites, such as flavonoids (Base et al.
Flavonoids act as antioxidants, antiinflammatory, antibacterial, anticancer and antiviral so they can play a role in the process of restoring damaged body tissue, especially in conditions where diabetic wounds occur (Rathnavelu et al.
, 2.
This is confirmed by research which states that ethanol extract of pineapple peel with a concentration of 50% produces an inhibitory zone with a diameter of 13 mm against Escherichia coli bacteria, 16 mm against Staphylococcus aureus (Omorotionmwan et al.
, 2.
Wounds that are moist can accelerate the rate of tissue epithelialization and speed up the process of tissue autolysis, thereby reducing the risk of wound infection (Angriani et al.
, 2.
Accelerating the wound healing process can also be helped by using medication in the form of a hydrogel patch.
Hydrogel patches are a form of dressing that possess beneficial properties and are often used as wound dressings in the healing process, particularly for diabetic ulcers (Harliantika dan Noval.
The coating on the hydrogel patch preparation has the ability to absorb and retain water moisture when it comes into contact with a wet wound, thereby preventing infection and accelerating skin regeneration (Hanistya & Samlan, 2.
This research was conducted in the context of testing and evaluating the use of pineapple peel extract in the preparation of hydrogel patches for healing diabetic ulcers.
This research aims to develop a formulation and test the effectiveness of a hydrogel patch preparation containing pineapple peel extract (Ananas comosus L.
) as a wound dressing for diabetic ulcers in Deutschland Denken Yoken (DDY) mice.
It is hoped that this research can provide innovative and effective solutions in treating diabetic ulcers and provide additional benefits in utilizing pineapple peel waste.
METHODS
This research was conducted at the Chemistry Laboratory.
Microbiology Laboratory.
Pharmaceutical Laboratory.
Animal House.
Faculty of Medicine.
University of Lampung, from April to June Materials and Tools The materials used in this research are pineapple peel, filter paper, distilled water, ethanol 96%, chloroform, dimethyl sulfoxide (DMSO).
Polyvinyl
Alcohol
(PVA),
Polyethylene Glycol
(PEG
Hydroxypropyl Methyl Cellulose (HPMC), aluminum foil, non- woven plaster , universal pH, mouse feed, male white mice (Mus musculu.
Deutschland Denken Yoken (DDY) strain, alloxan, razor, biopsy punch, glucometer strips.
Staphylococcus aureus bacterial culture.
Hydrocolloid Dressing.
Mueller Hinton Agar (MHA) media, and Gentamicin 10 g/disc paper disks, blank paper disk.
The tools used in this research Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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includes were analytical balance, blender, rotary evaporatory, desiccator, vernier caliper, incubator, oven, thermometer, hot plate, magnetic stirrer, glucometer, waterbath, injection syringe.
Plant Determination Test Plant taxonomic identification was carried out at the Biological Botany Laboratory.
FMIPA.
University of Lampung.
Making Pineapple Peel Simplisia Pineapple skin samples (Ananas comosus L.
) from this research were obtained from the Bambu Kuning Market.
Bandar Lampung City.
The pineapple skin that will be used is washed first using running water and then drained.
Clean pineapple skin is wet Next, drying was carried out using sunlight and an oven at a temperature of 50AC.
Simplify dry pineapple skin, blend until it becomes powder.
Making Pineapple Peel Extract Pineapple skin is extracted using the maceration method.
The stages of the maceration process include collecting materials, sorting, washing, drying, grinding, and soaking in 96% ethanol for 3 x 24 hours in a container protected from direct sunlight.
The filtrate was obtained by filtering using filter paper and evaporating using a rotary evaporator at a temperature of 40-50AC until a thick extract was formed.
Phytochemical Screening Test The thick extract obtained was subjected to a qualitative phytochemical screening test to identify secondary metabolite compounds in pineapple samples by observing color changes after adding The compounds analyzed included flavonoids, tannins, saponins, phenolics, alkaloids, steroids, and triterpenoids (Reiza et , 2.
For flavonoids, the extract was dissolved in 70% ethanol, heated, and then mixed with concentrated HCl and Mg a yellow-orange to dark red color indicated positive results.
Phenolics were tested by adding FeCl3 solution to the extract, with a blue or purple color indicating their The saponin test involved shaking the extract solution, and persistent foam indicated saponin content.
Tannins were tested using 1Cl3, with a blackish green or blackish blue color indicating positive Triterpenoid and steroid tests used chloroform, anhydrous acetic acid, and concentrated H2SO4, with a brownish or violet ring for triterpenoids and a bluish-black green color for steroids indicating positive The alkaloid test involved Mayer.
Wagner, and Dragendorff reagents, showing positive results through white, orange to brown precipitates in each test tube (Reiza et , 2.
Antibacterial Test The antibacterial test was carried out using the disc diffusion method on MHA media against gram-positive bacteria in the form of Staphylococcus aureus.
The extract concentrations tested were 10%, 20%, and 30% and Gentamicin as a positive control (Sitepu et al.
, 2.
This method involves absorbing antibacterials onto paper discs, which are then attached to a medium homogenized with bacteria.
After incubation, an inhibitory zone forms around the disc as an indication of an antibacterial reaction (Novita.
Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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Formulation of the Hydrogel Patch Table 1.
Pineapple Peel Extract Hydrogel Patch
Formulation Material Function
Control
Pineapple Peel
Extract
Active
PVA
Backing PEG 400
Plasticizer HPMC
Gel base
DMSO
Enhancer Aquadest .
Solvent Description: control = formula without pineapple peel extract.
F1 =
formula with 10% pineapple peel extract.
F2 = formula with 20% pineapple peel extract.
F3 = formula with 30% pineapple peel The formulation stage of this preparation begins with the process of weighing the ingredients, then all the ingredients are mixed for 30 minutes using a magnetic stirrer.
Next, the preparation was put into a baking dish and dried in the oven at 40AC for 24 hours.
Once solid, it is cut to 1 x 1 cm 2 then attached to a layer of non- woven Packaged in aluminum foil and stored in a cool place away from sunlight.
Application of Hydrogel Patch on Test Animals Selection of Test Animals In testing the effectiveness of the preparation, test animals were used in the form of male white mice ( Mus musculus ) of the DDY strain aged 6-8 weeks with a body weight of around 25-30 grams (Azizah & Qomariyah, 2.
Animal Model Testing of Diabetes Mellitus Before being given treatent and measuring initial blood glucose levels, all mice underwent fasting for 12 hours.
Then the mice were induced subcutaneously with 150 mg/kg alloxan.
On the third day after induction, fasting blood sugar is checked.
Mice with fasting blood sugar more than 180 mg/dl are considered diabetes model mice (Haryanto et al.
, 2.
Create Diabetic Ulcers in Test Animals Ulcer creation was carried out when the mice were already experiencing The creation of ulcers begins with shaving the hair on backs, then the mice are anesthetized with chloroform inhalation.
Then an ulcer was created by making a hole in the back of the mouse with a 5 mm biopsy punch.
The wound was then made into an infected wound model by administering Staphylococcus aureus bacteria.
After the wound model is formed, the wound is left for 3 days.
(Golmohammadi et al.
, 2.
Hydrogel Patch Preparation in Test Animals Mice that had become diabetic ulcer models were divided into 5 treatment groups.
The positive control group (K.
was given Hydrocolloid Dressing.
The negative control group (K.
was given a hydrogel patch without extract.
Treatment groups I (K.
II (K.
, and i (K.
were given pineapple peel extract hydrogel patches with concentrations of 10%, 20%, and 30%.
Observations were made of wound diameter closure with a pineapple peel extract hydrogel patch for 21 Replacement of pineapple peel extract hydrogel patch and documentation done every 2 days.
Evaluation of healing was carried out by measuring the wound diameter vertically and horizontally using ImageJ Data analysis Data analysis used included data normality tests using the Shapiro-Wilk test Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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and homogeneity tests using the Levene test.
The data were distributed normally and homogeneously, followed by the ANOVA ( Analysis of Variant ) test to test the significance of the differences in the averages of several groups, to determine whether there were differences between treatment groups, which was followed by the LSD ( Least Significant Difference ) test with a significant value of p<0, 05 states that there is a significant average difference (Mustika et al.
RESULT AND DISCUSSION
Plant Determination Plant determination tests were carried out at the Biological Botany Laboratory.
FMIPA.
University of Lampung.
Based on the identification results, it can be seen with certainty that the sample used in this research is Pineapple (Ananas comosus (L.
) Merr.
Pineapple plants belong to the Ananas genus and the Bromeliaceae family.
Pineapple Skin Extraction Results In the extraction process, 10 kg of pineapple skin was used.
Pineapple skin shrinks after drying by 89.
The net weight of pineapple peel simplicia after drying is 1032 grams.
Extraction of pineapple peel waste with 96% ethanol solvent produced a thick extract of 121.
grams, then the yield value was calculated.
The results of the yield calculation obtained a value of 11.
74%, while the requirement for the yield of thick extract was that the value was not less than 10% (Farmakope Herbal, 2.
So the yield of pineapple peel extract meets the requirements (Syamsul et al.
, 2.
Phytochemical Test Results Based on the results of phytochemical tests that have been carried out, the ethanol extract of pineapple peel positively contains several secondary metabolites, namely tannins, saponins, flavonoids, triterpenoids, phenolics and alkaloids in dragendorff and bounchardat reagents (Reiza Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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Table 2.
Results of Phytochemical Tests Results of Phytochemical Tests Compound Reagents Positive results Results Identifications Conclusions Tannin FeCl3 1% Dark blue or blackishgreen in color Green-black in color Positive Saponins HCl Foam forms and lasts for 5 minutes Foam lasts 5 minutes Positive Flavonoids Concentrated HCl Orange to red in color Deep red in color Positive Steroids Acetic anhydride and concentrated H2SO4 Bluish-green in color No color change Negative Terpenoids Acetic anhydride and concentrated H2SO4 Brownish and violet Brownish ring formed Positive Phenolics Iron.
chloride 1% Blue or purple in color Blue coloured Positive Alkaloids Mayer White precipitate Not formed Negative Bauchardat Formation of brown Brown precipitate Positive Dragendorff Formed brownish orange precipitation Formed brownish orange precipitation Positive Antibacterial Test Results In the antibacterial test, the inhibition zone was measured to determine the antibacterial activity of the hydrogel patch The Antibacterial inhibitory power is grouped into four groups.
weak (<5 m.
, moderate .
-10 m.
, strong .
-15 m.
, and very strong (>15 m.
(Davis and Stout, 1971 Masykuroh.
Table 3.
Antibacterial Test Results Concetration Control ( ) Control(-) Zone of inhibition .
Categorize Strong Weak Weak Weak Weak Information: D = disk diameter.
VD = vertical diameter.
HD = horizontal Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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Ethanol extract of pineapple peel has antibacterial activity that falls into the weak category in inhibiting the growth of Staphylococcus aureus bacteria.
In this study, the average value of the diameter of the inhibition zone formed at concentrations of 10%, 20%, and 30% was 0.
48 mm, 1.
mm, 0.
85 mm, respectively, and the negative control did not form an inhibition zone so it did not have an average, and the positive control had an average of 16.
34 mm.
Test Results of the Effectiveness of Pineapple Peel Extract Hydrogel Patch Preparation in Healing Diabetic Ulcers in Test Animals The healing process of diabetic ulcers in test animals treated with the pineapple peel extract hydrogel patch showed promising results over the course of the study .
On the first day, the skin around the wound treated with the pineapple peel extract hydrogel patch showed typical signs of inflammation, namely redness and This shows that an inflammatory process occurs which indicates increased blood flow and immune cell activity in response to injury (Andriyono Ilham Reandy, 2.
Furthermore, on the 6th day the redness around the wound was still clearly visible.
However, when entering the 10th day, the wound begins to dry out and a thin layer forms which indicates the proliferation process of forming new tissue to cover the wound.
This outcome is likely due to the moist environment provided by the hydrogel, the proteolytic activity of bromelain, and the antioxidant properties of the compounds in the pineapple peel extract, which contributed to the accelerated healing of the diabetic ulcer.
Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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Days toK1 Figure 1.
Evaluation of wound healing Description: K1: group 1 as positive control with hydrocolloid wound dressing.
K2: group 2 as negative control with formula without extract.
K3: group 3 with hydrogel patch 10% pineapple peel extract.
K4: group 4 with hydrogel patch 20% pineapple K5:
In the results of observations on days 6 to 14, the wound closure process was clearly especially in group 1 and group 4.
the negative control, the wound had not completely closed on day 21, but the wound closure process was still occurring.
From measuring the diameter of the wound, the percentage of wound closure calculated using the formula:
Initial wound area Ae Final wound area Wound closure percentage = x 100 % Initial wound areal (Handayani et al.
, 2.
Table 4.
Percentage of Wound Closure in Test Animals .
nits%) Days to Group (K) 68 A 0.
03 A 2.
01A3.
03 A 5.
09 A 1.
14 A 8.
52A10.
05A5.
77A20.
24 A 13.
17A16.
78A20.
47A12.
74A8.
00A6.
21A15.
64A15.
98A21.
89A15.
26A11.
74A17.
89A19.
87A5.
20A6.
87A25.
60A8.
67A10.
63A8.
67A7.
40A8.
61 A 10.
60A11.
30A19.
35A14.
47A9.
90A8.
96A9.
54A13.
22A10.
92A5.
37A2.
46A11.
71A17.
57A20.
64A8.
In testing the effectiveness of the pineapple peel extract hydrogel patch preparation on test animals with diabetic ulcers, it showed that there was a difference in wound closure Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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On the 8th day, the percentage of diabetic ulcer wound closure in group 4 5%, significantly greater than the positive group with a percentage of 58.
Meanwhile, the negative control experienced a decrease in percentage, this could be due to the condition of the ulcer becoming more The percentage of closure of the diameter of diabetic ulcers reaches 100% on days 16 to 21.
Figure 2.
Diabetic ulcer wound closure chart The results showed that group 4 had the fastest healing process compared to all other groups.
A decrease in wound diameter was observed in all groups, including the negative control group, which was given a hydrogel patch without pineapple peel Overall, using pineapple peel extract hydrogel patches as a wound dressing increased the percentage of diabetic ulcer The hydrogel patch with 20% extract provided the best results, outperforming the positive control, a market hydrocolloid Hydrocolloid dressings were chosen as the positive control due to their similar physical properties to hydrogel patches and their common use in diabetic ulcer treatment.
Wound diameter closure data on day 16 was tested for normality using the Shapiro-Wilk test obtained p>0.
05 so that it can be stated that the data is normally distributed, then a homogeneity test was carried out with the leneve method obtained the result p=0.
>0.
so it can be concluded that the data obtained is From these two tests the data is declared normally distributed and homogeneous so that it can be continued with the one way anova test to analyze the average difference of grouped data.
In the one way anova test, the results obtained were p = 0.
>0.
so it can be concluded that there is no difference in mean values between test groups.
In descriptive observations, it was found that the application of pineapple peel extract hydrogel patches had a trend of decreasing wound diameter in mice with diabetic ulcer.
Conclusion Based on the results of phytochemical screening, pineapple peel extract is positive for tannins, saponins, flavonoids, terpenoids, phenolics and alkaloids.
The antibacterial activity of pineapple peel extract with concentrations of 10%, 20%, and 30% against aureus is classified as weak with inhibition 85 mm respectively with 20% concentration superior to other Media Farmasi Indonesia Vol 19 No 2 | DOI 10.
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In the patch characteristic test of pineapple peel extract hydrogel, it was found that formula 0 and formula 1 were more stable than the other formulas.
In the effectiveness test of pineapple peel extract hydrogel patches on test animals, it was found that group 4 with the administration of hydrogel patches containing pineapple peel extract by 20% achieved the fastest percentage of diabetic ulcer closure compared to other treatment groups.
Descriptive tests showed that there was a trend of decreasing the diameter of diabetic ulcers in test animals given pineapple peel extract hydrogel It can be concluded that pineapple peel extract contains secondary metabolites that have the potential for wound closure.
Giving hydrogel patches containing pineapple peel extract by 20% effectively accelerates diabetic ulcer wound closure in test animals.
Bibliography