Extract Gel of Chinese Petai Leaf (Leucaena leucocephala) for Acute

Wounds in Rats (Rattusnorvegicus)

Anis Ika Nur Rohmah

, Edi Purwanto and Vicky Nurdiansyah

Department of Nursing, Faculty of Health Science, University of Muhammadiyah Malang, Jalan. Bendungan Sutami

No. 188A, Malang, East Java, Indonesia

Keywords:

Chinese petai leaf, acute wounds, rats.

Abstract:

The many occurrences of acute injuries due to the surgical process or accident have a considerable risk of

complications if not given appropriately. Chinese Petaileaves contains saponins, tannins, and flavonoids

are useful for accelerating through the process of acute wound healing. In this study, extract Chinese Petai

leaf made in gel preparations to be practical, sterile, and become the latest innovation. The purpose of this

study was examined the extract gel Chinese Petai leaf effectively for the treatment of acute wounds to rats.

This study used true-experimental design with a randomized control group posttest design as an approach.

The samples in intervention group (n = 7) and control group (n = 6) used simple random sampling and for

data was analyzed using Man Whitney test. The results indicated that the first group (Extract Gel Chinese

Petai Leaf) and the second group (Normal Saline) had significant differences in the percentage of wound

closure on the eighth day with the score (p = 0.014). Difference in the percentage of wound closure was

seen starting early in the epithelialization phase of acute wounds and more clearly on the eighth day.

Extract gel of Chinese Petai leaf effectively accelerated the closure of wound acute stage II.

1 INTRODUCTION

Wounds come from trauma to the skin which is a

condition where the continuity of the body’s tissue

continuation can disrupt to the normal functioning of

the body (Damayanti, Pitriani, & Ardhiyanti, 2013).

According to the duration of healing, wounds are

divided into 2 types, namely acute and chronic

wounds. Acute wounds are wounds that heal

according to the physiology of the wound healing

process. Acute wounds can be caused by trauma

suddenly or planned as during the surgical process.

A chronic wound is a complication of an acute

wound, which is where acute wound care is not

appropriate (Wijaya, 2018).

Acute wounds have a high risk of complications

if they do not get proper wound care. Complications

of acute injuries can occur within 24 hours after the

trauma. Several kinds of complications can occur in

acute wounds, including bleeding, hematoma or

tissue edema, dehiscence or not joining the wound

edges, and infection (Wijaya, 2018). Infection In

acute wounds can occur in the inflammatory phase.

This can occur because of bleeding that is long

enough (10-20 minutes), so that the blood supply in

the inflammatory phase is greatly reduced and white

blood cells do not reach the site of the wound (Peate

& Glencross, 2015). Complications of acute wounds

can also lead to amputation caused by an infection in

the wound and sufferers who have risk factors for

diabetes mellitus (Supriyadi, 2017).

Several factors influence wound healing,

including: vascularization, age, and concomitant

diseases. Vascularization affects because the wound

requires a good circulatory condition. The speed of

cell repair takes place in line with the growth or

maturity of a person, but the aging process can

reduce the speed of cell repair. While other diseases,

such as anemia, diabetes mellitus and kidney

disorders that can slow wound healing (Damayanti

et al., 2013).

The national wound prevalence in Indonesia is

8.2%. Comparison of the results of Riskesdas 2007

with Riskesdas 2013 showed an increase in wound

prevalence from 7.5% to 8.2% (Trihono, 2013). The

three most common types of injuries suffered were

70.9% abrasions; sprained 27.5%, and torn wounds

23.2%. Whereas in 2018 Riskesdas there was an

increase in wound prevalence from 8.2% in 2013 to

9.2% in 2018 results (Riskesdas, 2018).

Rohmah, A., Purwanto, E. and Nurdiansyah, V.

Extract Gel of Chinese Petai Leaf (Leucaenaleucocephala) for Acute Wounds in Rats (Rattusnorvegicus).

DOI: 10.5220/0009125701370142

In Proceedings of the 2nd Health Science International Conference (HSIC 2019), pages 137-142

ISBN: 978-989-758-462-6

137

Chinese Petai has a variety of chemical contents

both in the seeds or leaves. The chemical content of

old seeds in every 100 grams has 148 calories of

calories, 10.6 grams of protein, 0.5 grams of fat,

26.2 grams of charcoal hydrate, 155 grams of

calcium, 59 mg of phosphorus, 2.2 mg of iron ,

vitamin A 416 SI, vitamin B 1 0.23 mg, and vitamin

C 20 mg. In the results of a study found that Chinese

Petai leaf extract contains saponins, alkaloids,

tannins, and flavonoids (Widyantoro & Sugihartini,

2015). The saponin content contained in Chinese

Petai leaves can be antibacterial (Pratiwi, Soetjipto,

& Hartini, 2014). Flavanoid is a compound derived

from flavone compounds. Flavanoid is a protein

structure that plays a role in the process of wound

healing and can be a cleanser for wounds (Praja &

Oktarlina, 2016).

Chinese Petai leaves have a variety of chemical

contents, where the content can be used as an

ingredient in the treatment of new wounds and

swelling by pounding or chewing. Furthermore, the

results of leaves that have been smooth are

immediately affixed to the wound or swollen area

(Thomas, 2007). In another study showed that the

scouring of 30 grams of Chinese Petai leaves

provided significant healing results with the most

length of clean wound healing on day 5, while the

length of clean wound healing using povidone iodine

was 10% on the 8th day (Rahmawati, 2015).

The gel is a semi-solid preparation consisting of

organic and inorganic suspensions. Gel for topical

use can immediately melt if it comes into contact

with the skin and will form a layer. Absorption of

the skin in gel form is better than other ointment

preparations (Yanhendri & Yenny, 2012). Gel-

shaped ointments have finer consistency, are

generally liquid and contain little or no mucosa as a

base (Elmitra, 2017).

Treatment using Chinese leaves with crushed or

chewed cannot last long and must be made every

day. This can make people less interested in using it

and choosing a more practical way. Therefore, an

alternative is needed in the presentation of Chinese

Petaileaves so that it is easy to use. On this occasion,

the researchers wanted to know the effectiveness of

Chinese Petaileaf extract gel for the treatment of

incision wounds in rats. The presence of Chinese

Petaileaf extract gel is expected to simplify wound

care that can be used at any time, is more effective,

and is practical for storage as supplies.

2 METHODS

This study is an experimental study using a true

experimental research design and using a

randomized control group posttest design approach.

This study consisted of 2 groups, namely the

intervention group (n = 7) who were treated with

wound care using Chinese Petaileaf extracts, and the

control group (n = 6) who were given standard

treatment using 0.9% NaCl.

The sampling technique used is simple random

sampling. The method of taking the sample is by

drawing, so it is chosen randomly according to the

experimental design to place the rat in 2 groups

(Surahman, Rachmat, & Supardi, 2016).

White rat used in this study must meet the

following experimental animal requirements: (1)

White male rats (rattusnorvegicus), (2) 2-3 months

old, (3) White rat with normal body weight of 160

grams, (4) active (healthy) moves, and (5) have

never been used as experimental animals in other

studies.

In this study, ethical information was obtained

No.E.5.a / KEPK-UMM / II / 2019 from the UMM

Health Research Ethics Commission. This research

was conducted at the Pharmacology Laboratory of

the Faculty of Medicine, University of

Muhammadiyah Malang. When the research was

carried out on 05 to 12 April 2019.

The dose of Chinese Petaileaf extract gel was

0.006 g each time applying 1cm2 to the wound.

Measurement of gel dosage using micro scales that

can read the mass of objects under 1 gram and

adjusted to changes in wound area until the gel is

applied evenly. Giving Chinese Petaileaf extract gel

is done 2 times a day in the morning at 07.00 and in

the afternoon at 16.00 (Andrie & Sihombing, 2015).

Experimental animals adapted in rat cages were

given the same treatment for 7. During adaptation,

all try animals looked healthy, actively moving, and

nothing stressed. When intervening, animals try to

get food and drink according to their needs.

The data collection stage was carried out by

measuring the area of the wound on the 4th day and

the 8th day. Measuring the area of the wound using

image J software. The wound image is taken by

attaching mica paper over the wound, then draw by

following the edge of the wound using a marker and

after that, the picture is photographed and inserted

into the image J application.

The resulting wound area is then entered into the

formula for wound closure percentage. The formula

for measuring wound closure:

HSIC 2019 - The Health Science International Conference

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Percentage of wound closure= (AO - AD)/AO

x100% (1).

Information:

AO = area of the wound on day 0.

AD = area of the wound on the day after

treatment (Avinash et al., 2016).

3 RESULTS AND DISCUSSION

Based on the figure 1, the picture shows that all

white rat animals have been measured the

percentage of wound closure on day 4 in group 1

(intervention) with an average value of 14.6%, and

in group 2 (control) with a mean value an average of

9.7%. On the 8th day in group 1 with an average

value of wound closure was 81.2% and in group 2 it

was 62.9%. There was an increase in the percentage

of initial wound closure on day 4 by 14.6 in group 1

to 81.2% on day 8, whereas in group 2 the

percentage of wound closure initially on day 4 was

9.7% to 62.9% on the 8th day.

The results of the study on the percentage of

wound closure on day 4 obtained from the SPSS

output found that the significant value (p), in the

Man Whitney statistical test the significance value of

0.628 where the value is greater than the value of α

(0.05). The results of these calculations can be

concluded that there was no significant difference in

the percentage of wound closure in the intervention

group given Chinese Petai leaf extract gel and in the

control group given normal saline for acute wounds

in the rat on the 4th day.

The results of the study on the percentage of

wound closure on the 8th day obtained from the

SPSS output found that the significant value (p), in

the man whitney statistical test the significance

value of p = 0.014 where the value is smaller than

the value of α (0.05). The results of these

calculations can be concluded that there is a

significant difference in the percentage of wound

closure in the intervention group given Chinese

Petaileaf extract gel and in the control group given

normal saline for acute wounds in rats on the 8th

day.

Based on the results of research in the

intervention group that was given Chinese Petaileaf

extract gel showed that the wound healing process

went normal. None of the wound healing stages

were missed and there were no interruptions in each

process, which was proven in the intervention group

that there were no signs of wound infection. This

proves the theory revealed by Pratiwi (2014) and

Yuliarti (2009) that the saponin and tannin content

in Chinese Petai leaves that function as anti-bacterial

and anti-inflammatory properties can prevent

infection so that the inflammatory process is not

interrupted.

In the intervention group, the results of the study

showed the percentage of wound closure on day 4

with an average value of 14.6%, the condition of the

wound had begun to epithelialization phase, and no

signs of infection were found. Reassessment was

carried out on the 8th day which found that the

percentage of wounds had increased with an average

value of 81.2%, a good wound condition which was

proven that the scab (necrose) had been peeled off

on the 6th day so that a new layer of skin had been

seen.

In the control group, the results of the study were

the percentage of wound closure on day 4 with an

average value of 9.7%, the condition of the wound

had begun to epithelialization phase, and found signs

of infection in the form of edema. Reassessment was

conducted on the 8th day which found that the

percentage of wounds had increased with an average

value of 62.9%, found scabs (necrose) had only

begun to peel on the 8th day.

In the control group given normal saline showed

that the wound healing process proceeded normally,

even though on the 3rd and 4th day there were signs

of infection in the form of edema in some rats. That

is, not until there are severe complications that

become chronic wounds. Wounds in the control

group can heal according to the process of healing

acute wounds. In accordance with other studies

which state that irrigation wounds using normal

saline routinely can reduce the incidence of surgical

wound infections. Most liquids used in addition to

normal saline show inconsistencies as wound

irrigation fluid (Pieper et al., 2018).

During the study, the rat wound condition in the

intervention group healed faster, visible from the

polymerization phase faster than the control group

and visible signs of infection in the form of edema in

the wound area in 3 rats in the control group on days

3 and 4. In the intervention group, there was 6 rats

with a scab (necrose) covering the skin to peel off

earlier in the 6th day. Whereas in the control group

the new necrose peeled off on the 8th day.

Characteristics of scab (necrose) in the rat have a

clear color and when peeling the wound dries

immediately does not cause wound infection.

Extract Gel of Chinese Petai Leaf (Leucaenaleucocephala) for Acute Wounds in Rats (Rattusnorvegicus)

139

46.80%

18.40%

20.30%

0% 0% 0

80.80%

78.60%

60.50%

55.30%

41.90%

62.90%

0.00%

10.00%

20.00%

30.00%

40.00%

50.00%

60.00%

70.00%

80.00%

90.00%

A B C D F G

Day 4

Day 8

Rat

Day 8

Figure 1: Change in percentage of wound closure on day 4.

Figure 2: Change in percentage of wound closure on day 8.

According to (Aponno et at., 2014) in (Fitriani,

2016) explains that the speed of the formation of a

scab indicates the speed of wound healing. This

indicates the occurrence of the growth of new cells

in the skin so that it helps accelerate the release of

46.80%

18.40%

20.30%

0% 0% 0

80.80%

78.60%

60.50%

55.30%

41.90%

62.90%

0.00%

10.00%

20.00%

30.00%

40.00%

50.00%

60.00%

70.00%

80.00%

90.00%

A B C D F G

Day 4

Day 8

Rat

Day 8

HSIC 2019 - The Health Science International Conference

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scabs and accelerate the contraction of the wound

edges.

The above results prove that Chinese Petaileaves

which have useful ingredients for the wound healing

process is proven to accelerate the polymerisation

process as seen from the percentage of wound

closure with an average value of 81.2% on the 8th

day. By the conceptual framework expected from

the content of flavonoids, saponins, and tannins

which can accelerate proliferation. These results are

consistent with Tsala, Nga, Thiery, Bienvenue, dan

Theophile (2015) research which explains that the

content of flavonoids, tannins, and saponins can be

antioxidants and accelerate the process of

epithelialization of incision wounds. Widyantoro

dan Sugihartini (2015) stated that in each extract of

Chinese Petaileaf contains saponin, alkaloid, tannin,

and flavonoid compounds. Saponins that are

abundant in the roots and leaves can provide many

benefits because it has anti-bacterial and antiviral

(Mardiana, 2012). Flavonoids, tannins, and saponins

can also be antibacterial and can accelerate the

contraction of wound edges (Begashaw et al., 2017).

Flavonoids also function as antioxidants and

protect collagen tissue (Yuliarti, 2009). This is in

line with other studies which explain that the

flavonoid content in hesperidin can accelerate the

wound healing process in the rat (Li et al., 2018).

Other studies on latex jatropha curcas leaves which

contain flavonoids, tannins and saponins are

effective in accelerating the wound healing process

(Balqis et al., 2018). In another study also explained

the effectiveness of flavonoids from butea

monosperms showed significant results in the

process of wound healing (Muralidhar et al., 2013).

Chinese Petaileaf extract gel which was used for

wound care during the research proved that after it

was applied it could immediately melt and fill the

wound area which could make the active ingredient

of Chinese Petaileaf absorbed into rat skin. Wounds

given by Chinese Petaileaf extract gel can also be

protected from contamination. Following the theory

(Elmitra, 2017) which explains that gels for topical

use can immediately melt when in contact with the

skin and will form a layer.

The results of this study which aims to determine

the percentage of wound closure in line with other

studies show that the scouring of 30 gram Chinese

Petaileaves provides significant healing results with

the most length of clean wound healing on day 5,

while the length of clean wound healing using 10%

providone iodine at 8th day (Rahmawati, 2015). The

results of this study indicate a higher percentage of

wound closure with an average value of 81.2% on

the 8th day when compared to other studies of the

combination of water-oil ointment cork fish extract

which obtained a 50% percentage of open wound

closure stadium II on the 8th day (Andrie and

Sihombing, 2015).

During the study, a limitation that could not be

avoided from the research was that there was one

mouse that died in the control group. As per the code

of ethics of animal studies, dead rats are buried. In

addition, necrosa on some rat peel prematurely due

to friction in places around the cage.

4 CONCLUSIONS

In this study, a significant difference in the

percentage of wound closure (p = 0.014) using

Chinese Petaileaf extract gel compared with normal

saline on the 8th day. In the intervention group, there

were no signs of infection and necrose was more

rapid peeling. This proves that Chinese Petaileaf

extract gel can accelerate the process of closing the

acute wound in the rat. Further research is needed to

get accurate results on human samples so that the

Chinese Petaileaf extract gel can be proven to

accelerate the process of wound closure in humans.

Future studies are expected to be able to control the

limitations of this study in the form of the influence

of friction around the place which results in peeling

off scabs prematurely.

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striata ) pada Proses Penyembuhan Luka Akut

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Extract Gel of Chinese Petai Leaf (Leucaenaleucocephala) for Acute Wounds in Rats (Rattusnorvegicus)

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glauca , Benth ) dan Povidon Iodine 10 % Dalam

Mempercepat Penyembuhan Luka Bersih Pada

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pp. 48–60.

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Edited by B. Rini. Yogyakarta: ANDI.

Andrie, M. and Sihombing, D. (2015) ‘Efektivitas Sediaan

Salep yang Mengandung Ekstrak Ikan Gabus ( Channa

striata ) pada Proses Penyembuhan Luka Akut

Stadium II Terbuka pada Tikus Jantan Galur Wistar

The Effectiveness of Snakehead ( Channa striata )

Extract- Containing Ointment on Healing P’, Pharm

Sci Res, pp. 88–101.

Balqis, U. et al. (2018) ‘Angiogenesis Activity of Jatropha

Curcas L. Latex in Cream Formulation on Wound

Healing in Mice’, Veterinary World, 11(9).

Begashaw, B. et al. (2017) ‘Methanol Leaves Extract

Hibicus Micranthus Linn Exhibited Antibacterial and

Wound Healing Activities’, Complementary and

Alternative Medicine, 17(337).

Elmitra (2017) Dasar Dasar Farmasetika dan Sediaan

Semi Solid. 1st edn. Yogyakarta: Deepublish.

Fitriani, N. (2016) Uji Aktivitas Gel Etil P-

Metoksisinamat Terhadap Penyembuhan Luka

Terbuka Pada Tikus Putih (Rattus norvegicus) Jantan

Galur Sprague Dawley.

Li, W. et al. (2018) ‘Hesperidin, A Plant flavanoid

Accelarated The Cutaneous Wound Healing in

Streptozotocin-Induced Diabetics Rats: Role of TGH-

B/SMADS and ANG-1/TIE-2 Signaling Pathways’,

EXCLI Journal, 17, pp. 399–419.

Mardiana, L. (2012) Daun Ajaib Tumpas Penyakit.

Jakarta: Penebar Swadaya.

Muralidhar, A. et al. (2013) ‘Wound healing activity of

flavonoid fraction isolated from the stem bark of

Butea monosperma ( Lam ) in albino wistar rats’, J.

Exp. Bio, 3(6), pp. 1–6.

Pieper, D. et al. (2018) ‘The Role of Saline Irrigation Prior

to Wound Closure in The Reduction of Surgical Site

Infection: Protocol for a Systematic Reviev and Meta

Analysis’, 7(158).

Pratiwi, M., Soetjipto, H. and Hartini, S. (2014) Isolasi

Saponin Daun Petai Cina ( Leucaena leucocephala

(Lam.) De Wit.) dan Aplikasinya Sebagai Pembusa

Alami Serta Antibakteri Dalam Shampo.

Rahmawati, I. (2015) ‘Perbedaan Efek Perwatan Luka

Menggunakan Gerusan Daun Petai Cina ( Leucaena

glauca , Benth ) dan Povidon Iodine 10 % Dalam

Mempercepat Penyembuhan Luka Bersih Pada

Marmut ( Cavia porcellus )’, Jurnal Wiyata, 2, pp. 73–

78.

Tsala, D. E. et al. (2015) ‘Evaluation of the Antioxidant

Activity and the Healing Action of the Etanol Extract

of Calotropis Procera Bark Against Surgical Wounds’,

Intercultural Ethnopharmacology, 4(1).

Widyantoro, O. B. and Sugihartini, N. (2015) ‘Uji Sifat

Fisik dan Aktivitas Ekstrak Daun Petai Cina (

Leucaena glauca , Benth ) Dalam Berbagai Tipe Basis

Salep Sebagai Obat Luka Bakar’, Media Farmasi, 12,

pp. 48–60.

Yuliarti, N. (2009) A to Z Food Supplement. 1st edn.

Edited by B. Rini. Yogyakarta: ANDI.

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