THE EFFECT OF VARYING CONCENTRATIONS OF SAMBONG …

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1099

Satyam et al. World Journal of Pharmacy and Pharmaceutical Sciences

THE EFFECT OF VARYING CONCENTRATIONS OF SAMBONG

LEAVES (Blumea balsamifera) DECOCTION ON WOUND HEALING

Krishna Deo Das1, Satyam Prakash

2* and Khushbu Yadav

3

1Assistant Professor, Department of Community Medicine, Janaki Medical College Teaching

Hospital , Janakpur, Nepal.

2*Assistant Professor, Department of Biochemistry, Janaki Medical College Teaching

Hospital, Janakpur, Nepal.

3Medical Microbiologist, Department of Microbiology, National Institute of Science and

Technology, Kathmandu, Nepal.

ABSTRACT

Background: Blumeae balsamifera (Sambong) is an ancient medicinal

herb with a rich constituents of essential oils and widely used in the

Philippines for a long period of time before the introduction of modern

medicine for the treatment of septic wounds and other infections. The

dominance of commercially manufactured products is increasing day

by day due to the rapid development and progress. Drugs are first and

foremost need to cure ailments and different diseases for human

beings. Almost all the country has always been dependent on highly

priced commercial drugs giving little chance for the utilization of

locally grown medicinal herbs and plants. Therefore, the present study

was focussed to determine the effect of Blumea balsamifera on wound

healing as an alternative to expensive medicine such as betadine. Methods: The descriptive

method was used to find out the active constituents of the Sambong leaves through

phytochemical screening whereas experimental method was used to determine the

characteristics of the experimental mice for their treatment of wounds using Sambong leaves

decoction. Results: The treatments with 50, 100 and 150 grams of Sambong leaves decoction

in all three replications range from 11-12, 7-9 and 5-6 days for deep wound which was found

fair, good and very good respectively and with betadine from 3-4 days was very good to

excellent as positive control. The effect of Sambong leaves decoction and betadine in the

number of days and redness and swelling of wound healing in mice was found to be

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 5.210

Volume 5, Issue 2, 1099-1116 Research Article ISSN 2278 – 4357

Article Received on

08 Nov 2015,

Revised on 28 Dec 2015,

Accepted on 17 Jan 2016

*Correspondence for

Author

Satyam Prakash

Assistant Professor,

Department of

Biochemistry, Janaki

Medical College Teaching

Hospital, Janakpur, Nepal.


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statistically insignificant. Conclusion: The varying concentrations of Sambong leaves

decoction produce different effect on deep wounds of mice. The greater the concentration of

Sambong leaves decoction, the faster the wound healing was ensured.

KEYWORDS: Wound healing, Blumea balsamifera, Phytochemistry, Betadine, Decoction.

INTRODUCTION

Medicinal plants and herbal medicines have been used for many centuries as a source of

people’s drugs for the treatment and prevention of diseases, disorders and the promotion of

good health and still provide the first line of primary health-care even in the present age to

major segments of the population worldwide.[1]

Nowadays, herbal medicines are widely

consumed and their sales have been rising significantly all over the world. According to the

reports of the World Health Organization (WHO), to treat diseases over 80% of the

populations in developing countries mainly rely on herbs, which are considered to be safer

and more effective than synthetic drugs.[2,3]

Blumeae balsamiferae, also named as Sambong in some tropical countries, is an ancient

medicinal herb with a rich constituents of essential oils and is a remarkable medicinal plant

that grows wild in India to Southern China and throughout Southeast Asian nations such as

the Philippines.[4]

Sambong, the herbal medicines have been widely used in the Philippines

for a long period of time before the introduction of modern medicine. Traditionally, medicine

practitioners have described the therapeutic efficacies of many traditional and indigenous

plants against diseases.[5]

Wound healing is a process of restoring damaged cells and tissues.[6]

The phases of wound

healing occur in a precise and regulated order. The wound-healing process consists of four

highly integrated and overlapping phases: hemostasis, inflammation, proliferation and tissue

remodeling or resolution.[7,8]

These phases and their biophysiological functions must occur in

the proper sequence, at a specific time and continue for a specific duration at an optimal

intensity.[9]

The wound also undergoes physical contraction, which might be mediated by

contractile fibroblasts.[10]

Neuropeptide Substance P (SP) is a pro-inflammatory neuropeptide,

and modulates inflammatory responses of skin wounds. SP also promotes the synthesis and

metabolism of fibroblast and increases accumulation of collagen in the proliferative phase of

mesenchymal cell growth and dynamics.[11]

In addition, SP is an important medium in the

process of wound repair and scar healing.[12,13]


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With a consistent efficacy with B. balsamifera medicinal materials, which could induce

resuscitation, clear heat and relieve pain. Recently, extracts of its leaves have been verified to

display various new physiological activities, such as antitumor, antifungal[14,15]

, radical-

scavenging and anti-obesity properties.[16]

The main active compound is L-borneol, which

was characterized by a high volatility. Besides, essential oils, flavonoids and terpenoids with

several different biological activities were also reported.[17]

It’s leaves have been used for

healing many conditions including eczema, dermatitis, skin injury, skin bruises, beriberi,

lumbago, menorrhagia, rheumatism and some other diseases.[18]

Recently, the extracts of the

leaves have been verified to display physiological activities on plasmin-inhibitory,[19,20]

anti-

fungal,[14]

free radical scavenging and anti-obesity functions.[21,16]

Blumeae balsamiferae was used to treat snake bite injury and skin wounds and itch. It is

documented that external application of the mashed fresh leaves or leaf water washings

decoction could treat traumatic injury, carbuncle and skin pruritus.[22]

Natural products that

are safe and attain physiological properties are tremendous sources of new-fangled

therapeutics for the treatment of conditions like mechanical damage of the skin.[23]

As the world rapidly progresses and develops, there seems to be a dominance of

commercially manufactured products and one of which is drugs that man primarily need to

cure ailments and different diseases. The country has always been dependent on highly priced

commercial drugs giving little chance for the utilization of locally grown medicinal herbs and

plants. Therefore, the present study was focussed to determine the effect of a locally grown

plant, Sambong (Blumea balsamifera) on wound healing as an alternative to expensive

medicine such as betadine. The output of the study is the effect of varying concentrations of

Sambong leaves decoction on the wound healing of mice. The findings of this study could be

used as reference on the preparation of Sambong leaves as decoction when used to treat

wounds. Thus, this study was designed to create awareness of the importance of medicinal

Sambong plant and provide vital information regarding its utilization that can be used as an

alternative to high-cost manufactured drugs and preparation in the treatment of wounds.

MATERIALS AND METHODS

The descriptive and experimental research based study was conducted at Virgen Milagrosa

University Foundation (VMUF) Veterinary Laboratory in collaboration with pharmacy

laboratory at the Faculty of Graduate School, VMUF, San Carlos City, Pangasinan,

Phillipines from August 2013 to March 2014. The descriptive method was used to find out


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the active constituents of the Sambong leaves through phytochemical screening whereas

experimental method was used to determine the characteristics of the experimental mice for

their treatment of wounds using Sambong leaves decoction.

Study Population

This study comprises 24 mice, among them 6 white mice served as control and 18 mice

served as experimental groups.

Plant material

Sambong is a half woody, strongly aromatic shrub, densely and softly hairy, 1 to 4 meters

high. Stems grow up to 2.5 centimeters in diameter. Leaves are simple, alternate, elliptical to

oblong lanceolate, 7 to 20 centimeters long, toothed at the margins, pointed or blunt at the tip,

narrowing to a short petiole which are often appendaged.

Figure 1: Blumea balsamifera.

Sample collection

Fresh leaves of Blumea balsamifera as shown in fig. 1 were collected from the plants grown

in the campus of VMUF, San Carlos City, Pangasinan, Phillipines.

Preparation of the Extract

Extract of the dried plant material reduced to a moderate coarse powder was prepared by

refluxing 50 gms of the powdered plant material in a 500 ml Erlenmeyer flask with 300 ml of

80% ethanol for 1 hour in a boiling water bath. The flask was removed and the contents were

allowed to cool at room temperature and were then filtered. Sufficient ethanol was added

through the residue on the filter paper to make 500 ml of the extract.


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PHYTOCHEMICAL SCREENING

Screening for Alkaloids

Seventy milliliters of the ethanolic extract was evaporated via steam bath to get the residue.

This was dissolved in seven to fifteen milliliters of hydrochloric acid, aided by warming on

the steam bath for 1 or 2 minutes. It was then cooled, filtered and adjusted to a volume of

only seven milliliters by washing the residue on the filter paper with a sufficient quantity of

one percent hydrochloric acid. A few drops of grains of the powdered sodium chloride was

added to the filtrate, it was shaken and was filtered. One milliliter of the filtrate was placed

into each small test tube. To the first test tube, three drops of Modified Mayer’s Reagent was

added; to the next, 3 drops of Wagner’s Reagent (Iodine and Potassium iodide T.S.); then

three drops of Valser’s and to the last test tube, three drops Bouchardat’s reagent. Positive

indication was observed for the production of Precipitate.

Screening for Unsaturated Sterols and Triterpenes

Thirty milliliters of the ethanolic extract were evaporated via water bath. The residue was

allowed to cool at room temperature and fifteen milliliters of light petroleum ether were

added, mixed well and filtered. More volumes of petroleum ether were added as needed until

the last volume of petroleum ether became colorless then ethereal filtrates were all combined.

The defatted residue was set aside for screening for flavonoids and leucoanthocyanins. The

combined ethereal filtrates were evaporated and then the residue was dissolved in 15 ml of

chloroform. The chloroformic solution was dried over anhydrous sodium sulfate and filtered.

Lieberman – Burchard Test and Salkowski tests were essentially dehydration reactions and

therefore moisture was excluded in each of the experimental steps.

Screening for Flavonoids

The defatted residue was dissolved in 30 ml of 50% ethanol and was filtered. Bate – Smith –

Metcalf Test and Wilstater – Cyanidin Test were performed and observed for color

change.[24]

Screening for Steroid (Cardioactive) Glycosides

Libermann-burchard test, kedde reaction tests and keller-killiani test were done for screening

of steroids.[24]

Screening for Saponin

Screening of saponin was done by Froth and Hemolysis test which are as follows.


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Froth Test

Ten milliliters distilled water were added in 2 separate test tubes, test tube 1 containing two

milliliters of 10% gugo extract (control) and test tube 2 containing two milliliters of the

ethanolic extract. Both tubes were shaken vigorously for 30 seconds. It was observed over a

period of 30 minutes for the formation of Honeycom.

Hemolysis Test

A blood agar was obtained using small test tube; a mini cup of blood sugars which are

equidistant from one another, each agar cups was numbered from the bottom of the inverted

plate with a marking pencil, with a small pipette, enough plant extract and distilled water was

added in the agar cups. The plate was allowed to stand undisturbed after an hour. Then agar

plate was observed for the formation of halozone in the three agar cups.

Screening for Tannins and Polyphenolic Compounds

About 100 ml of the plant extract were taken and evaporated to incipient dryness over a

steam bath. It was cooled to room temperature; the residue was extracted with 25 ml of hot

distilled water. This was centrifuged for several minutes and the upper half from each tube

used was decanted. 3-4 drops of 10% sodium chloride solution were added to salt out

undesirable constituents through precipitation. Precipitates were filtered off. Gelatin Test,

Gelatin Block Test and Ferric chloride Test was applied for screening of Tannins and

Polyphenolic Compounds.[24]

Screening for Anthraquinones

Borntrager’s Test and Modified Borntrager’s Test were performed for anthraquinones

screening.[24]

Experimental Procedures

In gathering data regarding the effect of varying concentration of Sambong leaves extract on

the wound healing on the mice, mice’s legs were incised. After the incisions of wound,

treatment of the wound was done with three different treatments of Sambong leaves

decoction for the experimental groups. The treatment was done twice a day, every morning

and afternoon at 6:00 pm. After every treatment, each mice wound was covered with the use

of gauze and plaster to prevent foreign materials from contaminating the wound. The

observation was done before every treatment of the wound (Figure 2).


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Figure 2: Flowchart scheme of Experimental procedure.

Treatment of Experimental Group using varying concentrations of Sambong leaves

extract

The experimental group was divided into three groups as T1, T2 and T3. The fresh leaves of

Bulmea balsamifera were taken for T1, T2 and T3 groups. Fifty grams for T1, 100 grams for

T2 and 150 grams for T3 of Sambong leaves were weighed and cut into small pieces. About

150 ml of water was measured and was then put it in a pot together with the sambong leaves.

Moderate heat was applied until the leaves become tender. After cooling it was filtered using

filter paper. The filtrate was placed in a clean bottle to be ready for use. Different amounts of

Sambong leaves decoction on the replicates of mice legs wound by swabbing it on the

affected area.

Incisison of wound (2cm wide; 7 mm deep was

made vertically) on the mice To Betadine

Application of the different concentration of sambong leaves decoction on the

wound of mice (Betadine for the control group)

Application of the Gauze on the treated area

Gathering of Data

Treatment of Data

Observation

Replicate


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A (50gms) B (100gms) C (150gms)

Control Group

The wound was incised 2cm wide and 7 mm deep then betadine was applied on the mice

wound, gauze was put in the wound area also to prevent scratching of the wound by the mice

and to avoid contaminants in the wound.

DATA ANALYSIS

The Analysis of Variance (ANOVA) was used as the statistical tool for this study. This was

used to test the significant difference on the effect on wound healing using varying

concentrations of sambong leaves decoction.

RESULT AND DISCUSSION

Active constituents present in Sambong Leaves through Phytochemical Screening

During phytochemical screening, this study showed that Sambong leaves was positive for

alkaloids as manifested by formation of white precipitate after the addition of Mayer's

reagent, Wagner's reagent and Bourchardat's reagent. Saponins were detected using the Froth

test and Hemolysis test wherein persistent foam or honeycomb froth was formed when the

aqueous solution was agitated. Flavonoids were identified after the addition of 1% HCl

wherein there was red coloration and upon the addition of magnesium turnings still red

coloration was noted and lastly, tannins were detected by the presence of greenish color

solution, production of bluish black solution and the presence of precipitate by gelatin test,

ferric chloride test and gelatin salt block test respectively. Constituents were found to exert

astringent, emulsifying and cleansing effect on wounds. The results are as shown in table 1.


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Table 1. Constituents present in Sambong Leaves through Phytochemical Screening.

Screening Result Inference Indications

Alkaloids

Mayer’s reagent

Formation of white precipitate Presence of alkaloids

Valser’s reagent Formation of white precipitate Presence of alkaloids

Wagner’s reagent Formation of white precipitate Presence of alkaloids

Bouchardat’s reagent Formation of white precipitate Presence of alkaloids

Unsaturated Sterols &

Triterpenes

Leiberman- Burchard

No immediate change in the

color of the solution to blue-

green.

Absence of unsaturated

sterol and triterpenes

Salkowski test

No immediate change in the

color of the solution to blue-

green.

Absence of unsaturated

sterols and triterpenes

Flavonoid 1% HCl No strong red coloration Presence of flavonoids

1% HCl w/ Mg turnings No red coloration Persence of Flavonoids

Steroid (CARDIOACTIVE

Glycosides:Liebermann –

Burchard

No reddish brown precipitate Absence of steroids

Kedde Reaction No blue coloration of the

solution Absence of Steroids

Keller- Killiani test No precipitate Absence of Steroids

Saponins

Hemolysis test No Zone of hemolysis Presence of saponins

Emulsifying

effect

Froth test No Honeycomb froth

formation Absence of Saponins

Cleansing

property

Tannins

Gelatin test

Ferric chloride Test

Gelatin salt block test

Greenish blue color solution

Production of bluish black

solution

Production of precipitate

Presence of Tannin Astringent

effect

Anthraquinone Heterosides

Borntrager test

No Color change in the

solution

Absence of Anthraquinone

Heterosides

Effect on the number of days that wound heals

Treatments with 50% Sambong leaves decoction in all three replications range from 11 to 12

days for wound healing. Categorically described, this range of number of days it takes for

wound in mice to heal was labeled 2, indicating that the length of time for the deep wound to

heal was fair. The number of days it takes for deep wounds in mice to heal completely ranges

from 7 to 9 days, implying that the number of days of healing of wounds treated with 100

grams of Sambong leaves decoction was good throughout. And for deep wounds in mice

treated with 150 grams of Sambong leaves decoction, 5-6 days was needed for complete

healing. This number of days needed for complete healing of wounds in mice was considered

very good. The results are shown in table 2. This may be due to the concentration of


1108


Sambong leaves decoction increases, the number of days needed for healing of deep wounds

in mice decreases.

Meanwhile, the use of positive control Betadine shows the least number of days needed for

deep wound healing, ranging only from 3-4 days or from very good to excellent in terms of

the rates of wound healing. The healing of wounds in mice with the use either of the

experimental drugs in different concentrations or control drugs may be explained by the

antiseptic effects of both types of medication in wounds.

The healing of wounds brought about by the use of either types of treatment is rather indirect.

Either medication induces healing of wounds by preventing bacteria and other microorganism

from thriving and multiplying. Without microorganisms invading the wound areas, the faster

the healing takes place because the surge of hematological materials in the area will only

concentrate on replacing dead tissues or at least repairing damaged yet viable tissues and the

need to protect the wound from further damage by bacterial or other opportunistic infection

by sending more non-specific and immune defense cells are minimized, if not totally

eliminated.

Table 2. Effect of Sambong leaves decoction at various concentrations and of Betadine

on the period of healing of the wound in mice in number of days.

RE

PL

ICA

TIO

N

SAMBONG CONCENTRATIONS BETADINE

(CONTROL) 50 gm. 100 gm. 150 gm.

TREATMENT TREATMENT TREATMENT TREATMENT

1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6

Act

ual

Valu

es

1 12 12 12 11 12 12 8 8 7 9 7 8 6 5 6 6 6 5 3 3 4 3 3 3

2 11 12 11 12 12 11 9 8 8 7 8 7 5 5 5 5 6 5 4 3 3 4 3 3

3 12 12 12 11 12 11 7 7 8 8 9 9 6 6 5 5 5 6 4 4 3 3 4 3

Cate

gori

cal

Valu

es*

1 2 2 2 2 2 2 3 3 3 3 3 3 3 4 4 4 4 4 5 5 4 5 5 5

2 2 2 2 2 2 2 3 3 3 3 3 3 4 4 4 4 4 4 4 5 5 4 5 5

3 2 2 2 2 2 2 3 3 3 3 3 3 4 4 4 4 4 4 4 4 5 5 4 5

*Legend: 5 – Excellent; the deep wound takes 1- 3 days to heal; 4 – Very good; the deep

wound takes 4 – 6 days to heal; 3 – Good; the deep wound takes 7 – 9 days to heal; 2 – Fair;

the deep wound takes 10 -12 days to heal; 1 – Poor; the deep wound takes 13-15 days to heal.


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Difference in the Effect of Sambong Leaves as to Number of Days

The difference in the effect of Sambong leaves as to number of days takes for wounds to heal

was found to be significantly different. ANOVA was calculated. The result of this analysis is

presented in Table 3.

Table 3. Differences in Means in the number of days of Healing of wound in mice as to

treatment applied.

Source of Variation Sum of

Squares Df

Mean

Square F p-value

Between Groups 72.708 3 24.236

333.315 0.000 Within Groups 4.944 68 7.271E-02

Total 77.653 71

The ANOVA computation revealed a significant difference in the number of days deep

wounds heal with the use of Sambong leaves decoction at different concentrations and

Betadine. The computed p-value (0.000) was lower than the set alpha (0.01). The results are

shown in table 4. Pairwise comparison using Scheffe’s test showed that for those deep

wounds treated with 50 grams of Sambong leaves decoction, the number of days needed for

deep wound healing to occur was significantly different from those treated with 100 grams of

Sambong leaves decoction (p=.000). Likewise, when 50 gm of Sambong leaves decoction

was compared to 150 gm of Sambong decoction, the number of days of healing was also

significantly different (p=.000). Furthermore, when 50 grams of Sambong leaves treatment

was compared to Betadine in terms of the effect in wound healing measured in number of

days, significant difference was also evident (p=.000).

Comparing the other Sambong leaves concentrations and the positive control (Betadine) with

each other revealed similar significant differences (p=.000). This means that Betadine was

significantly different to the three concentration of Sambong leaves decoction in terms of

effecting deep wound healing in mice measured in number of days, none of the Sambong

leaves was comparable to its effect on wound healing.

Table 4. Post hoc test for the differences in Means in the Days of Healing of wound in

mice Classified as to their treatment applied (with sambong in three different

concentrations and Betadine as positive control) using Scheffe test.

Treatment I Treatment J Mean Difference (I-J) Sig.

50gm sambong 100 -1.06 .000

150 -2.00 .000


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Betadine(control) -2.67 .000

100gm sambong

50 1.06 .000

150 -0.94 .000


150gm sambong

50 2.00 .000

100 0.94 .000


Betadine (Control)

50 2.67 .000

100 1.61 .000

150 0.67 .000

Effect on varying concentrations of Sambong leaves decoction and using betadine as

control

The use of Sambong leaves decoction with 50 grams concentration, the number of days

needed for the resolution of redness takes 11-12 days and this period of regression of

swelling was considered fair in all replications. Using 100 gm concentration of Sambong

leaves decoction applied on deep wound resulted in resolution of redness in 8-9 days,

described as good all throughout the treatments in all three replications. As for the use of 150

gm of Sambong leaves decoction, the redness resolved between 4-6 days, which was a very

good indication as it was consistent all throughout the treatments in all three replications.

The number of days needed for swelling to subside with the use of 50 gm Sambong leaves

decoction was within 10-11 days, considered as fair in terms of its speed all throughout the

treatment in all three replications. Seven to eight days was needed for swelling to subside

with the use of 100 grams of Sambong leaves decoction and this rate of resolution of swelling

was considered good as it was consistent all throughout the treatments in all three

replications. It would take 3-5 days for swelling to subside with the use of 150 grams of

Sambong decoction, as evidenced by such occurrence in all treatments and replications and

this rate of resolution of swelling was considered 3-5 days, crossing midway between being

described as very good and excellent.

With the use of positive control (Betadine), 2-3 days was needed for swelling to subside and

this rate of resolution was considered excellent as it was consistent all throughout the

treatments in all its three replications. The results are shown in table 5.


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Table 5. The effects of Sambong leaves decoction at various concentrations and of

Betadine as control on the redness and swelling in the wound of mice.

VA

LU

ES

RE

PL

ICA

TIO

NS

PA

RA

ME

TE

RS

SAMBONG CONCENTRATIONS BETADINE

(CONTROL) 50 gm. 100 gm. 150 gm.

TREATMENT TREATMENT TREATMENT TREATMENT

1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6 1 2 3 4 5 6

AC

TU

AL

VA

LU

ES

1 Redness 12 11 12 11 11 12 8 9 9 8 8 9 4 6 5 5 6 4 3 4 3 4 3 3

Swelling 11 10 11 10 10 11 7 8 8 7 7 8 3 5 4 4 5 3 2 3 2 3 2 2

2 Redness 11 12 12 12 11 12 9 8 8 8 8 9 6 6 5 5 4 5 4 4 3 3 4 3

Swelling 10 11 11 11 10 11 8 7 7 7 7 8 5 5 4 3 4 3 3 3 2 2 3 2

3 Redness 12 11 11 12 12 11 8 8 9 9 8 9 5 4 6 6 5 3 3 3 4 3 4 3

Swelling 11 10 10 11 11 10 7 7 8 8 7 8 4 3 5 5 4 4 2 2 3 2 3 2

CA

RT

EG

OR

ICA

L V

AL

UE

S* 1

Redness 2 2 2 2 2 2 3 3 3 3 3 3 4 4 4 4 4 4 5 4 5 4 5 5

Swelling 2 2 2 2 2 2 3 3 3 3 3 3 5 4 4 4 4 5 5 5 5 5 5 5

2 Redness 2 2 2 2 2 2 3 3 3 3 3 3 4 4 4 4 4 4 4 4 5 5 4 5

Swelling 2 2 2 2 2 2 3 3 3 3 3 3 4 4 4 5 4 5 5 5 5 5 5 5

3 Redness 2 2 2 2 2 2 3 3 3 3 3 3 4 4 4 4 4 5 5 5 4 5 4 5

Swelling 2 2 2 2 2 2 3 3 3 3 3 3 4 5 4 4 4 4 5 5 5 5 5 5

*Legend: 5 – Excellent; the deep wound takes 1- 3 days to heal; 4 – Very Good; the deep

wound takes 4 – 6 days to heal; 3 – Good; the deep wound takes 7 – 9 days to heal; 2 – Fair;

the deep wound takes 10 -12 days to heal;1 – Poor; the deep wound takes 13-15 days to heal.

Resolution of Redness and Swelling

Although the relative descriptions of the effects of various concentrations of Sambong leaves

as compared with a positive control in terms of resolution of both redness and swelling were

concerned, the range of days involved was not quite similar. For instance, it can be noted that

while the redness subsides in 11-12 days with the use of 50 grams of Sambong leaves

decoction, its swelling resolved in 10-11 days, or that swelling starts resolving one day earlier

than redness. The same observation can be made with the other concentrations of Sambong

leaves extract and even with the use of Betadine as positive control.

Whenever there is cell and tissue damage, repair is initiated by the onset of inflammation.

There are four cardinal signs of inflammation: redness (rubor), heat (calor), swelling (tumor)

and pain (dolor).[25,26]

The onset of pain is due to the release of prostaglandins upon the

damage on plasma membranes of cells of tissues damaged by infliction of wound on the

mice. Damaged cells will also trigger the release of clotting factors to stop leakage of blood


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as well as of chemotactic factors that will attract platelets and thrombocytes in the area of cell

damage.[27]

These chemotactic substances will also trigger release of serotonin by the

attracted platelets or thrombocytes that will, in turn, cause vasoconstriction that will attempt

to prevent blood loss.[28]

Later, vasodilation occurs. Dilation of blood vessels in the wound

area results in increased flow of blood to the area, thus explaining for the redness. Cells in the

area of damage become more permeable to virtually all substances needed for cellular and

histological repair during these times, accounting for the swelling.[29]

Vasodilation will be

constant until repair of damaged cells and tissues is complete. Since mice blood is warm, the

surge of blood flow to the area results also in increased local temperature, explaining for the

occurrence of calor.[27]

This study found that swelling subsides before redness occurs as shown in table 6. This may

be swelling is caused directly by the increased permeability of the tissue damage area, while

the redness is directly caused by dilation of the blood capillaries in the area of tissue damage.

Therefore, serotonin secretion persists a little longer than the increased permeability of cells

and tissues in the damaged area.

Table 6. ANOVA Results for the Differences in Means in the number of days of

resolution of redness and swelling in mice as to treatment applied.

Parameter Sources of

Variation

Sum of

Squares Df

Mean

Square F Sig.

Days of Resolution of

Redness


376.429 .000 Within Groups 4.278 68 6.291E-02

Total 75.319 71

Days of Resolution of

Swelling


689.714 .000 Within Groups 3.111 68 4.575E-02

Total 97.778 71

Pairwise comparison using Scheffe’s test showed that for those deep wounds treated with 50

grams of Sambong leaves decoction, the number of days needed for resolution of redness and

swelling to occur was significantly different from those treated with 100 grams of Sambong

leaves decoction (p=.000). Likewise, when 50gm of Sambong leaves decoction was

compared to 150 gm of Sambong decoction, the number of days for redness and swelling to

subside was also significantly different (p=.000). Furthermore, when 50 grams of Sambong

leaves treatment was compared to Betadine in terms of the effect in resolution of redness and

swelling measured in number of days, significant difference was also evident (p=.000).

Comparing the other Sambong leaves concentrations and the positive control (Betadine) with


1113


each other reveal similar significant differences (p=.000). The results are shown in table 7.

This may be due to Betadine was significantly different to the three concentration of

Sambong leaves decoction in terms of resolution of redness in mice measured in number of

days, none of the Sambong leaves was comparable to its effect on wound healing in terms of

rate or speed of redness resolution.

Table 7. Post hoc test for the differences in Means in the resolution of redness and

swelling of wound in mice classified as to their treatment applied (with Sambong in

three different concentrations and Betadine as positive control) using Scheffe test.

(I) Concentration (J) Concentration Mean Difference

(I-J) Std. Error

Sig.

(pvalue)

50

100 -1.00 8.36E-02 .000

150 -2.00 8.36E-02 .000

Betadine (control) -2.61 8.36E-02 .000

100

50 1.00 8.36E-02 .000

150 -1.00 8.36E-02 .000


150

50 2.00 8.36E-02 .000

100 1.00 8.36E-02 .000

Betadine (control) -.61 8.36E-02 .000

Betadine control

50 2.61 8.36E-02 .000

100 1.61 8.36E-02 .000

150 .61 8.36E-02 .000

50

100 -1.00 7.13E-02 .000

150 -2.22 7.13E-02 .000


100

50 1.00 7.13E-02 .000

150 -1.22 7.13E-02 .000


150

50 2.22 7.13E-02 .000

100 1.22 7.13E-02 .000

Betadine (control) -.78 7.13E-02 .000

Betadine (control)

50 3.00 7.13E-02 .000

100 2.00 7.13E-02 .000

150 .78 7.13E-02 .000

CONCLUSION

The present study concluded that varying concentrations of Sambong (Blumea balsamifera)

leaves decoction bent different effects on deep wounds of mice. The greater the concentration

of Sambong leaves decoction, the faster the wound healing was found. There was significant

difference in the effect of Sambong leaves decoction and Betadine in the number of days of


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wound healing in mice. Also significant difference in the effect of the Sambong leaves

decoction and Betadine in the resolution of redness and swelling was found.

Wound healing in general and the cardinal signs of inflammation are resolved not solely by

application of antiseptics or by antiobiotic treatment, because the use of these are meant to

augment healing as well as prevent further complication of the pathological processes and do

not absolutely determine wound healing and resolution of cardinal signs of inflammation.

Moreover, wound healing and its signs and symptoms are not solely dependent on the use of

antiseptics and antibiotics, but also on other factors like food taken i.e. some nutrients found

in food speed up the healing process in several ways and metabolic rate among others.

In the light of the foregoing conclusions, parallel studies and clinical trials adding more

parameters on wound healing should be conducted to determine the predictors of wound

healing, redness and swelling resolution of the cardinal signs of inflammation together with

the use of Sambong leaves decoction.

ACKNOWLEDGEMENT

Authors extend their deepest gratitude and cordial thanks to Dr. Leo B. Solis, Advisor,

Virgen Milagrosa University Foundation, San Carlos City, Pangasinan, Phillipines for his

valuable suggestions, supervision, guidance and support for the completion of this research

work.

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