3851 5553 1 SM PDF
3851 5553 1 SM PDF
3851 5553 1 SM PDF
ABSTRACT
Injuries to the gingiva are often encountered because of abnormalities in the oral cavity. Gingival
wound healing is more complex because it lies in an open area, often contaminated and exposed to various
types of bacteria in the oral cavity. Gingiva wound healing is determined by increase of fibroblasts. Plants
widely used by humans as medicine is star fruit (Averrhoa Bilimbi Linn). This study aims to determine an
increase of gingival fibroblasts on wound healing of male white rats given various concentrations of
starfruit leaves extract topically. The study was conducted with a pretest-posttest control group design,
consisted of four groups: the control group with distilled water and the groups treated with ethanol extract
of starfruit leaves with a concentration of 10%, 20% and 40%. The result based on comparison test between
the groups with One Way Anova, showed that the average amount of fibroblast in four groups after
receiving the treatment was significantly different (p<0.01). The result of the test showed an increase in the
number of fibroblasts in groups concentration of 10% and 20%, there was a decline observed in group
concentration of 40%. It was concluded that application of ethanol extract of starfruit leaves with
concentration of 10% increase the highest number of fibroblasts in the rat gingival wound healing process.
Keywords: wound healing, leaves of starfruit (Averrhoa Bilimbi Linn), fibroblasts.
picture of an increase in inflammatory cells, Starfruit can be used as a drug to somatitis wound
vascularity, epithelial cell density, increased number of because it contains saponins which have the ability to
fibroblasts and collagen fibers. The wound healing clean and as an anti-septic, tannins and flavonoids act
as anti-inflammatory by inhibiting the enzyme
Correspondence: Hartini IGAA cyclooxigenase and lypoxygenase. While alkanoid able
Address: Faculty of Dentistry Mahasaraswati University,
Bali-Indonesia to relieve pain (analgesic) and vitamin C play a role in
the repair of oral mucosal tissues, which can help the
5
process of wound healing.
35
ISSN. 2085-4773. Indonesia Journal of Biomedical Science (2012), Volume 6, Number 1: 35-39
Research in some plants obtain flavonoid that have added 70% ethanol and stirred for 30 minuntes with
an anti-inflammatory activity through inhibition magnetic stirrer and allowed to stand for 24 hours.
reaction of some enzymes. Aglikon is a non polar This mixture was then filtered using Buchner funnel.
flavonoid, whereas glilosida is a polar flavonoid. The filtrate obtained was dried via evaporation using
Therefore, polar flavonoid can be isolated from star vacuum rotary evaporator. After evaporation, crude
7
fruit using water solvent or etanol 70%. Etanol 70% is extract was obtained and from this crude extract many
an effective solvent to produce an optimal amount of consentrations of starfruit, i.e 10%, 20%, and 40%
active ingredient, does not cause swelling of the cell were prepared based on method mentioned in Voigt,
6
membrane and improve the stability of the 1994.
6
components dissolved.
This research aims to investigate application of star Treatment of Rats
fruit (Averrhoa Bilimbi Linn) extracted using ethanol Rats for this experiment were adapted for one
for increasing fibroblast in wound healing of gingiva week in a plastic cage. To make the rats do not getting
injury of white male rat. stress, the environment should be clean, dry or moist,
odorless, and have pretty good air circulation,
0
METHODS temperature between 25-35 C, have a sufficient
Research Design intensity of light and avoid noise. Food composition to
This is an experimental study with Randomized reach nutrious food are compromise of vitamins A
8
Pre-Posttest Control Group Design. A number of 32 4000UI/kg, vitamin D 1000UI/kg, -tocopherol
white male rats were recruited and divided into 4 30mg/kg, 3g/kg linoleic acid, 4mg/kg thiamine,
groups. The first is control group treated only with riboflavin 3mg/kg , pantothenic 8mg/kg, vitamin B12
aquadest. The second is treatment group 1 treated 50g/kg, 10g/kg biotin, pyridoxine 40-300 g / kg,
with 10% of star fruit extract. The third is treatment and 1000mg/kg kaolin.
group 2 treated with 20% of star fruit extract. The last Before making wound, rats were anesthetized
group is treatment group 3 treated with 40% of star topically on their xylonor gingiva using pellets. The
fruit extract. wound was made on the labial gingiva below the
mandibular central incisors using a 2 mm punch biopsy
Time and Places and depth of the wound on the alveolar bone.
This reseasrch was carried out at Pharmacology Starfruit leaf ethanol extract 10%, 20% and 40%
Laboratory Faculty of Medicine Udayana University respectively were spread out on the group II, III and
and Veterinary Faculty of Udayana University. The IV, whereas in group I only distilled water was applied.
research was carried out during Mach June 2011. Each group was smeared for 1 minute using a sterile
cotton bud and taken twice a day, i.e. in the morning
Research variable and evening.
Independent variable After 7 days of treatment, the rats were
1. Topical administration of 10% star fruit extract. decapitated using chlorofom. Wound tissue and
2. Topical administration of 20% star fruit extract. surrounding tissue is cut and put into pots containing
3. Topical administration of 40% star fruit extract. 10% buffered formalin and sent to the Laboratory
Faculty of Veterinary Udayana University to make
Dependent variable microscopic preparations.
Number of fibroblast in gingiva.
Preparation of microscopic materials
Control variables Tissue fixation was performed with a 10%
1. Food and rat cages buffered formalin for 24 hours. Then, the tissue was
2. Rata ge (2 months) cut using a scalpel. Fixation improvement was carried
3. Sexes of rat (male) out using automatic tissue processor and dehydrated
4. Weight of rats (180-200 g) with alcohol 70% -100% to clean the remnants of
5. Humidity fixative. To clean up any residual alcohol, xylol
6. Temperature cleaning and paraffin infiltration at temperature of 57-
0
7. Light 59 C were carried out and forming a block of paraffin.
This block cooled briefly in a freezer. Each block of
Procedure paraffin was sliced to 3-4 m thick using a microtome.
Preparation of starfruit ethanol extract Staining process was initiated by paraffin
Ethanol extract of starfruit leaves were prepared clearance using xylol, followed by rehydration with
at Laboratory of Pharmacy Faculty of Medicine alcohol from high concentration to low concentration
Udayana University. Starfruit leaves obtained from to take out remaining xylol and to put water in to the
Bualu village, Nusa Dua, Bali-Indonesia. Starfruit tissue. Residual alcohol is removed by washing the
leaves were washed and dried in room, then crushed preparat under running water and then stain with
in a blender to form powder. This powder then was Harris Hematoxylin-Eosin.
36
ISSN. 2085-4773. Indonesia Journal of Biomedical Science (2012), Volume 6, Number 1: 35-39
RESULTS
There were 37 of male Rattus novergicus age of
2 months and weight between 180 200 g, used as
samples. A number of 5 rats were applied for
obtaining pretest data. The rest, 32 rats were grouped
into 4 groups for treatment, i.e. group 1 as a control
group treated with aquadest, group 2 treated with
10% of starfruit leaf extract, group 3 treated with 20%
of starfruit leaf extract, and the last is group 4 treated Figure 2
with 40% starfruit leaf extract. All fibroblast data Fibroblast Observed with H-E Staining (400 zoom)
obtained were normally distributed and their A. For group 1 (control), B. Group 2 with 10%
variances were also homogeneous. Starfruit Leaf Extract, C. Group 3 with 20%
In this study, average number of fibroblast Starfruit Leaf Extract, and D. Group 3 with 40%
before treatment was 25.000.51. Their Starfruit Leaf Extract
immunohistochemistry was presented in Figure 1.
DISCUSSION
The different test between four groups after
treatment was tested using Anova. The results
indicate that there was a significant different of
number of fibroblast between group after treatment
with p value of 0.001.
Fibroblast increase in some treatment groups
after treatment were due to bioactive contents of the
starfruit leaf extract, such as flavonoid, tanin, saponin,
alkaloid and antioksidant. Flavonoid, saponin and
tanin have an effect as antiinflamatory, antibacterial,
Figure 1
and increase of fibroblast ploriferation.
Immunohistochemistry of pretest data
Some of active ingredient of star fruit, such as
saponin improve extracellular matrix metabolism and
Treatment effect analysis was performed based
activate of TGF- synthesis that stimulate of collagen
on number of fibroblast between groups after treated 9
biosynthesis. Then, inflammatory processes was
with starfruit extract. The significant test was
decrease by antiinflamation effect of flavonoid and
determined using One Way Anova and the results was
saponin, therefore, wound can be healed faster.
presented on Table 1. This table reveals that average
Research by Sudarsono, et al. (2002), indicates
number of fibroblast in control group was
that natural chemistry incidence in starfruit that
25.000.67, and for the treatment groups, i.e group 1, suspected as an antiinflamatory is flavonoid and
group 2, and group 3 were 27.230.39, 26.011.14, saponin. Research by Fahrani (2009), indicates that
and 16.380.36, respectively. Anova test indicates that starfruit extract contain flavonoid, saponin and
there was a significant different between groups 11
tanin. Other research by Jannah et.al. (2010), stated
indicates by p = 0.001. that aquadest extract of starfruit leaf tested was
Table 1 potent as an antibacteria that caused fish decay, i.e.
Summary of Anova Test between Groups 12
M. Luteus and P. Flurescens. The active component
37
ISSN. 2085-4773. Indonesia Journal of Biomedical Science (2012), Volume 6, Number 1: 35-39
38
ISSN. 2085-4773. Indonesia Journal of Biomedical Science (2012), Volume 6, Number 1: 35-39
11. Faharani, B.G.R. 2009. Uji Aktivitas Antibakteri 16. Narayana, K. R., Reddy, M.R., Chaluvadi, M.R. 2001.
Daun Belimbing Wuluh (Averrhoa bilimbi L) Bioflavonoids Classification,Pharmacological,
terhadap Bakteri Staphylococcus Biochemical Effects and Therapeutic Potential.
aureus dan Escherichia coli secara Bioautografi. Indian.
(Skripsi). Yogyakarta: Jurusan Farmasi FMIPA UII. 17. Katzung, B.G. 2001. Farmakologi Dasar dan
12. Jannah A, Hayati E.K, Mukhlisoh W. 2010. Klinik.(Dripa, S,.Pentj). Jakarta: Salemba Medika.
Efektivitas Antibakteri Ekstrak Akuades dan Ekstrak 449-471.
Tanin Pada Daun Belimbing Wuluh 18. Volk, W.A and M.F Wheeler. 1993. Mikrobiologi
(Averrhoa bilimbi L) secara In Vitro. Seminar Dasar. (Markham, Pentj). Jakarta: PT.Gelora Aksara
Nasional Kimia Bahan Alam. Bandung: ITB. Pratama.
13. Ilham, K., Sunardi. 2007. Uji aktifitas antioksidan 19. Hermanto, E dan Taufiqurrahman, I. 2008.
ekstrak daun belimbing wuluh (Averrhoa Bilimbi Manfaat terapi oksigen hiperbarik dalam
Linn) terhadap 1,1-Diphenyl-2-Pycrylhidrozyl mempercepat proses penyembuhan luka. (cited
(DPPH). Tehnologi Farmasi Fakultas Tehnik : 2009. okt 4). Available from: http://www.pdgi
Universitas Setiabudi. online.com/v2/index.php? option=com_content&
14. Biworo, A., Suhartono, E., Setiawan, B. 2004. task=view&id=732.
Aktivitas antioksidan enzimatik infus daun
belimbing wuluh (Averrhoa Bilimbi Linn) dan
potensinya sebagai analgesik. Kalsel : univ
Lambung Mangkurat Banjar Baru.
15. Bashori, Y.M. 2008. Efek Antiinflamasi Ekstrak
Etanol Daun Belimbing Wuluh (Averrhoa Bilimbi
Linn) Pada Tikus Putih Jantan Galur Wistar. (cited
2009.Nov. 11). Available rom:http //etd.eprints.
ums.ac.id /1425/k100040021.pdf.
39