Effect of Green Tea Extract (Camellia Sinensis) with Nanocitosan Capsulation on Gingivitis Healing | Uningojati | Journal of Food and Pharmaceutical Sciences 27333 58526 1 SM

J.Food Pharm.Sci. 4 (2016) 1 – 4

Avalaible online at www. jfoodpharmsci.com

Research Article

Effect of Green Tea Extract (Camellia Sinensis) with Nanocitosan
Capsulation on Gingivitis Healing
Annisa Hidaratri Uningojati1 , Dilla Asriyani1, Urfa Tabtila1, Fathul Muin2, B. Nadya Kausara2, Indra
Bramanti3
1

Faculty of Dentistry, Gadjah Mada University, Yogyakarta
Faculty of Pharmacy, Gadjah Mada University, Yogyakarta
3
Pediatric Dentistry, Faculty of Dentistry, Gadjah Mada University, Yogyakarta
2

ARTICLE INFO
Received 16/04/2016
Received in revised form 17/05/2016

Accepted 20/06/2016
Available online 02/01/2017

ABSTRACT
Gingivitis is one of the high oral manifestations in Indonesia. Gingivitis can be
cured by using green tea extract containing flavonoids. This flavonoid can be
encapsulated with nanochitosan. This study aims to determine the effect of
topical application of green tea extract gel with chitosan capsulation on
gingivitis healing. The subjects of this study were 36 rats wistar white male
rats. Rat was divided into 4 groups. Group I was given chicaflo gel (chitosan
encapsulated flavonoid), group II was given green tea extract gel as
comparison, group III was given base gel as negative control and group IV
was given standard anti-inflammatory gel as positive control. The gel was
applied to wounds in gingival mice that had previously been injured using a
biopsy punch. On the 2nd, 5th and 7th days after gel application, 3 rats from
each group were decapitated and jawed for histologic preparations.
Furthermore, the number of inflammatory cells on the preparations of each
rat group is computed and compared in number. The results showed a
significant decrease in the number of inflammatory cells in chicaflo gel
applications compared to controls. Our conclusion is that the use of chicaflo

gel can improve the effectiveness of gingivitis healing.
Keywords: gingivitis, chitosan, green tea

1.

Introduction
Green tea (Camellia sinensis) is one of the plants
with the highest flavonoids[1]. The flavonoids proven to
have antiinflamatory, antibacterial and antioxidant
activities. Flavonoids in green tea evidently can inhibit
the growth of bacterial pathogens common periodontal
disease like Porphyromonas gingivalis, Provotella
intermedia and Rovotella nigrescens. Flavonoids green
tea also shown bactericidal effect against gram negative
anaerobic rod bacterial pigment black species P.
gingivalis and Provotella. Green tea has proven to have
anti-Streptococcus mutans activity. Gingivitis treatment
using green tea polyphenols (EGCG) reduces the ability
of P. gingivalis to attach oral epithelial cells[2].

Chitosan is one of polisaccharides derived from
Crustacean shells that mostly used as matrix of various
drugs and plant extracts. Chitosan began to applied
widely in the pharmaceutical, food and health industries.
Chitosan have some characteristics include antimicrobial, wound healing, non-toxic, biocompatibility,
degradable and also dissolves in water. Chitosan in
micro/nanoparticel form have so many benefit that is
non-toxic, stable during the use of high surface area, and
can be the matrix of various drugs and plant extracts[3].
Inflamation on gingivitis was an acute inflamation.
Early stage of gingivitis will increase vascular
permeability that can lead to migration of leukocyt
polymorphonuclear from vascular then caused edema.
During inflamation, monocyt will migrates from

A.H. Uningojati, et al./ J.Food Pharm.Sci 4 (2016), 1 - 4

peripheral blood vessel, then penetrates blood vessel
wall to reach inflamatory tissue. This migrate monocyt
will differentiate into macrophage in inflamatory tissue.

Lesions of gingivitis later can be changed from initially
only the presence of PMN leukocytes to the increase of
limfocyt and macrophage[4]. Gingivitis prevalence in
Indonesia ranks second, reach 96,58%[5].
Research using green tea extracts with nanochitosan drug carrier for gingivitis treatment never be
done before. This study used white male rats Wistar
strain. This study was intended to determine the effect
of green tea extract with nano-chitosan drug carrier in
gingivitis therapy on proliferation stage. In addition, the
results of this study was compared by Difflam Mouth Gel
as positive control.
2. Materials and Methods
2.1. Material and Samples
Materials: grinder, rotary evaporator, magnetic
stirrer, homogenizer, microscope, spectrophotometer,
pH stick, scatter test kits, sticking power test kit, mortar,
and stamper
Samples: dry green tea, chitosan, sodium
tripolyphosphate, acetic acid, 70% ethanol, 95% NaCl,
formalin buffer, CMC-Na, nipagin, nipasol, and aquadest.

2.2. Making Tea Extract
Preparation of tea extract was done using
maseration technique. The dried green tea is smoothed
using a grinder. The purpose of the treatment is to
increase the particle surface area to increase the powder
mixing with the solvent. Then, green tea powder is
macerated with 70% ethanol solvent for 24 hours with
stirring. The maserate is then filtered and the obtained
filtrate is concentrated with a rotary evaporator to
obtain a thick green tea extract.
2.3. Nanocitosan Capsule
The capsule of green tea extract into nanocitosan
was done by ionic gelation method. Preparation of
chitosan nanoparticles was done by dissolving chitosan
200 mg into 100 mL 1% acetic acid solution by using
magnetic stirrer. Then, 2 grams of green tea extract
dissolved in 100 mL chitosan solution with magnetic
stirrer. Preparation of sodium tripolyphosphate solution
was done by dissolving 40 mg of sodium
tripolyphosphate with 40 mg of aquadest. After that, 1%

sodium tripolyphosphate solution was poured gradually
with 5000 rpm stirring using homogenizer for 30
minutes to form a nanoparticle suspension. Sodium
tripolyphosphate will serve as a cross-linker.
Nanoparticles chitosan-extract green tea then separated
by way of centrifugation.
2.4. Gel Formulation
The green extract gel and CHICAFLO were
prepared using some additives. The standard gel formula
with CMC-Na base is as follows:

2

R / CMC-Na
2
Nipagin
0.2
Nipasol
0.06
Green tea extract 20 grams

Aquades add
60 mL
The first thing to do is that all the required
ingredients are weighed, then inserted CMC-Na into the
mortar, then added water and crushed until
homogeneous and fluffy. After mixing with homogeny
inserted nipagin and nipasol then re-crushed. The above
treatments were replicated 2 times with the addition of
the active ingredient, the first mortar inserted the green
condensed extract, the second mortar incorporated the
chitosan-encapsulated green tea extract (Chicaflo), and
the third mortar was not added the active ingredient.
Then each mortar is crushed to homogeny to form an
elastic gel mass.
Furthermore, gel quality test in the form of
homogeneity test, organoleptic test, spreading test,
sticky power test, Ph.
2.5. Wound Healing Test
2.5.1. Preparing Animal
Try: The experimental animals used in this study

were male white rats wistar strain aged 3 months heavily
200 grams. Total of 36 male white rats were divided into
four treatment groups, each group consisted of 9 tails.
2.5.2. Animal Injury
Try: First try the animal injected ketamine. The
gingivitis openings were performed with a 2 mm biopsy
punch. The treatment group was divided into 4 groups:
1. Group I: Given application of green tea extract gel
with nanocitosan capsulation (Chicaflo Gel)
2. Group II: Application of green tea extract gel
3. Group III: Application of gel without active
substance as a negative control
4. Group IV: Application of Difflam gel as a positive
control.
Drug and Gel administration in experimental
animals: Dosage is applied evenly on the wound area
every 2 times a day for seven days.
Observation of wounds. Wound observations were
performed on days 2, 5, and 7 by observing changes in
the number of inflammatory cells and the proliferation

process in wound healing including angiogenesis,
epithelialization, and fibrosis.
Observation of wounds. Wound observations were
performed on the 2nd, 5th, and 7th days by observing
changes in the number of inflammatory cells and the
proliferation process in wound healing including
angiogenesis, epithelization, and fibrosis.
3. Results and Discussion
3.1. Research Result
In this study the data obtained will be tabulated
and analyzed statistically with SPSS using two-way
ANOVA method.

A.H. Uningojati, et al./ J.Food Pharm.Sci 4 (2016), 1 - 4

3

Table 1. Result of Gel Quality Test

Chracteristics

Color
Smell
Taste
Consistency

Gel type
Green tea
Gel without active
extract gel
substance
Brownish black
transparent
Green tea
No smell
Green tea
typical CMC-Na
Homogeneous
Homogeneous

Chicaflo gel

Brownish black
Green tea
Green tea
Homogeneous

Difflam mouth-gel
transparent
No smell
No taste
Homogeneous

Table 2. Results of Counting of Angiogenesis

Gel type
Chicaflo gel

Green tea extract gel
Gel without active
substance
Difflam mouth-gel

Day
2
5
7
2
5
7
2
5
7
2
5
7

Angiogenesis
6,11
3,33
2,78
2
1
1,44
3,11
2,00

Table 3. Dependent variable: Angiogenesis

Gel type

Gel type

Chicaflo
gel

Green tea
extract gel
Gel without
active substance
Difflam mouth
gel
F
2.170

Mean
difference

Std. error

Sig.

1.6650

0.97406

0.351

95% Convidence
interval
Lower
Upper
bound
bound
-1.1218
4.4518

3.4433

0.97406

0.013

0.6565

6.2301

2.2233

0.97406

0.144

-0.5635

5.0101

df1
7

3.2. Discussion
Based on the results of the study, it was shown
that green tea extract gel with nanocitosan capsulation
(Chicaflo) was able to stimulate the proliferative phase in
wound healing process shown by the increasing amount
of fibroblast, angiogenesis and epithelization. The
proliferative phase consists of fibroplasia, granulation,
epithelialization and angiogenesis that begins 24 hours
after the wound[6]. During this process proliferation of
fibroblasts and endothelial cells plays an important role
in the occurrence of angiogenesis. This process is
controlled by FGF, TGF-β and VEGF[7].
The process of healing the wound on the oral
mucosa runs faster than the wound on the skin. This is
due to injury to the oral mucosa containing immune
mediators, blood vessels and less profibrotic mediators
but has higher rates of re-epithelization and faster
fibroblast proliferation than in skin[8].
Wound healing process is a process of restoring the
structure of cells and tissue layers damaged in a complex
and dynamic[9]. When injured, the physiological response
of the body will automatically perform the process of

df2
16

Sig.
0.095

wound healing and cell regeneration automatically[10, 11].
Although this process runs natural and natural, but
Required the existence of certain conditions that can
accelerate the healing process, one of which is
nutrition[12, 13]. Treatment of gingivitis using Chicaflo gel
showed increased proliferative phase stimulation caused
by nutrients contained in this gel have an influence on
wound healing process.
Chicaflo gel made from green tea extract
containing polyphenols flavonoids have the ability as
antiinflamasi[14]. There are two ways of flavonoids in
inhibiting inflammation, by inhibiting capillary
permeability and inhibit acid metabolism arakidonat[15, 16].
Gingivitis treatment using the chicaflo gel proved able to
stimulate wound more effectively. The results showed
the increase of thickness of the epithelium in gingivitis
treatment using a gel Chicaflo significant compared to
the control treatment of gingivitis. This shows that
Chicaflo gel can increase epithelialization.
Epithelialization thickness resulting in gingivitis
therapy using Chicaflo gel showed epithelialization and
wound closure process that is faster and shorter

A.H. Uningojati, et al./ J.Food Pharm.Sci 4 (2016), 1 - 4

inflammatory phase. Chicaflo gel contains flavonoids
from green tea which works to improve the process of
mitogenesis, cell interaction and adhesion molecules
play an important role in the proliferative phase and
epithelialization terhada healing process of scar tissue.
4. Conclusion
The conclusion of this study is that green tea
extract with nanocitosan capsulation / Chicaflo gel
(Chitosan encapsulated flavonoids) can improve the
effectiveness of gingivitis healing.

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