International Journal of Materials Science and Applications

2017; 6(6): 284-289

http://www.sciencepublishinggroup.com/j/ijmsa
doi: 10.11648/j.ijmsa.20170606.13
ISSN: 2327-2635 (Print); ISSN: 2327-2643 (Online)

FGF-2, MMP-8 and Integrin α2β1 Expression in Periodontal

ligament Remodelling Tension Area with Nanopowder
Stichopus hermanii Application to Prevent Orthodontic
Relapsing
Noengki Prameswari1, *, Puguh Bayu Prabowo2
1
Orthodontics Department, Dentistry Faculty, Hang Tuah University, Surabaya, Indonesia
2
Dental Material Department, Dentistry Faculty, Hang Tuah University, Surabaya, Indonesia

Email address:
[email protected] (N. Prameswari), [email protected] (P. B. Prabowo)
*
Corresponding author

To cite this article:

Noengki Prameswari, Puguh Bayu Prabowo. FGF-2, MMP-8 and Integrin α2β1 Expression in Periodontal ligament Remodelling Tension Area
with Nanopowder Stichopus hermanii Application to Prevent Orthodontic Relapsing. International Journal of Materials Science and
Applications. Vol. 6, No. 6, 2017, pp. 284-289. doi: 10.11648/j.ijmsa.20170606.13

Received: October 19, 2017; Accepted: October 23, 2017; Published: November 16, 2017

Abstract: Background: Orthodontic relapse tendency occurs 33–90 per cent after at least 10 years post-treatment. Orthodontic
Relapsing is the return, following orthodontic correction, at the end of treatment. Relapse, a return to the original teeth position,
caused by periodontal, occlusal, soft tissue factor and growth. Nanopowder Stichopus hermanii have various active ingredient
such as protein, collagen, flavonoid, chondroitin sulphate, cell growth factor, EPA DHA, that might have role to FGF-2, MMP-8,
Integrin α2β1 in periodontal ligament remodelling tension area in reducing orthodontic relapsing. Objectives: The aim of this
study is to investigate application of Nanopowder Stichopus hermanii to prevent relaps orthodontic by periodontal ligament
remodelling tension area through FGF-2, MMP-8, and Integrin α2β1 parameter. Material and Method: The experiment was held
by Post Test Only Group design. Twenty four male Cavia Cobaya were divided into three groups. K(-) group as negative control
group (without treatment), K(+) group as positive control group which were applied with relaps orthodontic forces, and the other
groups P, were applied with relaps orthodontic forces and nanopowder Stichopus hermanii 3%. After treatment the cavia cobaya
were sacrificed. FGF-2, MMP-8, and Integrin α2β1 expression is examined with immunohistochemistry. Results: Application
nanopowder Stichopus hermanii can reduce relaps until 30%. This study showed FGF-2 expresssion are 13,9±2,6; 6,5±1,4;
21±4,6; MMP-8 expression means and SD in K(-), K(+), P are 3.75±1.49; 14.88±2.64; 9.13±1 and Integrin α2β1 expression are
5.25±1.28; 12.88±1.36; 16.50±2.33. There was significantly differences in group P compare to K(-) and K(+). Conclusion:
Application of Nanopowder Stichopus hermanii have role in periodontal ligament remodeling tension area through increasing
FGF-2, decreasing MMP-8, increasing Integrin α2β1 and Collagen type 1 parameter to prevent relaps orthodontic until 30%.
Keywords: Nanopowder Stichopus hermanii, FGF-2, MMP-8, Integrin α2β1, Relaps Orthodontic

advised of potential for relapse prior treatment and the need to

1. Introduction stay in long-term retention [2]. Beside retention, fiberotomy is
Relapse occurred at the end of active orthodontic appliance. also known for diminished relapse [3].
There was a rapid relapse initially after appliance removal but Many technique used to prevent orthodontic relapsing but
after 3 days, and began to gradually decrease. After appliance there is no natural has been used for relapse orthodontics.
removal, the teeth began to relapse in the direction of their Stichopus hermanii belong to the phylum Echinodermata,
original position [1]. The teeth began moved to tension area under the class Holothuridea. It was further divided into three
and produce periodontal remodeling. Instability or a tendency subclasses namely Dendrochirotacea, Aspidochirotacea, and
toward relapse should be anticipate. Patients should be Apodacea. There are six orders under these subclasses,
 International Journal of Materials Science and Applications 2017; 6(6): 284-289 285

named as Aspidochirotida, Apodida, Dactylochirotida, separator was removed for 7 days for producing orthodontic
Dendrochirotida, Elasipodida and Molpadiida [4]. Stichopus relapse orthodontic. Separator forces was 0,0474 kN,
hermanii have been well recognized as traditional medicine in measured by autograph. Biometric orthodontic relapsing was
Chinese The Chinese have long regarded sea cucumbers as a measured by caliper.
general health tonic, beneficial for treating tendonitis and
arthritis, and as an aphrodisiac, among many other medicinal
claims. The Chinese also consider Stichopus hermanii as
culinary [5]. Malaysian value Stichopus hermanii for
hypertension, asthma, rheumatism, burns, impotence and
constipation treatment [6] and also for wound healing,
treatment of stomach ulcers and as a painkiller, and as culinary
delicacies [5]. The Japanese eat Stichopus hermanii row,
nibble on their pickled intestines over drinks and eat their
dried gonads as a special treat [7].
Previous research showed that Stichopus hermanii can
increase Collagen type 1 expression in relaps orthodontic.
Figure 1. Biometric orthodontic relapsing measure.
Stichopus hermanii can act as anticandidal [8],[9] Stichopus
hermanii treated wounds and stimulation tissue regeneration. 2.3. Preparation and Applied Stichopus hermanii Gel
[10]. The other study show that studies have shown that the
extract of Stichopus species also affects viability or Stichopus hermanii gel 3% was made from 0,3 gr Stichopus
proliferation of human fibroblasts and osteoclast cells in a hermanii powder was diluted with NaCMC 2% in DMSO 5%
negative manner [10]. until 10 ml. Stichopus hermanii gel was applied in gingival
sulcus with insulin syringe 0,025 ml twice per day.
2. Material and Methods The procedure of this study was began with acclimatization
of animals for 48 hours. Next, Guinea pigs were divided into 4
The study is true experimental laboratories with completely groups. K(-) group as negative control group (without
randomized control group post test only design. Ethical treatment), K(+) group as positive control group which were
permission was obtained from Ethics and Scientific Research triggered orthodontic tooth movement by using elastic
Committee of Experimental Animal Use in Dentistry Faculty separator, the force was 0,0474 kN, measured with gauge
Airlangga University. Twenty four male guinea pigs (Cavia during experiment and the other groups P1, were applied with
cobaya) aged 2,5 months and weighed 200-300 grams. The both orthodontic forces and Stichopus hermanii 3% in 14 days
guinea pigs, fed with a standard pellet diet and tap water ad and released the forces for 7 days. Stichopus hermanii gel was
libitum, were randomly divided into three equal groups. The applied in gingival sulcus with insulin syringe 0,025 ml once
materials used were 3% and 3,5% Stichopus hermannii, 10% per day.
ketamine injection as anesthetic drug, a dose of 0.1-0.2 ml/kg The research was conducted in Biochemistry Laboratory
for acepromazine 0.5 ml, 10% buffered formalin, and cotton. Medical Faculty of Airlangga University. The guinea pigs
were monitored during the experiment, and all of the groups
2.1. Nanopowder Stichopus hermanii Preparation and were sacrificed on the twenty first day of the experiment. The
Quantitative Analysis Content maxillary insisivi teeth were dissected and placed in 10%
Stichopus hermanii were used in this study from coastal buffered formalin. Afterwards, histological section were
regions around Sumenep, East Java Indonesia. Stichopus prepared with FGF-2, MMP-8, integrin α2β1
hermanii was cleaned by making a longitudinal incision 3-5 immunohistochemistry as periodontal remodeling marker, and
cm on the ventral side of Stichopus hermanii without then observed by using a microscope.
damaging the internal organs using scalpel. Stichopus The photos were taken to measure the FGF-2, MMP-8,
hermanii was dried by ovens, blender, made nanopowder by integrin α2β1 expression in periodontal ligament tension area
using High Energy Milling method. SEM and TEM pictures was observed seen on the microscope with an enlargement
reveal the morphology and particle size of nanopowder 400x. Each histological section was observed and calculated
Stichopus hermanii. Quantitatives analysis of Stichopus as many as three times in the field of view.
hermanii active ingredients use spectrophotometry to examine Finally, the data were statistically measured by using
flavonoid, gas chromatography to examine EPA and DHA, Statistical Package for the Social Science (SPSS) version 20
and HPLC to examine chondroitin sulphate. program. The research data result tabulated and planned to
analyze by descriptive statistic test, normality distribution test
2.2. Preparation of Orthodontic Relapsing to know if the data that obtained come from population with
normal distribution, ANOVA test (analysis of varians) to
Orthodontic relapsing forces was produced by using analyze the difference of each variable compared with control.
separator rubber application with separating plier in mesial Then the data were tested with LSD Test (p<0.05).
left insisivus maxilla cavia cobaya 14 days and after day 15
286 Noengki Prameswari and Puguh Bayu Prabowo: FGF-2, MMP-8 and Integrin α2β1 Expression in Periodontal ligament
Remodelling Tension Area with Nanopowder Stichopus hermanii Application to Prevent Orthodontic Relapsing

3. Result normality test, homogenity test and show the data was
homogen and have a normal distribution. ANOVA test
The aim of this study is to investigate application of (p=0.05) for the biometric orthodontic relapsing Cavia
Nanopowder Stichopus hermanii to prevent relaps orthodontic Cobaya applied with Stichopus hermanii showed significantly
by periodontal ligament remodelling tension area through differences.
FGF-2, MMP-8, and Integrin α2β1 parameter.
3.1. Biometric Orthodontic Relapsing

Table 1 and figure 2 below show the biometric orthodontic

relapsing.

Table 1. Descriptive Mean and Stardard Deviation of Biometric orthodontic

relapsing (mm).

Group Mean ± Standard Deviation

Figure 2. Line chart mean biometric orthodontic relapsing.
K(-) 0.00 ±0.00
K(+) 0.09 ±0.02
P 0.06 ±0.02 3.2. FGF-2 Expression

Table 1 shows means and SD in K(-), K(+), P are 0,00±0,00 ; FGF-2 expression mean and standard deviation as seen as
0,09±0,02; 0,006±0,02. Then the data were tested with table 2 and figure 3 below.

Figure 3. FGF-2 expression in K(-) group (A), K(+) group (B), P group (C).

Table 2. Descriptive Mean and Stardard Deviation of FGF-2 expression expression of FGF-2 in orthodontics relapsing. Cavia Cobaya
(cell/view of field). applied with Nanopowder Stichopus hermanii showed
significantly differences. With the Tukey HSD test, showed
Group Mean ± Standard Deviation
K(-) 13,9±2,65
that MMP-8 expression was significantly increased in P
K(+) 6,5±1,41 compare to K(+) and K(-).
P 21±4,61
3.3. MMP-8 Expression
Table 2 show FGF-2 means and SD in K(-), K(+), and P are
MMP-8 expression mean and standard deviation as seen as
13,9±2,65 ; 6,5±1,41; 21±4,61, P group have the highest
table 3 and figure 4 below.
FGF-2 expression. Then the data were tested with normality
test, homogenity test and show the data was homogen and
have a normal distribution. ANOVA test (p=0.05) for the

Figure 4. MMP3 expression in K(-) group (A), K(+) group (B), P group (C).

Table 3. Descriptive Mean and Stardard Deviation of MMP-8 expression Table 3 show MMP-8 means and SD in K(-), K(+), P are
(cell/view of field). 3,75±1,49 ; 14,88±2,64; 9,13±1,55. Then the data were tested
with normality test, homogenity test and show the data was
Group Mean ± Standard Deviation
K(-) 3.75 ±1.49
homogen and have a normal distribution. ANOVA test
K(+) 14.88 ±2.64 (p=0.05) for the expression of MMP-8 in relaps orthodontics
P 9.13 ±1.55 Cavia Cobaya applied with Nanopowder Stichopus hermanii
 International Journal of Materials Science and Applications 2017; 6(6): 284-289 287

showed significantly differences. With the Tukey HSD test, 3.4. Integrin α2β1 Expression
showed that MMP-8 expression was significantly increased in
P compare to K(+) and K(-). Integrin α2β1 expression mean and standard deviation as
seen as table 4 and figure 5 below

Figure 5. Integrin α2β1 expression in K (-) group (A), K(+) group (B), P group (C).

Table 4. Descriptive Mean and Stardard Deviation of Integrin α2β1 activation of fibroblasts and osteoblasts in the periodontal
expression (cell/view of field). ligament and osteocytes within bone. Combination of
remodelling of the periodontal and remodelling of the alveolar
Group Mean ± Standard Deviation
K(-) 5,25±1,28
bone makes the tooth to move [16]. Differently when relaps
K(+) 12,88±1,36 occurred and tension area losing strain and implied decreasing
P 16,55±2,33 electrical conductivity. Remodelling of the periodontal
ligament is essential for orthodontic tooth movement [17],
Table 4 show the highest Integrin α2β1 expression means [14]. The periodontal contribution to the equilibrium that
and SD in K(-), K(+), P is in P1 16,55±2,33. After the data normally controls tooth position [12].
were tested with normality test, homogenity test and show the Relapse in cavia cobaya models occurs rapidly. The left first
data was homogen and have a normal distribution, then insisivus compressed towards the distal side during 14 days
ANOVA test (p=0.05) was used for the expression of Integrin orthodontic tooth movement and relapsed toward the mesial
α2β1 in relaps orthodontics Cavia Cobaya applied with side. There was a rapid relapse initially following 7 days
Nanopowder Stichopus hermanii. The result showed appliance removal. Increasing FGF-2 expression by applying
significantly differences in all groups. With the Tukey HSD Stichopus hermanii during relapse period means there is
test, showed that Integrin α2β1 expression was significantly increasing fibroblast that manner as a collagen production that
increased in P compare to K(+) and K(-). is important for periodontal ligament remodeling begins.
Periodontal ligament plays a central role for remodeling
4. Discussion periodontal ligament dan alveolar bone.
Periodontal Ligament is a fibrous structure with tensile
One major reasons is the gingival and periodontal tissues strength and elasticity, connecting the tooth to the alveolar
are affected by orthodontic tooth movement and require time bone. When mechanical forces load, the Periodontal ligament
for reorganization when appliance are removed [12], [2]. fibers will be repelled from one another and then rebound as
Stretching of supraalveolar gingival fibres, the transseptal simulating an elastic effect. When the initial phase of
fibres, in particular, has been suggested as the cause of relapse orthodontic tooth movement in which the tooth is displaced
[13]. within the periodontal ligament space, the inflammation
The aim to this study was to investigate the periodontal occurred and affect the viscoelastic properties of both the bone
ligament remodeling by Stichopus hermanii showed FGF-2 and the periodontal ligament. There is decreasing tensile
means and SD in K(-), K(+), P are 13,9±2,65 ; 6,5±1,41; strength of the collagen bundles as a result of release of matrix
21±4,61. It means in relaps orthodontic, there are increasing metalloproteinases (MMPs) and other catabolic agents which
fibroblast activity in P compare to K (-) and K (+). By giving disrupt the cross-linkages and molecular integrity of the
Nanopowder Stichopus hermanii 3% concentration in gingival extracellular matrix, and the hydrodynamic damping effect of
sulcus tension site, FGF-2 activity was significant increase. the periodontal ligament is decreased, while the elasticity of
When orthodontic tooth movement occurs, cells in the the bone is increased [15]. The same way occurred when the
periodontal ligament response differently to tensile and to tooth move back to the original position or orthodontic
compressive strains, which predominantly catabolic tissue relapsing.
changes at sites under compression and predominantly Stichopus hermanii contain various active ingredient such
anabolic activity at sites under tension area. Tensional forces as hyaluronic acid, chondroitin sulphate, cell growth factor,
of sufficient magnitude will stretch the plasma membrane EPA DHA, flavonoid [4]. In this research showed that there is
increasing the channel diameter and the flow of ions implied decreasing MMP-8 when we apply Nanopowder Stichopus
the electrical conductivity of the membrane is increased. This hermanii it means increasing the tensile strength of the
activates intracellular signalling eliciting cellular responses collagen increase and implied bone remodeling. Nanopowder
[14], [15].. This causes cell deformation, which produce
288 Noengki Prameswari and Puguh Bayu Prabowo: FGF-2, MMP-8 and Integrin α2β1 Expression in Periodontal ligament
Remodelling Tension Area with Nanopowder Stichopus hermanii Application to Prevent Orthodontic Relapsing

Stichopus hermanii 3% contain flavonoid are known as strong expression, bone protein that important for bone
inhibitors of metaloproteinases degrading matrix proteins. The remodeling that can prevent orthodontic relapsing
increased level of collagen could result from inhibition of
metaloproteinases activity[18]. Acknowledgements
The applied orthodontic forces compress when orthodontic
tooth movement, stretching occured in tension area, alter the The research was supported by a grant from Unggulan
collagen fibers and fluid within the periodontal ligament space, Research Program funded by Ministry of Research
thus disrupting the configuration of the extracellular matrix Technology and higher Education Indonesia 2015-2016.
proteins, induce FGF-2 that can activate the fibroblasts via
integrins and focal adhesion[12]. Some of these activate
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