Vol. 10 No.

1 Februari 2016 ISSN : 1907-5987

RESEARCH ARTICLE

The Effect Of Sea Cucumber Powder And

Hyperbaric Oxygen Therapy On The Expression Of
Tumor Necrosis Factor Alfa In Rats With Diabetes
Mellitus Induced By Porphyromonas gingivalis
Caroline Prajnaparamitha Anggraini*, Kristanti Parisihni**, Widyastuti***
*Undergraduate Dentistry Faculty Hang Tuah University Surabaya
***Oral Biology Faculty Hang Tuah University Surabaya
****Periodontia Faculty Hang Tuah University Surabaya

ABSTRACT

Background: Periodontitis have a bidirectional relationship with diabetes mellitus, both can
increase oxidative stress that trigger an increase of pro-inflammatory cytokines, such as
TNF-α. Sea cucumbers have anti-inflammatory component that act to inhibit the release of
inflammatory mediators. Hyperbaric oxygen therapy reduce oxidative stress. Purpose: The
aim of this study is to investigate the effect of the sea cucumber powder and hyperbaric
oxygen therapy on the expression of TNF-α in rats with diabetes mellitus induced by
P.gingivalis bacteria. Materials and Methods: The study was an experimental laboratories
research with factorial design. Twenty wistar rats were divided into 5 groups. K0 negative
control, K1 positive control. K1-K4 groups were induced with streptozotocin for diabetes
condition and P.gingivalis bacteria for periodontitis condition. K2 group was treated with sea
cucumber 3% for 7 days, K3 with OHB 2.4 ATA 3 x 30’ interval 5’ for 7 days, and K4 group
was treated with combination of sea cucumber and OHB. At the 51st day all rats were
sacrified, then the expression of TNF-α on periodontal macrophages were examined by
immunohistochemistry stain. Data were analized by Kruskal Wallis followed by Mann-
whitney. Result: Expression of TNF-α were increased in K1 (11.50±1.291) compare K0
(p<0.05). Sea cucumber treatment, OHB, and combination treatment decreased expression of
TNF-α significantly in amount of 2.50±0.577 (K2), 8.25±2.217 (K3), 3.00±0.816 (K4).
Conclusion: Sea cucumber powder and OHB therapy affected the expression of TNF-α in rats
with diabetes mellitus induced by P. gingivalis bacteria.

Keyword: Sea cucumber powder, TNF-α, periodontitis, diabetes mellitus, OHB therapy

Correspondence: Kristanti Parisihni, Department Oral Biology, Faculty of Dentistry, Hang

Tuah University, Arif Rahman Hakim 150, Surabaya, Phone 031-5945864, Email:
[email protected]

79
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

BACKGROUND subgingival flora.7 Many bacteria

associated with periodontal disease
The relationship between including Porphyromonas gingivalis one
periodontitis and diabetes mellitus has of the most important periodontal
long been discussed, both diseases have pathogens. At the time of periodontal
a relatively high incidence. Diabetes is a infection, the balance of bacterial flora
risk factor which increasing the severity in the oral cavity become changed, so
of periodontitis. Instead, periodontitis the species of pathogenic bacteria is
may be a risk factor for glycemic grow and survive, even when confronted
control in people with diabetes.1,2 with the immune response.6
Approximately 177 million people Lipopolysaccharide (LPS) is one of
worldwide have diabetes mellitus and most important microbial virulence
diabetes has been confirmed as a major factors in generating a host-mediated
risk factor for periodontitis. The risk of periodontal tissue destruction than that,
periodontitis increased approximately lipopolysaccharide is a potent stimulator
three-fold in individuals with diabetes of various inflammatory mediators,
compared with non-diabetic including IL-1β and TNF-α.8
individuals.3,4 Tumor necrosis factor (TNF α) is
Individuals with diabetes have reported have a role as a key to the
high glucose levels in the gingival fluid pathogenesis of diabetes mellitus type 2.
and blood more than individuals without Research in animals and in vitro human,
diabetes. Increased glucose levels in the TNF-α inhibits the action of insulin. In
gingival fluid and blood in individuals animal and human cell models in vitro,
with diabetes can change microflora TNF-α inhibits insulin. In one study, an
environment, induce qualitative changes increase in circulating levels of TNF-α
of bacteria that contribute to the is associated with insulin resistance and
severity, periodontal disease in patients type 2 diabetes. Individuals with
with uncontrolled diabetes. Poor periodontitis disease have high TNF-α
glycemic control led to increasing of response, monocytes from individuals
oxidative stress, advanced glycation end with diabetes and periodontitis release
products (AGEs), hyperresponsive massive amounts of TNF-α.9,8
innate immune system that contribute to Treatment of periodontitis with
damage factor diabetic periodontitis. In diabetes mellitus did not only include
addition, the function of the normal local factors, but also systemic factors
immune cells, such as chemotaxis, cell such as handling blood sugar.7
adherence, phagocytosis, cytokine Researchers showed that the local
production and secretion, are all affected periodontitis treatment such as scaling
by hyperglycemia.5,6 and root planing histologically not
Diabetics have more severe indicate new connective tissue
periodontal disease because of the attachment.5 Periodontitis treatment of
presence of glucose which induce diabetes that has been done is the tissue
accumulation of advanced glycation end surrounding dental treatments,
products (AGEs) and affect the activity antibiotics, and blood sugar control.
of migration and phagocytic activity of While the primary pathological
mononuclear and polymorphonuclear, condition in people with diabetes
so the formation of a more pathogenic

80
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

mellitus is a disorder of the healing patients with diabetes mellitus who use
process.10,11 OHB therapy there is decreased levels
Sea cucumbers have long been of malondialdehyde (MDA), which is
used as a traditional medicine. One one biomarker of oxidative stress.17,18 In
species of sea cucumbers which used as addition, OHB therapeutic effects on
medicine is Stichopus hermanii that monocytes and macrophages are causing
contains nutrients in wound healing, as disruptions to production of
an anticoagulant and antithrombotic, proinflammatory cytokines, resulting in
decrease cholesterol level and blood fat, a reduction in pro-inflammatory
anti-cancer, anti-tumor, cytokines in conditions of oxidative
immunostimulant, antirheumatic, stress.19
antimalarials, antiviral, antifungal and The research results of Fitria et al
antibacterial.12 The content of omega-3 (2013),20 cucumbers therapy for 7 days
(EPA and DHA) in Stichopus hermanii effectively decrease blood sugar levels
can decrease cholesterol levels because as well as the golden sea cucumber is
infection of mix periodontopatogen used as an antibacterial and antifungal in
bacteria and served to accelerate the inhibiting the growth of bacteria. The
healing of wounds and also as an anti- research results of Prabowo et al
inflammatory that inhibits the (2014),21 OHB therapy 100% 2.4 ATA 3
production of TNF-α and IL-1β. 13,14
x 30 minutes a day with a pause of 5
According to research of Rizal (2012)15, minutes for 7 days decreased blood
Stichopus hermanii is contained by sugar levels in diabetes mellitus type 2.
flavonoids. The content of flavonoids as It is necessary to investigate the
antioxidants that indirectly supporting combination of sea cucumber powder
anti-inflammatory effects by inhibiting gold and OHB therapy against TNF-α
the release of inflammatory expression in diabetes mellitus induced
mediators.15,16 P. Gingivalis bacteria.
Sea cucumbers are one of the
marine life that used in the health field.
Sea cucumbers have shown a beneficial MATERIALS AND METHODS
effect on periodontal therapy as well as
patients with diabetes mellitus. In The research is true experimental
addition to cucumbers, one therapy that research with factorial design study.
is emerging today is hyperbaric oxygen Parameters were examined in this study
therapy (OHB). was the expression of TNF α in diabetes
Hyperbaric Oxygen Therapy mellitus and periodontitis. The samples
(OHB) is a treatment in patients with were 20 Wistar rats (Rattus novergicus
giving 100% pure oxygen by inhalation Wistar strain) were divided into five
method in hyperbaric hyperoxia at the groups, where the selected criteria is the
air pressure of more than 1 atmosphere type males, aged 3-4 months with a
Absolute (ATA) and has been used as body weight of 150-200 grams.
adjuvant therapy in various diseases.10 The tools used in this research is
Hyperbaric Oxygen Therapy the syringe 5 cc and syringes 3cc,
(OHB) shown a beneficial effect on syringes insulin 1 mL, blood glucose
periodontal tissues by increasing oxygen test strips, gauges blood sugar, animal
pressure in the pocket. In periodontitis cages, food and drink rat, scales,

81
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

measuring cups and a stirrer, test tubes, sodium carboxymethylcellulose

22
test tube rack, micropipette, micro (CMCNa) 2%.
brush, glass, glass slide, surgical
scissors, microscope, chamber of Diabetes Induction procedures for
animals, immunohistochemistry kit. Wistar rats. Day 7 after adaptation,
Materials used nicotinamide 230 induction of streptozotocin (STZ). DM
mg/kg, 200 mg canamycin, 200 mg induction is done by giving
ampicillin, 0.6% chlorhexidine nicotinamide (NAD) of approximately
gluconate, streptozotocin (to make the 230 mg / kg, dissolved in the liquid PBS
rats diabetes mellitus), 10% dextrose, 10 minutes and then given STZ intra-
citric acid buffer 0.05M pH 4.5, peritoneal for 15 minutes injected a
aluminum foil, bacteria Porphyromonas single dose of 65 mg / kg in rat that had
gingivalis ATCC 33277, powder golden fasted overnight between 8-12 hours.
sea cucumber (Stichopus hermanii), Then rats were incubated for 7 days.
phosphate buffered saline (PBS), 4.13% Otherwise DM rats when blood glucose
EDTA, xylol, TNF-α antibody (sc-52 levels over 220 mg / dl.23
746-Santa Cruz Biotechnology, Inc.,
Santa Cruz, CA, USA ), H2O2, Procedure Premedication before
aquadest, Wistar rats, Wistar rats induction of bacteria. All the rats were
standard food, beverages Wistar rats given canamycin (20 mg) and ampicillin
(ordinary tap water), 100% pure oxygen (20 mg) daily for 4 days in drinking
in a hyperbaric chamber. water, and oral cavity swabbed with
0.12% chlorhexidine gluconate as
The division of research subjects. antiseptic.24
Those wistar rats were divided into five
group: first, negative control group Procedure Preparation and Induction
(K0); second, groups of diabetic Bacteria. Induction of bacteria using
periodontitis rats (K1); third, group of ATCC P.gingivalis imposing 33277
diabetic periodontitis rats with treatment with 2 ml of 1 x 109 cells / ml of live
golden sea cucumber (K2); fourth, bacteria in PBS, and peroral
groups of diabetic periodontitis rats with administration. Most bacteria applied
treatment of hyperbaric oxygen (K3); using cotton bud along the gingival
and five, groups of diabetic periodontitis sulcus and inserted of anal to the
rats with treatment combination golden colorectal region with syringe canula.
sea cucumber and hyperbaric oxygen Then incubation for 3 weeks since
(K4). giving bacteria counted first.25
Making the golden sea cucumber After the rats in the induction of
powder. Dissected sea cucumber to STZ and P.gingivalis bacteria, rats were
remove all viscceral organs, then blow- given a therapeutic treatment. For K2
dry sea cucumber using freeze dryer at a and K4 given group therapy treatment
temperature of 2-8 ° C with a pressure gel golden sea cucumber powder 3%
of 5 mTorr. The dried sea cucumbers are applied topical as much as 0.1 mg / day
weighed and then blended into a for 7 days using microbrush on inflamed
powder. Making the golden sea sulcus gingival. K3 and K4 group
cucumber powder gel using a mixture of treated OHB therapy, administration of
100% oxygen 2.4 ATA 3 x 30 minutes a

82
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

day with a pause of 5 minutes for 7 compared (comparing the expression of

days.26.21 TNF-α in each treatment group).
On day 8 after therapy gel golden
sea cucumber and OHB, all groups were
sacrificed then performed euthanasia by RESULT
the neck (cervical) dislocation, the
dosage is taken on the part of the Before doing hypothesis test
mandible. After that, do the between groups, first of all each group
decalcification phase part of the tested the normality by using statistical
mandible for 4 weeks using 4.13% test of Shapiro-Wilk, because in this
EDTA. Furthermore, the tissue study sample size <50 and the results
processing and staining by obtained is Shapiro-Wilk test has a
immunohistochemistry (IHC) was significant value of p> 0.05, so that the
performed using the streptavidin-biotin- distribution of data on this research is
peroxidase labeled streptavidin-biotin abnormal.
(Dako, Carpinteria, USA), and then
observed using a microscope with Table 1. The average and expression
1000x magnification and calculated as deviation standard of TNF-α in each group
much as 20x field view. Average ± Deviation
Data were analyzed using Group
Std.
parametric statistical test one way K0 5.00 ± 0.816
ANOVA, with a test for normality using K1 11.50 ± 1.291
the Shapiro-Wilk test, then continued K2 2.50 ± 0.577
homogeneity statistic test using Levene K3 8.25 ± 2.217
test. Then the calculation results K4 3.00 ± 0.816

Average dan Deviation Standard of

TNF-α expression
Tnf alfa expression

15

10

5
Tnf alfa
0 expression
K0 K1 K2 K3 K4
Group

Figure 1. Average and Deviation Standard of TNF-α expression in each group

83
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

A B C

D E

Figure 2. TNF-α expression in macrophages in the periodontal ligament rat (Rattus norvegicus)
with 400x magnification. A. K0 (normal); B. K1 (diabetes + bacteria); B. K2 (diabetes + bacteria
+ sea cucumbers); D. K3 (diabetes + bacteria + OHB); E. K4 (diabetes + bacteria + OHB + sea
cucumber). Expression of TNF-α are marked with brown color in macrophages (arrows).

It was found that the expression of

TNF-α have variances were not Tabel 3. Mann-Whitney test Result
homogenous because p = 0.041 K0 K1 K2 K3 K4
(p<0.05). Therefore, it can be concluded K0 0.020* 0.019* 0.028* 0.027*
that there is a difference of variance K1 0.019* 0.059 0.020*
between sets of data are compared. K2 0.019* 0.343
The research data were not
K3 0.020*
normally distributed, and found that the
data are not homogeneous, then doing *there is significant difference
non parametric Kruskal Wallis
hypothesis test to determine the
expression of TNF-α. DISCUSION

Table 2. Kruskal Wallis test Result Nicotinamide (NAD) is an

Sig. antioxidant which exerts protective
TNF-α expression 0.002
effect on the cytotoxic action of STZ by
scavenging free radicals and causes only
minor damage to pancreatic beta cell
In the table 2. obtained mass producing type 2 diabetes.23
significance value of 0.002 (p <0.05). It Streptozotocin is a drug that has a
shows a significant difference in each specific toxicity in pancreatic β cells.
group under these conditions, followed Streptozotcin cause DNA alkylation and
by Mann-Whitney test to find out where β cell death because STZ enter cells
is the difference in each group with a through the glucose transporter GLUT2,
significance level P <0.05. toxic mechanism is not specific for ß

84
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

cells and can cause damage to other (11:50 ± 1.291) increased compared
tissues including the liver and with normal rats (5.00 ± 0816), this is
kidneys.27,28 Damage to the pancreatic due to streptozotocin-induced rats to
beta cells causes the body can not make samples of rats became diabetic
produce insulin, causing blood glucose and bacteria capable P.gingivalis
levels to rise (a state of hyperglycemia). inducing the sample into periodontitis.
Conditions of hyperglycemia can result Some research it is known that the
in the formation of reactive oxygen golden sea cucumber (Stichopus
species (ROS) which would then trigger hermanii) as a potential therapy in the
an active nuclear factor kappa β (NFƙβ) field of dentistry, besides the golden sea
that play a role in the increased cucumber (Stichopus hermanii) itself
production of pro-inflammatory has long been used as a traditional
cytokines. 23,29 According to research medicine. Golden sea cucumber powder
from Swaroop et al (2012) 30 showed used on research using the golden sea
that TNF-α can play a potentially cucumber (Stichopus hermanii) which
important pathophysiological role in the are found in waters of Indonesia.33
development of insulin resistance. Based on the results of the
Results of research on groups of research showed that there were
bacterial-induced diabetic rats found significant differences on the expression
increased expression of TNF-α of TNF-α between groups of normal
compared with normal rats, it was rats, groups of bacterial-induced
shown that the group of bacterial- diabetic rats and rats treated groups of
induced diabetic rats occurred diabetes. sea cucumbers (see table 5). The test
Porphyromonas gingivalis is an results showed that the expression of
anaerobic gram-negative rod bacteria TNF-α in groups of rats treated
and include black-pigmented bacteria, cucumbers decreased compared to the
these bacteria cause inflammation and group of normal rats and a group of
tissue damage in diseases diabetic rats induced bacteria, because
periodontal.31,32 P.gingivalis get into the content of the golden sea cucumber
crevices using fimbriae and attached that is therapeutic as it helps wound
gingival epithelial cells digingiva, healing, antibacterial, antifungal,
proteases have the ability to destroy antitumor, antianaphylactic, anti-
periodontal tissue directly or indirectly, inflammatory, antinociceptive and
and LPS can cause inflammatory antioxidant.34
responses and resulting in the Teripang gold (Stichopus
production of cytokines in periodontal hermanii) examined contains the active
tissue, cause the activation of genes and ingredient antibacterial, antifungal
cause bone damage alveolar.31,6 Based (antifungal), antitumor and
on the results of the research showed anticoagulant (anti-clotting) and it also
that there were significant differences contains compounds that can reduce
(p=0.020) on TNF-α expression cholesterol and lipid, anticancer and
between groups of normal rats and a antitumor compounds, as well as
group of bacterial-induced diabetic rats antibacterial compounds.35 Golden sea
(see Table 5). The test results showed cucumber has a various kinds of
that the expression of TNF-α in the content, one of the active ingredients of
group of bacterial-induced diabetic rats the golden sea cucumber is Triterpenoid

85
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

strong role as an immunostimulatory. In anti-inflammatory effects by inhibiting

general Triterpenoid group capable of the release of mediators
damaging the cell membrane, deactivate inflamasi.16Penurunan expression TNF-
enzymes and proteins denature so that α at 2.4 ATA hyperbaric oxygen 100%
the cell wall is damaged due to a oxygen 3x30 minutes with 5 minute
decrease in permeability. Changes intervals for 7 days clinically
permeability of the cytoplasmic recommended, because it can lower
membrane allows ions important blood sugar levels, mRNA expression
organic entry into the cell resulting in and activity of antioxidant enzymes
poor growth and even to kill cells.36.35 In significantly.21,40,26
addition, the gold content of sea
cucumbers that PUFAs (DHA and EPA)
may block the formation of CONCLUSSION
prostaglandins and proinflammatory
cytokines (IL-6, IL-1β and TNF-α).37,38 According to the research, it can
Hyperbaric oxygen increases the be concluded that there is the influence
formation of oxygen free radicals, then of the golden sea cucumber powder 3%
oxidize proteins and lipid membranes of and hyperbaric oxygen therapy 3 x 30
bacteria, destroys DNA and inhibits minute interval of 5 minutes for 7 days
bacterial metabolic functions. The on the expression of TNF-α in rats with
enzyme superoxide dismutase, catalase, diabetes mellitus induced by the
glutathione and glutathione reductase bacterium Porphyromonas gingivalis.
inhibiting free radical formation, until
the oxygen concentration exceeded
REFERENCE
enzyme concentration.39 Based on the
results of the research showed that there 1. Malik G, Lehl G, Talwar M. 2011.
were significant differences between Association Of Periodontitis With Diabetes
groups of normal rats and rats treated Mellitus: A Review. Journal of Medical. 1
(1): 14-10. Available from
group OHB, but there is no significant http://gmch.nic.in/journalgmch/Archives/5
difference between groups of bacterial- %20Review%20article-
induced diabetic rats and rats treated %20Periodontitis%20with%20Diabetes%2
group OHB. The test results obtained by 0mellitus.pdf. Diakses 7 April 2015.
2. Akhila S and Malaiappan S. 2014.
the expression of TNF-α in groups of Periodontitis and Diabetes- A Two Way
rats treated group OHB increased Connection. Journal of Dental and Medical
compared to normal rats, but decreased Sciences e-ISSN: 2279-0853, p-ISSN:
the expression of TNF-α in groups of 2279-0861, 13 (1): 61-58.
3. Petersen PE and Ogawa H. 2005.
rats treated OHB compared with Strengthening the Prevention of Periodontal
bacterial-induced diabetic rats. Disease: The WHO Approach. J
Based on the results of the study Periodontal, 76(12): 2193-2187.
showed that there were significant 4. Preshaw PM, Alba AL, Herrera D, Jepsen
S, Konstantinidis A, Makrilakis K, Taylor
differences between groups of bacterial- R. 2012. Periodontitis and Diabetes: A
induced diabetic rats and mice Two-Way Relationship. Diabetolodia.
combination treatment group due to the PMCID: PMC3228943, 55(1): 8-1.
content of one of the golden sea 5. Newman MG, Takei HH, Takei HH,
Klokkevold P, Carranza F. 2012. Caranza’s
cucumber are flavonoids that act as Clinical Periodontology, 11th ed.,
antioxidants and indirectly support the

86
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

Philadelphia : WB Saunders Company. P. Proses Pulp Healing. Skripsi, Fakultas

43-41. Kedokteran Gigi Universitas Hang Tuah
6. Morran MP, Alexander LA, Slotterbeck Surabaya. Indonesia.
BD, McInerney MF. 2009. Dysfunctional 16. Hidayati NA, Listyawati S and Setyawan
Innate Immune Responsiveness to AD. 2008. Kandungan Kimia dan Uji
Porphyromonas Gingivalis Antiinflamasi Ekstrak Etanol Lantana
Lipopolysaccharide in Diabetes. Oral camara L. padaTikus Putih (Rattus
Microbiol Immunology, 24: 339–331. norvegicus) Jantan. ISSN: 0216-6887.
7. Anil S, Varma SV, Preethanath RS, Anand Bioteknologi, 5(1): 17-10. Available from
PS and Al Farraj A. 2012. The Emerging http://biosains.mipa.uns.ac.id/C/C0501/C05
Concepts on The Impact of Periodontitis on 0102.pdf. Diakses 2 April 2015.
Systemic Health. Periodontal Disease – A 17. Nogueira-Filho GR, Rosa BT, and David-
Clinician’s Guide. ISBN : 978-953-307- Neto JR. 2010. Effects of Hyperbaric
818-2. P. 164-131. Oxygen Therapy on The Treatment of
8. Jiang Z, Cui Y, Gao R, Lia Y, Fuc Z, Severe Cases of Periodontitis. Undersea
Zhang B, andGuan C. 2013. Study of TNF- and Hyperbaric Medical Society, Inc:
α, IL-1β and LPS Levels in The Gingival 37(2): 114-107.
Crevicular Fluid of A Rat Model of 18. Mulawarmanti D and Widyastuti. 2008.
Diabetes Mellitus and Periodontitis, 34: Effect of Oxygen Hyperbaric Therapy on
304-295. Malondialdehyde Levels in Saliva of
9. Engebretson S, Chertog R, Nichols A, Hey- Periodontitis Patients With Type 2 Diabetes
Hadavi J, Celenti R and Grbic J. 2007. Mellitus. Dental Jurnal Majalah Kedokteran
Plasma Levels of Tumour Necrosis Factor- Gigi, 41(4): 156-151.
α in Patients With Chronic Periodontitis 19. Thom SR. 2011. Hyperbaric Oxygen-Its
and Type 2 Diabetes. J Clin Periodontol, Mechanism and Efficacy. PlastReconstr
34: 24-18. Surg, 127(1): 141s-131s. Available at
10. Mulawarmanti D. 2008. Profil cAMP, NF- http://www.ncbi.nlm.nih.gov/pmc/articles/P
κβ, HIF-1α, i-NOS, dan MMPs Jaringan MC3058327/. Diakses 21 Maret 2015.
Periodontal Tikus Wistar Pada Kondisi 20. Fitriah, Theodorus dan Kamaluddin MT.
Hiperglikemia yang Dipapar Hiperoksia 2013. Efek pemberian serbuk teripang
Hipebarik. Disertasi. Surabaya : Program (Stichopus Variegatus) jangka panjang
Pascasarjana Universitas Airlangga 2008. terhadap kadar gula darah tikus Putih
11. Gurdol F, Cimsit M, Oner-Iyidogan Y, Jantan Galur Wistar Model Hiperglikemik.
Kocak H, Sengun S, and Yalcinkaya- MKS, 45(1): 10-5.
Demirsoz S. 2010. Collagen Synthesis, 21. Prabowo Sulistiana, Nataatmadja Maria,
Nitric Oxide and Asymmetric Poernomo Hadi Janto, Dikman Irmawati,
Dimethylarginine in Diabetic Subjects Handajani Fitri, Tehupuring Sihning E. J.,
Undergoing Hyperbaric Oxygen Therapy. Soetarso1 Iswahyudi,
Physiol, 59: 429-423. Suryokusumo Mohammad Guritno,
12. Farouk AE, Ghouse FAH and Ridzwan BH. Aulanni’am Aulanni’am, Herawati Anita,
2007. New Bacterial Species Isolated from West Malcolm. 2014. Hyperbaric Oxygen
Malaysian Sea Cucumbers with Optimized Treatment in a Diabetic Rat Model Is
Secreted Antibacterial Activity. ISSN Associated with a Decrease in Blood
1553-3468. American Journal of Glucose, Regression of Organ Damage
Biochemistry and Biotechnology, 3(2): 65- and Improvement in Wound Healing.
60. Health, 6, 1958-1950.
13. Sari RP and Revianti S. 2011. The activity Availabe from
of Stichopus hermanii extract on http://dx.doi.org/10.4236/health.2014.6152
triglyceride serum level in periodontitis. 28
Dental Jurnal Majalah Kedokteran Gigi, 22. Mulawarmanti D, Parisihni K, Wedarti YR.
44(2): 110-106. 2014. Catalase Activitty of Sea Cucumber
14. Calder PC. 2006. Long-chain Extract (Stichopus hermanii) to
Polyunsaturated Fatty Acids and Periodontitis induced by Porphyromonas
Inflammation. Scandinavian Journal of Gingivalis. Regional Oral Biology Sientific
Food and Nutrition, 50(S2): 61-54. Meeting 2014. Central Library, Universitas
15. Rizal MB. 2012. Komposisi Senyawa Indonesia. P. 30-26.
Organik dan Anorganik Teripang Pasir dan 23. Srinivasan K and Ramarao P. 2007. Animal
Teripang Emas yang Berperan dalam models in type 2 diabetes research: An

87
Vol. 10 No. 1 Februari 2016 ISSN : 1907-5987

overview. Indian J Med Res. 2007 March; Sea Cucumbers for Functional Foods—A
125: 472-451. Review.Mar. Drugs. 2011 October 10; 9:
24. Kesavalu L, Sathishkumar S, 1761-1805. doi:10.3390/md9101761. ISSN
Bakthavatchalu V, Matthews C, Dawson D, 1660-3397.
Steffen M, and Ebersole JL. 2007. Rat 34. Shahrulazua A, Samsudin AR, Iskandar
Model of Polymicrobial Infection, MA, Amran AS. 2013. The In-Vitro
Immunity, and Alveolar Bone Resorption Effects of Sea Cucumber (Stichopus sp1 )
in Periodontal Disease. American Society Extract on Human Osteoblast Cell Line.
for Microbiology. INFECTION AND Malaysian Orthopaedic Journal, 7(1): 41-8.
IMMUNITY. 75(4): 1712-1704. Doi:
doi:10.1128/IAI.00733-06. http://dx.doi.org/10.5704.MOJ.1303.015.
25. Praptiwi. 2008. Inokulasi Bakteri dan 35. Roihanah S, Sukoso, Andayani S. 2012.
Pemasangan Cincin Ligatur untuk Induksi Aktivitas Antibakteri Ekstrak Teripang
Periodontitis padaTikus. Majalah Holothuria sp. Terhadap Bakteri Vibrio
Kedokteran Gigi, 15(1) : 84-81. harveyi Secara In vitro. Fakultas Perikanan
26. Harnanik T. 2008. Efek Oksigenasi dan Ilmu Kelautan, Universitas Brawijaya,
Hiperbarik Terhadap Peningkatan Aktivitas Malang.
Antioksi dan pada Penderita Diabetes 36. Winarni D, Handono CD, Sugiharto. 2013.
Mellitus Tipe 2. Tesis. Surabaya: Program Efek Imunostimulatori Beberapa Fraksi
Pasca Sarjana. Universitas Airlangga. P. 87. Teripang Lokal Phyllophorussp terhadap
27. Lenzen S. 2007. The mechanism of Histopatologi Limpa Mencit
Alloxan and Streptozotocin induced (Musmusculus) Yang Diinfeksi
Diabetes. Diabetologia (2008), 51 : 226- Mycobacterium tuberculosis. Available
216. from
28. Deeds MC, Anderson JM, Armstrong http://journal.unair.ac.id/filerPDF/Cipto%2
AS, DA Gastineau, HJ Hiddinga, A 0Dwi%20Handono.PDF. Diakses 13 April
Jahangir, NL Eberhardt, and YC Kudva. 2015.
2011. Single Dose Streptozotocin Induced 37. Kordi MGH. 2010. A to Z Budidaya Biota
Diabetes: Considerations for Study Design Akuatik untuk Pangan, Kosmetik dan Obat-
in Islet Transplantation Models. Lab Anim, obatan. Yogyakarta: Lily Publisher. H. 18.
45(3): 131– 38. Heba El Bialy, Ola MG, and Khaled SA.
140.doi: 10.1258/la.2010.010090. 2011. Conversion of oil waste to valuable
PMCID: PMC3917305. Available from fatty acids using Oleaginous yeast. World J
http://www.ncbi.nlm.nih.gov/pmc/articles/P Microbiol Biotechnol, 27: 2791-98.
MC3917305/. Diakses 11 Januari 2016. Available from
29. Mealey BL and Thomas WO. 2006. AAP- http://www.academia.edu/723079/Conversi
Commissioned Review Diabetes Mellitus on_of_oil_waste_to_valuable_fatty_acids_
and Periodontal Diseases. Journal using_Oleaginous_yeast. Diakses 4
Periodontol, 77 (8): 1303-1289. doi: November 2015.
10.1902/jop.2006.050459. 39. Wibowo A. 2015. Oksigen Hiperbarik:
30. Swaroop JJ, Rajarajeswari Dand J.N. Terapi Penyembuhan Luka. Int. J. Unila, 5
Naidu. 2012. Association of TNF-α with (9): 128-124. Available from
insulin resistance in type 2 diabetes http://download.portalgaruda.org/article.ph
mellitus. Indian J Med Res, 135: 130-27. p?article=328322&val=5503&title=Oksige
31. Kimura S, Ohara-Nemoto Y, Shimoyama n%20Hiperbarik:%20Terapi%20Percepatan
Y, Ishikawa T and Sasaki M. 2012. %20Penyembuhan%20Luka. Diakses 12
Pathogenic Factors of P. gingivalis and the Desember 2015.
Host Defense Mechanisms. ISBN 978-953- 40. Matsunami T, Sato Y, Yuki H, Satomi A,
307-924-0. P. 17-1. Haruka K, Takuya S, Masayoshi Y. 2011.
32. Hussain M, Stover Cordula M, and Dupont Original Article Enhancement of reactive
A. 2015. P.Gingivalis in periondontal oxygen species and induction of
disease and atherosclerosis–sense of action apoptosis in streptozotocin-induced diabetic
for antimicrobial peptides and complement. rats under
Mini Review Aticle. 2015 February; 6(45): hyperbaric oxygen exposure. Int J Clin Exp
5-1. Pathol, 4(3) : 266-255. Available from
33. Bordbar S, Anwar F, and Saari N. High- http://www.ijcep.com/IJCEP1101009.
Value Components and Bioactives from Diakses 14 Januari 2016.

88