Banun Kusumawardani
Fakultas Kedokteran Gigi Universitas Jember

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Placental Trophoblast Responses to Porphyromonas gingivalis Mediated by Toll-like Receptor-2 and -4 Kusumawardani, Banun; Soesatyo, Marsetyawan HNE; Dasuki, Djaswadi; Asmara, Widya
Journal of Dentistry Indonesia Vol 20, No 2 (2013): August
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (935.461 KB) | DOI: 10.14693/jdi.v20i2.150

Abstract

Trophoblast participates in preventing allorecognition and controlling pathogens that compromise fetal wellbeing. Toll-like receptors recognize conserved sequences on the pathogens surface and trigger effector cell functions. Porphyromonas gingivalis is thought to spread to the umbilical cord and cause fetal growth restriction. Objective: To characterize expression and function of TLR-2 and TLR-4 in trophoblast cells from Porphyromonas gingivalisinfected pregnant rats. Methods: Live Porphyromonas gingivalis were challenged into the maxillary first molar subgingival sulcus of female rats before and/or during pregnancy and sacrified on gestational day (GD) 14 and 20. Porphyromonas gingivalis was detected by API-ZYM system in the maternal blood of the retro-orbital venous plexus and the umbilical cord. TLR-2 and TLR-4 expressions in trophoblast cells was detected by immunohistochemistry. Results: Porphyromonas gingivalis was first detected in the maternal blood and finally spread to the umbilical cord. Syncytiotrophoblast, spongitrophoblast and trophoblastic giant cell in treated groups had significantly higher expression of TLR-2 and TLR-4 than control group (p<0.05). Conclusion: Syncytiotrophoblast, spongitrophoblast and trophoblastic giant cell are able to recognize Porphyromonas gingivalis through TLR-2 and TLR-4 expression. The ligation of TLR-2 and TLR-4 promoted cytokine production and induced trophoblast cell death. These findings strengthen links between periodontal disease and fetal growth restriction.DOI: 10.14693/jdi.v20i2.150
Gestational Day-Dependent Expression of Interleukin-10 and Tumor Necrosis Factor-alpha in Porphyromonas gingivalis-infected Pregnant Rats Kusumawardani, Banun; Soesatyo, Marsetyawan HNE; Dasuki, Djaswadi; Asmara, Widya
Journal of Dentistry Indonesia Vol 20, No 3 (2013): December
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (926.193 KB) | DOI: 10.14693/jdi.v20i3.199

Abstract

Fetal growth restriction remains a major cause of neonatal morbidity and mortality. Porphyromonas gingivaliscan induce placental inflammatory response resulting in fetal growth restriction. Objective: This study aimed to evaluate the potential utility of the pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine IL-10 in rat placental tissues to understand whether these events were causally related. Methods: Female rats were infected with live-Porphyromonas gingivalis at concentration of 2x109 cells/ml into subgingival sulcus area of the maxillary first molar before and/or during pregnancy. They were sacrificed on gestational day (GD)-14 and GD20. The expression of TNF-α and IL-10 in macrophages and trophoblast cells were detected by immunohistochemistry. Results: A higher expression of TNF-α was found in spongiotrophoblast of the Pg-BD group on GD14 (6.30±1.16), and in trophoblastic giant cells of Pg-D group on GD20 (5.50±1.35). Furthermore, a higher expression of IL-10 was found in trophoblastic giant cells of the Pg-BD group on GD14 (4.50±1.51) and in syncytiotrophoblasts of Pg-BD group on GD20 (8.70±2.67). Conclusion: The expression of TNF-α on GD14 and GD20 were accompanied by increased expression of IL-10. The placental pathologic conditions induced by Porphyromonas gingivalis can be inhibited by elevated expression of IL-10 in macrophages and trophoblast cells.DOI: 10.14693/jdi.v20i3.199
Regulation of caspase-3 expression to maintain fetal growth in Porphyromonas gingivalis-infected pregnant rats Kusumawardani, Banun; Arina, Yuliana MD; Purwandhono, Azham
Journal of Dentomaxillofacial Science Vol 1, No 1 (2016)
Publisher : DiscoverSys Inc

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15562/jdmfs.v1i1.16

Abstract

Periodontal disease has been involved in a variety of systemic disorders and suspected as a potential risk factor for fetal growth restriction. Periodontal pathogenic bacteria may actively regulate embryonic development, implantation and placental trophoblast cell invasion. This study aimed to analyze the role of TNF-α, IL-10 and caspase-3 to maintain fetal growth in Porphyromonasgingivalis-infected pregnant rats. Female rats were infected with live-Porphyromonas gingivalis at concentration of 2x109 cells/ml into subgingival sulcus area of the maxillary first molar before and during pregnancy. They were sacrificed on gestational day (GD)-14 and GD20. The weight and length of placentas and fetuses were evaluated. The expression of TNF-α, IL-10 and caspase-3 in macrophages and trophoblast cells were detected by immunohistochemistry. On GD14, TNF-α (R2=0.416;P=0.000) and IL-10 (R2=0.187;P=0.012) had an important role to increase expression of caspase-3 in the placenta, but only TNF-α (R2=0.393;P=0.000 ) was able to increase the expression of caspase-3 on GD20. TNF-α and caspase-3 also had an important role (P<0.000) to decrease fetal weight, fetal length and placental weight on GD14 and GD20, but it was not the case with IL-10 (P>0.000). The increasing expressions of TNF-α and IL-10 did not only enhance immune protection, but also maintained the trophoblast cells survival by regulating expression of caspase-3. Porphyromonas gingivalis infection in maternal periodontal tissue can lead to decrease in placental weight, fetal weight and fetal length which mediated by increasing expression of TNF-α, IL-10 and caspase-3 in the placenta. 
Ekspresi Protein p53 pada Kultur Sel Fibroblas Gingiva yang Dipajan Lipopolisakarida Bakteri Gram-Negatif Kusumawardani, Banun; Utoro, Totok; Supartinah S, Al
Journal of Dentistry Indonesia Vol 12, No 3 (2005): December
Publisher : Faculty of Dentistry, University of Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1793.642 KB) | DOI: 10.14693/jdi.v12i3.878

Abstract

Bacterial lippolysaccharide (LPS) is implicated in the etiology of inflammatory periodontal disease. Aside from immunopathologic reactions which may be involved in the pathogenesis of the disease, the possibility exist that direct cytotoxic effect on cultured human gingival fibroblasts may be equally destructive. The expression of p53 protein can be one of markers to examine the state of impaired DNA. The purpose of this study was to investigate the effect of LPS toward expression of p53 protein on cultured human gingival fibroblasts. Cultured human gingival fibroblasts were exposed to LPS in concentrations of 50 and 200 µg/ml and untreated medium for a period of 24 and 48 hours. Cells were harvested and prepared for immunohistochemical evaluation. After exposure for 24 hours, the fraction of p53-positive cells was 81.7% in case of 50 µg/ml LPS, and 88.8% in case of 200 µg/ml LPS. After exposure for 48 hours, the fraction of p53-positive cells was 32.2% in case of 50 µg/ml LPS, and 21.1% in case of 200 µg/ml LPS. None of untreated group showed p53-positive cells. Up-regulation of p53 protein during the initial logarithmic phase of growth may be a consequence of on-going DNA damage.
PENGARUH PENYAKIT PERIODONTAL SELAMA MASA KEHAMILAN TERHADAP JUMLAH TOTAL LEUKOSIT DAN HITUNG JENIS LEUKOSIT Nuarita, Rusliyana; Praharani, Depi; Kusumawardani, Banun
STOMATOGNATIC- Jurnal Kedokteran Gigi Vol 9, No 3 (2012)
Publisher : Fakultas Kedokteran Gigi Universitas Jember

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Abstract

The changes of hormonal during pregnancy can increase the hormones estrogen and progesterone. Increased estrogen and progesterone can cause excessive response in periodontal tissues so easily happen inflammation of the periodontal tissues. The most important aspects of the inflammation is the leukocyte reaction. The purpose of this study was to determine the effect of periodontal disease during pregnancy to the total number of leukocytes and leukocyte counts. Research procedures performed each trimester which includes examining periodontal index (PI) to assess the severity of periodontal disease and total leukocyte counts and leukocyte counts were taken from venous blood. The data obtained were analyzed with linear regression test. Statistical analysis showed that the higher the score PI will increase the total number of leukocytes. The conclusion of this study is periodontal disease during pregnancy may increase the total number of leukocytes and leukocyte counts.
Efek Stresor Rasa Sakit Renjatan Listrik terhadap Limfosit dan Makrofag pada Gingiva Tikus Sprague Dawley (The Effect of Electrical Shock Stressor on Lymphocytes and Macrophages in Gingival Tissue of Sprague Dawley Rats) Kantasa, Vananda Duanta; Kusumawardani, Banun; ., Herniyati
Pustaka Kesehatan Vol 4, No 1 (2016)
Publisher : Pustaka Kesehatan

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Abstract

Background: Stress has negative impact for oral health and can cause periodontal disease, because the secretion of cortisol increase when stress occur. The excessive secretion of cortisol assumed to decrease the number of lymphocytes and macrophages in gingiva which play a role on immunity of oral health. Objective: To determine the effect of electrical shock stressor on the number of lymphocytes and macrophages in gingival tissue of Sprague Dawley rats. Methods: Sprague Dawley rats were given electrical shock for 7 days (S1), 14 days (S2), and 28 days (S3). Lymphocytes and macrophages observed on histological preparation in interdental gingiva between first molar and second molar used binocular microscope, 1000x magnification. Result: The results showed that the number of lymphocytes in S1 group (3,67±0,816), S2 (3,83±0,752), and S3 (3,17±0,527), also macrophages in S1 group (0,83±0,752), S2 (1,00±0,894), and S3 (0,67±0,816) were lower than the number of lymphocytes (0,67±0,816) and macrophages (2,00±0,632) in control group. The number of lymphocytes (p = 0,002) and macrophages (p = 0,033 ) had significant difference between groups. Conclusion: The electrical shock stressor decreased the number of lymphocytes and macrophages in gingival tissue of Sprague Dawley rats that could cause the oral cavity risk to periodontal infection.   Keywords: electrical shock stressor, gingiva, lymphocytes, macrophages, stress
Maternal endotoxin-induced fetal growth restriction in rats: Fetal responses in toll-like receptor Kusumawardani, Banun; Soesatyo, Marsetyawan HNE.; Dasuki, Djaswadi; Asmara, Widya
Dental Journal (Majalah Kedokteran Gigi) Vol 45, No 3 (2012): (September 2012)
Publisher : Faculty of Dental Medicine, Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (639.284 KB) | DOI: 10.20473/j.djmkg.v45.i3.p144-149

Abstract

Background: Porphyromonas gingivalis as a major etiology of periodontal disease can produce virulence factor, lipopolysaccharide/LPS, which is expected to play a role in the intrauterine fetal growth. Trophoblast at the maternal-fetal interface actively participates in response to infection through the expression of a family of natural immune receptors, toll-like receptor (TLR). Purpose: the aims of study were to identify endotoxin concentration in maternal blood serum of Porphyromonas gingivalis-infected pregnant rats, to characterize the TLR-4 expression in trophoblast cells, and to determine its effect on fetal growth. Methods: Female rats were infected with live-Porphyromonas gingivalis at concentration of 2 x 109 cells/ml into subgingival sulcus area of the maxillary first molar before and/or during pregnancy. They were sacrified on 14th and 20th gestational day. Fetuses were evaluated for weight and length. Endotoxin was detected by limulus amebocyte lysate assay in the maternal blood serum. The TLR-4 expression in trophoblast cells was detected by immunohistochemistry. <span style="font-family: Tribune-Bold; font-size:
PENYAKIT PERIODONTAL DAN KOMPLIKASI KEHAMILAN Kusumawardani, Banun
STOMATOGNATIC- Jurnal Kedokteran Gigi Vol 10, No 3 (2013)
Publisher : Fakultas Kedokteran Gigi Universitas Jember

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Abstract

Periodontal disease affects oral health in the mast people. Some researches have found and review the relationship between periodontal disease and systemic disease, such as cardiovascular disease, diabetes, stroke, and pregnancy complication. However, there is detailed explanation about the mechanism that analysis how the relationship between periodontal disease and pregnancy complication. The objective of this study was to review biological mechanism that correlated periodontal disease and pregnancy complication. Periodontal disease was able to cause alteration abnormal systemic immune which underlies pregnancy complication. Periodontal pathogen bacteria were directly colonized in placenta, and they caused local inflammation respond that caused pre-maturity and other pregnancy complications.
Daya Hambat Ekstrak Daun Tembakau terhadap Pertumbuhan Mikroba Rongga Mulut Putri, Riria Hendarto; Barid, Izzata; Kusumawardani, Banun
STOMATOGNATIC- Jurnal Kedokteran Gigi Vol 11, No 2 (2014)
Publisher : Fakultas Kedokteran Gigi Universitas Jember

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Abstract

Oral cavity diseases can be caused by the oral microbes  such as Streptococcus mutans, Porphyromonas gingivalis and Candida albicans. The oral microbes growth can be inhibited by chlorhexidine gluconate mouthwash, but its long term use have adverse effects, therefore it is important to do a further research to find effective antibacterial and antifungal as alternative substance mouthwash. Flavonoid compounds in tobacco leaf have potential as an antibacterial and antifungal. The objective was to analyze the inhibition effect of tobacco leaf extract to the growth of Streptococcus mutans, Porphyromonas Gingivalis and Candida albicans. This research used 20%, 40%, 60%, 80% and 100% ethanol extract of tobaco leaf,  1% DMSO as negative control and 0.2% chlorhexidine gluconate as positive control. Antibacterial and antifungal activities were tested by diffusion disc method with four repetitions. The observation was conducted by measuring the diameter of inhibition zone after incubating for 24 and 48 hours. The largest inhibition zone of Streptococcus mutans growth were producted by ethanol extract of tobacco leaf with 80% concentration, the largest inhibition zone on Porphyromonas gingivalis and Candida albicans growth were produced by ethanol extract of tobaco leaf with 100% concentration. Diameter of inhibition zone ethanol extract of tobacco leaf was equal with the diameter of inhibition zone of 0.2 % chlorhexidine gluconate. The conclusion described that ethanol extract of tobacco leaf can inhibit the growth of Streptococcus mutans, Porphyromonas gingivalis and Candida albicans.
THE LEVELS OF TNF-Α IN GINGIVAL CREVICULAR FLUID (GCF) OF OSING TRIBE WOMEN WITH OCCLUSAL DISHARMONY Suhartini, Suhartini; Kusumawardani, Banun
UNEJ e-Proceeding 2016: Proceeding of 1st International Conference on Medicine and Health Sciences (ICMHS)
Publisher : UNEJ e-Proceeding

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Abstract

The Osing tribal region is demographically adjacentto the island of Java, Madura and Bali. The proximityof this demographic regions affects multiple systemsof their organization, culture, and arts. The Osing’sfamilial and societal patterns are similar to the otherJavanese tribes, including housing, food, lifestyle andhealth. Osing tribe is frequently compared toBalinese culture, such as traditional clothing,wedding dresses, and others [1]. One of the culturesassociated with the teeth is Pangur or asa i.e.reducing/flattening the occlusal surfaces of the teethespecially for Osing bride women. In addition, thereis also a tradition of Osing women called Nginang orbetel chewing [2]. This situation may lead to animbalance of dental occlusion and disruption of thenormal flora in the oral cavity alleging effect on oralhealthDental and oral health can be achieved if there is arelationship of harmony and balance between theorgans and tissues in the oral cavity. This is becausethe oral cavity is an integral stomatognatic complexsystem.Stomatognatic system is unitary organ that hasmutual-related functions. These organs include jawbones, temporo mandibular joint (TMJ), teeth, andother supporting structures e.g. masticationmuscles, facial muscles, head and neck. Thisstomatognatic system plays an active role in theprocess of mastication. In normal conditions, itoccurs a harmonious relationship of all componentsof the masticatory system i.e. teeth, muscles, TMJ,lips, cheeks, palate, tongue and salivary secretion[3]. The conditions of normal tooth anatomy andwell-structured on curved teeth will put bothcondyle joints at the center of the articular disc. Thissituation will create an effective mastication process[4].Abnormal condition of tooth structure will causemalocclusion [3]. Occlusion is an interocclusalrelation (intercups) between the teeth of each jaw inaccordance with the position and movement of themandible [5]. Occlusion pathological state is knownas occlusal disharmony. Occlusal disharmony can becaused by several factors such as the loss of teeth,caries, attrition, tooth anomalies, bruxism, and thehabit of chewing on one side [6].Occlusal disharmony is suspected to causeinflammation in periodontal tissues. Periodontaltissue disease is one of the dental and oral diseasethat most commonly occurs. Periodontal diseasescan be caused by bacteria or mechanical theoverloads. The overloads can be caused by theocclusal disharmony. Thus, occlusal disharmony issuspected to cause inflammation in the periodontaltissues. Abnormal oocclusion condition in thisocclusion disharmony leads into occlusal traumawhich will increase the load received by theperiodontal tissue [7]. If this condition is persistent,it will cause periodontal tissue inflammation.Inflammation undergoing locally to the periodontaltissues will induce the immune system to increasethe activity of the inflammatory cytokine secretion,one of which is TNF-α (tumor necrosis factor-alpha).TNF-α will play a role in activating osteoclasts, whichcan result in the destruction of the alveolar bone [8].This inflammatory cytokines can be found in gingivalcrevicular fluid (GCF), which are inflammatoryexudates collected in the gingival sulcus [9].TNF-α is a cytokine that plays a role in theoccurrence of periodontal disease through itsactivities, which triggers proliferation, differentiationand activity of osteoclasts resulting in boneresorption, and inducing production of proteinase inthe cells of the mesenchyme that is responsible inbond tissue destruction. In patients withperiodontitis it occurs increased levels of TNF-α,which will be decreased after periodontal treatment.Based on the description above, it is necessary toknow the levels of TNF-α in GCF Osing women as anindicator of periodontal tissue inflammation due toocclusion disharmony. It is important for preventionand treatment of abnormalities of occlusiondisharmony that will improve oral health and qualityof life, especially in the Osing community in Kemirenvillage, Glagah District, Banyuwangi Regency.