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The role of Hsp0, CD-8 and IFN-γ in immunopathobiogenesis of periapical granuloma in dental caries Risya Cilmiaty; Mandojo Rukmo
Dental Journal (Majalah Kedokteran Gigi) Vol. 47 No. 1 (2014): March 2014
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Background: The incidence of dental caries with periapical granulomas in Indonesia is quite high. However, the mechanism of the formation of periapical granulomas in dental caries caused by bacterial infection in immunopathobiogenesis cannot be explained completely. Thus, this explanation is necessary in order to be used as a basis for diagnostic, preventive and therapeutic measures. Purpose: This research was aimed to determine the role of Hsp60, CD-8 and IFN-γ in immunopatobiogenesis of periapical granuloma in dental caries. Methods: This research was an analytic observational study with cross sectional approach. Samples of this research were 36 teeth of patients with dental caries, consisting of 18 caries teeth with periapical granulomas and 18 caries teeth without periapical granulomas. The variables observed in this research were Hsp60, CD-8 and IFN-γ. Measurements were conducted by using immunohistochemical methods on periapical tissue. Results: The mean of Hsp60, CD-8 and IFN-γ in granuloma group was significantly higher than those in non granuloma group (p<0.05). The positive role of IFN-γ on the incidence of granulomas appeared to be more prominent. Conclusion: The study suggested that in immunopathobiogenesis of periapical granuloma in dental caries, Hsp60, CD-8 and IFN-γ played important roles, but the role of IFN-γ was found to be more prominent.Latar belakang: Angka kejadian gigi karies dengan granuloma periapikal di Indonesia cukup tinggi, Namun mekanisme terbentuknya granuloma periapikal pada gigi karies yang disebabkan oleh infeksi bakteri secara imunopatobiogenesis belum dapat dijelaskan secara tuntas. Adanya penjelasan ini diperlukan agar dapat digunakan sebagai dasar pengembangan diagnosis, langkah preventif dan terapinya. Tujuan: Penelitian ini bertujuan untuk mengetahui peran Hsp60, CD-8 dan IFN-γ dalam immunopatobiogenesis dari granuloma periapikal karies gigi. Metode: Penelitian ini merupakan penelitian observasional analitik dengan pendekatan cross sectional. Sampel dari penelitian ini adalah 36 gigi pasien dengan karies, yang terdiri dari 18 karies gigi dengan granuloma periapikal dan 18 karies gigi tanpa granuloma periapikal. Variabel yang diamati adalah Hsp60, CD-8 dan IFN-γ. Pengukuran dilakukan dengan menggunakan metode imunohistokimia pada jaringan periapikal. Hasil: Rerata Hsp60, CD-8 dan IFN-γ pada kelompok granuloma secara signifikan lebih tinggi dibanding kelompok non granuloma (p <0,05). Peran positif dari IFN-γ terhadap kejadian granuloma tampaknya lebih menonjol. Simpulan: Studi ini menunjukkan bahwa dalam imunopatobiogenesis dari granuloma periapikal karies gigi, Hsp60, CD-8 dan IFN-γ memainkan peran penting, tetapi peran IFN-γ ditemukan lebih menonjol.

Prevotella intermedia and Porphyromonas gingivalis in dental caries with periapical granuloma Risya Cilmiaty; Afiono Agung Prasetyo; Khilyat Ulin Nur Zaini; Mandojo Rukmo; Suhartono Taat Putra; Widya Asmara
Dental Journal (Majalah Kedokteran Gigi) Vol. 46 No. 4 (2013): December 2013
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Background: Dental caries with necrotic pulp is a multifactorial disease that attacks enamel involving tooth pulp. The anaerobic bacteria infection in the pulp chamber could induce the formation of periapical granuloma. However, the presence of the most frequently anaerobic bacteria identified in apical periodontitis, Porphyromonas gingivalis and Prevotella intermedia, in periapical granuloma have not been confirmed. Purpose: The aims of study were to determine the presence of Porphyromonas gingivalis and Prevotella intermedia in dental caries with necrotic pulp and to determine its relation to periapical granuloma. Methods: Thirty-six patients of dental caries with necrotic pulp in Dr. Moewardi General Hospital in Surakarta, Indonesia were involved and classified into two groups, the group of patients with periapical granuloma and the group of patients without periapical granuloma. The caries tooth was extracted, and the chronic periapical tissue was swabbed and cultured on blood agar medium in anaerobic condition. The bacterial DNA was extracted from the positive cultures and subjected for Polymerase Chain Reaction (PCR). Results: Periapical granuloma was more likely found in women (OR 5.5, 95% CI=1.277-23.693; RR 2.5, 95% CI= 1.025-6.100). Black colonies bacteria were associated with periapical granuloma (OR 2.2, 95% CI=0.517-9.594; RR 1.5, 95% CI=0.655-3.623). Porphyromonas gingivalis and Prevotella intermedia were detected in group with or without periapical granuloma, however, only Prevotella intermedia was associated with periapical granuloma (OR 1.6, 95% CI=0.418-5.903; RR 1.3, 95% CI=0.653-2.393). Conclusion: The presence of Porphyromonas gingivalis and Prevotella intermedia in periapical granuloma were confirmed, however, only Prevotella intermedia were associated with periapical granuloma.Latar belakang: Karies gigi dengan pulpa nekrosis adalah penyakit multifaktorial yang menyerang enamel hingga ruang pulpa gigi. Infeksi bakteri anaerob di pulpa nekrosis dapat menginduksi pembentukan granuloma periapikal. Namun, keberadaan bakteri anaerob yang paling banyak ditemukan di periodontitis apikal, Porphyromonas gingivalis dan Prevotella intermedia, di granuloma periapikal masih perlu diteliti . Tujuan: Penelitian ini bertujuan meneliti keberadaan bakteri Porphyromonas gingivalis dan Prevotella intermedia, di karies gigi dengan pulpa nekrosis dan menganalisis kaitannya dengan granuloma periapikal. Metode: Tiga puluh enam pasien karies gigi dengan pulpa nekrosis di Rumah Sakit Umum Dr. Moewardi di Surakarta Indonesia dilibatkan dan diklasifikasikan dalam dua kelompok yaitu kelompok pasien dengan granuloma periapikal dan kelompok pasien tanpa granuloma periapikal. Gigi karies diekstraksi dan jaringan periapikal kronis diusap dan dikultur di media agar darah dalam kondisi anaerob. DNA bakteri diekstrak dari kultur yang positif dan dilakukan pemeriksaan dengan Polymerase Chain Reaction (PCR). Hasil: Granuloma periapikal lebih banyak ditemukan pada wanita (OR 5,5, 95% CI= 1,277-23,693; RR 2,5, 95% CI= 1,025-6,100). Koloni bakteri berwarna hitam diasosiasikan dengan granuloma periapikal (OR 2,2, 95% CI= 0,517-9,594; RR 1,5, 95% CI= 0,655-3,623). Porphyromonas gingivalis dan Prevotella intermedia terdeteksi di jaringan karies dental dengan atau tanpa granuloma periapikal, namun hanya Prevotella intermedia yang diasosiasikan dengan granuloma periapikal (OR 1,6, 95% CI= 0,418-5,903; RR 1,3, 95% CI= 0,653-2,393). Simpulan: Porphyromonas gingivalis dan Prevotella intermedia ditemukan di granuloma periapikal, namun hanya Prevotella intermedia yang terkait langsung dengan granuloma periapikal.

The effect of nickel as a nickel chromium restoration corrosion product on gingival fibroblast through analysis of BCl-2 FX Ady Soesetijo; Mandojo Rukmo
Dental Journal (Majalah Kedokteran Gigi) Vol. 45 No. 4 (2012): December 2012
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Background: Restoration of NiCr may undergo corrosion process in artificial saliva. Corrosion product is soluble Ni substances in salivary electrolytes. Ni2+ may freely enter the cells through passive transport DMT-1. Ni2+ in the cell causes initiation of the ROS formation,which subsequently can conduct the redoxs reactions leading to DNA damage. The damage DNA affects the genetic expression, especially bcl-2, and even triggers apoptosis. Purpose: The aim of this study was to reveal the mechanism of Ni toxicity as a corrosion product of NiCr restoration on gingival fibroblasts through expression analysis of Bcl-2. Methods: Cells with a density of 105 planted on each coverslip in 72 wells to the treatment group and 24 wells to the control group (24 hours incubation). In the treatment groups, each well exposed with 20 μL artificial saliva containing Ni concentration results immerse each restoration, whereas the control group was exposed to 20 μL artificial saliva (incubation 1, 3, and 7 days). The data collected were subsequently analyzed using two-ways ANOVA, followed by one-way ANOVA. Comparing between experimental groups after one-way ANOVA was conducted using Fisher’s LSD. Whereas, the calculation and documentation of Bcl-2 expression was performed camera of Olympus Microscope BX-50 Japan. Results: Statistical analysis of two-ways ANOVA showed the presence of interaction between the increasing Ni concentration and exposure duration on the expression of Bcl-2 gingival fibroblasts (p=0.021<a=0.05). Conclusion: It can be concluded that the higher concentration of Ni exposed to gingival fibroblasts, and the longer incubation time will decreased Bcl-2 expression.Latar belakang: Restorasi NiCr dapat mengalami proses korosi di dalam saliva artificial. Produk korosi yang dihasilkan adalah substansi Ni yang terlarut di dalam elektrolit saliva. Ni2+ bebas dapat memasuki sel (fibroblas gingiva) melalui transport pasif DMT-1. Ni2+ di dalam sel menginisiasi pembentukan ROS, yang selanjutnya dapat menjalankan reaksi redoks dan dapat menimbulkan kerusakan DNA. DNA yang rusak mempengaruhi ekspresi genetik, terutama Bcl-2 dan bahkan dapat memicu apoptosis. Tujuan: Tujuan penelitian ini adalah untuk mengungkap mekanisme toksisitas Ni sebagai suatu produk korosi restorasi NiCr pada fibroblas gingiva melalui analisis ekspresi Bcl-2. Metode: Sel dengan kepadatan 105 ditanam pada tiap-tiap coverslip di dalam 72 well untuk kelompok perlakuan dan ditanam pada tiap-tiap coverslip di dalam 24 well untuk kelompok kontrol (inkubasi selama 24 jam). Pada kelompok perlakuan, masing-masing well dipapar dengan 20 μL saliva artificial yang mengandung konsentrasi Ni hasil perendaman tiap-tiap restorasi, sedangkan pada kelompok kontrol dipapar 20 μL saliva artificial (inkubasi 1,3 dan 7 hari). Data yang terkumpul selanjutnya dianalisis menggunakan ANOVA dua arah dan ANOVA satu arah. Perbandingan antar kelompok eksperimental setelah analisis ANOVA satu arah menggunakan uji Fisher’s LSD. Penghitungan jumlah sel yang mengekspresikan Bcl-2, kemudian dilanjutkan dengan dokumentasi dengan menggunakan kamera Olympus Microscope BX-50 Japan. Hasil: Analisis statistik ANOVA dua arah menunjukkan adanya interaksi antara peningkatan konsentrasi Ni dan lama paparan terhadap ekspresi Bcl-2 fibroblas gingiva > (p = 0,021 < á = 0,05). Kesimpulan: Dapat disimpulkan bahwa semakin tinggi konsentrasi paparan Ni pada fibroblas gingiva dan semakin lama masa inkubasi, maka akan menurunkan ekspresi Bcl-2.

The expression of nuclear factor of activated T cell c1 and receptor activator of nuclear factor kappa β induced by Enterococcus faecalis in osteoclastogenesis (laboratory experiment on Wistar rats) Nirawati Pribadi; Rosita Rahmawati; Mandojo Rukmo; Adelina Kristanti Tandadjaja; Hendy Jaya Kurniawan; Ratna Puspita Hadi
Dental Journal (Majalah Kedokteran Gigi) Vol. 52 No. 4 (2019): December 2019
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/j.djmkg.v52.i4.p172-176

Background: Enterococcus faecalis (E. faecalis) is the most common bacteria species in persistent endodontic infection of teeth undergoing root canal treatment at a prevalence of 38%. The virulence factor of this bacterium is Lipoteichoic acid (LTA) which can be recognized by Toll-like receptors-4 (TLR-4) that produce a stimulus and provoke an immune response. Inflammation results in bone defects that feature multiple cytokines and interactions between different cell types. Bone loss within a periapical tooth is characterized by osteoclast formation (osteoclastogenesis) in the bone. Purpose: This study aimed to determine the expression of nuclear factor of activated T cell c1 (NFATc1) and receptor activator of nuclear factor kappa β (RANK) which played a role in osteoclastogenesis at different time intervals. Methods: 36 upper molar teeth of the research subjects were induced with 106 CFU Enterococcus faecalis and subsequently observed for 7 and 21 days with the NFATc1 and RANK being counted microscopically at 1000X magnification across 20 viewing fields. Thereafter, the data was examined and analyzed by means of an independent T test using SPSS. Results: NFATc1 and RANK expression were higher in the group including E. faecalis on days 7 and 21 than in the control group. There were significant differences between the treatment group and control group with regard to NFATc1 and RANK expression (p<0.05). Conclusion: The study showed that the expression of NFATc1 and RANK, which plays a role in osteoclastogenesis, was higher in periapical bone defects in Wistar rats induced by E. faecalis than those which were not induced.

Contrasting efficacy of cocoa POD HUSK extract and 8% propolis extract in maintaining of root canal wall cleanliness Tamara Yuanita; Uli Sasi Andari; Mandojo Rukmo; S. Sukaton; Deavita Dinari
Dental Journal (Majalah Kedokteran Gigi) Vol. 52 No. 3 (2019): September 2019
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/j.djmkg.v52.i3.p159-162

Background: The existence of the smear layer, which can be produced during root canal instrumentation, may compromise the bond between filling material and the root canal walls. Therefore, the use of an effective root canal irrigation solution, a commonly employed form of which is sodium hypochloride (NaOCl), is important. Sodium hypochloride has several positive properties including effectiveness as a disinfectant agent and its ability to promote tissue-dissolution, although it is ineffective at cleaning the smear layer. There have been numerous recent studies of the application of phytomedicines in endodontics due to their advantages such as minimum toxicity and cost effectiveness. The saponin contained in both the propolis and cocoa pod husk acts as a surfactant that may lower surface tension and dissolve debris containing organic and anorganic materials. Purpose: The study aimed to provide evidence of the differences between root canal wall cleanliness when treated with 8% propolis extract and different concentrations of cocoa pod husk extract. Methods: 25 extracted teeth with single straight root canals were randomly divided into five categories (n=5). Sample preparation was performed using a rotary file and irrigated with different solutions. The first group was administered 2.5% NaOCl, the second group 8% propolis, the third group 3.12% cocoa pod husk extract, the fourth group 6.25% cocoa pod husk extract, and the fifth group 12.50% cocoa pod husk extract. The samples were then dissected into two sections at the apical third and their cleanliness scores subjected to a Mann-Whitney test with a significance level of p=0.05. Results: A significant difference was identified between all groups (p<0.05) and on the median control test, the highest value of 1.6 was recorded by the 6.25% cocoa pod husk extract, compared to the other four groups Conclusion: Cocoa pod husk extract demonstrates greater efficacy at cleaning root canal walls compared to 8% propolis extract.

UJI SITOTOKSISITAS EKSTRAK DAUN MIMBA (Azadirachta indica) TERHADAP SEL FIBROBLAS BHK 21 Qurni Restiani; Mandojo Rukmo; Devi Eka Juniarti
Conservative Dentistry Journal Vol. 7 No. 1 (2017): January - June
Publisher : Universitas Airlangga

Background: The leaves of neem (Azadirachta indicia) is one of herbal medicine that recommended as an alternative material of root canal irrigants. The active ingredients of neem leaves such as alkaloids, tannins, saponins and flavonoids has been proven to have antibacterial activity against E. faecalis. The ideal properties of an alternative material of root canal irrigants is not only have antibacterial activity but also is not toxic to the tissues, but the toxicity of neem leaves remains unclear until now. Objective: The aims of this study to determine the toxicity effect of neem leaves extract in specific concentration. Methods: This research was an experimental laboratory in vitro study of baby hamster kidney fibroblast (BHK-21). The neem leaves extract was made by maceration method using ethanol 96% and certain dilution performed to obtain various concentrations. Cytotoxicity test was conducted by MTT assay and the optical density was measured using ELISA reader at wavelength of 620 nm. Then, the optical density values were calculated using the formula for determining the number of survival fibroblasts after tested. Results: The percentage of survival fibroblast at concentration of 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95, and 100% in sequence were 71.3%, 64, 2%, 62%, 60.2%, 40.1%, 36.3%, 36.7%, 29%, 28.4%, 27.5%, and 25.6% . Conclusion: The extract of neem leaves (Azadirachta indica) has cytotoxic effects at concentration of 70% up to 100%.

EKSPRESI TNF-α DAN CALCINEURIN PADA ASIMTOMATIS APIKAL PERIODONTITIS AKIBAT INDUKSI Enterococcus faecalis Randy Carlos Sietho; Mandojo Rukmo; Edhie Arif Prasetyo; Tamara Yuanita
Conservative Dentistry Journal Vol. 7 No. 2 (2017): July - December
Publisher : Universitas Airlangga

Background. Gram positive bacteria strain are the major cause of endodontic failure as asymptomatic apical periodontitis. One of the dominant group of bacteria is Enterococcus faecalis that still persistent in root canal system post endodontic therapy procedures. This bacteria has lipoteichoic acid on its membrane that can cause induction of cytokines expression such as Tumor Necrosing Factor-α (TNF-α) and Calcineurin Purpose. This experiment to demonstrated asymptomatic apical periodontitis that induced with Enteroccus faecalis produce raising amount of TNF-α and Calcineurin expression cells in pericapical tissue of wistar rat. Method. The upper right molar teeth of the rat was drilled until perforation then exposed by BHIB 10µl (control positive group), E.faecalis 106 CFU in BHIB 10µl (experimental group) and without drilling (control negative group) then observed until 21th days and counting the amount of TNF-α and Calcineurin expression cells. Conclusion.The results show that asymtomatic apical periodontitis that was induced E.faecalis produce increasing amount of TNF-α and Calcineurin expression cells in periapical tissue wistar rat.

Antibakteri Ekstrak Kulit Buah Kakao (Theobroma cacao) 6,25% dan NaOCl 2,5% Terhadap Bakteri Streptococcus sanguinis Sepdhyo Wahyu Nugroho; Mandojo Rukmo; Edhie Arief Prasetyo; Tamara Yuanita
Conservative Dentistry Journal Vol. 9 No. 1 (2019): January - June
Publisher : Universitas Airlangga

Background: Streptococcus sanguinis is a gram-positive bacterium that infects and penetrates into dentinal tubules from a depth of 150 μm up to 792 μm. Chemicals, namely 2.5% NaOCl, are used to mitigate the growth of these bacteria via irrigation of the root canals, but 2.5% NaOCl still has disadvantages including irritating periradicular tissue, having an unpleasant odor, and being toxic. Due to these shortcomings, natural materials are expected to be used as alternatives. Cocoa peel extract has active tannin compounds, flavonoids, alkaloids, terpenoids, and saponins which have antibacterial attributes; a concentration of 6.25% is counted as the minimum bactericidal concentration (MBC) of Streptococcus sanguinis. Aim: To compare the difference of antibacterial power of 6.25% cocoa peel extract (Theobroma cacao) and 2.5% NaOCl against Streptococcus sanguinis. Method: This research is an in vitro experimental laboratory with post-test only control group design. The diffusion method was used on Streptococcus sanguinis planted in tubes containing BHIB, then cultured on petri dishes containing nutrient agar and divided into 3 parts, namely 6.25% cocoa peel extract, 2.5% NaOCl, and negative controls, then each Petri dishes were given a paper disc that had been dripped by 0.01 ml of each ingredient, then incubated in an incubator for 2x24 hours at 37°C; the diameter of the inhibitory zone formed was then observed using a caliper. Results: The average diameter of the inhibition zone formed on 6.25% cocoa peel extract was 19.2000 mm and 2.5% NaOCl was 17.2813 mm against Streptococcus sanguinis. Conclusion: The antibacterial power of 6.25% cocoa peel extract (Theobroma cacao) is higher than 2.5% NaOCl against Streptococcus sanguinis.

Antibiofilm activity of flavonoid mangosteen pericarp extract against porphyromonas gingivalis bacteria Aprillia Sonya Federika; Mandojo Rukmo; Setyabudi Setyabudi
Conservative Dentistry Journal Vol. 10 No. 1 (2020): January-June
Publisher : Universitas Airlangga

Background. Necrosis teeth need a procedure called the root canal treatment to eliminate microorganisms in root canal, so the inflammation process did not continue to the periapical.One of the microbes that exist in thenecrosis teethis Porphyromonas gingivalis, that capable of forming biofilm because it has the ability to communicate with bacteria signaling (quorum sensing). It is necessary to discover another antibiofilm agent as an irrigation material in root canal preparation deriving from nature. Flavonoidmangosteen pericarp extract can disrupt acyl-homoserine lactones (AHLs) thus quorum sensing bacteria will be disrupted and causing permanent attachment and the establishment of colonies on the surface of the teeth any barrier. Purpose. The aim of this study found out the antibiofilmactivity of flavonoid mangosteen pericarp extract against Porphyromonas gingivalis bacteria. Method. This studyhasa laboratory experimental with post-test only group design. Porphyromonas gingivalis ATCC 33277 was diluted according to Mc. Farland standard 106CFU/ml in Tryptic Soy Broth (TSB) medium and inserted into microtitterplate flexible U bottom. Porphyromonas gingivalis was incubatedfor 6x24 hour and checked with a simple staining to see the formation of biofilm. Flavonoid mangosteen pericarp extract was added in various concentrations and OD (optical density) readings done with a wavelength of 595 nm. Result. Flavonoid mangosteen pericarp extractshowed a decrease of OD value from control in all concentrations.Conclusion.Flavonoid mangosteen pericarp extract has antibiofilm activity against Porphyromonas gingivalis bacteria.

The effects of propylene glycol addition in the combination of calcium hydroxide and propolis on compressive and flexural strength Mandojo Rukmo; Nirawati Pribadi; Elisa Kusuma Wardani; Sukaton; Belinda Belinda Meilani Perdana Putri
Conservative Dentistry Journal Vol. 12 No. 1 (2022): January-June
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/cdj.v12i1.2022.32-37

Background: The mechanical properties of pulp-capping materials may affect their resistance to fracture during placement of a final restorative material or while supporting an overlying restoration over time when the mastication process is carried out. The combination of calcium hydroxide and propolis as a pulp capping material has the weakness on mechanical properties so it is necessary to add another material to improve its mechanical properties. Propylene glycol is used as a vehicle because it improves its handling property. Purpose: To explain how the compressive and flexural strength of the pulp capping material in the combination of calcium hydroxide, propolis, and propylene glycol compared without adding of propylene glycol. Methods: The study used 4 treatment groups with each group consisting of 7 replications. Control group is a combination of calcium hydroxide-propolis with a ratio of 1: 1.5, group 1 is a combination of calcium hydroxide-propolis-propylene glycol 20% with a ratio of 1:1.5:0.375, group 2 is a combination of calcium hydroxide-propolis-propylene glycol 30% with a ratio of 1:1.5:0.375, and group 3 is a combination of calcium hydroxide-propolis-propylene glycol 30% with a ratio of 1:1.5:0.375. Materials were mixed according to comparison and printed on a cylindrical mold with the size of 4 mm x 6 mm and a block mold with the size 65 mmx10 mmx6 mm. Then, the compressive and flexural strength was tested using an Autograph test instrument. Results: The statistical analysis was performed with ANOVA and Tukey HSD’s post-hoc test. There were statistically significant differences on compressive and flexural strength between groups (P<0.05). Conclusion: Addition of propylene glycol in the combination of calcium hydroxide and propolis have an influence of increasing of compressive and flexural strength, especially in the addition of 40% propylene glycol.

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