Heni Susilowati
Bagian Biologi Mulut, Fakultas Kedokteran Gigi, Universitas Gadjah Mada, Yogyakarta, Indonesia

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Ekspresi COX-2 dan Jumlah Neutrofil Fase Inflamasi pada Proses Penyembuhan Luka Setelah Pemberian Sistemik Ekstrak Etanolik Rosela (Hibiscus sabdariffa) (studi in vivo pada Tikus Wistar) Endah Kusumastuti; Juni Handajani; Heni Susilowati
Majalah Kedokteran Gigi Indonesia Vol 21, No 1 (2014): August
Publisher : Faculty of Dentistry, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (909.588 KB) | DOI: 10.22146/majkedgiind.8778

Abstract

Inflamasi merupakan respon alami tubuh terhadap adanya kerusakan jaringan. Salah satu medikamen untuk mengatasi inflamasi adalah antiinflamasi non steroid (AINS). Penggunaan AINS mempunyai beberapa efek samping dan dalam beberapa hal penggunaan tanaman obat dinilai lebih aman. Rosela merupakan salah satu tanaman obat yang mempunyai potensi sebagai antiinflamasi. Penelitian ini bertujuan untuk mengetahui efek pemberian sistemik ekstrak etanolik rosela terhadap ekspresi COX-2 dan jumlah neutrofil fase inflamasi pada proses penyembuhan luka. Bunga rosela didapatkan dari perkebunan di Dusun Bulusari Desa Pojok Kecamatan Tarokan Kabupaten Kediri Jawa Timur. Pembuatan ekstrak rosela dilakukan di LPPT unit I UGM Yogjakarta dengan cara perkolasi. Tikus putih galur Wistar sebanyak 36 ekor diberi perlukaan dengan punch biopsi ɵ 3 mm pada mukosa bukal. Subjek dibagi menjadi 3 kelompok, masing-masing kelompok 12 ekor tikus. Pembagian kelompok terdiri dari kontrol negatif (saline), kontrol positif (ibuprofen 20 mg/kg BB) dan perlakuan (ekstrak rosela 500 mg/kg BB). Pemberian minum sesuai kelompoknya sehari sekali selama 4 hari. Pada hari ke-1, ke-2, ke-3 dan ke-4 tikus dikorbankan lalu jaringan mukosa yang mengalami perlukaan dibuat preparat histologis. Pewarnaan Hematoksilin Eosin (HE) dilakukan untuk mengamati jumlah neutrofil. Ekspresi COX-2 diamati pada preparat dengan pewarnaan imunohistokimia menggunakan rabbit polyclonal antibody COX-2 (Lab Vision, USA). Jumlah neutrofil dan ekspresi COX-2 dihitung di bawah mikroskop cahaya lalu data dianalisi menggunakan ANAVA dan LSD. Hasil penelitian menunjukkan bahwa ekspresi COX-2 dan jumlah neutrofil lebih rendah pada kelompok perlakuan dibanding kontrol. Pengamatan klinis pada hari ke-4 juga tampak luka seluruh subjek telah menutup sempurna setelah pemberian minum rosela. Disimpulkan bahwa ekstrak etanolik rosela mempunyai kemampuan menghambat ekspresi COX-2 dan menurunkan jumlah neutrofil sehingga dapat digunakan sebagai bahan anti-inflamasi. Expression of COX-2 and The Number of Neutrophil in Inflammation stage of Wound Healing Process after Systemic Administration of Ethanolic Extract Rosela. Inflammation is an initial stage of body’s natural response to tissue damage.The use  empirically plants often used for traditional medicine because it is easily found in the community and fewer side effects. Flavanoid presence of roselle (Hibiscus sabdariffa) is thought to have anti inflammatory effects. This study aimed to know the effect of systemic administration of Roselle ethanolic extract toward COX-2 expression and neutrophils number in the inflammatory phase of wound healing processes. Roselle was obtained from plantations in Bulusari hamlet, Tarokan, Kediri, EastJava. Making roselle extract was performed in LPPT unit 1 UGM Yogyakarta by percolation ways. Wistar rats were given a total of 36 injuries with ɵ 3 mm punch biopsy of the buccal mucosa. Subjects were divided into three groups, each group of 12 rats. The division consists of the negative control group (saline), positive control (ibuprofen 20 mg/kg) and treatment (roselle extract 500 mg/kg). Giving drink suitable group once daily for four days. On day 1, the 2nd, 3rd and fourth rats were sacrificed, and mucosal tissue injury was made histological preparat. Hematoxylin eosin staining (HE) was performed to observe the number of neutrophils. COX-2 expression was found in preparations for immunohistochemical staining using rabbit polyclonal COX-2 antibody (Lab Vision, USA). The number of neutrophils and expression of COX-2 is calculated under a light microscope data were analyzed using Two-way ANOVA and LSD. The results showed that the expression of COX-2 and neutrophil number were least in the treatment group compared to the control. Clinical observation on day four also appears around the wound has completely closed the subject after administration of roselle drink. It was concluded that the ethanolic extract of roselle can inhibit COX-2 expression and decrease the number of neutrophils that can be used as an anti-inflammatory ingredient. 
Perubahan morfologi sel HeLa setelah paparan ekstrak etanolik Curcuma longa Suryani Hutomo; Heni Susilowati; Yanti Ivana Suryanto; Chandra Kurniawan
Majalah Kedokteran Gigi Indonesia Vol 2, No 1 (2016): April
Publisher : Faculty of Dentistry, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (990.092 KB) | DOI: 10.22146/majkedgiind.11248

Abstract

Cell morphological changes on HeLa cells after Curcuma longa etanolik extract exposure. Curcuma is mostly found in the areas with tropical and sub-tropical climate, and is one of original plants of Southeast Asia. In Indonesia, curcuma can be found in almost all regions and areas. Curcumin, which is curcuma’s main constituent, is a potent anti oxidant. Previous studies reported that curcuma longa extract may decrease the growth of cancer cells by interfering with cell proliferation, and by causing the cell apoptosis; however, the mechanism of apoptosis remains unclear. The purpose of this study was to investigate the effect of Curcuma longa extract on the morphological change of HeLa cells, indicating the cell damage. HeLa cells (5x10⁴ cells/well) were cultured in complete RPMI 1640 overnight before stimulation. Etanol extract of Curcuma longa (50 µg/ml, 100 µg/ml, 150 µg/ml) were added to the culture of HeLa cells and were incubated for 24 hours in antibiotic-free of culture medium. HeLa cells morphological analysis was performed under phase contrast microscope after haematoxilent eosin staining. Docsorubisin (0,5625 mg/ml) was used as positive control in this study. The results demonstrated that Curcuma longa extract caused cell morphological changes on HeLa cells indicated by cell shrinkage, lost contact with neighboring cells as the alteration of apoptotic cell death in most of cell population. The nuclei were dark as a result of their capability to absorb haematoxylene dye. Statistical analysis showed a significant difference between controls and treatment groups. It was then concluded that Curcuma longa extract induced cell damage on HeLa cells in a way of cell shrinkage.ABSTRAKKunyit (Curcuma longa) merupakan tanaman yang dapat tumbuh di daerah tropis dan sub tropis, serta merupakan tanaman asli Asia Tenggara. Di Indonesia, kunyit menyebar secara merata di seluruh daerah. Kurkumin yang merupakan unsur utama kunyit, merupakan antioksidan yang kuat. Beberapa penelitian juga menunjukkan bahwa kunyit mampu menghambat pertumbuhan beberapa tipe sel kanker. Mekanisme anti-kanker kurkumin adalah dengan menghambat proliferasi sel. Penelitian terdahulu melaporkan bahwa ekstrak Curcuma longa menginduksi apoptosis pada sel HeLa, tetapi mekanisme kematian sel tersebut belum jelas. Tujuan penelitian ini adalah untuk mengkaji pengaruh ekstrak Curcuma longa pada perubahan morfologi sel HeLa, dimana perubahan morfologi merupakan parameter kerusakan sel. Sel HeLa (5x104 sel/well) dikultur dalam RPMI 1640 semalam sebelum stimulasi. Ekstrak etanol kunyit (50 µg/ml, 100 µg/ml, 150 µg/ml) ditambahkan pada kultur HeLa dan diinkubasi selama 24 jam dalam medium tanpa antibiotik. Analisis morfologi sel HeLa dilakukan dengan menggunakan mikroskop fase kontras setelah pewarnaan haematoksilen eosin. Doksorubisin (0,5625 µg/ml) digunakan sebagai kontrol positif induksi apoptosis. Hasil penelitian menunjukkan bahwa ekstrak Curcuma longa menyebabkan perubahan morfologi sel yang ditandai dengan semakin mengecilnya ukuran sel, hilangnya prosesus sitoplasmik sehingga sel berbentuk bulat, serta hilang kontak dengan sel lain yang merupakan ciri apoptosis pada sebagian besar sel HeLa. Nukleus tampak berwarna gelap karena peningkatan kapasitas penyerapan zat haematoksilen. Analisa statistik menunjukkan perbedaan yang bermakna antara kelompok kontrol positif dan negatif dengan kelompok stimulasi dalam jumlah sel yang mengalami perubahan morfologi menuju apoptosis. Disimpulkan bahwa ekstrak Curcuma longa mampu menginduksi perubahan morfologi sel HeLa yaitu berupa cell shrinkage.
Peran Kalsium Intraseluler pada Respon Seluler terhadap Intermedilysin Bakteri Komensal Oral Streptococcus Intermedius Heni Susilowati
Majalah Kedokteran Gigi Indonesia Vol 18, No 1 (2011): August
Publisher : Faculty of Dentistry, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (5100.735 KB) | DOI: 10.22146/majkedgiind.16493

Abstract

Latar Belakang. Bakteri komensal oral Streptococcus intermedius mampu memproduksi intermedilysin (ILY), toksin yang berpotensi menyebabkan terbentuknya pori pada membran, yang termasuk dalam golongan cholesterol-dependent cytolysin. Toksin ini dikenal sebagai sitolisin yang unik karena sifatnya yang spesifik hanya menimbulkan respon pad a sel-sel manusia. Tujuan dari studi literatur ini adalah untuk memahami mekanisme sel-sel tubuh manusia dalam merespon paparan ILY. Ringkasan Pembahasan. Penelitian-penelitian terdahulu menunjukkan keterlibatan S. intermedius dalam infeksi purulen oral maupun nonoral karena kemampuannya memproduksi ILY. Intermedilysin diketahui meny.ebabkan ketidakseimbangan metabolisme sel dan kematian sel pada beberapa sel tubuh manusia seperti hepatosit, sel polimorfonuklear, bile duct cell, dan eritrosit. Spesifisitas ILY pada sel-sel tubuh manusia ditentukan oleh ikatan yang eksklusif hanya dengan reseptor membran human CD59. Pada beberapa kasus, ILY menimbulkan respon seluler melalui perubahan konsentrasi kalsium intraseluler ([Ca2+] i). Peningkatan konsentrasi [Ca2+]i mengakibatkan degranulasi sitoplasma pada sel PMN serta aktivasi kalsineurin yang diikuti dengan aktivasi faktor transkripsi NFAT1 pada sel HuCCT1 dan NF-KB pada sel HepG2. Kesimpulan. Studi literatur ini dapat disimpulkan bahwa peningkatan konsentrasi [Ca2+]i berperan penting dalam mekanisme respon seluler sel-sel tubuh manusia terhadap ILY yang diproduksi oleh S. intermedius.
The effect of in vitro royal jelly provision on adhesion of Pseudomonas aeruginosa Sifra Kristina Hartono; Tetiana Haniastuti; Heni Susilowati; Juni Handajani; Alma Linggar Jonarta
Majalah Kedokteran Gigi Indonesia Vol 5, No 1 (2019): April
Publisher : Faculty of Dentistry, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (243.249 KB) | DOI: 10.22146/majkedgiind.30221

Abstract

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic bacterium, which could aggressively infect immunocompromised patients and thus, cause high mortality rate. In addition, P. aeruginosa in oropharynx could be aspirated and cause ventilator associated pneumonia. Royal jelly is one of bee’s products that has been used for therapeutic needs including antibacteria. Adherence factor of P. aeruginosa were flagelum, pili and lectin. The aim of the study was to determine the effect of royal jelly to P. aeruginosa adhesion. Suspension of P. aeruginosa (ATCC® 27853) was incubated at 37 °C for 18 h. Treatment groups were exposed to royal jelly with several concentrations, 2%, 4%, 6%; while distilled water was being used as negative control. Bacterial adhesion test was determined using spectrophotometer λ = 600 nm to measure optical density values of adhered bacterial suspension in tubes. The result of one-way ANOVA showed significant differences (p<0.05) of optical density values among groups indicating that royal jelly affected the bacterial adhesion. LSD results showed significant difference of optical density values between 2%, 4%, and 6% royal jelly compared to distilled water. Six percent of royal jelly had the least optical density value compared to the other groups. In conclusion, royal jelly has the ability to decrease adhesion of P. aeruginosa. Six percent of royal jelly has better ability to decrease adhesion of P. aeruginosa than other concentrations.
The effect of bitter gourd (Momordica charantia) ethanolic extract on inflammatory infiltrates and NF-κB activation in periodontitis Aryudhi Armis; Tetiana Haniastuti; Heni Susilowati
Majalah Kedokteran Gigi Indonesia Vol 5, No 1 (2019): April
Publisher : Faculty of Dentistry, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (80.726 KB) | DOI: 10.22146/majkedgiind.31579

Abstract

Periodontitis is a periodontal disease involving the gingiva, periodontal ligament, cementum, and alveolar bone due to an inflammatory process. Virulence factors of periodontopathogens and inflammatory responses in periodontitis can stimulate nuclear factor-kappa B (NF-κB) activity. Charantoside c and Momordicosides g in bitter gourd prevent NF-κB activation stimulated by TNF-α in HepG2 cells. This study aims to determine the effect of bitter gourd fruit (Momordica charantia) ethanolic extract as an anti-inflammatory substance on the level of inflammatory infiltrates and the number of cells that experience NF-κB activation in the periodontitis model. Eighty male Wistar rats were divided into 5 groups. The mandibular incisors were ligated for 14 days to induce periodontitis. Each group was given Momordica charantia extract of 500 mg/kg BW, 250 mg/kg BW, and 100 mg/kg BW; ibuprofen 100 mg/kg BW; and aquades orally using oral gavage on the day 14. Rat necropsy was carried out on day 1, 3, 5, and 7 after giving the substances. Taking out the lower jaw was done to make tissue preparations followed by staining them with hematoxylin eosin (HE). Immunohistochemicalanalysis was performed to observe cells that were positive for NF-κB activation. The results showed a decrease in the density of inflammatory infiltrates in all groups, except for those given aquades. The number of inflammatory cells ofneutrophils, macrophages, and lymphocytes that experienced NF-κB activation showed the most effective decrease in the group of Momordica charantia 500 mg/kg BW, on the 7th day. The conclusion of this study is that ethanolic extractof Momordica charantia has an anti-inflammatory effect and prevents the activation of NF-kB in rat gingival induced by periodontitis. The highest effect was found at a dose of 500 mg/kg BW on day 7 after giving of extracts.
Dealing with the high-risk potential of COVID-19 cross-infection in dental practice Heribertus Dedy Kusuma Yulianto; Nunuk Purwanti; Trianna Wahyu Utami; Anne Handrini Dewi; Dyah Listyarifah; Intan Ruspita; Asikin Nur; Heni Susilowati
Majalah Kedokteran Gigi Indonesia Vol 6, No 1 (2020): April
Publisher : Faculty of Dentistry, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/majkedgiind.56588

Abstract

The World Health Organization reported that the SARS-CoV-2 virus has infected more than 5 million people around the world. Dental care providers and health care professionals need to be aware of the high-risk potential of crossinfection since the routes of virus transmission commonly happen through droplets and aerosols. This review aimed at collecting essential knowledge about the COVID-19 needed by dental practitioners. The review focused on the oral involvement in COVID-19, the role of oral transmission as the high-risk potential of cross-infection and recommended strategies to minimize the risk of cross-infection in dental practice. We searched all the published clinical features from PubMed, Google Scholar, Scopus and hand searched library online databases, from January 2015 until May 2020. Keywords used were “COVID-19”, “Dentistry”, “Dental protection”, “Cross-contamination”, “Aerosol and non aerosol”, and ”Povidone-iodine” with their combinations. We identified 52 articles to review after the initial selection with inclusion and exclusion criteria. Results showed use of topical applications of povidine-iodine and viricidal mouthwash could significantly reduce the high-risk of cross-infection from dentistry patients who are asymptomatic with COVID-19 infection. Further safeguards include suspending all non-emergency procedures temporarily and closely screening patients for symptoms which may be suspected to be COVID-19 infection.
RONGGA MULUT SEBAGAI RESERVOIR POTENSIAL UNTUK INFEKSI Pseudomonas aeruginosa Puspita Hajardhini; Heni Susilowati; Heribertus Dedy Kusuma Yulianto
ODONTO : Dental Journal Vol 7, No 2 (2020): December 2020
Publisher : Faculty of Dentistry, Universitas Islam Sultan Agung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.30659/odj.7.2.125-133

Abstract

Introduction : Pseudomonas aeruginosa (P. aeruginosa) is Gram-negative bacteria which is normally inhabiting in environment, however it tends to be an opportunistic pathogen within oral cavity. It utilizes the oral cavity as potential reservoir to infect either lungs or oral cavity itself. Both planktonic and biofilm forms can mediate its infection in oral cavity so that making its difficulties to eradicate since its broad resistance to antibiotics.Method : We review the pathogenesis of oral infection and host defense mechanism to P. aeruginosa as well.Discussion: Several prevention strategies, both chemical and nonchemical, are elaborated to avoid oral bacterial contamination.Conclusion : Prevention strategies in dental aspect to P. aeruginosa is highly needed as its occurrence are susceptible to cross-infection.
Effect of Red Onion (Allium cepa var ascalonicum) Skin Ethanolic Extract on the Motility and the Adhesion Index of Pseudomonas aeruginosa and Macrophage Phagocytosis Index Irma Prasety Ayu Nugraheni; Derana Widyastika; Sofia Maulida; Heni Susilowati; Alma Linggar Jonarta
Majalah Obat Tradisional Vol 24, No 1 (2019)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (361.726 KB) | DOI: 10.22146/mot.45532

Abstract

Red onion skin (Allium cepa var ascalonicum) contains various ingredients that may function as antibacterial agents against microorganisms, as well as anti-inflammatory and immunomodulator agents for host cells, such as macrophages. Pseudomonas aeruginosa found in the oral cavity is commensal bacteria that may turn into opportunistic pathogen by utilizing its virulence factors such as motility and adhesion to the host cell. The purpose of this study was to investigate the effect of red-onion-skin ethanolic extract towards P. aeruginosa ATCC 9027 on the motility and adhesion ability, furthermore, to know its effect on the macrophage phagocytosis. The research was conducted into three parts of experiment using red-onion-skin ethanolic extract. Extract-induced bacterial motility test was carried out on semi-solid media, stained using 0.1% crystal violet, then the radial length of the bacterial movement was measured. The bacterial adhesion index to buccal cells was calculated after incubated for two hours and stained with Gram stain. Phagocytic activity of the host cells on P. aeruginosa was done by exposing the extract to the mouse peritoneal macrophages, then the phagocytosed bacteria were counted after Giemsa staining. Statistical test results from the three experiments showed significant differences between the test groups compared to the control groups (p <0.05). It was concluded that the red onion-skin ethanolic extract not only affects P. aeruginosa by reducing swarming motility and preventing bacterial adhesion to buccal epithelial cells, but also induces the host cells by increasing the ability of macrophage phagocytosis to these bacteria.
Analysis of anti-Streptococcus sanguinis IgY ability to inhibit Streptococcus sanguinis adherence Suryani Hutomo; Heni Susilowati; Dewi Agustina; Widya Asmara
Dental Journal (Majalah Kedokteran Gigi) Vol. 51 No. 1 (2018): March 2018
Publisher : Faculty of Dental Medicine, Universitas Airlangga https://fkg.unair.ac.id/en

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2021.962 KB) | DOI: 10.20473/j.djmkg.v51.i1.p33-36

Abstract

Background: Streptococcus sanguinis (S. sanguinis), an oral commensal bacterium, is often implicated in infective endocarditis. Its adherence to the tooth surface is the initial step in dental plaque formation. In addition to the important role of S. sanguinis in systemic disease and antimicrobial resistance, it is necessary to develop methods to control dental plaque formation. Immunoglobulin Y (IgY) has been used to prevent bacterial infection. Purpose: The purpose of this study is to analyze the ability of anti-S. sanguinis IgY antibodies to inhibit S. sanguinis adherence to hydroxyapatite (HA) discs as a model of the tooth surface. Methods: Antibodies were produced by immunizing hens with S. sanguinis suspension. Boosters were given three times following the first injection. An agar gel precipitation test (AGPT) was used to detect the presence of anti-S. sanguinis IgY. A bacterial adherence assay was performed twice to analyze the ability of IgY and the optimal concentration required to inhibit bacterial adherence. Results: The formation of a precipitation line using AGPT confirmed the presence of the antibody. In addition, it was shown that the anti-S. sanguinis IgY antibody could inhibit bacterial adherence to HA. Statistical analysis using One-way ANOVA revealed a significant difference in the optical density (OD) value between the groups (p<0.05). The results of electron microscopy scanning confirmed the quantitative analysis by means of a bacterial adherence test. Conclusion: Anti-S. sanguinis IgY has the ability to inhibit adherence of S. sanguinis to HA discs at an optimal concentration of 30%. The inhibitive effect was stronger in the presence of saliva.
Cyclosporine A and FK506 as Potent Inhibitors of Streptococcus intermedius Intermedilysin-Induced NFAT-1 Activation Heni Susilowati; Hirohiko Okamura; Katsuhiko Hirota; Kaya Yoshida; Atsushi Tabata; Hideaki Nagamune; Tatsuji Haneji; Yoichiro Miyake
Indonesian Journal of Cancer Chemoprevention Vol 1, No 2 (2010)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev1iss2pp67-73

Abstract

Cyclosporine A (CsA) and tacrolimus (FK506), a member of calcineurin inhibitors, inhibit inflammation process as part of immune response. Nuclear activated T cells subfamily NFAT1 is a trascription factor responsible for the regulation of immune response genes. Streptococcus intermedius, an oral commensal bacterium, has been shown to strongly associate with liver abscess.  The S. intermedius strains produce intermedilysin (ILY), which is responsible for the bacterial virulence. Cyclosporine A and FK506 have been widely used to control NFAT activation in most of cell types, however the ability of CsA and FK506 to inhibit ILY-induced NFAT1 activation remains to be investigated. The aim of this study was to investigate the effect of CsA and FK506 on NFAT1 activation caused by ILY. Human cholangiocellular cell line HuCCT1 was stimulated with various concentrations of ILY. The cell and nuclear morphological change was observed by microscopy analysis. The NFAT1 nuclear translocation that indicates its activation was detected by immunocytochemistry. The inhibitory effect of CsA and FK506 was tested after 30 min application before ILY treatment by using immunofluorescence microscope. The results showed cell and nuclear shrinkage in ILY-treated cells. The NFAT1 was translocated to the nuclei in HuCCT1 cells, and observed in dose dependent manner.  Cyclosporine A and FK506 inhibited ILY-induced NFAT1 nuclear translocation.  In conclusion, CsA and FK506 may act as potent inflammation control agents in S. intermedius ILY-infected cells.Keywords: Cyclosporine A, FK506, NFAT1, intermedilysin