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Analisis Bioaktivitas Bahan Irigasi Ekstrak Buah Lerak (Sapindus Rarak Dc) Terhadap Patogenesis Fusobacterium Nucleatum (Kajian

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Analisis Bioaktivitas Bahan Irigasi Ekstrak Buah Lerak (Sapindus Rarak Dc) Terhadap Patogenesis Fusobacterium Nucleatum (Kajian ANALISIS BIOAKTIVITAS BAHAN IRIGASI EKSTRAK BUAH LERAK (SAPINDUS RARAK DC) TERHADAP PATOGENESIS FUSOBACTERIUM NUCLEATUM (KAJIAN UJI ANTI-ADHESI, UJI BIOMASSA, DAN UJI POROSITAS DINDING SALURAN AKAR GIGI) (PENELITIAN IN VITRO) TESIS Oleh JIHAN RAHMADIAN FITRIA 177160006 PROGRAM PENDIDIKAN DOKTER GIGI SPESIALIS ILMU KONSERVASI GIGI FAKULTAS KEDOKTERAN GIGI UNIVERSITAS SUMATERA UTARA MEDAN 2020 Universitas Sumatera Utara ANALISIS BIOAKTIVITAS BAHAN IRIGASI EKSTRAK BUAH LERAK (SAPINDUS RARAK DC) TERHADAP PATOGENESIS FUSOBACTERIUM NUCLEATUM (KAJIAN UJI ANTI-ADHESI, UJI BIOMASSA, DAN UJI POROSITAS DINDING SALURAN AKAR GIGI) (PENELITIAN IN VITRO) TESIS Diajukan Sebagai Salah Satu Syarat Untuk Memperoleh Spesialis Konservasi Gigi (Sp.KG) Pada Fakultas Kedokteran Gigi Universitas Sumatera Utara Oleh JIHAN RAHMADIAN FITRIA 177160006 PROGRAM PENDIDIKAN DOKTER GIGI SPESIALIS ILMU KONSERVASI GIGI FAKULTAS KEDOKTERAN GIGI UNIVERSITAS SUMATERA UTARA MEDAN 2020 Universitas Sumatera Utara Universitas Sumatera Utara PERNYATAAN ANALISIS BIOAKTIVITAS BAHAN IRIGASI EKSTRAK BUAH LERAK (SAPINDUS RARAK DC) TERHADAP PATOGENESIS FUSOBACTERIUM NUCLEATUM (KAJIAN UJI ANTI-ADHESI, UJI BIOMASSA, DAN UJI POROSITAS DINDING SALURAN AKAR GIGI) (PENELITIAN IN VITRO) TESIS Dengan ini saya menyatakan bahwa dalam tesis ini tidak terdapat karya yang pernah diajukan untuk memperoleh gelar kesarjanaan di suatu perguruan tinggi, dan sepanjang pengetahuan saya juga tidak terdapat karya atau pendapat yang pernah ditulis atau diterbitkan oleh orang lain, kecuali yang secara tertulis diacu dalam naskah ini dan disebutkan dalam daftar pustaka. Medan, Februari 2020 Jihan Rahmadian Fitria Universitas Sumatera Utara DAFTAR ISTILAH 1. µl : Mikroliter 2. Adh : Adhesin 3. AS : Agregation substance 4. ATCC : American Tissue Culture Collection 5. BHI : Brain Heart Infusion 6. C : Celcius 7. CEJ : Cemento Enamel Junction 8. CFU : Colony forming unit 9. CMCP : Camphoroted Monochlorophenol 10. DNA : Deoxyribonucleic acid 11. EDTA : Ethylenediamine Tetrracacetic Acid 12. ELISA : Enzyme-Linked Immuno Sorbent Assay 13. EPS : Extra Polimeric Substance 14. GCMS ; Gas Chromatography Mass Spectrometry 15. HSP : Heat Shock Protein 16. Ig A : Imunoglobulin A 17. IL : Interleukin 18. LPS : Lipopolisakarida 19. LTA : Lipoteichoic acid 20. m RNA : Messenger RNA 21. MHB : Mueller Hinton Broth 22. ml : Milliliter 23. MMP : Matrix Metallproteinase 24. NaOCl : natrium hipoklorit 25. nm : Nanometer 26. OD : Optical Density 27. OMPs : Outer Membrane Protein 28. PBS : Phosphate Buffer Saline 29. PSA : Perawatan Saluran Akar 30. Pz : Aktivitas Fosfolipase 31. SEM : Scanning Electron Microscope 32. TLR : Toll-Like Receptor i Universitas Sumatera Utara ABSTRAK Salah satu bakteri pada biofilm yang terbentuk pada infeksi endodontik adalah Fusobacterium nucleatum. Dari aspek mikrobiologis, infeksi endodonti dapat dirawat dengan prosedur kemikal dan mekanikal (cleaning and shaping). Irigasi kombinasi EDTA dan NaOCl adalah metode yang paling sering direkomendasikan. Namun kelemahan utama NaOCl adalah sitotoksisitasnya ketika ekstrusi ke dalam jaringan periradikuler dan kombinasi tersebut menghasilkan erosi yang kuat pada dentin. Akibat kelemahan bahan irigasi tersebut, penggunaan bahan alami sebagai alternatif bahan irigasi saluran akar diharapkan lebih biokompatibel dan salah satu bahan alami yang dapat dikembangkan sebagai bahan irigasi adalah buah lerak (Sapindus rarak DC). Penelitian ini menggunakan sampel koloni bakteri Fusobacterium nucleatum dan dibiakkan dengan media Crystal Violet Erythtromycin (CVE) Agar. Ekstrak buah lerak dengan konsentrasi 6,25, 12,5%, 25% dan dibandingkan dengan NaOCL 2,5% dan EDTA 17% dengan waktu 24 jam, 48 jam, 72 jam untuk diuji antiadhesi, biomassa dan porositas setelah diinteraksikan dengan bakteri F.Nucleatum. Berdasarkan analisis statistik kruskal wallis dan one way anova serta uji lanjutan nya dengan LSD dan mann whitney test, hasil penelitian ini menunjukkan bahwa ekstrak buah lerak dari berbagai konsentrasi (6,25%, 12,5%, 25%) sama baiknya dengan larutan NaOCl 2,5%+EDTA 17% yang menjadi gold standar dalam irigasi saluran akar gigi dalam mencegah adhesi dan menghambat perkembangan bakteri Fusobacterium nucleatum. Secara klinis NaOCl 2,5%+EDTA 17% memiliki tingkat porositas paling tinggi dan berbeda signifikan dengan ekstrak lerak 6,25%, 12,5% dan 25%. Sehingga dapat di asumsikan bahwa ekstrak lerak dalam berbagai konsentrasi (6,25%, 12,5%, 25%) tidak menyebabkan tingkat porositas yang berlebihan dibandingkan dengan NaOCl 2,5%+EDTA 17% yang menjadi gold standar irigasi saluran akar gigi. Ekstrak buah lerak memiliki kemampuan dalam mencegah adhesi dan menghambat perkembangan bakteri F.nucleatum serta secara klinis tidak menyebabkan porositas yang berlebihan pada dinding saluran akar gigi. Kata kunci : Ekstrak buah lerak, antiadhesi, Fusobacterium nucleatum, irigasi saluran akar, porositas saluran akar ii Universitas Sumatera Utara ABSTRACT One of the bacteria in biofilms formed in endodontic infections is Fusobacterium nucleatum. From the microbiological aspect, endodontic infections can be treated with chemical and mechanical procedures (cleaning and shaping). Combination of EDTA and NaOCl irrigation is the most commonly recommended method. However, the main disadvantage of NaOCl is its cytotoxicity when extrusion into periradicular tissue and this combination results in strong erosion of dentin. As a result of the weakness of the irrigation material, the use of natural material as an alternative to root canal irrigation is expected to be more biocompatible and one of the natural materials that can be developed as irrigation is lerak fruit (Sapindus rarak DC). This study used a sample of the bacterial colony Fusobacterium nucleatum and was cultured with Crystal Violet Erythtromycin (CVE) agar. Lerak fruit extract with a concentration of 6.25, 12.5%, 25% and compared with 2.5% NaOCL and 17% EDTA with 24 hours, 48 hours, 72 hours to be tested for anti-adhesion, biomassa and porosity after being interacted with F.Nucleatum bacteria. Based on the statistical analysis of kruskal wallis and one way anova and their follow-up tests with LSD and mann whitney test, the results of this study showed that lerak extract from various concentrations (6.25%, 12.5%, 25%) was as good as NaOCl solution 2.5% + 17% EDTA which became the gold standard in irrigation of root canals in preventing adhesion and inhibiting the development of the bacterium Fusobacterium nucleatum. Clinically, 2.5% NaOCl + 17% EDTA has the highest porosity level and is significantly different with 6.25%, 12.5% and 25% lerak extracts. So it can be assumed that lerak extract in various concentrations (6.25%, 12.5%, 25%) does not cause excessive porosity compared to 2.5% NaOCl + 17% EDTA which is the gold standard of tooth root canal irrigation. Lerak fruit extract has the ability to prevent adhesion and inhibit the development of F.nucleatum bacteria and clinically does not cause excessive porosity in the root canal wall of the tooth. Keywords: Lerak fruit extract, anti-adhesion, Fusobacterium nucleatum, root canal irrigation, root canal porosity iii Universitas Sumatera Utara KATA PENGANTAR Puji dan syukur penulis panjatkan kepada Allah SWT atas Rahmat dan Karunia-Nya sehingga penulis dapat menyelesaikan tesis ini sebagai salah satu syarat untuk memperoleh gelar Spesialis Konservasi Gigi dari Universitas Sumatera Utara. Ucapan terima kasih yang sebesar-besarnya penulis sampaikan kepada kedua orang tua terkasih, yaitu ayahanda Aliasmi, SOS dan ibunda Syafrida Ridwan yang telah membesarkan, memberikan kasih sayang, doa, semangat dan dukungan baik moril dan materi yang tak terbalas dan terucapkan. Penulis menyampaikan terima kasih kepada bapak mertua Zulkismen, S.Pd dan ibu mertua Defirina, S.Pd yang telah membantu dan mendukung baik doa dan semangat bagi penulis, dan adik penulis dr. Habib Budiman Agung, drh. Zena Fisdiora, Atika Karina Asmi dan Dandila Diko serta segenap keluarga yang memberikan dukungan dan doa kepada penulis. Penulis juga menyampaikan terimakasih kepada suamiku tercinta drg. Desmedio Deno Merinda yang telah mendukung penulis dengan penuh kasih sayang, doa, kesabaran, pengorbanan dan semangat yang telah diberikan selama ini. Semoga apa yang kita capai dapat memberikan kebahagiaan dan diberkati oleh Allah SWT. Dalam pelaksanaan penelitian dan penulisan tesis ini, penulis telah banyak mendapatkan bimbingan, bantuan dan doa dari berbagai pihak. Pada kesempatan ini, dengan segala kerendahan hati dan penghargaan yang tulus, penulis ingin menyampaikan rasa terima kasih yang sebesar-besarnya kepada: iv Universitas Sumatera Utara 1. Dr. Trelia Boel, drg, Sp.RKG(K), M.Kes selaku Dekan Fakultas Kedokteran Gigi Universitas Sumatera Utara. 2. Cut Nurliza, drg., Sp.KG., M.Kes selaku Ketua Departemen Ilmu Konservasi Gigi Fakultas Kedokteran Gigi Universitas Sumatera Utara dan pembimbing kedua yang banyak meluangkan waktu, memberikan petunjuk, arahan, semangat serta dukungan kepada penulis sehingga tesis ini dapat diselesaikan dengan baik. 3. Prof. Trimurni Abidin, drg., M.Kes., Sp.KG (K) selaku Ketua Program Studi Pendidikan Dokter Gigi Spesialis Konservasi Gigi Fakultas Kedokteran Gigi USU dan selaku anggota panitia penguji yang telah banyak meluangkan waktu, memberikan bimbingan, arahan, semangat serta dukungan kepada penulis sehingga tesis ini dapat diselesaikan dengan baik. 4. Drg. Nevi Yanti, M.Kes, Sp.KG(K) selaku Sekretaris Departemen Ilmu Konservasi Gigi Fakultas Kedokteran Gigi USU dan pembimbing pertama
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