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"Latar Belakang: Bahan irigasi endodontik yang sering digunakan saat ini memiliki sifat toksik terhadap sel punca yang berperan penting pada perawatan endodontik regeneratif. Sehingga dibutuhkan pemilihan bahan irigasi alami yang tidak toksik, seperti ekstrak jeruk purut yang juga memiliki efek antibakteri terhadap bakteri gram positif. Tujuan: Mendapatkan perbandingan sitotoksisitas larutan ekstrak daun jeruk purut berbagai konsentrasi pada waktu observasi 24 jam terhadap hDPSCs. Metode: HDPSCs diisolasi dari gigi molar tiga, diberikan media perlakuan berupa larutan ekstrak daun jeruk purut dengan konsentrasi 2,5%, 5%, 10% dan 20%, NaOCl 2,5% dan DMEM sebagai kontrol. Pengamatan proliferasi hDPSCs dengan uji MTT. Hasil dibaca dengan microplate reader dengan panjang gelombang 570 nm. Nilai absorbansi larutan ekstrak daun jeruk purut 2,5%, 5%, 10%, 20% dan NaOCl 2,5% dihitung persentasinya dibandingkan dengan nilai absobansi kelompok kontrol menggunakan rumus perhitungan viabilitas sel. Hasil: Nilai rerata viabilitas sel kelompok larutan ekstrak daun jeruk purut diatas 90%, menandakan larutan ekstrak daun jeruk purut tidak toksik terhadap hDPSCs. Kesimpulan: Larutan ekstrak daun jeruk purut tidak memiliki efek sitotoksisitas terhadap hDPSCs, dengan nilai viabilitas sel paling tinggi pada konsentrasi 20%."

"Latar Belakang: Obat berbahan dasar alami di Indonesia harus terstandar, memiliki pembuktian keamanan dan khasiatnya secara ilmiah melalui uji preklinik. Uji yang dapat dilakukan untuk mengetahui keamanan suatu bahan yaitu uji sitotoksisitas. Tujuan: Mengetahui sitotoksisitas ekstrak asam jawa 2,5%, 5%, dan 10% terhadap human dental pulp stem cells. Metode: Identifikasi komponen senyawa kimia dengan uji fitokimia dan GC-MS. Uji sitotoksisitas dengan uji viabilitas diukur menggunakan MTT assay untuk mengetahui aktivitas selular. Pengukuran menggunakan microplate reader panjang gelombang 570 nm. Semakin tinggi nilai OD yang didapat, maka nilai viabilitas sel akan semakin tinggi. Sel ditanam pada 96 well dengan densitas 5x103 sel/well. Analisa data menggunakan Kruskal-Wallis dan Post Hoc Mann-Whitney. Hasil: Dari nilai median viabilitas sel, kelompok larutan ekstrak asam jawa 2,5% memiliki nilai viabilitas yang paling tinggi, sedangkan larutan ekstrak asam jawa 10% memiliki nilai viabilitas yang paling rendah. Tidak terdapat perbedaan yang bermakna antara kelompok larutan ekstrak asam jawa 2,5% dan 5%. Kesimpulan: Larutan ekstrak asam jawa dari desa babakan kecamatan Darmaga Kabupaten Bogor dapat diidentifikasi komponen senyawa flavonoid dan saponin dan memiliki komponen senyawa kimia yang berpotensi sebagai antibakteri, antiinflamasi, anti virus, antioksidan, larutan khelasi, dan solvent. Larutan ekstrak asam jawa 2,5% dan 5% tidak toksik terhadap hDPSCs.

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Background: Medicines made from natural ingredients in Indonesia must be standardized, having proven safety and efficacy through preclinical trials. The safety test that can be carried out is cytotoxicity test. Objective: To determine the cytotoxicity of 2,5%,5% and 10% Tamarindus indica extract on hDPSCs. Methods: identification of components chemical compound groups was measured by phytochemical test and GC-MS. Cytotoxicity test with viability test was measured using the MTT assay to determine cellular activity. Measured using microplate reader with a wavelength of 570nm. The higher OD value obtained, the higher the cell viability value. Cells inserted in 96 wells with a density of 5x103 cells/well. Data analysis using Kruskal-Wallis and Post Hoc Mann-Whitney. Results: From the median cell viability value, 2,5% Tamarindus indicia extract solution group had the highest viability value, while 10% Tamarindus indica etract had the lowest viability value. There was no significant difference between the 2,5% and 5% Tamarindus indica extract solution groups. Conclusion: Tamarindus indica extract from Babakan village, Darmaga Bogor can be identified as components of flavonoids and saponins, has chemical components that have potential as antibacterial, antiinflammation, antiviral, antioksidan, chelation solution, and solvents. Tamarindus indica extract of 2,5% and 5% were not toxic to hDPSCs."

"Latar Belakang: Ekstrak jintan putih Cuminum cyminum memiliki potensi efektivitas antibakteri dan anti jamur serta tidak toksik terhadap sel fibroblas tikus. Belum terdapat penelitian yang meneliti toksisitas ekstrak jintan putih terhadap Dental Pulp Stem Cells DPSCs . Tujuan: Mengetahui efek ekstrak jintan putih konsentrasi 0,1 mg/ml, 0,4 mg/ml, 0,7 mg/ml, dan 1,0 mg/ml terhadap viabilitas DPSCs. Metode: Menggunakan uji MTT dengan menghitung nilai absorbansi menggunakan microplate reader, dengan hasil akhir berupa nilai optical density OD yang dipersentasekan terhadap kelompok kontrol. Hasil: Terdapat perbedaan viabilitas DPSCs yang bermakna

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Introduction The extract of cumin Cuminum cyminum has the potential antibacterial and antifungal activity and it was not toxic for mouse fibroblasts. However, there have been no research investigating the toxicity of cumin extract on Dental Pulp stem Cells DPSCs . Aims To compare viability DPSCs of Cuminum cyminum extract 0,1 mg ml, 0,4 mg ml, 0,7 mg ml, and 1.0 mg ml . Methods Cell viability was analyzed using MTT Assay by calculating absorbance value using microplate reader, with optical density OD as the final result. Results There were significant differences statistically in viability on DPSCs p"

"Latar Belakang: Fokus disinfeksi saluran akar telah berubah dari disinfeksi agresif menjadi seleksi protektif dalam prosedur regeneratif endodontik. Larutan irigasi sintetik yang digunakan hingga saat ini toksik terhadap sel punca pulpa, salah satunya yang memiliki kemampuan proliferasi dan transdiferensiasi tinggi adalah hDPSCs. Oleh sebab itu, penelitian terkait disinfeksi berbahan alami yang mampu mempertahankan viabilitas sel punca terus berkembang pesat. Salah satu larutan irigasi alami yang bersifat antimikrobial dan agen kelator adalah larutan cuka apel. Untuk menjadikannya obat herbal terstandar hingga fitofarmaka, perlu diidentifikasi kelompok senyawa kimia dan uji viabilitas hDPSCs.Tujuan: Menganalisis pengaruh larutan cuka apel berbagai konsentrasi terhadap viabilitas hDPSCsMetode: hDPSCs ditambahkan DMEM+FBS10% (kontrol negatif), EDTA 17% (kontrol positif), larutan cuka apel dengan konsentrasi 2,5%, 5%, dan 10% dengan enam kali pengulangan. Selanjutnya, persentase viabilitas hDPSCs didapat dari MTT assays melalui microplate reader dalam nilai absorbansi. Data kemudian diolah statistik melalui uji parametrik One-way ANOVA.Hasil: Nilai rerata viabilitas sel hDPSC pada semua kelompok perlakuan bernilai diatas 70% sehingga tidak toksik menurut standar ISO dengan rerata viabilitas tertinggi pada kelompok 2,5% dan terendah pada kelompok EDTA 17% diikuti kelompok 10%.Kesimpulan: Larutan cuka apel dapat diidentifikasi kelompok senyawa kimia dan nilai viabilitas sel paling tinggi pada konsentrasi 2,5%.

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Background: Focus on root canals disinfection have shift from aggressive to protective selection in regenerative endodontic procedures. Synthetic root canals irrigation that had been used until now are toxic toward pulp stem cells, one of them, hDPSCs which have higher proliferation and transdifferentiation ability. Therefore, research on natural disinfection which maintain stem cell viability keep developing rapidly. One of the natural disinfection that has antimicrobial effect and chelating agent is apple cider vinegar. To standardized it as modern medicine, need to identify group of chemical compounds and analyzing the viability percentage of hDPSCs.

Objective: Analyze the impact of apple cider vinegar solution in various concentrations on viability of hDPSCs.

Methods: hDPSCs were given DMEM+FBS10% (negative control), 17% EDTA (positive control), apple cider vinegar solution in 2.5%, 5% and 10% concentrations with six repetitions. Percentage viability of hDPSCs were analyze from MTT assays with microplate reader in absorbance value. Then, data were proccessed statictically with parametric One-way ANOVA.

Results: The average viability of hDPSCs were above 70% which considered non-toxic according to ISO, with the highest cells viability in 2.5% and the lowest cells viability in 17% EDTA followed by 10% groups.

Conclusion: Apple cider vinegar solution’s chemical compounds can be identified with the highest cells viability were at 2.5%."

"Latar Belakang: Radikal bebas memiliki sifat destruktif pada perawatan kedokteran gigi, salah satunya pada prosedur perawatan saluran akar. Pada perawatan saluran akar, bahan irigasi yang selama ini digunakan dapat menyebabkan kerusakan pada saluran akar karena memiliki senyawa radikal bebas. Oleh karena itu, perlu ditemukan bahan alami herbal yang dapat menjadi sumber antioksidan sebagai penangkal radikal bebas yang dapat berperan positif dalam perawatan saluran akar. Penelitian sudah banyak dikembangkan oleh para peneliti untuk menjadikan larutan ekstrak daun jeruk purut dan larutan cuka apel Malang sebagai obat herbal terstandar dari Indonesia hingga fitofarmaka. Salah satu pengujian yang harus dilakukan adalah pengujian antioksidan karena pada kedua bahan alami herbal tersebut ditemukan senyawa antioksidan berupa fenol dan flavonoid pada uji fitokimia dan GCMS. Tujuan: Penelitian ini bertujuan untuk melihat perbedaan aktivitas dan kadar antioksidan pada larutan ekstrak daun jeruk purut dan larutan cuka apel Malang pada berbagai konsentrasi. Metode: Penelitian ini merupakan eksperimental laboratorik dengan metode DPPH. Sampel uji berupa larutan 10%, 5%, dan 2,5% ekstrak daun jeruk purut dan cuka apel Malang yang dibuat serial konsentrasi dengan pengenceran. Uji aktivitas antioksidan dilakukan dengan melihat perubahan warna pada larutan, dari warna ungu menjadi warna kuning apabila terdapat aktivitas antioksidan di dalam larutan uji. Selanjutnya dilakukan pengukuran kadar antioksidan dengan melihat tingkat absorbansi menggunakan Spektrofotometer UV/VIS dengan panjang gelombang 517 nm dan kemudian dilakukan pengulangan sebanyak 3 kali. Setelah itu, dilakukan perhitungan persentase inhibisi dan didapatkan kadar IC50 antioksidan. Kemudian, perbedaan kadar antioksidan keduanya dilakukan uji statistik Independent T-Test. Hasil: Terdapat aktivitas antioksidan pada larutan ekstrak daun jeruk purut dan larutan cuka apel Malang. Larutan ekstrak daun jeruk purut dapat dideteksi dengan larutan induk konsentrasi 0,1% dengan kadar antioksidan 347,691 µg/ml dan larutan cuka apel Malang dapat dideteksi dengan larutan induk 5% dan 2,5% dengan kadar antioksidan 8375,25 µg/ml dan 8021,162 µg/ml. Keduanya memiliki perbedaan kadar antioksidan secara statistik. Kesimpulan: Terdapat perbedaan kadar antioksidan pada larutan ekstrak daun jeruk purut dan larutan cuka apel Malang dengan berbagai konsentrasi. Larutan ekstrak daun jeruk purut efektif menangkal radikal bebas pada konsentrasi sekitar 0,03%, sedangkan larutan cuka apel Malang efektif pada konsentrasi sekitar 0,8%.

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Background: Free radicals have destructive properties in dental treatments, one of which is in root canal procedures. In root canal treatment, irrigation materials that have been used can cause damage to the root canal because they have free radical compounds. Therefore, it is necessary to find natural herbal ingredients that can be a source of antioxidants as an antidote to free radicals that can play a positive role in root canal treatment. Many studies have been developed by researchers to make kaffir lime leaf extract solution and Malang apple vinegar solution as standardized herbal medicines from Indonesia to phytopharmaceuticals. One of the tests that must be done is antioxidant testing because the two natural herbal ingredients are found antioxidant compounds in the form of phenols and flavonoids in phytochemical and GCMS tests. Objective: This study aims to see the differences in antioxidant activity and levels in kaffir lime leaf extract solution and Malang apple vinegar solution at various concentrations. Methods: This research is a laboratory experiment with DPPH method. Test samples in the form of 10%, 5%, and 2.5% kaffir lime leaf extract solution and Malang apple vinegar solution were made in serial concentrations by dilution. Antioxidant activity test was conducted by looking at the color change in the solution, from purple to yellow color if there is antioxidant activity in the test solution. Furthermore, the antioxidant content was measured by looking at the absorbance level using UV/VIS Spectrophotometer with a wavelength of 517 nm and then repeated 3 times. After that, the percentage of inhibition was calculated and the IC50 value of antioxidant was obtained. Then, the difference in antioxidant levels between the two was carried out Independent T-Test statistical test. Results: There is antioxidant activity in kaffir lime leaf extract solution and Malang apple vinegar solution. Kaffir lime leaf extract solution can be detected with 0.1% concentration mother liquor with antioxidant levels of 347.691 µg/ml and Malang apple vinegar solution can be detected with 5% and 2.5% mother liquor with antioxidant levels of 8375.25 µg/ml and 8021.162 µg/ml. Both have statistically different antioxidant levels. Conclusion: There are differences in antioxidant levels in kaffir lime leaf extract solution and Malang apple vinegar solution with various concentrations. Kaffir lime leaf extract solution is effective against free radicals at a concentration of about 0.03%, while Malang apple vinegar solution is effective at a concentration of about 0.8%."

"Latar Belakang: L-arginin merupakan asam amino semiesensial yang produksinya tidak mencukupi kebutuhan dalam kondisi stres oksidatif akibat inflamasi. L-arginin adalah satu-satunya substrat bagi enzim nitric oxide synthase (NOS) yang memproduksi nitric oxide (NO) yang dapat mengaktivasi focal adhesion kinase (FAK) pathwaydan memicu terjadinya proses migrasi sel. Tujuan: Mengetahui potensi media kultur asam amino L-arginin terhadap laju kecepatan migrasi hDPSCs. Metode: Evaluasi media kultur asam amino L-arginin konsentrasi 300, 400, 500 ¼mol/L, serta DMEM sebagai kontrol terhadap laju kecepatan migrasi hDPSCs menggunakan uji scratch assay menggunakan uji scratch assay yang dihitung dengan rumus laju kecepatan migrasi setelah 24 jam. Analisis statistic menggunakan Paired T-Test dan Oneway ANOVA dengan post hoc LSD. Hasil: Terdapat perbedaan bermakna potensi L-arginin 500 μmol/L dibandingkan konsentrasi 300 dan 400 μmol/L, serta kontrol. Kesimpulan: Media kultur asam amino L-arginin 500 ¼mol/L memiliki potensi laju kecepatan migrasi yang lebih baik dibandingkan konsetrasi 300, 400 ¼mol/L dan kontrol.

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Background: L-arginine is semiessential amino acid which the production is insufficient under oxidative stress due to inflammation. L-arginine is the only substrate of nitric oxide synthase (NOS) enzyme that produces nitric oxide (NO) which activates focal adhesion kinase (FAK) pathway to stimulate cell migration.

Objective: To understand potential of L-arginine amino acid culture media towards speed rate of hDPSCs migration.

Methods: Evaluation of 300, 400, 500 ¼mol/L of L-arginin amino acid culture media and DMEM as control towars speed rate of hDPSCs migration using scratch assay and calculation of migration speed rate after 24 hours. Statistical analysis using Paired T-Test and Oneway ANOVA with post hoc LSD.

Results: Significant result was shown between 500 ¼mol/L of L-arginin amino acid culture media compared with 300 and 400 ¼mol/L concentration and control towards migration speed rate after 24 hours.

Conclusion: 500 ¼mol/L of L-arginin amino acid culture media has a better migration rate compared with lower concentrations and control."

"Latar Belakang: Pada kondisi inflamasi pulpa, terdapat penurunan dari jumlah protein dan asam amino. L-Arginine adalah asam amino semi-esensial karena berperan penting pada kondisi tertentu, gangguan imun berat dan luka bakar, yang membutuhkan asupan tambahan L-Arginine eksternal. Asam amino L-Arginine menjadi satu-satunya substrat sintesis nitric oxide (NO) dan poliamina berasal dari konversi L-Arginine menjadi ornithinen melalui arginase. NO dan poliamina merangsang proliferasi sel dan memiliki efek positif pada perkembangan melalui siklus sel. Tujuan: Mengetahui potensi asam amino L-Arginine terhadap proliferasi hDPSCs. Metode: Evaluasi asam amino L- Arginine konsentrasi 300, 400, 500 μmol/L, serta DMEM sebagai kontrol terhadap proliferasi hDPSCs menggunakan uji cell count setelah 24 jam. Analisis statistic menggunakan Oneway ANOVA dengan post hoc Bonferroni. Hasil: Terdapat perbedaan bermakna potensi L-Arginine 300,400 dan 500 μmol/L dibandingkan kontrol, dan L- Arginine 500 μmol/L memiliki rerata proliferasi hDPSCs paling tinggi sebesar 436.666 sel/ml. Kesimpulan: Asam amino L-Argininee memiliki potensi terhadap proliferasi hDPSCs dan proliferasi tertinggi pada asam amino L-Arginine konsentrasi 500 μmol/L.

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Background: In the inflammatory condition of the pulp, there is a decrease in the amount of protein and amino acids. L-Arginine is a semi-essential amino acid because it plays an important role in certain conditions, severe immune disorders and burns, which require additional intake of external L-Arginine. The amino acid L-Arginine is the sole substrate for the synthesis of nitric oxide (NO) and polyamines derived from the conversion of L- Arginine to ornithine via arginase. NO and polyamines stimulate cell proliferation and have a positive effect on progression through the cell cycle. Objective: To determine the potential of L-Arginine amino acid on the proliferation of hDPSCs. Methods: Evaluation of L-Arginine amino acid with concentrations of 300, 400, 500 μmol/L, and DMEM as a control for hDPSCs proliferation using cell count test after 24 hours. Statistical analysis using Oneway ANOVA with Bonferroni post hoc. Results: There was a significant difference in the potency of L-Arginine 300,400 and 500 μmol/L compared to control, and L-Arginine 500 mol/L had the highest average proliferation of hDPSCs of 436.666 cells/ml. Conclusion: The amino acid L-arginine has the potential to proliferate hDPSCs and the highest concentration at 500 μmol/L."

"Latar belakang: Jeruk memiliki khasiat untuk kesehatan karena megandung vitamin, antioksidan, dan senyawa lain. Jeruk purut (Citrus Hystrix) merupakan jenis jeruk yang memiliki senyawa fenol yang tinggi. Jeruk purut memilki potensi antibakteri terhadap bakteri Gram-positif dan Gram-negatif. Daun jeruk purut mengandung senyawa bioaktif seperti flavonoids, fenolik, tannin dan minyak esensial. Pada fase vegetative, kandungan flavonoid jeruk purut tertinggi pada daun tua. Efek antibakteri flavanoids adalah dengan mekanisme menghambat sintetik asam nukleat, menghambat fungsi membran sitoplasma sel, dan merubah permeabilitas membran sehingga memengaruhi sifat patogenitas bakteri Tujuan: Mendapatkan perbedaann efek antibakteri berbagai konsentrasi larutan ekstrak daun jeruk purut terhadap biofilm Enterococcus faecalis. Mendapatkan perbedaan efek antibakteri larutan ekstrak daun jeruk purut konsentrasi 2,5%, 5%, 10% dan 20% dan NaOCl 2,5% terhadap bakteri Enterococcus faecalis. Metode: Empat kelompok sampel diuji dengan larutan ekstrak daun jeruk purut masing masing 2,5%, 5%, 10%, dan 20%. Kelompok kontrol positif dilakukan pemaparan NaOCl 2,5%, dan kelompok kontrol negatif tanpa perlakuan. Efek antibakteri dilihat dari jumlah koloni pada media BHI agar. Hasil: Rerata koloni bakteri Enterococcus faecalis dari masing masing kelompok dengan nilai p=0,00 (berbeda bermakna). Nilai koloni tertinggi pada kelompok kontrol negatif dan larutan ekstrak daun jeruk purut 2,5% dan terendah pada kelompok kontrol positif dan larutan ekstrak daun jeruk purut 20%. Kelompok ekstrak daun jeruk purut dengan konsentrasi 2,5%, 5%, 10% dan 20% menunjukan perbedaan bermakna dengan kompok positif NaOCl 2,5% dan kelompok kontrol negatif. Kelompok ekstrak daun jeruk purut dengan konsentrasi 2,5% juga memiliki perbedaan bermakna dengan konsentrasi 10% dan 20%. Kesimpulan: Konsenstrasi larutan ekstrak daun jeruk purut 20% memiliki efek antibakteri Enterococcus faecalis yang paling baik dibandingkan pada konsentrasi 10%, 5%, 2,5%. Efek antibakteri larutan ekstrak daun jeruk purut 2,5%, 5%,10% dan 20% terhadap bakteri Enterococcus faecalis lebih rendah dibandingkan dengan larutan NaOCl 2,5%.

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Citrus contains vitamin, antioxidant, and other compounds that beneficial to the health. Lime (Citrus Hystrix) contains high concentration of phenol that has antibacterial potential against gram-positive and gram-negative bacterias. Lime leaf contains bioactive compounds such as flavonoids, phenolic, tannin, and essential oils. In vegetative state, old lime leaf contains the highest concentration of flavonoids. Flavonoids inhibit synthetic of nucleatic acid and citoplasmic cell membrane's function of bacteria, and affect bacterial pathogenetic by altering its membrane permeability.Objective: To obtain the difference of antibacterial effects of various lime extract concentration (2,5%; 5%; 10%; and 20%) and 2,5% of NaOCl against Enterococcus faecalis biofilm. Methods: Four sample groups tested using 2,5%; 5%; 10%; and 20% concentration of lime extract. 2,5% concentration of NaOCl was used as positive control group and no treatment was used as negative control group. Antibacterial effects were observed by the amount of bacterial colonies in BHI agar. Results: The mean of Enterococcus faecalis in each group with p=0.00 (significant). Negative control group and 2.5% lime extract concentration group had the highest amount of bacterial colonies. Positive control group and 20% lime extract concentration group had the lowest amount of bacterial colonies. All sample groups showed significant difference with positive and negative control group. 2.5% lime extract group had significant difference with group of 10% and 20% lime extract concentration. Conclusion: 20% lime extract concentration showed higher potential of antibacterial against Enterococcus faecalis than 2,5%; 5%; and 20% concentration. Antibacterial effects of lime extract in every concentration groups were lower than 2,5% NaOCl"

"Triethylene glycol dimethacrylate (TEGDMA) is a common component of the bonding agents and resin composites used in dentistry for restorative dentistry. However, TEGDMA could be released from composite resins following incomplete polymerization and degradation processes by salivary enzymes in the mouth. Subsequently, TEGDMA is available in saliva and diffuses toward and affects the dental pulp which contains various cells, and thus may cause severe cytotoxic effects. Objectives: To determine the total protein concentration of human dental pulp cells following exposure to TEGDMA. Materials & methods: Dental pulp cells were isolated from the pulp of the freshly extracted teeth and cultured in DMEM for 48 h (37°C, 5% CO2). Then, 2 mM, 4mM and 8 mM TEGDMA were added to these cells and incubated for 24 h. The total protein was measured by Bradford Protein Assay. Results: The total protein concentration of dental pulp cell after expsured to 4 mM, 8mM, and 12 mM TEGDMA were statistically lower (22762.27 ug/ml ± 3385.87; 20268.44 ug/ml ± 1701.14; 23706.51 ug/ml ± 3214.52; respectively) than the control group (24253.77 ug/ml ± 3072.88). Furthermore, the total protein concentration of culture medium after exposured to 4 mM, 8mM, and 2 mM TEGDMA, were statustically higher (28635 ug/ml ± 2373.4; 35288.41 ug.ml ± 3469.48; 38199.79 ug/ml ± 2752.47; respectively) when compared with the controls (27073.85 ug/ml ± 2772.47). Conclusion: 2 mM, 4 mM, and 8 mM TEGDMA caused cytotoxicity to human dental pulp cells showed by decreasing the total protein of cells and increasing total protein of the culture medium."

"ABSTRAKbelakang: Sambiloto Andrographis paniculata Ness. telah digunakan secara luas sebagai obat tradisional. Tanaman ini mempunyai potensi sebagai antikanker. Andrografolida sebagai senyawa aktif utama sambiloto telah terbukti bersifat sitotoksik terhadap berbagai jenis sel kanker. Evaluasi toksisitas penting dilakukan untuk memastikan keamanan andrografolida dan tanaman sambiloto. Penelitian ini bertujuan menganalisis efek andrografolida dan ekstrak/fraksi sambiloto terhadap viabilitas, siklus, serta faktor transkripsi diferensiasi sel punca mesenkimal asal sumsum tulang manusia bone marrow mesenchymal stem cells/BMMSC menjadi osteoblas. Metode: Penilaian efek andrografolida dan fraksi etil asetat sambiloto FEAS terhadap viabilitas, siklus sel serta tingkat ekspresi mRNA CDK4 dan p21 dilakukan dengan memaparkan andrografolida konsentrasi 5, 10 dan 15 g/mL dan FEAS konsentrasi 20, 40 dan 60 g/mL selama 12, 24 dan 36 jam. Viabilitas sel diperiksa berdasarkan prinsip reduksi garam formazan WST-8, analisa siklus sel menggunakan flow cytometer dan tingkat ekspresi mRNA dengan quantitative RT-PCR. Penilaian terhadap diferensiasi osteoblas dilakukan dengan menginduksi BMMSC dengan medium osteogenik disertai pemberian andrografolida konsentrasi 5 dan 10 g/mL serta FEAS konsentrasi 20 dan 40 g/mL selama 7, 14 dan 21 hari, kemudian dinilai intensitas pewarnaan alizarin red AR , kadar deposit kalsium ekstraseluler serta tingkat ekspresi runx2 dan osterik. Hasil: Andrografolida dan FEAS menurunkan viabilitas BMMSC sesuai tingkatan konsentrasi dan waktu paparan. Kedua bahan uji tersebut menghambat proliferasi BMMSC dengan meningkatkan secara bermakna persentase populasi sel pada fase G1 dan menurunkan populasi sel yang memasuki fase S dan G2 siklus sel pada paparan 24 jam. Tidak terdapat efek terhadap tingkat ekspresi mRNA CDK4 namun ekspresi mRNA p21 meningkat bermakna. Andrografolida dan FEAS menurunkan intensitas warna merah AR, kadar matriks kalsium ekstraseluler dan ekspresi mRNA runx2 secara bermakna, namun meningkatkan ekspresi mRNA osterik pada proses diferensiasi BMMSC menjadi osteoblas Kesimpulan: Andrografolida konsentrasi 5, 10 dan 15 g/mL maupun fraksi etil asetat sambiloto konsentrasi 20, 40 da 60 g/mL mempunyai efek toksik terhadap viabilitas, proliferasi dan diferensiasi osteoblas pada BMMSC secara in vitro. Kata kunci : Andrografolida, Andrographis paniculata, BMMSC, siklus sel, osteoblas.

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Background: Sambiloto Andrographis paniculata Ness./AP , has been used extensively as a traditional medicine. Andrographolide as the main active component of this plant showed cytotoxic activity in vitro on various type of cancer cells line. Toxicity evaluation is important to ensure the safety of andrographolide and this bitter plant. The objective of this study was to investigate the effects of andrographolide and the extract of AP on viability, cell cycle, and transcription factors of osteoblast differentiation on human bone marrow mesenchymal stem cells BMMSC Methods: BMMSC were treated with andrographolide at 5, 10 and 15 ?g/mL and ethyl acetate fraction of AP EAFA at 20, 40 and 60 ?g/mL for 12, 24 and 36 hours. The cells viability was assessed using tetrazolium salt WST-8 assay, the cell cycle was evaluated using flow cytometer with propidium iodide DNA?binding fluorescent dyes and the expression of CDK4 mRNA and p21 was analyzed by RT-PCR. Further examination was investigated the effects of the compounds on the osteogenic differentiation of BMMSC. The cells were cultured on osteogenic medium and treated with andrographolide at 5 and 10 ?g/mL and EAFA at 20 and 40 ?g/mL for 7, 14 and 21 days. The matrix mineralization was assessed by alizarin red-s staining AR , the semi-quantification of calcium was determined by acetic acid extraction of calcium binding AR and the expression of runx2 and osterix were analysed by RT-PCR Results: This research was revealed that andrographolide and EAFA decreased the cell viability, arrested the cell cycle at G1 phase, and up regulated the expression of mRNA p21. Moreover andrographolide and EAFA supplementation decreased the intensity of AR and calcium deposition on cell culture. The expression of transcriptor factors runx2 was down regulated while osterix was up regulated. Conclusion: Andrographolide at 20, 40 and 60 ?g/mL and EAFA at 20, 40 and 60 ?g/mL showed potentially toxic on cell viability, arrested cell cycle and impaired osteoblast differentiation of BMMSC in vitro. "