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"Latar belakang: xylitol adalah gula alkohol berantai karbon lima (polyol) yang banyak digunakan sebagai pemanis alami dalam bentuk permen karet untuk mencegah karies gigi. Xylitol memiliki efek antikaries karena dapat menghambat pertumbuhan S. mutans yang merupakan salah satu agen utama penyebab karies gigi, menurunkan pembentukan plak dan meningkatkan remineralisasi gigi. Pulpa gigi berperan penting bagi vitalitas gigi. Pada pulpa gigi yang terbuka, xylitol dapat berpenetrasi dan menimbulkan efek biologik pada sel. Tujuan: untuk mendeteksi efek xylitol terhadap viabilitas dan profil protein sel-sel pulpa gigi (in vitro). Metode: sel-sel pulpa gigi didapat dari gigi sehat yang baru diekstraksi, dan dikultur dalam medium kultur DMEM (37°C, 5% CO2) hingga confluent. Selanjutnya sel-sel tersebut disubkultur pada kondisi yang sama selama semalam di 24-wellplate. Setelah itu kelompok perlakuan dipaparkan xylitol dengan konsentrasi 2%, 4%, 8% dan 16%. Sedangkan pada kelompok kontrol tidak diberi xylitol. Viabilitas sel diukur dengan MTT assay. Sedangkan profil protein dianalisis dengan SDS PAGE. Hasil: rerata optical density (OD) kelompok xylitol 2% (1,784 ± 0,052), 4% (2,465 ± 0,057), 8% (2,168 ± 0,162), dan 16% (1,912 ± 0,148) lebih tinggi dibandingkan dengan kelompok kontrol (1,566 ± 0,069). Uji statistik Oneway ANOVA menunjukkan bahwa seluruh kelompok perlakuan berbeda bermakna dengan kontrol (p<0,05). Persentase viabilitas sel diperoleh dari rerata optical density. Viabilitas sel kelompok xylitol 2% (113,92%), 4% (157,40%), 8% (138,44%), dan 16% (122,09%) lebih tinggi dibandingkan dengan kelompok kontrol (100%). Dari hasil SDS PAGE, tampak perubahan profil protein sel-sel pulpa gigi. Simpulan: terdapat peningkatan viabilitas sel dan perubahan profil protein sel-sel pulpa gigi setelah pemaparan xylitol.

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Background: xylitol is five carbon sugar alcohol (polyol) which is used as natural sweetener in chewing gum to prevent dental caries. Xylitol has anticaries effect as it can inhibit the growth of S. Mutans, one of the main etiology of dental caries, decrease plaque formation, and increase tooth remineralization. Dental pulp has an important role in dental vitality. In exposed dental pulp, xylitol can penetrate and induce biological response of the cells. Objective: to detect the effects of xylitol to cell viability and protein profile of dental pulp cells (in vitro). Method: dental pulp cells were obtained from healthy and freshly extracted teeth, and were cultured in DMEM (37°C, 5% CO2) until confluent. Subsequently, they were subcultured in same condition overnight on 24-well plate. Afterwards, the treatment groups were exposed by 2%, 4%, 8%, and 16% xylitol. Whilst, the control group was not exposed by xylitol. Cell viability was measured by MTT assay. Whereas, the protein profile was analized by SDS PAGE. Results: the mean of optical density of treatment group with xylitol 2% (1,784 ± 0,052), 4% (2,465 ± 0,057), 8% (2,168 ± 0,162), and 16% (1,912 ± 0,148) were higher than control group (1,566 ± 0,069). Statistical test Oneway ANOVA showed that all the treatment groups were significantly different compared with the control (p<0,05). The percentage of cell viability was obtained from the mean of optical density. The cell viability of xylitol 2% (113,92%), 4% (157,40%), 8% (138,44%), dan 16% (122,09%) were higher than control group (100%). From SDS PAGE, there was protein profile alteration. Conclusion: there was an increased of cell viability and the alteration of protein profile of dental pulp cells after treated with xylitol."

"Latar belakang: Trietylene glycol dimethacrylate (TEGDMA) merupakan salah satu monomer yang terkandung dalam resin komposit. Jika polimerisasi resin komposit tidak sempurna, TEGDMA dapat terlepas ke dalam rongga mulut dalam beberapa menit hingga jam dan dapat berpenetrasi mencapai pulpa. TEGDMA dilaporkan bersifat toksik terhadap sel dan jaringan rongga mulut. Tujuan: Mengetahui efek TEGDMA terhadap sel-sel pulpa gigi ditentukan berdasarkan viabilitas dan profil protein sel pulpa (in vitro). Metode: Sel-sel pulpa berasal dari jaringan pulpa gigi sehat yang baru diekstraksi, kemudian dikultur dalam DMEM (37o C, 5% CO2) sampai confluent (± 2 malam). Selanjutnya dilakukan subkultur dengan kondisi yang sama selama 1 malam pada 24-wellplate. Kemudian pada kelompok perlakuan dipaparkan TEGDMA dengan konsentrasi 4 mM, 8 mM dan 12 mM selama 24 jam; sedangkan pada kelompok kontrol tidak dipaparkan TEGDMA. Viabilitas sel diukur dengan menggunakan MTT assay dan hasilnya dibaca dengan microplate reader (490 nm), sedangkan gambaran profil protein dideteksi dengan menggunakan SDS-PAGE dan diinterpretasikan dengan menggunakan Gel Doc. Hasil: Rerata optical density (OD) ± SD kelompok perlakuan TEGDMA 4 mM (1,71 ± 0,08); 8 mM (1,59 ± 0,11); dan 12 mM (1,50 ± 0,16) lebih rendah dibandingkan dengan kelompok kontrol (1,81 ± 0,11). Uji statistik One Way ANOVA menunjukkan bahwa nilai rerata OD kelompok TEGDMA 8 mM dan 12 mM berbeda bermakna dengan kelompok kontrol (p<0.05). Profil protein sel mengalami perubahan setelah pemaparan TEGDMA. Kesimpulan: Pada penelitian ini viabilitas sel menurun dan terjadi perubahan profil protein sel setelah pemaparan TEGDMA.

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Background: Trietylene glycol dimethacrylate (TEGDMA) is one of monomer contained in composite resin. If the polimerized was incomplete TEGDMA could bereleased into oral cavity in minutes to hours and could penetrate to the dental pulp. Itwas reported that TEGDMA has cytotoxic effects to cells and tissues in oral cavity.

Objectives: To determine the toxic effect of TEGDMA on dental pulp cells culture based on cell viability and Protein Cell Profile.

Methods: The pulp cells were isolated from the pulp tissue of the freshly extracted teeth, cultured in DMEM (37o C, 5% CO2) until confluent (± 2 nights). Afterwards, subcultured with the same condition overnight in 24-wellplate. Then, the treatment groups were treated with TEGDMA 4 mM, 8 mM, dan 12 mM for 24 hours, whereas in control group without TEGDMA exposure. The optical density of cell viability was measured by MTT assay then it was read with microplate reader in 490 nm. The protein cell profile was identified by SDS-PAGE method and analyzed by Gel Doc.

Results: Mean optical density ± SD of TEGDMA treatment group 4mM (1,71 ± 0,08), 8mM (1,59 ± 0,11), and 12 mM (1,50 ± 0,16) were lower than the control group (1,81 ± 0,11). One Way ANOVA analysis showed that TEGDMA treatment group 8 mM and 12 mM had significant differences compared with the control group (p<0,05). The protein profile of cells was altered after TEGDMA exposure.

Conclusion: In this research the cell viability was decreased and the protein profile of cells was altered after TEGDMA exposure."

"ABSTRAKLatar Belakang Giant cell tumor (GCT) merupakan tumor jinak yang bersifat lokal agresif destruksif. Tumor ini memiliki rekurensi yang tinggi sebanyak 25-50% setelah tindakan pembedahan. Berbagai macam pemberian zat kimia lokal sebagai terapi ajuvan, telah digunakan pada tatalaksana pembedahan. Namun perbandingan efektifitas untuk masing-masing zat kimia ini belum diketahui. Studi mengenai sitotoksisitas dan mekanisme kematian sel dengan membandingkan pemberian etanol dan H2O2 pada sel GCT tulang secara in vitro masih sedikit dan belum ada di Indonesia.Metode Penelitian ini merupakan studi in vitro eksperimental dengan mengambil empat sampel jaringan tumor dari pasien yang didiagnosis GCT tulang dan dilakukan isolasi-kultur sel. Cell line yang didapatkan dikarakterisasi melalui analisis morfologi serta pemeriksaan ekspresi penanda gen Nanog dan Oct 4 dengan Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Sel yang telah 80% konfluens dilakukan terapi dengan H2O2 1%, 3%, 5% dan etanol 75%, 85%, 95% selama10 menit serta dosis in vitro H2O2 (0,003%, 0,005%, 0,01%, 0,03%, 0,1%, 0,3%) selama 5 menit serta inkubasi selama 24 jam. Morfologi sel dievaluasi dibawah mikroskop cahaya dengan membandingkan kontrol dan setelah pemberian zat kimia, viabilitas sel dihitung menggunakan automatic cell counter serta toksisitas sel dinilai dengan uji Annexin V dan Propidium Iodida (PI) pada flow cytometry.Hasil Kultur jaringan sel GCT dengan metode eksplan dan kolagenase mempunyai angka keberhasilan yang sama dalam mendapatkan cell line GCT. Namun metode eksplan membutuhkan waktu yang lebih cepat dan memiliki jumlah sel yang lebih banyak. Sel yang tumbuh dari jaringan GCT terkarakterisasi dengan analisis morfologi serta ekspresi gen Oct 4 dan Nanog. Viabilitas sel GCT menurun secara signifikan setelah paparan terhadap dosis klinis H2O2 1% (p = 0,046), H2O2 3% (p = 0,043), dan H2O2 5% (p = 0,043) selama 10 menit dibandingkan dengan kontrol. Tidak ada perbedaan yang bermakna untuk viabilitas sel antara konsentrasi H2O2 1%, 3% dan 5%. Sementara pada konsentrasi in vitro (0,003%, 0,005%, 0,01%, 0,03%, 0,1%, 0,3%), konsentrasi H2O2 0,3% (p < 0,001) selama 5 menit memiliki efektivitas paling baik dalam sterilisasi GCT secara in vitro. Terdapat fenomena fiksasi sel setelah pemberian etanol pada semua konsentrasi. Dari uji RT-PCR didapatkan penurunan ekspresi gen Oct 4 dan Nanog seiring dengan peningkatan konsentrasi H2O2 pada dosis in vitro. Flow cytometry dengan marker Annexin V dan propidium iodide (PI) didominasi oleh marker PI yang menunjukkan kematian sel akibat nekrosis dengan persentase terbesar pada konsentrasi 0,3%.Kesimpulan Eksplan merupakan metode terbaik dalam isolasi dan kultur sel GCT. Semua sel hasil isolasi dan kultur terkarekterisasi sebagai GCT. Pemberian ajuvan kimia lokal dengan dosis klinis H2O2 konsentrasi 1%, 3%, dan 5% selama 10 menit secara in vitro mempunyai efektivitas yang sama dalam membunuh sel GCT. Sedangkan konsentrasi H2O2 0,3% selama 5 menit merupakan terapi optimal dalam sterilisasi GCT secara in vitro dengan mekanisme kematian nekrosis sel.

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ABSTRACTBackground Giant cell tumor (GCT) is a benign, aggressive local tumor with high tendency to recur after surgery. Various chemicals have been used as an adjuvant treatment for GCT. However, the comparative effect of these chemicals remains unclear. To date, there are no studies about the cytotoxicity and mechanism of injury to etanol and H2O2 in GCT in Indonesia especially in vitro experiment. The present study aims to find the best method to isolation and culture of GCT from primary human patients, the optimal treatment of etanol and H2O2 for reducing GCT recurrence.Methods This is an experimental in vitro study that took four tumor tissue samples from patients diagnosed with bone GCT and conducted cell-culture isolation. Cell line characterized by morphology, gene markers Nanog and Oct 4 expression with Polymerase Chain Reaction (RT-PCR) Reverse Transcriptase was obtained. Cells that had 80% confluence were treated with H2O2 1%, 3%, 5% and etanol 75%, 85%, 95% for 10 minutes and in vitro doses of H2O2 (0.003%, 0.005%, 0.01%, 0.03 %, 0.1%, 0.3%) for 5 minutes and were incubated for 24 hours. Cell morphology was evaluated under a light microscope by comparing the morphology of controls and after exposure a chemical agents, cell viability was calculated using automatic cell counter and cell toxicity was assessed by Annexin V and Propidium Iodida (PI) on flow cytometry.Results Collagenase and explant methods had the same success rate in obtaining GCT cell line characterized by morphology, the gene expression Oct 4 and Nanog. But explants need a less time and had more cell than collagenase method. Viability of GCT cells decreased significantly after exposure to the clinical dose of H2O2 1% (p = 0,046), H2O2 3% (p = 0,043), and H2O2 5% (p = 0,043) for 10 minutes compared to controls. There was no significant difference for cell viability between 1%, 3% and 5% H2O2 concentrations. While in in vitro doses (0,003%, 0,005%, 0,01%, 0,03%, 0,1%, 0,3%), 0.3% H2O2 concentration for 5 minutes has the best effectivity in sterilizing GCT in vitro. There was a phenomenon of cell fixation after exposure of GCT cells to etanol in various concentrations, in which all cells die and its viability could not be analyzed. From the RT-PCR test it was found that there was a decrease in the expression of Oct 4 and Nanog genes along with an increase in the concentration of H2O2 in vitro. Flow cytometry using Annexin V in conjunction with propidium iodide (PI) was dominated with PI marker detection which showed cell death due to necrosis, with the highest concentration amounted to 0.3%Conclusion Explant was the best method for isolation and GCT cell culture. All of the cell from isolation and culture result had a characterization of GCT. Giving local a chemical adjuvants with clinical doses of H2O2 concentrations of 1%, 3%, and 5% for 10 minutes in vitro had the same effectiveness in killing GCT cells. While the concentration of 0.3% H2O2 for 5 minutes is the optimal therapy in GCT sterilization in vitro with necrosis cell death mechanism."

"ABSTRAK Latar Belakang: Berbagai material kaping pulpa berbahan dasar kalsium silikatterus dikembangkan, diantaranya semen berbasis kalsium silikat denganpenambahan steroid. Tujuan: Menganalisis efek penambahan steroid pada semenberbasis kalsium silikat terhadap viabilitas sel fibroblas. Metode: Sel fibroblasembrio ayam direndam dalam ekstrak larutan semen berbasis kalsium silikatdengan penambahan steroid dan MTA. Viabilitas sel dihitung dengan menggunakan uji MTT. Hasil: Terdapat perbedaan bermakna (p ≤ 0,05)viabilitas sel pada kelompok semen berbasis kalsium silikat dengan penambahansteroid dibandingkan kelompok kontrol dan MTA. Kesimpulan: Penambahansteroid menurunkan viabilitas sel. Terdapat peningkatan pada 72 jam, yang menandakan terjadinya proliferasi sel.ABSTRACT Background: Calcium silicate based materials are being developed continuously,one of them is calcium silicate based cement containing steroid. Objective: Toanalyze the effect of steroid addition in calcium silicate based cement onfibroblast cells viability. Methods: Chicken embryonic fibroblast cells wereimmersed in extract solution of calcium silicate based cement containing steroidand MTA. Viability was analyzed by MTT Assay. Results: Significant difference (p ≤ 0,05) of viability on calcium silicate based cement containing steroid groupwas found, compared to control and MTA group. Conclusion: Steroid additiondecrease viability. There was an increase in 72 hours, marking cells proliferation.;Background: Calcium silicate based materials are being developed continuously,one of them is calcium silicate based cement containing steroid. Objective: Toanalyze the effect of steroid addition in calcium silicate based cement onfibroblast cells viability. Methods: Chicken embryonic fibroblast cells wereimmersed in extract solution of calcium silicate based cement containing steroidand MTA. Viability was analyzed by MTT Assay. Results: Significant difference (p ≤ 0,05) of viability on calcium silicate based cement containing steroid groupwas found, compared to control and MTA group. Conclusion: Steroid additiondecrease viability. There was an increase in 72 hours, marking cells proliferation.;Background: Calcium silicate based materials are being developed continuously,one of them is calcium silicate based cement containing steroid. Objective: Toanalyze the effect of steroid addition in calcium silicate based cement onfibroblast cells viability. Methods: Chicken embryonic fibroblast cells wereimmersed in extract solution of calcium silicate based cement containing steroidand MTA. Viability was analyzed by MTT Assay. Results: Significant difference (p ≤ 0,05) of viability on calcium silicate based cement containing steroid groupwas found, compared to control and MTA group. Conclusion: Steroid additiondecrease viability. There was an increase in 72 hours, marking cells proliferation."

"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"

"Pit Fissure Sealant berbahan resin merupakan salah satu produk pencegahan karies. Pada penelitian sebelumnya, ditunjukkan adanya pelepasan komponen dan material tersebut ke lingkungannya yang menimbulkan respon hipersensitifitas. Tujuan : untuk mengetahui biokompatibilitas dari Resin Pit Fissure Sealant terhadap sel keratinosit kulit yang dicerminkan dari viabilitas sel HaCaT. Material dan Metode: Spesimen Resin Pit & Fissure Sealant dibuat pada cetakan akrilik (N=18; diameter 15mm; ketebalan 1mm) menurut ISO 4049 dan dipolimerisasi dengan UV dari QTH (Quartz Tungsten Halogen) ( = 400 nm). Spesimen dipersiapkan dan disterilisasi untuk menghindari kontaminasi dari bakteri atau jamur. Setelah itu, spesimen direndam dalam DMEM (5mL) dan disimpan dalam inkubator (370C) selama 1, 2, dan 7 hari. Kultur sel dipersiapkan pada 96 well dan diinkubasi selama 24 jam. Rendaman spesimen dipaparkan ke setiap well dan diuji tingkat viabilitas selnya menggunakan MTT assay. Tingkat viabilitas sel diukur dengan microplate reader = 490 nm. Signifikansi diukur dengan metode analisis ragam satu arah Anova. Hasil : Viabilitas sel menurun pada hari pertama dan setelah hari kedua. Kesimpulan : Waktu perendaman mempengaruhi viabilitas sel, tetapi masih cukup aman untuk digunakan untuk perawatan gigi.

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Resin based Pit Fissure Sealant is one of dental caries prevention product. Previous research of resin showed that some components leached into aqueous environment and cause hypersensitivity responses.

Objectives: To observe the biocompatibility of Resin Pit Fissure Sealant due to skin keratinocytes which is determined by viability of HaCaT Cell lines.

Material & Methods : Resin based Pit Fissure Sealant were made in acrylic mould (N= 18; diameter 15mm; thickness 1mm) according to ISO 4049 and polymerized by UV light from QTH (Quartz Tungsten Halogen) ( = 400 nm). Specimen were prepared and sterilized to avoid contamination from bacterial or germs. After that, Specimens were immersed in DMEM (5mL) and stored in incubator (370C) for 1, 2, and 7 days. Cell Culture were prepared into 96 well and stored in incubator for 24 hours. The elution of specimens was exposed into every well, and examined the viability of cells by MTT assay. Viability Cell were counted in 490 nm microplate reader. Significance were measured by One Way Anova.

Results : The viability of HaCaT Cell Lines were decreased in first and after second days.

Conclusion : The elution time of Resin based Pit Fissure Sealant affect the viability of HaCat Cell line, but still safe to be used in dental clinic."

"Ekstrak mangostin sebagai biomaterial baru yang ditambahkan pada scaffold karbonat apatit / alginat / kitosan diharapkan dapat membunuh sisa sel kanker pada tulang setelah dilakukan tindakan pembedahan. Semua material yang akan digunakan pada tubuh harus memenuhi syarat untuk dapat diterima jaringan dan tidak menimbulkan reaksi toksik pada tubuh. Tujuan penelitian ini untuk memperoleh informasi mengenai viabilitas sel terhadap scaffold komposit karbonat apatit / alginat / kitosan dengan ekstrak mangostin. Scaffold dibuat dari bahan serbuk karbonat apatit / alginat / kitosan dengan ekstrak mangostin menggunakan metode freeze drying. Karakterisasi dilakukan dengan XRD, FTIR, SEM dan dilanjutkan dengan viabilitas sel menggunakan MTT assay pada konsentrasi 10%, 7.5%, 5%, 2.5%, 1% atau 0.5%. Hasil penelitian diperoleh porositas scaffold komposit karbonat apatit / alginat / kitosan dengan ekstrak mangostin berkisar 50 ? 300 μm. Diperoleh viabilitas tertinggi dari scaffold komposit karbonat apatit / alginat / kitosan dengan ekstrak mangostin pada konsentrasi 0.5 %. Terdapat perbedaan bermakna antara viabilitas sel dari sampel scaffold karbonat apatit / alginat / kitosan dibandingkan terhadap viabilitas sel dari sampel scaffold karbonat apatit / alginat / kitosan dengan ekstrak mangostin pada konsentrasi 10 % dan 7.5 % (P<0.05) tetapi tidak memberikan perbedaan bemakna pada konsentrasi 5%, 2.5%, 1% dan 0.5% (P>0.05). Dapat disimpulkan scaffold komposit karbonat apatit / alginat / kitosan dengan ekstrak mangostin mempengaruhi viabilitas sel sehingga bersifat sitotoksik.

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Mangosteen Extract as a new biomaterial to be added to carbonate apatite scaffold / alginate / chitosan is expected to remove any remaining cancer cells in the bones after surgery. All materials which will used on the body must eligible to be accepted by system and does not cause toxic reactions in the body. The purpose of this study was obtaining information on cell viability against carbonate apatite composite scaffold / alginate / chitosan with mangosteen extract. The scaffold is made of carbonate apatite powder / alginate / chitosan with extract mangosteen using freeze drying method. Characterization are performed by XRD, FTIR, SEM and followed by cell viability using MTT assay at concentrations of 10%, 7.5%, 5%, 2.5%, 1% or 0.5%. The results were obtained composite scaffold porosity carbonate apatite / alginate / chitosan mangosteen extract range 50-300 lm. Obtained the highest viability of carbonate apatite composite scaffold / alginate / chitosan with mangosteen extract at a concentration of 0.5%. There is a significant difference between the cells viability of composite of scaffold carbonate apatite / alginate / chitosan samples compared to the cells viability of composite of scaffold carbonate apatite / alginate / chitosan with mangostin extract samples at concentrations of 10% and 7.5% (p<0.05), but no difference significant in the concentration 5%, 2.5%, 1% and 0.5% (p>0:05). It can be concluded composite of scaffold carbonate apatite / alginate / chitosan with mangostin extract affect the viability of cells that are cytotoxic."

"Latar Belakang: Human platelet lysate (HPL) telah menjadi alternatif pengganti fetal bovine serum (FBS) dalam kultur sel. Penggunaan platelet yang telah melewati masa simpan belum diketahui dalam kultur HUVEC. Tujuan: Mengetahui pengaruh masa simpan platelet pada HPL terhadap efektivitas pengganti FBS dalam kultur HUVEC. Metode: Sel HUVEC dikultur dengan FBS, HPL fresh, dan HPL extended dan diuji dengan uji MTT dan Bradford. Hasil: Suplementasi HPL menghasilkan viabilitas dan protein total yang lebih tinggi dibandingkan dengan FBS. Viabilitas tertinggi dihasilkan oleh HPL extended dan protein total tertinggi dihasilkan HPL fresh. Simpulan: HPL extended dapat digunakan sebagai pengganti FBS dalam kultur HUVEC.

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Background: Human platelet lysates (HPL) has been proved to be FBS alternatives in cell culture. Platelet that passes its shelf-life in HUVEC culture has not been studied previously.

Objective: To evaluate the effectiveness of platelet shelf-life to HPL as FBS substitute in HUVEC culture.

Method: HUVEC cultured with FBS, HPL fresh, and HPL extended were tested by MTT and Bradford assay.

Result: Cultures supplemented with HPL has higher cell viability and total protein than FBS supplemented. HUVEC cultured with HPL extended presents the highest cell viability. However, HPL fresh has the highest total protein concentration.

Conclusion: HPL extended can be used as a substitute to FBS in HUVEC culture."

"Lingkungan mikro tumor berperan penting dalam meregulasi sifat kepuncaan, proliferasi, ketahanan terhadap apoptosis, dan metabolisme sel punca kanker. Penelitian ini bertujuan untuk menganalisis efek modulasi lingkungan ekstraseluler melalui kondisi hipoksia dan alkalinisasi pada metabolisme glukosa dan ketahanan hidup sel punca kanker CSC payudara manusia CD24-/CD44 . Pada penelitian in vitro eksperimental ini, CSC payudara manusia dikultur pada kondisi hipoksia dan kondisi alkali. Kultur sel diinkubasi selama 30 menit, 4, 6, 24, dan 48 jam pada suhu 37 C kemudian dilakukan analisis status metabolisme glukosa, regulasi pH, ketahanan hidup, dan penanda kepuncaan serta pluripotensi CSC payudara menggunakan berbagai teknik yaitu qRT-PCR, kolorimetri, fluorometri, dan aktivitas enzimatik. Kondisi hipoksia menyebabkan peningkatan ekspresi mRNA dan konsentrasi HIF1? sehingga mengaktivasi gen-gen yang berada di bawah regulasinya. Hipoksia juga menyebabkan penekanan proliferasi namun meningkatkan ketahanan terhadap apoptosis. Alkalinisasi menyebabkan peningkatan pH ekstraseluler pHe yang menstimulasi peningkatan aktivitas dan ekspresi mRNA gen regulator pH seluler. Status metabolisme menunjukkan peningkatan aktivitas glikolisis anaerobik disertai peningkatan ekspresi transporter GLUT1. Alkalinisasi menyebabkan penekanan proliferasi CSC payudara bahkan kematian sel. Sebagai kesimpulan, modulasi lingkungan ekstraseluler baik melalui hipoksia maupun alkalinisasi dapat meningkatkan aktivitas glikolisis yang selanjutnya mempengaruhi ketahanan hidup dan kepuncaan CSC payudara CD24-/CD44.

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This study was aimed to analyze the effect of extracellular pH and O2 level modulation on glucose metabolism and survival of the human CD24 CD44 breast cancer stem cells BCSCs . The primary BCSCs CD24 CD44 cells were cultured under hypoxia 1 O2 or under supplementation of sodium bicarbonate 100 mM for various periods. After each incubation periods, the pH regulation, glucose metabolism, survival, stemness and pluripotency markers were analyzed using various techniques including qRT PCR, colorimetry, fluorometry, dan enzymatic reactions. This study demonstrated that hypoxia caused an increase of HIF1 mRNA expression and protein level, and shifted metabolic states to be more glycolytic. Hypoxia also promoted the suppression of cell proliferation and induced the apoptosis evasion. Alkalinization caused a high pHe then stimulated an increase of mRNA and activity of cellular pH regulator that lead to upregulation of anaerobic glycolysis. Alkalinization inhibited BCSCs proliferation and promoted apoptosis. To conclude, modulation of the extracellular environment of human BCSCs through hypoxic condition and alkalinization could shift the metabolic state toward the anaerobic glycolysis which in turn affected the proliferation and survival."

"Latar Belakang: Efektivitas Human Platelet Lysate (HPL) tanpa penambahan faktor pertumbuhan (GF) sebagai suplemen dalam medium HUVEC belum diketahui. Tujuan: Menilai pengaruh HPL tanpa penambahan GF dalam medium kultur HUVEC terhadap viabilitas sel dan konsentrasi protein total. Metode: HUVEC dikultur dengan HPL dengan dan tanpa GF dan FBS serta diuji dengan MTT dan Bradford. Hasil: Viabilitas sel dan konsentrasi protein kelompok HPL tanpa GF lebih tinggi daripada kelompok FBS. Viabilitas sel kelompok HPL 2% tanpa GF lebih rendah daripada kelompok HPL 2% dengan GF. Konsentrasi protein total kelompok HPL 5% tanpa GF lebih tinggi daripada kelompok HPL 2% tanpa GF. Simpulan: HPL tanpa penambahan GF dapat menggantikan suplemen medium standar pada kultur HUVEC.

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Background: Effectiveness of Human Platelet Lysate (HPL) without additional growth factors (GF) as supplement for HUVEC culture medium was unknown.

Objective: To evaluate the effect of HPL without additional GF on cell viability and total protein of HUVEC.

Method: HUVEC cultured in medium supplemented with HPL with or without GF and FBS were examined by MTT and Bradford assay.

Results: Cell viability and total protein of HUVEC cultured in HPL without GF was higher than in FBS supplemented. Cell viability of HUVEC cultured in HPL 2% without GF was lower than HPL with GF supplemented. Total protein of HUVEC cultured in HPL 5% without GF was higher than HPL without GF supplemented.

Conclusion: HPL without GF can be used as a supplement substitute in HUVEC culture medium."