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"Latar Belakang: Terjadinya regenerasi pada proses penyembuhan luka pulpa yang mengalami cedera akan menggantikan struktur dan fisiologis jaringan sama dengan aslinya. Proses ini dimulai dengan sel punca pulpa bermigrasi ke tempat cedera dan berfungsi. Ketika ada invasi bakteri, lingkungan pulpa terinflamasi melepaskan berbagai sinyal termasuk sinyal yang memicu migrasi sel punca pulpa. Pentingnya proses migrasi pada penyembuhan jaringan pulpa yang terinflamasi, maka pada penelitian ini mengamati perbedaan kemampuan migrasi pada hDPSCs normal dan terinflamasi lipopolisakarida (LPS) bakteri E. coli dengan waktu observasi 6 jam dan 24 jam. Tujuan: Mengetahui perbedaan kemampuan migrasi pada hDPSCs normal dan terinflamasi yang dilihat dari laju kecepatan migrasi dan lebar luka hDPSCs pada hDPSCs normal dibandingkan dengan hDPSCs terinflamasi dengan waktu observasi 6 jam dan 24 jam. Metode: Penelitian ini merupakan penelitian eksperimental laboratorik in vitro dengan pengamatan migrasi menggunakan metode scratch assay. Hasil: Terdapat perbedaan bermakna laju kecepatan migrasi antara hDPSCs normal dan terinflamasi pada waktu observasi 6 dan 24 jam (p<0.05). Terdapat perbedaan bermakna lebar luka hDPSCs normal dan inflamasi pada waktu observasi 6 dan 24 jam (p<0.05). Kesimpulan: Hasil penelitian ini menunjukkan pulpa tetap memiliki potensi alamiah dalam menginduksi migrasi pada kondisi terinflamasi LPS bakteri E. coli pada periode waktu 24 jam.

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Background: Regeneration in the injured pulp wound healing process will replace its structure and tissue physiology to be the same as the original. It begins with hDPSCs migrating to the injured site and functioning. When there is a bacterial invasion, the inflamed pulp environment releases various signals stimulating hDPSCs migration. Due to the importance of the migration process in inflamed pulp tissue wound healing, this research observed the differences in migration capability of the normal and inflamed-with lipopolysaccharide (LPS) bacteria E. coli- hDPSCs. Objective: To discover the differences in migration capability between normal and inflamed hDPSCs observed from differences in migratory speed rate and wound width of normal and inflamed hDPSCs at 6 and 24 hours observation time. Methods: This research was an experimental laboratory in vitro using the scratch assay. Results: There were significant differences in migratory speed rate between normal and inflamed hDPSCs at 6 and 24 hours (p<0.05). There were significant differences in wound width in each group of normal and inflamed hDPSCs at 6 and 24 hours (p<0.05). Conclusion: These research results show that pulp remains have the natural potential to induce migration in conditions inflamed by LPS bacteria E. coli for 24 hours."

"Latar Belakang: Rekayasa jaringan tulang memerlukan tiga komponen utama, yaitu sel punca, scaffold, dan faktor pertumbuhan. IGF-1 merupakan salah satu faktor pertumbuhan yang berperan dalam proliferasi dan diferensiasi sel osteoblast. IGF-1 akan berikatan dengan reseptornya, yaitu IGF-1R untuk mengaktivasi jalur hilir. Dalam sirkulasi tubuh manusia, IGF berikatan dengan IGFBP-3 yang dapat memperpanjang waktu paruh serta menghambat IGF-1 berikatan dengan IGF-1R. Pada penelitian sebelumnya, tercatat bahwa tidak ada perbedaan kemampuan proliferasi dan diferensiasi antara DPSC subjek normal dan subjek CLP, namun ada perbedaan signifikan dalam jumlah ekspresi IGF-1. OCT-4, SOX-2 dan NANOG merupakan faktor transkripsi utama pluripotensi yang telah diteliti dapat mengatur pluripotensi, pembaruan diri, proliferasi, serta diferensiasi DPSC. Penelitian terbaru mencatat peningkatan ekspresi ketiga gen tersebut pasca dilakukan penghambatan jalur GSK-3 dan m-TOR yang merupakan jalur hilir dari aksi IGF-1 pada sel DPSC. Namun, belum diketahui secara pasti ekspresi ketiga gen tersebut pada DPSC subjek normal dan CLP setelah dilakukannya penghambatan IGF-1 menggunakan anti IGF-1R dan IGFBP-3. Tujuan: Menganalisis pengaruh anti IGF-1 dan IGFBP-3 terhadap ekspresi gen OCT4, SOX2, dan NANOG pada DPSC subjek normal dan CLP. Metode: Sampel RNA DPSC subjek normal (n=4) dan DPSC subjek CLP (n=3), sebelum dan setelah diberikan perlakuan anti IGF-1R atau IGFBP-3, diperoleh dari bahan biologis tersimpan di Laboratorium Oral Biologi Fakultas Kedokteran Gigi Universitas Indonesia. Selanjutnya, ekspresi gen OCT4, SOX2, NANOG, dan housekeeping gene GAPDH diuji dengan two step Real-Time PCR (RT-PCR). Hasil: Tidak terdapat perbedaan ekspresi gen OCT4, SOX2, dan NANOG, baik antara DPSC subjek normal dan CLP sebelum dan setelah diberikan perlakuan anti IGF-1R dan IGFBP-3 (p³0,05). Kesimpulan: Perlakuan anti IGF-1R dan IGFBP-3 tidak memengaruhi tingkat ekspresi gen OCT4, SOX2, dan NANOG sel punca pulpa gigi permanen subjek normal dan subjek celah bibir dan palatum

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Background: Bone tissue engineering requires three main components, namely stem cells, scaffold, and growth factors. IGF-1 is a growth factor that plays role in osteoblast proliferation and differentiation. IGF-1 will bind to its receptor, namely IGF-1R, to activate the downstream pathway. In the human body circulation, IGF binds to IGFBP-3 which can inhibit IGF-1 from binding to IGF-1R. Previous studies noted that there were no differences in the ability to proliferate and differentiate between DPSC from normal subjects and CLP subjects, yet there were significant differences in the level of IGF-1 expression. OCT-4, SOX-2 and NANOG are core pluripotency factors which regulate pluripotency, self-renewal, proliferation and differentiation of DPSC. Recent study has noted an increase in the expression of these three genes after inhibition of GSK-3 and m-TOR pathways, which are the downstream pathways of IGF-1 on DPSC cells. However, the expression of these three genes in DPSC from normal and CLP subjects after inhibition of IGF-1 using anti IGF-1R and IGFBP-3 is still unknown. Objective: To analyze the effect of anti IGF-1 and IGFBP-3 on OCT4, SOX2, and NANOG gene expression in DPSC of normal and CLP subjects. Methods: RNA samples of DPSC from normal and CLP subjects, before and after being treated with anti-IGF-1R or IGFBP-3, were obtained from Laboratory of Oral Biology, Faculty of Dentistry, Universitas Indonesia. Furthermore, the expression of OCT4, SOX2, NANOG, and housekeeping gene GAPDH were tested using two step Real-Time PCR (RT-PCR). Results: There was no difference between the expression of the OCT4, SOX2, and NANOG in DPSC from normal and CLP subjects before and after anti IGF-1R and IGFBP-3 treatment (p≥0.05). Conclusion: Anti-IGF-1R and IGFBP-3 did not affect the expression level of OCT4, SOX2, and NANOG in dental pulp stem cells of normal subjects and cleft lip and palate subjects.

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"Latar Belakang: Subjek celah bibir dan palatum membutuhkan perawatan rekonstruksi tulang berbasis rekayasa jaringan dengan menggunakan sel stromal mesenkim. Sel stromal mesenkim merupakan sel yang banyak digunakan untuk regenerasi tulang karena mempunyai kemampuan proliferasi tinggi. Sel tersebut dapat berasal dari pulpa gigi sulung (SHED) danpulpa gigi permanen (DPSCs) yang dapat berdiferensiasi menjadi osteoblas. Pada penelitiansebelumnya telah ditemukan beberapa karakteristik DPSCs dan SHED pada subjek celah bibir dan palatum, namun kemampuan diferensiasi dari sel stromal pulpa subjek celah bibir dan palatum belum diketahui. Tujuan: Mengevaluasi kemampuan diferensiasi osteogenik dari sel stromal pulpa gigi permanen dan sulung pada subjek celah bibir dan palatum melaluiekspresi gen Collagen Type I Alpha I (COL1A1). Metode : Sampel RNA yang diperoleh dari kultur RNA DPSCs dan SHED subjek celah bibir dan palatum, dengan Real-Time Polymerase Chain Reaction (RT-PCR) menggunakan primers Collagen Type I Alpha I (COL1A1), serta 18S sebagai housekeeping gene. Hasil : Tidak terdapat perbedaan ekspresi relatif gen COL1A1 antara sel stromal pulpa gigi permanen dan sel stromal pulpa gigi sulung pada subjek celah bibir dan palatum. Kesimpulan : SHED memiliki kemampuan diferensiasi osteogenik yang sama dengan DPSCs karena keduanya dapat mengekspresikan gen marker osteogenik COL1A1.

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Background: Cleft lip and palate subject need bone reconstruction based tissue engineering treatment with mesenchymal stromal cells (MSC). One of the most mesenchymal stromal cells that can be used is derived from dental pulp tissues, such as primary tooth pulp or stem cells from human deciduous teeth (SHED) and dental pulp stem cells (DPSCs) which can differentiate into osteoblasts. In previous studies, several characteristics of DPSCs and SHED of the cleft lip and palate subjects have been found. However, osteogenic differentiation ability of dental pulp stromal cells from cleft lip and palate subject is unknown.

Objective: To determine the osteogenic differentiation ability of DPSCs and SHED of cleft lip and palate subjects through the expression of the Collagen Type I Alpha I (COL1A1) gene.

Methods: RNA samples obtained from the culture of DPSCs and SHED of lip and palate cleft subjects, with Real-Time Polymerase Chain Reaction (RT-PCR) using primers Collagen Type I Alpha I (COL1A1) and 18S as a housekeeping gene.

Results: There was no difference in the relative expression of COL1A1 gene between DPSCs and SHED of CLP subjects.

Conclusion: SHED has the same osteogenic differentiation ability as DPSCs because they can express osteogenic marker genes COL1A1."

"Latar Belakang: Rekayasa jaringan merupakan alternatif untuk perawatan rekonstruksi tulang alveolar pasien celah bibir dan palatum. Alternatif tersebut menghubungkan penggunaan sel punca, biomaterial/scaffolds, dan molekul sinyal. Sumber sel yang ideal untuk rekayasa jaringan adalah sel autologous karena tidak bersifat immunogenik. Sel stromal pulpa gigi permanen (DPSC) menarik untuk terapi klinis karena akses perolehannya yang mudah, morbiditas yang sangat rendah, menunjukkan kapasitas imunoregulasi yang menguntungkan, dan dapat berdiferensiasi menjadi banyak tipe sel, termasuk osteoblas. Pada penelitian sebelumnya, DPSCs pasien celah bibir dan palatum ditemukan memiliki potensi kemampuan osteogenik. Namun, kemampuan diferensiasi osteogeniknya belum diketahui. Kemampuan diferensiasi osteogenik tersebut dapat diamati dari ekspresi marker osteogenik, salah satunya sclerostin yang diekspresikan pada tahap akhir diferensiasi osteoblas. Tujuan: Membandingkan kemampuan diferensiasi osteogenik DPSCs pasien celah bibir dan palatum dengan DPSCs subjek normal melalui pengamatan ekspresi gen sclerostin. Metode: DPSCs dikultur hingga mencapai 70%-80% confluent. Sampel RNA dari sel diperoleh dengan melakukan prosedur ekstraksi RNA. Ekspresi gen sclerostin diamati menggunakan Real-Time PCR menggunakan primer sclerostin dan 18s sebagai housekeeping gene. Hasil: DPSCs pasien celah bibir dan palatum memiliki nilai rata-rata ekspresi relatif gen sclerostin yang lebih tinggi 1,9 kali lipat dibandingkan dengan DPSCs subjek normal dan secara statistik berbeda bermakna dengan p = 0,013. Kesimpulan: DPSCs pada pasien celah bibir dan palatum mengekspresikan gen sclerostin sebagai marker diferensiasi osteogenik yang lebih tinggi dibandingkan DPSCs pada subjek normal secara in vitro.

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Background: Tissue engineering is an alternative for alveolar bone reconstruction treatment in cleft lip and palate (CLP) patients. The alternative links the use of stem cells, biomaterials/scaffolds, and signaling molecules. The ideal cell source for tissue engineering is autologous cells because they are not immunogenic. Dental pulp stromal cells (DPSC) are interesting for clinical therapy because of their easy accesses, very low morbidity, exhibit favorable immunoregulatory capacities, and can differentiate into many cell types, including osteoblasts. In a previous study, DPSCs in CLP patients were found to have a potential osteogenic ability. However, its osteogenic differentiation ability is not yet known. The ability of osteogenic differentiation can be observed from the expression of osteogenic markers, one of which is sclerostin, a marker that is expressed in the final stage of osteoblast differentiation. Objective: To compare osteogenic differentiation ability of DPSCs in CLP patients with DPSCs in normal subjects through the expression of sclerostin gene. Methods: DPSCs were cultured to reach 70%-80% confluent. RNA samples from cells were obtained by carrying out RNA extraction procedure. Sclerostin gene expression was assessed using Real-Time PCR using sclerostin primer and 18s as a housekeeping gene. Results: DPSCs from CLP patients have mean relative expression of sclerostin gene 1.9 times higher compared to DPSCs in normal subjects and it is statistically different with p = 0.013. Conclusions: DPSCs in CLP patients express the sclerostin gene as marker of osteogenic differentiation higher than DPSCs in normal subjects in vitro."

"Tujuan dari penelitian ini adalah untuk menganalisis pengaruh Platelet-Rich Plasma (PRP) Eksosom terhadap potensi regenerasi jaringan pulpa gigi dengan evaluasiin vitro viabilitas sel, aktivitas migrasi, dan ekspresi Vascular Endothelial Growth Factor-A (VEGF-A) sel punca pulpa gigi manusia (hDPSCs). HDPSC diambil darisembilan gigi molar tiga dari sembilan donor sesuai kriterian inklusi, dan isolasi serta kultur dilakukan dengan metode enzyme digestion (ED) yang dipanen antara P3 dan P4. Setelah starvatation, hDPSCs dikultur di dalam enam media, yaitu sebagai berikut: Dulbecco's Modified Eagle Medium (DMEM) dan 10% PRP sebagai kelompok kontrol, dan 0,5%, 1%, dan 5% PRP eksosom sebagai kelompok eksperimental. Semua kelompok memiliki tiga rangkap biologis (Triplo). Uji viabilitas sel dievaluasi dengan MTT assay, aktivitas migrasi sel dengan Scratch Assay dan Transwell Migration Assay, dan ekspresi VEGF-A dengan Enzyme- Linked Lmmunosorbent Assay (ELISA). Analisis data dilakukan dengan uji One Way ANOVA (p <0,05) serta uji Kruskal-Wallis dan post hoc Mann-Whitney (p<0,05). Nilai rata-rata viabilitas hDPSCs tertinggi pada 24, 48 dan 72 jam observasi pada kelompok PRP-Eksosom 5% (p <0,05). PRP Eksosom 5% menunjukkan aktivitas migrasi yang lebih tinggi dibandingkan dengan kelompok lain, meskipun tidak terdapat perbedaan bermakna dengan kontrol PRP 10% (p> 0,05). Ekspresi VEGF-A hDPSCs tertinggi terdapat pada kelompok PRP Eksosom 5% pada 72 jampengamatan. Dapat disimpulkan bahwa eksosom PRP 5% berpotensi menginduksi regenerasi pupa gigi manusia.

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The purpose of this study was to analyze the effect of Exosome Platelet-Rich Plasma (PRP) on their potential for human Dental Pulp regeneration by evaluating in vitro cell viability, migration activity, and expression of Vascular Endothelial Growth Factor-A (VEGF-A) of human Dental Pulp Stem Cells (hDPSCs). hDPSCs was taken from nine third molars from nine donors thatfit to the inclusion criteria of this study, isolation and culture were carried out by the enzyme digestion (EZ) method harvested between P3 and P4. After starvatation,hDPSCs were cultured in six media, namely as follows: Dulbecco's Modified EagleMedium (DMEM) and 10% PRP as the control group, and 0.5%, 1%, and 5% PRP exosomes as experimental groups. All groups had a biological triplication (Triplo). Cell viability test was evaluated by MTT assay, cell migration activity with Scratch Assay and Transwell Migration Assay, and VEGF-A expression by Enzyme- Linked Immunosorbent Assay (ELISA). Data analysis was performed using One Way ANOVA (p <0.05) and Kruskal-Wallis (p <0.05) and Pearson/Spearman Correlation test (p<0.05). The highest mean hDPSCs viability was at 24, 48 and 72 hours of observation in the PRP-exosome group 5% (p <0.05). Exosome PRP 5% showed higher migration activity compared to other groups, although there was no significant difference with PRP control 10% (p> 0.05). The highest expression of VEGF-A hDPSCs was found in the PRP exosome group 5% at 72 hours of observation. It can be concluded that the PRP 5% exosome have the highest potential ability in inducing pulp regeneration."

"Latar belakang: Inflamasi adalah respon imun untuk mempertahankan inang terhadap patogen atau kerusakan. Namun, inflamasi yang tidak diatur dapat menyebabkan banyak penyakit. Peradangan terutama dimediasi oleh faktor kekebalan bawaan, termasuk sitokin, kemokin, dan sel kekebalan bawaan. Sitokin adalah mediator peradangan. Lipopolisakarida (LPS) adalah stimulator pulpitis yang kuat yang telah ditemukan pada jaringan pulpa yang meradang, menyebabkan pelepasan sitokin inflamasi termasuk IL-6, TNF-α dan IL-1β. Eksosom muncul sebagai strategi terapi mutakhir untuk mengobati sistem kekebalan tubuh akibat inflamasi. Sebagian besar sel mengeluarkan eksosom, yang dapat memiliki aktivitas imunomodulator yang kuat, tergantung pada jenis sel tempat mereka berasal. Sel punca mesenkim Wharton’s Jelly (hWJ-MSC) yang berasal dari tali pusar telah disarankan untuk mengurangi peradangan, sebagian melalui pelepasan eksosom.Tujuan: Untuk menyelidiki efek eksosom Wharton's Jelly pada konsentrasi 0,5%, 1% dan 5% pada sel punca pulpa gigi manusia yang terpapar LPS (hDPSC) pada waktu pengamatan 24, 48 dan 72 jam (analisis ekspresi IL-6)Metode: Menghitung jumlah ekspresi IL-6 pada human dental pulp stem cell (hDPSCs) yang terpapar LPS yang telah dikultur terlebih dahulu kemudian diberikan eksosom Wharton’s jelly dengan masing-masing konsentrasi 0,5%, 1%, 5% dan dibandingkan dengan kelompok kontrol tanpa pemberian eksosom pada waktu pengamatan 24 jam, 48 jam, dan 72 jam dengan analisis ELISA. Data dianalisis menggunakan analisis varian (ANOVA) satu arah dengan uji lanjut LSD.Hasil : Terdapat penurunan ekspresi IL-6 pada human dental pulp stem cells (hDPSCs) yang dipapar LPS menggunakan eksosom Wharton's Jelly dengan konsentrasi 1% dan 5% pada waktu pengamatan 48 jam.Kesimpulan: Eksosom Wharton's Jelly dapat menurunkan ekspresi IL-6 pada HDPSC yang terpapar LPS

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Background: Inflammation is an immune response that defends the host against pathogens or damage. However, unregulated inflammation can cause many diseases. Inflammation is primarily mediated by innate immune factors, including cytokines, chemokines, and innate immune cells. Cytokines are the mediators of inflammation. Lipopolysaccharide (LPS) is a potent stimulator of pulpitis that has been found in inflamed pulp tissue, causing the release of inflammatory cytokines including IL-6, TNF-α dan IL-1β. Exosomes are emerging as a state-of-the-art therapeutic strategy for treating an overactive immune system. Most cells secrete exosomes, which can have a potent immunomodulatory activity, depending on the type of cell from which they originate. Human Wharton’s jelly mesenchymal stem cells (hWJ-MSC) derived from the umbilical cord have been suggested to reduce inflamation, in part through the release of extracellular vesicle-like exosomes.

Objective: To Investigate the effect of Wharton's Jelly exosomes at concentrations of 0.5%, 1% and 5% on LPS exposed human dental pulp stem cell (hDPSCs) at observation times of 24, 48 and 72 hours (IL-6 expresion analysis)

Methods: Calculating the amount of IL-6 expression in lps exposed human dental pulp stem cells that have been cultured first and then given wharton jelly exosomes with each concentration of 0.5%, 1%, 5% and compared with the control group without exosome administration at observation time 24 hours 28 hours and 72 hours using ELISA analysis. Data were analyzed using a one‐way analysis of variance (ANOVA) with LSD’S post‐test.

Result : There is a decrease in IL-6 expression in lps exposed human dental pulp stem cells (hDPSCs) using Wharton's Jelly exosomes with a concentration of 1% and 5% at 48 hours observation time.

Conclusions: Wharton's Jelly Exosome decrese IL-6 expresion on LPS‐exposed HDPSCs."

"Latar Belakang: Sumber sel stromal yang paling ideal digunakan dalam rekayasa jaringan adalah sel stromal pulpa gigi permanen (DPSC) dan sel stromal pulpa gigi sulung (SHED) dikarenakan sifat proliferasinya yang tinggi. Pada penelitian sebelumnya, dinyatakan bahwa terdapat peningkatan ekspresi gen homeobox salah satunya yaitu gen ALX4 sebagai pada pasien celah bibir dan palatum dengan subjek normal. Gen ALX4 adalah gen homeobox dibawah famili Alx dan memiliki peran langsung dalam perkembangan dan pembentukan kepala serta wajah serta mentranslasi protein yang meregulasi perkembangan dan proliferasi sel, pendewasaan dan diferensiasi sel, pergerakan sel, dan pertahanan sel. Namun, karakteristik DPSC dan SHED dilihat dari ekspresi gen ALX4 pada subjek normal dan pasien celah bibir dan palatum belum diketahui. Tujuan: Mengevaluasi karakteristik DPSC dan SHED subjek normal dan pasien CLP berdasarkan ekspresi gen ALX4. Metode: DPSC subjek normal, DPSC pasien celah bibir dan palatum, dan SHED pasien celah bibir dan palatum diperoleh dari bahan biologis tersimpan Laboratorium Oral Biologi Fakultas Kedokteran Gigi Universitas Indonesia. Selanjutnya ekspresi gen ALX4 dan housekeeping gene GAPDH diuji dengan two step quantitative RT-PCR (RT-PCR). Hasil: Tidak terdapat perbedaan ekspresi gen ALX4 baik diantara DPSC subjek normal dengan DPSC CLP (p=0,407) maupun DPSC CLP dengan SHED CLP (p=0,145). Kesimpulan: Tidak terdapat perbedaan karakteristik sel stromal pulpa gigi permanen dan sel stromal pulpa gigi sulung pada subjek normal dengan pasien celah bibir dan palatum berdasarkan ekspresi gen ALX4.

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Background: The most ideal sources of stromal cells used in tissue engineering are dental pulp stem cells (DPSC) and stem cells from human exfoliated deciduous teeth (SHED) due to their high proliferative properties. In previous studies, it was stated that there was an increase in the expression of homeobox genes (differentially expressed genes (DEGs), one of which was the ALX4 gene as in cleft lip and palate patients with normal subjects. The ALX4 gene is a homeobox gene under the Alx family and has a direct role in the development and formation of the skull and human face, along with the ALX4 proteins that regulate cell development and proliferation, cell maturation and differentiation, cell movement, and cell defence. However, the characteristics of ALX4 gene expression in DPSC and SHED in normal and cleft lip and palate patients are not known. Objective: To evaluate and compare the characteristics of Dental Pulp Stromal Cells (DPSC) and Stromal Cells from Human Exfoliated deciduous teeth (SHED) in cleft lip and palate and normal subjects by the expression of the ALX4 homeobox gene. Methods: DPSC of normal subjects, DPSC of CLP patients, SHED of CLP patients were obtained from stored biological material in the Oral Biology Laboratory, Faculty of Dentistry, University of Indonesia. Then, the examination of ALX4 gene expression was tested by Real-Time Polymerase Chain Reaction (RT-PCR) Results: There was no difference in ALX4 gene expression between DPSC in normal subjects and DPSC in cleft lip and palate subjects (p=0,407) and between DPSC in cleft lip and palate subjects and SHED in cleft lip and palate subjects (p=0,145). Conclusion: There were no differences in the characteristics of the pulp stromal cells of permanent and primary teeth in normal subjects with cleft lip and palate subjects through the expression of the ALX4 gene."

"Latar Belakang: Sumber sel stromal yang paling ideal digunakan dalam rekayasa jaringan adalah sel stromal pulpa gigi permanen (DPSC) dan sel stromal pulpa gigi sulung (SHED) dikarenakan sifat proliferasinya yang tinggi. Pada penelitian sebelumnya, dinyatakan bahwa terdapat peningkatan ekspresi gen homeobox salah satunya yaitu gen ALX4 sebagai pada pasien celah bibir dan palatum dengan subjek normal. Gen ALX4 adalah gen homeobox dibawah famili Alx dan memiliki peran langsung dalam perkembangan dan pembentukan kepala serta wajah serta mentranslasi protein yang meregulasi perkembangan dan proliferasi sel, pendewasaan dan diferensiasi sel, pergerakan sel, dan pertahanan sel. Namun, karakteristik DPSC dan SHED dilihat dari ekspresi gen ALX4 pada subjek normal dan pasien celah bibir dan palatum belum diketahui. Tujuan: Mengevaluasi karakteristik DPSC dan SHED subjek normal dan pasien CLP berdasarkan ekspresi gen ALX4. Metode: DPSC subjek normal, DPSC pasien celah bibir dan palatum, dan SHED pasien celah bibir dan palatum diperoleh dari bahan biologis tersimpan Laboratorium Oral Biologi Fakultas Kedokteran Gigi Universitas Indonesia. Selanjutnya ekspresi gen ALX4 dan housekeeping gene GAPDH diuji dengan two step quantitative RT-PCR (RT-PCR). Hasil: Tidak terdapat perbedaan ekspresi gen ALX4 baik diantara DPSC subjek normal dengan DPSC CLP (p=0,407) maupun DPSC CLP dengan SHED CLP (p=0,145). Kesimpulan: Tidak terdapat perbedaan karakteristik sel stromal pulpa gigi permanen dan sel stromal pulpa gigi sulung pada subjek normal dengan pasien celah bibir dan palatum berdasarkan ekspresi gen ALX4

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Background: The most ideal sources of stromal cells used in tissue engineering are dental pulp stem cells (DPSC) and stem cells from human exfoliated deciduous teeth (SHED) due to their high proliferative properties. In previous studies, it was stated that there was an increase in the expression of homeobox genes (differentially expressed genes (DEGs), one of which was the ALX4 gene as in cleft lip and palate patients with normal subjects. The ALX4 gene is a homeobox gene under the Alx family and has a direct role in the development and formation of the skull and human face, along with the ALX4 proteins that regulate cell development and proliferation, cell maturation and differentiation, cell movement, and cell defence. However, the characteristics of ALX4 gene expression in DPSC and SHED in normal and cleft lip and palate patients are not known. Objective: To evaluate and compare the characteristics of Dental Pulp Stromal Cells (DPSC) and Stromal Cells from Human Exfoliated deciduous teeth (SHED) in cleft lip and palate and normal subjects by the expression of the ALX4 homeobox gene. Methods: DPSC of normal subjects, DPSC of CLP patients, SHED of CLP patients were obtained from stored biological material in the Oral Biology Laboratory, Faculty of Dentistry, University of Indonesia. Then, the examination of ALX4 gene expression was tested by Real-Time Polymerase Chain Reaction (RT-PCR) Results: There was no difference in ALX4 gene expression between DPSC in normal subjects and DPSC in cleft lip and palate subjects (p=0,407) and between DPSC in cleft lip and palate subjects and SHED in cleft lip and palate subjects (p=0,145). Conclusion: There were no differences in the characteristics of the pulp stromal cells of permanent and primary teeth in normal subjects with cleft lip and palate subjects through the expression of the ALX4 gene."

"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 : Sel stromal yang dapat digunakan untuk regenerasi pada defek tulang diantaranya adalah Dental Pulp Stromal Cells (DPSC) dan Stromal Cells from Human Exfoliated deciduous teeth (SHED). Pada penelitian yang sudah ada, ditemukan bahwa terdapat differentially expressed genes (DEGs) pada beberapa gen homeobox salah satunya gen CUX1 yang mengalami penurunan ekspresi pada pasien celah bibir dan palatum dibandingkan subjek normal. Gen CUX1 atau Cut-like-homeobox 1 merupakan faktor transkripsi yang berperan dalam kontrol proliferasi dan diferensiasi. Validasi DEGs perlu dilakukan untuk memahami bagaimana gen diekspresikan dalam subjek yang sehat dan sakit serta dapat digunakan untuk memperoleh wawasan mengenai suatu penyakit. Oleh karena itu penelitian ini ditujukan untuk memvalidasi gen homeobox CUX1 sehingga dapat mengetahui karakteristiknya pada sel DPSC dan SHED pada pasien celah bibir dan palatum serta membandingkannya dengan DPSC subjek normal. Tujuan: Mengevaluasi karakteristik sel stromal gigi permanen (DPSC) dan sel stromal pulpa gigi sulung (SHED) pasien celah bibir dan palatum dan pasien normal melalui ekspresi gen homeobox CUX1. Metode: Sampel RNA DPSC subjek normal, DPSC CLP, SHED CLP diperoleh dari bahan biologis tersimpan Laboratorium Biologi Oral Fakultas Kedokteran Gigi Universitas Indonesia. Selanjutnya dilakukan uji ekspresi gen CUX1 dengan quantitative reverse-transcription PCR (RT-qPCR). Hasil : Tidak terdapat perbedaan ekspresi gen CUX1, baik antara DPSC subjek normal dengan DPSC CLP (p = 0,839) dan antara DPSC CLP dengan SHED CLP (p = 0,411). Kesimpulan: Tidak ada perbedaan karakteristik sel stromal pulpa gigi permanen dan gigi sulung pada subjek normal dengan subjek celah bibir dan palatum melalui ekspresi gen homeobox CUX1 sehingga dapat digunakan untuk perawatan rekayasa jaringan menggantikan autologous bone graft.

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Background : Stromal cells that can be used to regenerate bone defects include Dental Pulp Stromal Cells (DPSC) and Stromal Cells from Human Exfoliated deciduous teeth (SHED). In existing studies, it was found that there are differentially expressed genes (DEGs) in several homeobox genes, one of which is the CUX1 gene, which has decreased expression in cleft lip and palate patients compared to normal subjects. The CUX1 or Cut-like-homeobox 1 gene is a transcription factor that plays a role in the control of proliferation and differentiation. It is necessary to validate DEGs to understand how genes are expressed in healthy and diseased subjects and can be used to gain insight into a disease. Therefore, this study aimed to validate the homeobox gene CUX1 to determine its characteristics on DPSC and SHED in cleft lip and palate patients and compare them with DPSC from normal subjects. Objective : To evaluate the characteristics of Dental Pulp Stromal Cells (DPSC) and Stromal Cells from Human Exfoliated deciduous teeth (SHED) in cleft lip and palate and normal subject through the expression of the CUX1 homeobox gene. Methods : RNA samples from normal subject’s DPSC, cleft lip and palate subject’s DPSC and cleft lip and palate subject’s SHED were obtained from stored biological material in the Oral Biology Laboratory, Faculty of Dentistry, University of Indonesia. Then, the CUX1 gene expression test was performed using quantitative reverse-transcription PCR (RT-qPCR). Result : There was no difference in CUX1 gene expression, both between DPSC in normal subjects and DPSC in cleft lip and palate subjects (p = 0.839) and between DPSC in cleft lip and palate subjects and SHED in cleft lip and palate subjects (p = 0.411). Conclusion : There were no differences in the characteristics of the dental pulp stromal cells and Stromal Cells from Human Exfoliated deciduous teeth between normal subjects and cleft lip and palate subjects through the expression of the CUX1 homeobox gene so that it can be used for tissue engineering treatment to replace autologous bone graft."