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"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."

"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: Gen Homeobox adalah gen pengatur perkembangan antara lain morfogenesis sel dengan menyandikan faktor transkripsi pada tahap awal embriogenesis dan diferensiasi sel. Gen EN1 adalah gen Homeobox yang berperan dalam proses pembentukan tulang. Penelitian terbaru menunjukkan bahwa gen EN1 mengalami overexpression signifikan pada sel stromal pulpa gigi permanen pasien celah bibir dan palatum. Namun, pengaruh gen EN1 pada karakteristik sel stromal pulpa gigi sulung dan permanen subjek normal dan pasien celah bibir dan palatum belum diketahui secara pasti. Tujuan: Melakukan verifikasi karakteristik sel stromal gigi permanen pasien celah bibir dan palatum dan subjek normal serta sel stromal gigi sulung pasien celah bibir dan palatum melalui ekspresi gen EN1. Metode: Sampel RNA DPSC subjek normal (n=2), DPSC CLP (n=2), SHED CLP (n=2) diperoleh dari bahan biologis tersimpan Laboratorum Oral Biologi Fakultas Kedokteran Gigi Universitas Indonesia. Kemudian, dilakukan sintesis cDNA dan standarisasi konsentrasi sampel hasil sintesis cDNA. Selanjutnya, ekspresi gen EN1 dan gen referensi GAPDH diuji dengan quantitative reverse-transcription PCR (RT-qPCR). Hasil: Tidak terdapat perbedaan bermakna ekspresi gen EN1, antara DPSC subjek normal dengan DPSC CLP (p≥0,05) sedangkan terdapat perbedaan bermakna ekspresi gen EN1 antara sel DPSC CLP dengan sel SHED CLP (p≤,05). Kesimpulan: Tidak terdapat perbedaan karakteristik sel stromal pulpa gigi permanen antara subjek normal dan pasien celah bibir dan palatum, sedangkan terdapat perbedaan antara sel stromal pulpa gigi sulung dan sel stromal pulpa gigi permanen pada pasien celah bibir dan palatum.

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Background: Homeobox gene is a group of master regulatory developmental genes which are responsible for encode transcription factor in the early phase of embryogenesis and for cell differentiation. EN1 gene is a Homeobox gene that has a role in bone formation. The latest research discovered that EN1 gene was significantly overexpressed in Permanent Teeth’s Stromal Cell of Cleft Lip and Palate Subjects. However, the effect of EN1 gene on the characteristics of Permanent and Deciduous Teeth’s Stromal Cell of Normal Subjects and Cleft Lip and Palate Subjects still remain unknown. Objective: To Verify on the characteristic of the Permanent Teeth’s Stromal Cell between Cleft Lip and Palate Patients and Normal Subjects as well as the characteristic between the Permanent Teeth’s Stromal Cell of Cleft Lip and Palate Patients and Deciduous Teeth’s Stromal Cell of Cleft Lip and palate Patients. Methods: DPSC of normal subjects’ RNA sample (n=2), DPSC of CLP Patient’s RNA sample (n=2), SHED of CLP Patients’ RNA sample (n=2) obtained from Archived Biological Materials in Laboratorium. Subsequently, synthesis the RNA sample into cDNA sample and standardize the cDNA concentration sample. Afterwards, perform RT-PCR assay to validate EN1 and GAPDH reference gene expression. Results: No statistically significant difference of the EN1 gene expression between the Permanent Teeth’s Stromal Cell between Cleft Lip and Palate Patients and Normal Subjects (p≥0,05) and there is statistically significant difference of the EN1 gene expression between the Permanent Teeth’s Stromal Cell of Cleft Lip and Palate Patients and Deciduous Teeth’s Stromal Cell of Cleft Lip and palate Patients. (p ≤05) Conclusion: There is no characteristic difference between the Permanent Teeth’s Pulp Stromal Cell between Cleft Lip and Palate Patients and Normal Subjects, Meanwhile There is characteristic difference between the Deciduous Teeth’s Pulp Stromal Cell of Cleft Lip and Palate Patients and the Permanent Teeth’s Pulp Stromal Cell of Cleft Lip and Palate Patients."

"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: Celah bibir dan palatum (CLP) adalah kegagalan fusi prosesus frontonasal dan maksilaris yang menghasilkan celah yang meluas ke bibir, alveolus, dasar hidung, dan palatum keras maupun lunak. Diperlukan perawatan melalui teknik rekayasa jaringan dengan menggunakan sel stromal mesenkim yang dapat ditemukan pada dental pulp stromal cells (DPSC). Kemampuan osteogenik DPSC dapat dilihat melalui deteksi marker osteogenik seperti osteopontin (OPN). Osteopontin merupakan salah satu marker utama diferensiasi osteogenik yang diproduksi oleh osteoblas dalam proses pembentukan dan mineralisasi tulang. Pada pasien CLP diketahui perbedaan ekspresi gen yang dapat memengaruhi sintesis matriks ekstraseluler. Penelitian ini dilakukan untuk mengetahui kemampuan diferensiasi osteogenik melalui ekspresi marker osteopontin saat diferensiasi osteoblas pada pasien celah bibir dan palatum. Tujuan: Membandingkan kemampuan diferensiasi osteogenik sel stromal pulpa gigi subjek normal dengan sel stromal pulpa gigi pasien celah bibir dan palatum melalui ekspresi gen osteopontin. Metode: Sampel RNA yang diperoleh dari kultur sel pulpa gigi subjek normal dan pasien celah bibir dan palatum diuji dengan Real-Time Polymerase Chain Reaction (RT-PCR) dengan primer osteopontin (OPN) dan 18S sebagai housekeeping gene. Hasil: Tidak terdapat perbedaan antara ekspresi relatif gen OPN sel stromal pulpa gigi subjek normal dan pasien celah bibir dan palatum. Kesimpulan: Kemampuan diferensiasi osteogenik sel stromal pulpa gigi pada pasien celah bibir dan palatum ekuivalen dengan sel stromal pulpa gigi pada subjek normal.

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Background: Cleft lip and palate (CLP) occurs due to the failure of fusion of the frontal and maxillary process that results in a cleft that extends to the lip, alveoli, nose floor, and hard and soft palate. One of the potentially alternative treatment for CLP cases is tissue engineering technique using Mesenchymal Stromal Cell (MSC). MSC can be found in dental pulp stromal cells (DPSC). Osteogenic ability can be seen through the detection of osteogenic markers such as osteopontin (OPN). Osteopontin is one of the main markers of osteogenic differentiation produced by osteoblast in the process of bone formation and mineralization. Osteopontin is expressed by preosteoblasts in early bone formation and mature osteoblasts at bone remodelling sites. Osteopontin expression as one of osteogenic markers in cleft lip and palate patients is unknown. This study was conducted to determine the ability of osteogenic differentiation through the expression of osteopontin in cleft lip and palate patients. Objective: To compare osteogenic differentiation ability of mesenchymal stromal cells in cleft lip and palate patients and normal subject through the expression of osteogenic marker osteopontin. Methods: RNA sample that was obtained through RNA extraction from dental pulp stromal cells of cleft and lip palate patients and normal subjects were tested with Real-Time Polymerase Chain Reaction (RT-PCR) using osteopontin and 18S as housekeeping gene. Results: There was no difference between the relative expression of OPN gene in DPSC from normal subject and cleft lip and palate patients. Conclusion: Osteogenic differentiation ability of dental pulp stromal cells from cleft lip and palate patients is equivalent with dental pulp stromal cells from normal subjects."

"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."

"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 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 : DPSC dan SHED merupakan sumber sel stromal yang dapat digunakan untuk rekayasa jaringan sebagai alternatif perawatan pasien dengan CLP. Penelitian terdahulu menunjukkan ekspresi gen Homeobox pada DPSC pasien dengan CLP dibandingkan subjek normal. Gen Homeobox merupakan sekelompok gen yang mengkodekan serangkaian domain protein yang berperan dalam proses awal perkembangan dan diferensiasi sel saat embriogenesis. Dalam kelompok homeobox ini, terdapat gen SHOX yang berperan dalam pembentukan kerangka tulang pada tahap embriogenesis. Penelitian ini dilakukan untuk memvalidasi perbedaan ekspresi gen pada kelompok sampel DPSC dan SHED subjek normal dan pasien CLP. Tujuan : Melakukan evaluasi karakteristik sel pada sampel DPSC subjek normal dengan pasien CLP; DPSC dengan SHED pasien CLP. Metode : Menggunakan template RNA dari 3 kelompok sampel yaitu DPSC subjek normal, DPSC pasien CLP, dan SHED pasien CLP. Lalu, sintesis cDNA dan dilakukan metode RT-qPCR untuk melihat ekspresi gen SHOX dari setiap kelompok sampel. Hasil : Tidak terdapat perbedaan ekspresi gen SHOX pada perbandingan kelompok sampel DPSC normal dengan pasien CLP, dan kelompok DPSC CLP dengan SHED CLP. Kesimpulan : DPSC dan SHED subjek normal dan pasien CLP memiliki karakteristik gen SHOX yang sama.

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Background : DPSC and SHED are the sources for tissue engineering as an alternative treatment for patients with CLP. Previous studies showing expression of homeobox genes in DPSC of normal compared to CLP patients. Homeobox genes encode a series of protein domains that is involved in the process of development and cell differentiation. There is a SHOX gene involved in the bone skeleton formation during embryogenesis. This study was conducted to validate the differences in gene expression between the sample groups of DPSC and SHED of normal and CLP subjects. Objective : To evaluate the cell characteristics in sample groups of DPSC in normal and CLP subjects; DPSC and SHED in CLP subjects. Methods : Using RNA template of 3 sample groups, namely DPSC of normal subjects, DPSC of CLP subjects. cDNA was synthesized and the RT-qPCR method was used to see the SHOX gene expression of each group. Result : There was no differences in SHOX gene expression in the comparison of DPSC normal with CLP patients, and the DPSC and SHED in subjects with CLP. Conclusion : DPSC and SHED in normal subjects and CLP patients have the same characteristics of SHOX gene."

"Latar Belakang: Celah bibir dan palatum merupakan salah satu kelainan bawaan yang menyebabkan defek jaringan keras sehingga dikembangkan perawatan rekonstruksi tulang berbasis teknik rekayasa jaringan sebagai alternatif perawatan. Sumber sel stromal mesenkim dapat diperoleh dari pulpa gigi sulung dan gigi permanen. Kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen sudah banyak dilaporkan. Pada pasien celah bibir dan palatum, terdapat gen-gen yang diekspresikan berbeda dan kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum belum diketahui. Tujuan: Mengevaluasi perbandingan kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum melalui deposisi kalsium. Metode: Sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum dikultur menggunakan medium osteogenik selama 21 hari kemudian dilakukan pewarnaan Alizarin Red dan kuantifikasi terhadap deposisi kalsium. Hasil: Sel stromal pulpa gigi sulung dan gigi permanen yang dikultur menggunakan medium osteogenik menunjukkan adanya deposisi kalsium yang tinggi. Sel stromal pulpa gigi sulung dan gigi permanen tidak menunjukkan perbedaan nilai rerata absorbansi, intensitas pewarnaan, dan area pewarnaan yang bermakna secara statistik (p ≥ 0,05). Kesimpulan: Sel stromal pulpa gigi sulung pasien celah bibir dan palatum memiliki kemampuan diferensiasi osteogenik yang ekuivalen dengan sel stromal pulpa gigi permanen pasien celah bibir dan palatum.

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Background: Cleft lip and palate is one of the most common congenital anomalies resulting in hard tissue defects therefore tissue engineering is currently developed as an alternative treatment. The source of mesenchymal stromal cells can be obtained from human exfoliated deciduous teeth (SHED) and dental pulp (DPSCs). Osteogenic differentiation abilities of SHED and DPSCs have been widely studied. In cleft lip and palate patients, there are several differentially expressed genes and the osteogenic differentiation abilities of SHED and DPSCs in cleft lip and palate patients have not yet been known. Purpose: To compare the osteogenic differentiation abilities of SHED and DPSCs in cleft lip and palate patients by calcium deposition. Methods: SHED and DPSCs isolated from cleft lip and palate patients were cultured using osteogenic medium for 21 days then added Alizarin Red staining and the calcium deposition were quantified. Result: Both SHED and DPSCs that cultured in osteogenic medium demonstrated high calcium deposition. SHED and DPSCs did not show any statistically significant differences in the average absorbance values, staining intensity, and staining areas (p ≥ 0,05). Conclusion: SHED and DPSCs in cleft lip and palate patients have equivalent ability of osteogenic differentiation by calcium deposition."