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"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 : 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: 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 : 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: Celah bibir dan palatum (CLP) merupakan salah satu kelainan kongenital yang menghasilkan defek jaringan lunak maupun jaringan keras dan membutuhkan perawatan rekonstruksi tulang alveolar dan palatum. Celah bibir dan palatum dianggap berasal dari anomali proliferasi sel akibat faktor genetika. Autologous bone graft adalah baku emas untuk memperbaiki defek tulang palatum pada pasien CLP. Namun demikian, perawatan tersebut membutuhkan prosedur yang invasif. Perawatan melalui rekayasa jaringan dapat menjadi alternatif perawatan. Rekonstruksi tulang alveolar melalui rekayasa jaringan membutuhkan jumlah sel yang banyak sehingga kapasitas proliferasi sel punca merupakan aspek penting dalam penerapan klinis. Sel punca pulpa gigi sulung (SHED) dan sel punca pulpa gigi permanen (DPSCs) dapat menjadi sumber sel yang ideal karena memiliki kapasitas proliferasi yang tinggi, kemampuan diferensiasi ke berbagai tipe sel, isolasi yang mudah, dan aksesibilitas yang baik. Namun, kapasitas proliferasi SHED dan DPSCs pasien CLP belum diketahui.Tujuan: Penelitian ini bertujuan membandingkan kapasitas proliferasi SHED dan DPSCs pasien celah bibir dan palatum.Metode: SHED dan DPSCs dari pasien CLP dikultur hingga mencapai 70%-80% confluent. Kapasitas proliferasi sel setelah dikultur selama 24 jam, 48 jam, dan 72 jam dianalisis melalui uji MTT.Hasil: SHED setelah dikultur 24 jam menunjukkan nilai rata-rata optical density yang lebih tinggi secara signifikan (p<0,05). SHED dan DPSCs setelah dikultur 48 jam dan 72 jam tidak menunjukkan perbedaan nilai rata-rata optical density secara statistik (p>0,05).Kesimpulan: SHED pasien CLP memiliki kapasitas proliferasi lebih tinggi secara signifikan hanya pada 24 jam pertama. Pada 48 jam dan 72 jam pertama, SHED dan DPSCs pasien CLP memiliki kesamaan kapasitas proliferasi.

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Background: Cleft lip and palate (CLP) is one of orofacial congenital malformations that results in both soft tissue and hard tissue defect. It requires reconstruction of the maxillary alveolar cleft. Cleft lip and palate is thought to be came from anomalies of cell proliferation caused by genetic factors. Autologous bone graft have been the gold standard treatment to repair maxillary alveolar and palate clefts. However, such treatment needs an invasive procedure that may induce pain. To overcome those disadvantages, tissue engineering has received attention to be new alternative treatment.

Reconstruction of maxillary alveolar cleft requires huge number of stem cells so that proliferative capacity is important traits before clinical application. Stem Cells from Exfoliateed Deciduous Teeth (SHED) and Dental Pulp Stem Cells (DPSCs) can be ideal sources of stem cell since they are known to have high proliferative capacity, multilineage differentiation, ease of isolation, and well accesibility. However, proliferative capacity of SHED and DPSCs isolated from CLP patients have not yet known.

Objective: The aim of this study was to compare proliferative capacity between cultured stem cells from exfoliated deciduous teeth and dental pulp stem cells isolated from cleft lip and palate patients.

Methods: SHED and DPSCs isolated from cleft patient were cultured until it reached 70%-80% confluency. Proliferative capacity after culturing for 24 hours, 48 hours, and 72 hours were analyzed using MTT Assay.

Results: SHED after culturing for 24 hours showed higher optical density average value significantly (p<0,05). SHED and DPSCs after culturing for 48 hours and 72 hours has no difference optical density average value significantly (p>0,05).

Conclusions: SHED from cleft patients showed higher proliferative capacity significantly only on first 24 hours culturing. SHED and DPSCs have similar proliferative capacity on 48 hous and 72 hours culturing."