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Abstrak :
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. ......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.

Abstrak :
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 ......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.

Abstrak :
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 ......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.

Abstrak :
Latar Belakang: Pengembangan teknologi rekayasa jaringan sebagai terapi CLP berpotensi untuk menggantikan terapi autologous bone graft. Rekayasa jaringan terdiri dari tiga komponen yang dikenal sebagai triad rekayasa jaringan, yaitu sumber sel punca, biodegradable scaffold, dan faktor pertumbuhan. DPSC merupakan salah satu sumber sel punca yang diketahui efektif dalam memperbaiki defek CLP dengan metode isolasi sel yang relatif lebih mudah, tidak invasif, dan efek samping minimal. Pada penelitian sebelumnya DPSC yang diisolasi dari pasien CLP menunjukkan ekspresi gen IGF-1 yang berlebih. Faktor pertumbuhan tersebut diketahui berperan dalam proliferasi dan diferensiasi sel, namun ekspresi berlebih IGF-1 pada DPSC pasien CLP tidak diikuti oleh peningkatan kemampuan proliferasi dan diferensiasinya. Dalam sistem sirkulasi, IGF-1 berikatan dengan IGFBP-3 yang dapat memperpanjang waktu paruhnya. IGFBP-3 memiliki afinitas yang lebih tinggi terhadap IGF-1 dibanding dengan IGF-1R, sehingga dapat meregulasi dan menghambat peran IGF-1. Fungsi IGF-1 dijalankan dengan berikatan dengan IGF-1R untuk mengaktifkan jalur pensinyalan hilir, salah satunya adalah jalur MAPK/ERK1/2. ERK1 dan ERK2 diketahui meregulasi fungsi proliferasi dan diferensiasi sel, namun belum diketahui secara pasti bagaimana ekspresi gen ERK1 dan ERK2 pada DPSC subjek normal dan CLP. Tujuan: Menganalisis pengaruh anti IGF-1R dan IGFBP-3 terhadap ekspresi gen ERK1 dan ERK2 pada DPSC subjek normal dan CLP.Metode: Sampel RNA DPSC pasien normal (n=4) dan CLP (n=3) sebelum dan sesudah perlakuan anti IGF-1R atau IGFBP-3 diperoleh dari bahan biologis tersimpan di Laboratorium Biologi Oral Fakultas Kedokteran Gigi Universitas Indonesia. Dilakukan analisis ekspresi relatif gen ERK1, ERK2, dan GAPDH sebagai housekeeping gene dengan two-step Real-Time PCR (RT-PCR)Hasil: Tidak terdapat perbedaan ekspresi gen ERK1 dan ERK2 pada DPSC pasien CLP dibanding pasien normal, baik pada perlakuan anti IGF-1R maupun IGFBP-3. Kesimpulan: Inhibisi IGF-1 dengan anti IGF-1R dan IGFBP-3 tidak memengaruhi ekspresi gen ERK1 dan ERK2. ......Background: The development of tissue engineering as a therapy for CLP have potential to replace the current autologous bone graft that is considered not ideal in repairing the bone defect in CLP patients. Tissue engineering consists of three parts known as the tissue engineering triad: stem cell source, biodegradable scaffold, and growth factors. DPSC is one such stem cell source that is known to effectively repair CLP defects with a relatively easy cell isolation, less invasive, and minimal patient compromise. Recent studies have found that DPSC isolated from CLP patients display a higher expression of IGF-1 gene expression. IGF-1 is known for its role in cell proliferation and differentiation, however the overexpression of IGF-1 gene in CLP patient’s DPSC is not followed by the increase of proliferation and differentiation capability. In the circulation system, IGF-1 binds to IGFBP-3 to extend its half time in the system. IGFBP-3 displays a higher affinity towards IGF-1 than IGF-1R, thus acting as a regulator and inhibitor to IGF-1 activity. IGF-1 functions by binding with IGF-1R and activating the downstream signalling pathway. One such pathway is the MAPK/ERK1/2 signalling pathway. ERK1 and ERK2 are both known for its role in regulating the proliferation and differentiation function in cells, but the exact gene expression characteristics in both normal and CLP subject’s DPSC are not known. Objective: To analyze the effect of anti IGF-1R and IGFBP-3 to ERK1 and ERK2’s gene expression in normal and CLP subject’s DPSC. Methods: RNA samples of DPSC of normal (n=4) and CLP subjects (n=3) before and after treated with anti IGF-1R and IGFBP-3 were obtained from the Oral Biology Laboratory of Faculty of Dentistry Universitas Indonesia. Relative gene expression of ERK1, ERK2, and GAPDH as the housekeeping gene were analyzed using two-step Real-Time PCR (RT-PCR) Results: There was no difference in both ERK1 and ERK2 gene expression between normal and CLP subject following anti IGF-1R or IGFBP-3 treatment. Conclusion: anti IGF-1R and IGFBP-3 treatment did not influence ERK1 and ERK2 gene expression

Abstrak :
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. ......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.

Abstrak :
Latar Belakang: Rekayasa jaringan merupakan perawatan alternatif autologous bone graft pada rekonstruksi tulang alveolar pasien celah bibir dan palatum (CLP). Potensi klonogenik dan proliferatif yang baik serta kemudahan aksesibilitas membuat sel stromal pulpa gigi permanen (DPSC) dan gigi sulung (SHED) menjadi sel yang ideal untuk rekonstruksi tulang alveolar. Gen HOXC9 merupakan gen homeobox di bawah famili Hox, yang mengatur pola perkembangan skeletal. Penelitian terbaru menyatakan gen Hox tetap terekspresikan saat dewasa dan ditemukan dalam regenerasi jaringan. Namun, karakteristik ekspresi gen HOXC9 pada DPSC dan SHED subjek normal dan pasien celah bibir dan palatum belum diketahui secara pasti. Tujuan: Mengevaluasi karakteristik DPSC dan SHED subjek normal dan pasien CLP melalui ekspresi gen HOXC9. Metode: Sampel RNA DPSC subjek normal (n=2), DPSC CLP (n=3), SHED CLP (n=2) diperoleh dari bahan biologis tersimpan Laboratorium Oral Biologi Fakultas Kedokteran Gigi Universitas Indonesia. Selanjutnya ekspresi gen HOXC9 dan housekeeping gene GAPDH diuji dengan two step Real-Time PCR (RT-PCR). Hasil: Tidak terdapat perbedaan ekspresi gen HOXC9, baik antara DPSC subjek normal dengan DPSC CLP (p>0,05) ataupun DPSC CLP dengan SHED CLP (p>0,05). Kesimpulan: Sel stromal pulpa gigi permanen dan gigi sulung subjek normal dan pasien celah bibir dan palatum memiliki karakteristik yang sama melalui ekspresi gen HOXC9. ......Background: Tissue engineering is an alternative treatment of autologous bone graft in alveolar bone reconstruction for cleft lip and palate (CLP) patients. The clonogenic and proliferative capacity as well as the ease of accessibility make DPSC and SHED ideal cells for alveolar bone reconstruction. HOXC9 is a homeobox gene under the Hox family, which regulates the development of skeletal patterns. Recent research suggests that the Hox gene remains expressed in adulthood and is found in tissue regeneration. However, the characteristics of HOXC9 gene expression in DPSC and SHED of normal subjects and cleft lip and palate patients are unknown. Objective: To evaluate the characteristics of DPSC and SHED in normal subjects and CLP patients through HOXC9 gene expression. Methods: RNA samples from DPSC of normal subjects (n=2), DPSC of CLP patients (n=3), SHED of CLP patients (n=2) were obtained from the Laboratory of Oral Biology, Faculty of Dentistry, Universitas Indonesia. HOXC9 gene expression and housekeeping gene GAPDH were tested by two-step Real-Time PCR (RT-PCR). Results: There was no difference in HOXC9 gene expression, either between DPSC of normal subjects and DPSC of CLP patients (p>0.05) or DPSC and SHED of CLP patients (p>0.05). Conclusion: DPSC and SHED of normal subjects and cleft lip and palate patients have the same characteristic through HOXC9 gene expression.

Abstrak :
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. ......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.

Abstrak :
Latar Belakang: Celah bibir dan palatum merupakan salah satu kelainan kongenital yang paling sering terjadi. Kelainan ini dapat menyebabkan kendala dalam berbicara, abnormalitas telinga tengah, masalah psikologis, serta kelainan dental seperti anodontia parsial dan supernumerary teeth. Perawatan autologous alveolar bone grafting yang diambil dari tulang ilium pasien merupakan standar perawatan bagi pasien celah bibir dan palatum. Namun, pengambilan tulang tersebut bersifat invasif dan memiliki tingkat morbiditas dan mortalitas yang tinggi. Teknik rekayasa jaringan yang terdiri dari scaffold, faktor pertumbuhan, dan sel punca dapat menjadi solusi untuk masalah tersebut. Sumber donor sel punca yang tidak invasif bisa didapatkan dari sel punca pulpa gigi sulung (SHED) dan sel punca pulpa gigi permanen (DPSCs). Salah satu syarat dapat digunakannya suatu sel punca adalah memiliki kapasitas proliferasi yang baik. Perbandingan antara kapasitas proliferasi SHED dan DPSCs pada pasien normal telah diketahui, namun pada pasien celah palatum belum pernah diteliti. Tujuan: Penelitian ini bertujuan untuk mengetahui perbandingan kapasitas proliferasi sel punca pulpa gigi sulung dan sel punca pulpa gigi permanen pasien celah bibir dan palatum. Metode: SHED dan DPSCs pasien celah bibir dan palatum dikultur hinga tingkat confluency 70-80%, setelah itu sel dipanen dan dilakukan Uji PDT pada sel yang telah dikultur selama 7 hari. Hasil: SHED pasien celah bibir dan palatum menunjukkan nilai PDT yang lebih tinggi dibandingkan dengan DPSCs, namun secara statistik perbedaan tersebut tidak berbeda bermakna. Kesimpulan: SHED dan DPSCs penderita celah bibir dan palatum memiliki kapasitas proliferasi yang sama baiknya. ......Background: Cleft lip and palate is one of the most common congenital abnormalities. This disorder can cause speech impediments, middle ear abnormalities, psychological problems, and dental abnormalities such as partial anodontia and supernumerary teeth. Treatment of autologous alveolar bone grafting taken from the patient's ilium bone is the standard of care for cleft lip and palate patients. However, bone removal is invasive and carries a high rate of morbidity and mortality. Tissue engineering techniques consisting of scaffolds, growth factors, and stem cells can be a solution to the problem that. Sources of non-invasive stem cell donors can be obtained from primary dental pulp stem cells (SHED) and permanent dental pulp stem cells (DPSCs). One of the conditions for the use of a stem cell is to have a good proliferative capacity. Comparison between the proliferative capacity of SHED and DPSCs in normal patients known, but in cleft palate patients it has not been studied. Objective: This study was aimed to compare the proliferative capacity of pulp stem cells of primary teeth and pulp stem cells of permanent teeth in cleft lip and palate patients. Methods: SHED and DPSCs of cleft lip and palate patients were cultured to level 70-80% confluency, after that the cells were harvested and PDT test was performed on cells that had been cultured for 7 days. Results: SHED of cleft lip and palate patients showed a higher PDT value than DPSCs, but statistically the difference was not significantly different. Conclusion: SHED and DPSCs patients with cleft lip and palate have the same good proliferative capacity.

Abstrak :
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. ......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.

Abstrak :

Latar Belakang: Celah bibir dan palatum merupakan kelainan kongenital yang paling sering terjadi pada regio orofasial. Pasien celah bibir dan palatum menunjukkan sejumlah permasalahan seperti defek tulang alveolar yang lebar, kehilangan gigi kongenital, supernumerary teeth, hipoplasia dan gigi impaks. Autologous alveolar bone grafting dianggap sebagai perawatan gold standard untuk rekonstruksi tulang alveolar dengan menggunakan tulang kanselus dari puncak ilium anterior. Namun, pengambilan tulang ilium bersifat invasif dan memiliki potensi terjadinya komplikasi. Mengingat hal tersebut, teknik rekayasa jaringan yang memanfaatkan scaffolds, faktor pertumbuhan, dan sel punca dipertimbangkan sebagai pilihan perawatan yang baru. Sel punca mesenkim bisa didapatkan dari jaringan pulpa, yang disebut dengan sel punca pulpa gigi sulung atau stem cells from exfoliated deciduous teeth (SHED) dan sel punca pulpa gigi permanen atau dental pulp stem cells (DPSC). Salah satu kriteria yang harus dimiliki sel punca mesenkim adalah mengekspresikan surface marker CD73, CD90, dan CD105. Pada pasien normal, penelitian yang membandingkan karakteristik antara SHED dan DPSC telah membuktikan bahwa keduanya merupakan sel punca mesenkim dengan mengekspresikan surface markersesuai dengan kriteria. Namun, pada pasien celah bibir dan palatum belum banyak diteliti. Tujuan: Menganalisis perbedaan persentase sel yang mengekspresikan surface marker (CD73, CD90, dan CD105) pada SHED dan DPSC pasien celah bibir dan palatum. Metode: Sel punca pulpa gigi sulung dan gigi permanen diisolasi dari jaringan pulpa pasien celah bibir dan palatum. Persentase sel yang mengekspresikan surface marker (CD73, CD90, dan CD105) dianalisis dengan uji flow cytometry. Hasil: Analisis flow cytometry menunjukkan bahwa baik SHED maupun DPSC mengekspresikan masing-masing surface marker dalam persentase yang tinggi (>90%). Setelah dilakukan uji Independent T-test untuk membandingkan ekspresi masing-masing surface markerpada kedua grup, didapatkan hasil >0,05. Kesimpulan:  Tidak terdapat perbedaan bermakna antara ekspresi masing-masing surface marker pada SHED dan DPSC pasien celah bibir dan palatum.

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Background: Cleft lip and palate is the most common congenital anomaly in the orofacial region. Cleft lip and palate patients present with a number of complaints such as wide alveolar bone defects, congenitally missing teeth, supernumerary teeth, hypoplastic dan impacted teeth. Autologous bone grafting is considered to be the gold standard for alveolar bone reconstruction using the cancellous bone harvested from the anterior iliac crest. However, the procedure is invasive and carries a risk of complications. Bearing all that in mind, tissue engineering that utilizes scaffolds, growth factors, and stem cells arises as a new therapeutic option. Mesenchymal stem cells can be obtained from dental pulp, which are called stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC). One of the criterias to define mesenchymal stem cells is the expression of surface markers CD73, CD90, and CD105. In normal patients, both SHED and DPSC have been known to express those surface markers. However, the expression of CD73, CD90, and CD105 in SHED and DPSC from cleft lip and palate patients has not been fully explored. Objective: To analyze the difference in the percentage of cells that express CD73, CD90, and CD105 in SHED and DPSC from cleft lip and palate patients. Methods: SHED and DPSC were isolated from dental pulp. The expression of surface markers were analyzed with flow cytometry. Results: Flow cytometry analysis showed that SHED and DPSC from cleft lip and palate patients highly express (>90%) surface markers that are associated with mesenchymal stem cells such as CD73, CD90, and CD105. The Independent T-Test was then performed to see a comparison between the expression of each surface marker in both groups and the value was >0.05. Conclusions: There is no significant difference between the percentage of cells that express each surface marker in SHED and DPSC from cleft lip and palate patients.