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"Latar Belakang: Dalam rangka mengembangkan terapi alternatif rekonstruksi tulang alveolar pada celah bibir dan palatum (CLP), teknologi rekayasa jaringan menjadi alternatif yang menjanjikan dengan menggunakan komponen sel stromal dan faktor pertumbuhan. Penelitian terbaru mengenai DPSC pada pasien CLP menghasilkan penyembuhan tulang yang memuaskan. Selain itu, ditemukan ekspresi berlebih dari IGF-1 pada DPSC pasien CLP. AKT dan MTOR merupakan downstream dari jalur pensinyalan IGF-1. AKT memiliki peran sebagai pengatur kelangsungan hidup dan proliferasi sel. Pensinyalan MTOR mengatur transkripsi gen dan sintesis protein untuk mengatur proliferasi sel dan diferensiasi sel. Namun, karakteristik DPSC subjek normal dan pasien celah bibir dan palatum berdasarkan ekspresi gen AKT dan MTOR dengan penghambat IGF-1 belum diketahui secara pasti. Tujuan: Mengevaluasi efek anti IGF-1R dan IGFBP-3 terhadap karakteristik DPSC subjek normal dan pasien CLP melalui ekspresi gen AKT dan MTOR. Metode: RNA DPSC subjek normal (n=4) yaitu kelompok sebelum perlakuan, kontrol negatif tanpa FBS, kontrol negatif dengan FBS, perlakuan anti IGF-1R. RNA DPSC CLP (n=3) yaitu kelompok sebelum perlakuan, kontrol negatif tanpa FBS, kontrol negatif dengan FBS, perlakuan anti IGF-1R. Analisis ekspresi gen AKT dan MTOR menggunakan GAPDH sebagai housekeeping gene dengan Real time PCR. Hasil: Tidak terdapat perbedaan ekspresi gen AKT dan MTOR, antara DPSC subjek normal dengan CLP sebelum perlakuan (p0,05) ataupun DPSC subjek normal dengan CLP setelah perlakuan anti IGF-1R (p0,05). Kesimpulan: Sel stromal pulpa gigi permanen subjek normal dan pasien celah bibir dan palatum pada kelompok sebelum perlakuan dan sesudah perlakuan anti IGF-1R memiliki karakteristik yang sama melalui ekspresi gen AKT dan MTOR.

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Background: In order to develop alternative therapies for alveolar bone reconstruction in cleft lip and palate (CLP), tissue engineering technology is a promising alternative using stromal cell and growth factors. Recent studies regarding DPSC in CLP patients have resulted in satisfactory bone healing. In addition, overexpression of IGF-1 was found in the DPSC of CLP patients. AKT and MTOR are downstream of the IGF-1 signaling pathway. AKT has a role as a regulator of cell survival and proliferation. MTOR signaling regulates gene transcription and protein synthesis to regulate cell proliferation and cell differentiation. However, the characteristics of DPSC normal subjects and cleft lip and palate patients based on AKT and MTOR gene expression with IGF-1 inhibitors are unknown. Objective: To evaluate the effect of anti-IGF-1R and IGFBP-3 on DPSC normal and CLP subject through AKT and MTOR gene expression. Methods: RNA from DPSC normal subject (n=4) with pretreatment, negative control without FBS, negative control with FBS, anti IGF-1R treatment dan DPSC CLP subject (n=3) with pretreatment, negative control without FBS, negative control with FBS, anti IGF-1R treatment. Analysis of AKT and MTOR gene expression using GAPDH as a housekeeping gene with Real time PCR test. Results: There was no difference in AKT and MTOR gene expression, either between DPSC of normal subjects and CLP patients before treatment (p0.05) or DPSC of normal and CLP patients after anti IGF-1R treatment (p0.05). Conclusion: DPSC of normal subjects and cleft lip and palate patients have the same characteristic with anti IGF-1R treatment through AKT dan MTOR gene expression."

"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.

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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"

"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: 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.

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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."

"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 adalah kelainan bawaan yang mempengaruhi regio orofacial. Perawatan yang menjadi baku emas untuk pasien celah bibir dan palatum adalah autologous bone graft. Namun, perawatan ini masih invasif dan ada beberapa kekurangannya sehingga perlu teknik rekayasa jaringan dengan sel stromal. Sel stromal mesenkim yang terdapat dalam rongga mulut adalah sel stromal pulpa gigi sulung (SHED) dan sel stromal pulpa gigi permanen (DPSC). Kemampuan diferensiasi osteogenik SHED dan DPSC pada subjek normal sudah diketahui. Namun, kemampuan diferensiasi osteogenik dengan ekspresi gen RUNX-2 pada DPSC dan SHED pasien celah bibir dan palatum belum diketahui secara pasti. Tujuan: Membandingkan kemampuan diferensiasi osteogenik sel stromal pulpa gigi permanen pasien celah bibir dan palatum dengan sel stromal pulpa gigi sulung pasien celah bibir dan palatum melalui ekspresi gen RUNX-2. Metode: DPSC celah bibir dan palatum dan SHED celah bibir dan palatum dikultur dengan medium osteogenik dan tanpa medium osteogenik selama 21 hari. Sampel RNA diperoleh kultur sel stromal pulpa gigi permanen (DPSC) dan sel stromal pulpa gigi sulung (SHED) pasien celah bibir dan palatum. Selanjutnya diuji ekspresi gen RUNX-2, dan housekeeping gene 18S dengan Real-Time Polymerase Chain Reaction (RT-PCR). Hasil: Tidak ada perbedaan kemampuan diferensiasi sel stromal pulpa gigi permanen pasien celah bibir dan palatum dengan sel stromal pulpa gigi sulung pasien celah bibir dan palatum melalui ekspresi gen RUNX-2. Kesimpulan: Kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung pasien celah bibir dan palatum ekuivalen dengan sel stromal pulpa gigi permanen pasien celah bibir dan palatum.

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Background: Cleft lip and palate are congenital anomalies that affect the orofacial region including lips, alveolar ridge, hard palate, and soft palate. Patients with cleft lip and palate have impaired esthetic and stomatognathic functions. The gold standard treatment for cleft lip and palate patients is an autologous bone graft. However, this treatment is still invasive and has some limitations therefore requires tissue engineering techniques by using stromal cells. Mesenchymal stromal cells that are found in the mouth are stromal cells from human exfoliated deciduous teeth (SHED) and dental pulp stromal cells (DPSC). The osteogenic differentiation of SHED and DPSC normal subjects are well known. Nevertheless, the osteogenic differentiation capacity by RUNX-2 mRNA expression in DPSC and SHED cleft lip and palate patients is still need to be elucidated. Objective: To compare the osteogenic differentiation capacity of stromal cells from human exfoliated deciduous teeth and dental pulp stromal cells in cleft lip and palate patients through RUNX-2 gene expression. Methods: DPSC and SHED cleft lip and palate patients were cultured with and without osteogenic medium for 21 days. RNA sample were collected from cell culture followed by the examination of RUNX-2 and 18S gene expression were tested by Real-Time Polymerase Chain Reaction (RT-PCR). Result: There was no difference in osteogenic differentiation capacity between DPSC and SHED cleft lip and palate patients through RUNX-2 gene expression. Conclusion: The osteogenic differentiation capacity of SHED was equivalent to DPSC of cleft lip and palate patients."

"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."