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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 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. ......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.

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