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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: Rekayasa jaringan merupakan alternatif untuk perawatan rekonstruksi tulang alveolar pasien celah bibir dan palatum. Alternatif tersebut menghubungkan penggunaan sel punca, biomaterial/scaffolds, dan molekul sinyal. Sumber sel yang ideal untuk rekayasa jaringan adalah sel autologous karena tidak bersifat immunogenik. Sel stromal pulpa gigi permanen (DPSC) menarik untuk terapi klinis karena akses perolehannya yang mudah, morbiditas yang sangat rendah, menunjukkan kapasitas imunoregulasi yang menguntungkan, dan dapat berdiferensiasi menjadi banyak tipe sel, termasuk osteoblas. Pada penelitian sebelumnya, DPSCs pasien celah bibir dan palatum ditemukan memiliki potensi kemampuan osteogenik. Namun, kemampuan diferensiasi osteogeniknya belum diketahui. Kemampuan diferensiasi osteogenik tersebut dapat diamati dari ekspresi marker osteogenik, salah satunya sclerostin yang diekspresikan pada tahap akhir diferensiasi osteoblas. Tujuan: Membandingkan kemampuan diferensiasi osteogenik DPSCs pasien celah bibir dan palatum dengan DPSCs subjek normal melalui pengamatan ekspresi gen sclerostin. Metode: DPSCs dikultur hingga mencapai 70%-80% confluent. Sampel RNA dari sel diperoleh dengan melakukan prosedur ekstraksi RNA. Ekspresi gen sclerostin diamati menggunakan Real-Time PCR menggunakan primer sclerostin dan 18s sebagai housekeeping gene. Hasil: DPSCs pasien celah bibir dan palatum memiliki nilai rata-rata ekspresi relatif gen sclerostin yang lebih tinggi 1,9 kali lipat dibandingkan dengan DPSCs subjek normal dan secara statistik berbeda bermakna dengan p = 0,013. Kesimpulan: DPSCs pada pasien celah bibir dan palatum mengekspresikan gen sclerostin sebagai marker diferensiasi osteogenik yang lebih tinggi dibandingkan DPSCs pada subjek normal secara in vitro. ......Background: Tissue engineering is an alternative for alveolar bone reconstruction treatment in cleft lip and palate (CLP) patients. The alternative links the use of stem cells, biomaterials/scaffolds, and signaling molecules. The ideal cell source for tissue engineering is autologous cells because they are not immunogenic. Dental pulp stromal cells (DPSC) are interesting for clinical therapy because of their easy accesses, very low morbidity, exhibit favorable immunoregulatory capacities, and can differentiate into many cell types, including osteoblasts. In a previous study, DPSCs in CLP patients were found to have a potential osteogenic ability. However, its osteogenic differentiation ability is not yet known. The ability of osteogenic differentiation can be observed from the expression of osteogenic markers, one of which is sclerostin, a marker that is expressed in the final stage of osteoblast differentiation. Objective: To compare osteogenic differentiation ability of DPSCs in CLP patients with DPSCs in normal subjects through the expression of sclerostin gene. Methods: DPSCs were cultured to reach 70%-80% confluent. RNA samples from cells were obtained by carrying out RNA extraction procedure. Sclerostin gene expression was assessed using Real-Time PCR using sclerostin primer and 18s as a housekeeping gene. Results: DPSCs from CLP patients have mean relative expression of sclerostin gene 1.9 times higher compared to DPSCs in normal subjects and it is statistically different with p = 0.013. Conclusions: DPSCs in CLP patients express the sclerostin gene as marker of osteogenic differentiation higher than DPSCs in normal subjects in vitro.