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"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: Celah bibir dan palatum merupakan salah satu kelainan bawaan yang menyebabkan defek jaringan keras sehingga dikembangkan perawatan rekonstruksi tulang berbasis teknik rekayasa jaringan sebagai alternatif perawatan. Sumber sel stromal mesenkim dapat diperoleh dari pulpa gigi sulung dan gigi permanen. Kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen sudah banyak dilaporkan. Pada pasien celah bibir dan palatum, terdapat gen-gen yang diekspresikan berbeda dan kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum belum diketahui. Tujuan: Mengevaluasi perbandingan kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum melalui deposisi kalsium. Metode: Sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum dikultur menggunakan medium osteogenik selama 21 hari kemudian dilakukan pewarnaan Alizarin Red dan kuantifikasi terhadap deposisi kalsium. Hasil: Sel stromal pulpa gigi sulung dan gigi permanen yang dikultur menggunakan medium osteogenik menunjukkan adanya deposisi kalsium yang tinggi. Sel stromal pulpa gigi sulung dan gigi permanen tidak menunjukkan perbedaan nilai rerata absorbansi, intensitas pewarnaan, dan area pewarnaan yang bermakna secara statistik (p ≥ 0,05). Kesimpulan: Sel stromal pulpa gigi sulung pasien celah bibir dan palatum memiliki kemampuan diferensiasi osteogenik yang ekuivalen dengan sel stromal pulpa gigi permanen pasien celah bibir dan palatum.

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Background: Cleft lip and palate is one of the most common congenital anomalies resulting in hard tissue defects therefore tissue engineering is currently developed as an alternative treatment. The source of mesenchymal stromal cells can be obtained from human exfoliated deciduous teeth (SHED) and dental pulp (DPSCs). Osteogenic differentiation abilities of SHED and DPSCs have been widely studied. In cleft lip and palate patients, there are several differentially expressed genes and the osteogenic differentiation abilities of SHED and DPSCs in cleft lip and palate patients have not yet been known. Purpose: To compare the osteogenic differentiation abilities of SHED and DPSCs in cleft lip and palate patients by calcium deposition. Methods: SHED and DPSCs isolated from cleft lip and palate patients were cultured using osteogenic medium for 21 days then added Alizarin Red staining and the calcium deposition were quantified. Result: Both SHED and DPSCs that cultured in osteogenic medium demonstrated high calcium deposition. SHED and DPSCs did not show any statistically significant differences in the average absorbance values, staining intensity, and staining areas (p ≥ 0,05). Conclusion: SHED and DPSCs in cleft lip and palate patients have equivalent ability of osteogenic differentiation by calcium deposition."

"Latar belakang: Celah bibir dan palatum (CLP) adalah kegagalan fusi prosesus frontonasal dan maksilaris yang menghasilkan celah yang meluas ke bibir, alveolus, dasar hidung, dan palatum keras maupun lunak. Diperlukan perawatan melalui teknik rekayasa jaringan dengan menggunakan sel stromal mesenkim yang dapat ditemukan pada dental pulp stromal cells (DPSC). Kemampuan osteogenik DPSC dapat dilihat melalui deteksi marker osteogenik seperti osteopontin (OPN). Osteopontin merupakan salah satu marker utama diferensiasi osteogenik yang diproduksi oleh osteoblas dalam proses pembentukan dan mineralisasi tulang. Pada pasien CLP diketahui perbedaan ekspresi gen yang dapat memengaruhi sintesis matriks ekstraseluler. Penelitian ini dilakukan untuk mengetahui kemampuan diferensiasi osteogenik melalui ekspresi marker osteopontin saat diferensiasi osteoblas pada pasien celah bibir dan palatum. Tujuan: Membandingkan kemampuan diferensiasi osteogenik sel stromal pulpa gigi subjek normal dengan sel stromal pulpa gigi pasien celah bibir dan palatum melalui ekspresi gen osteopontin. Metode: Sampel RNA yang diperoleh dari kultur sel pulpa gigi subjek normal dan pasien celah bibir dan palatum diuji dengan Real-Time Polymerase Chain Reaction (RT-PCR) dengan primer osteopontin (OPN) dan 18S sebagai housekeeping gene. Hasil: Tidak terdapat perbedaan antara ekspresi relatif gen OPN sel stromal pulpa gigi subjek normal dan pasien celah bibir dan palatum. Kesimpulan: Kemampuan diferensiasi osteogenik sel stromal pulpa gigi pada pasien celah bibir dan palatum ekuivalen dengan sel stromal pulpa gigi pada subjek normal.

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Background: Cleft lip and palate (CLP) occurs due to the failure of fusion of the frontal and maxillary process that results in a cleft that extends to the lip, alveoli, nose floor, and hard and soft palate. One of the potentially alternative treatment for CLP cases is tissue engineering technique using Mesenchymal Stromal Cell (MSC). MSC can be found in dental pulp stromal cells (DPSC). Osteogenic ability can be seen through the detection of osteogenic markers such as osteopontin (OPN). Osteopontin is one of the main markers of osteogenic differentiation produced by osteoblast in the process of bone formation and mineralization. Osteopontin is expressed by preosteoblasts in early bone formation and mature osteoblasts at bone remodelling sites. Osteopontin expression as one of osteogenic markers in cleft lip and palate patients is unknown. This study was conducted to determine the ability of osteogenic differentiation through the expression of osteopontin in cleft lip and palate patients. Objective: To compare osteogenic differentiation ability of mesenchymal stromal cells in cleft lip and palate patients and normal subject through the expression of osteogenic marker osteopontin. Methods: RNA sample that was obtained through RNA extraction from dental pulp stromal cells of cleft and lip palate patients and normal subjects were tested with Real-Time Polymerase Chain Reaction (RT-PCR) using osteopontin and 18S as housekeeping gene. Results: There was no difference between the relative expression of OPN gene in DPSC from normal subject and cleft lip and palate patients. Conclusion: Osteogenic differentiation ability of dental pulp stromal cells from cleft lip and palate patients is equivalent with dental pulp stromal cells from normal subjects."

"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.

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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."

"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: Subjek celah bibir dan palatum membutuhkan perawatan rekonstruksi tulang berbasis rekayasa jaringan dengan menggunakan sel stromal mesenkim. Sel stromal mesenkim merupakan sel yang banyak digunakan untuk regenerasi tulang karena mempunyai kemampuan proliferasi tinggi. Sel tersebut dapat berasal dari pulpa gigi sulung (SHED) danpulpa gigi permanen (DPSCs) yang dapat berdiferensiasi menjadi osteoblas. Pada penelitiansebelumnya telah ditemukan beberapa karakteristik DPSCs dan SHED pada subjek celah bibir dan palatum, namun kemampuan diferensiasi dari sel stromal pulpa subjek celah bibir dan palatum belum diketahui. Tujuan: Mengevaluasi kemampuan diferensiasi osteogenik dari sel stromal pulpa gigi permanen dan sulung pada subjek celah bibir dan palatum melaluiekspresi gen Collagen Type I Alpha I (COL1A1). Metode : Sampel RNA yang diperoleh dari kultur RNA DPSCs dan SHED subjek celah bibir dan palatum, dengan Real-Time Polymerase Chain Reaction (RT-PCR) menggunakan primers Collagen Type I Alpha I (COL1A1), serta 18S sebagai housekeeping gene. Hasil : Tidak terdapat perbedaan ekspresi relatif gen COL1A1 antara sel stromal pulpa gigi permanen dan sel stromal pulpa gigi sulung pada subjek celah bibir dan palatum. Kesimpulan : SHED memiliki kemampuan diferensiasi osteogenik yang sama dengan DPSCs karena keduanya dapat mengekspresikan gen marker osteogenik COL1A1.

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Background: Cleft lip and palate subject need bone reconstruction based tissue engineering treatment with mesenchymal stromal cells (MSC). One of the most mesenchymal stromal cells that can be used is derived from dental pulp tissues, such as primary tooth pulp or stem cells from human deciduous teeth (SHED) and dental pulp stem cells (DPSCs) which can differentiate into osteoblasts. In previous studies, several characteristics of DPSCs and SHED of the cleft lip and palate subjects have been found. However, osteogenic differentiation ability of dental pulp stromal cells from cleft lip and palate subject is unknown.

Objective: To determine the osteogenic differentiation ability of DPSCs and SHED of cleft lip and palate subjects through the expression of the Collagen Type I Alpha I (COL1A1) gene.

Methods: RNA samples obtained from the culture of DPSCs and SHED of lip and palate cleft subjects, with Real-Time Polymerase Chain Reaction (RT-PCR) using primers Collagen Type I Alpha I (COL1A1) and 18S as a housekeeping gene.

Results: There was no difference in the relative expression of COL1A1 gene between DPSCs and SHED of CLP subjects.

Conclusion: SHED has the same osteogenic differentiation ability as DPSCs because they can express osteogenic marker genes COL1A1."

"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."