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"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.

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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."

"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: 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: Celah bibir dan palatum adalah keadaan dimana terdapat gangguan fusi atau celah abnormal bawaan pada daerah bibir atas, alveolar, dan palatum serta dapat menimbulkan masalah pada penderita seperti gangguan estetika dan masalah saat berbicara. Perawatan rekonstruksi tulang dengan autologous bone graft merupakan baku emas pada perawatan pasien celah bibir dan palatum, tetapi perawatan ini memiliki kekurangan sehingga dikembangkan alternatif perawatan seperti teknik rekayasa jaringan. Sumber sel stromal mesenkim yang digunakan dapat berasal dari jaringan pulpa gigi seperti sel stromal pulpa gigi sulung dan sel stromal pulpa gigi permanen. Kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan permanen pasien celah bibir dan palatum merupakan salah satu pertimbangan untuk penggunaan sel autologous dalam perawatan teknik rekayasa jaringan, sedangkan kemampuan diferensiasi osteogenik dari sel stromal pulpa gigi pasien CLP belum diketahui. Tujuan: Membandingkan kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum melalui ekspresi gen ALP. Metode: Sampel yang diisolasi dari jaringan pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum dikultur pada medium osteogenik, dilakukan ekstraksi RNA dan diuji dengan Real-Time Polymerase Chain Reaction (RT PCR) menggunakan primers alkaline phosphatase (ALP) dan 18s housekeeping gene. Hasil: Ekspresi relatif gen ALP pada sel stromal pulpa gigi sulung pasien celah bibir dan palatum setelah dilakukan uji statistik tidak memiliki perbedaan bermakna bila dibandingkan dengan sel stromal pulpa gigi permanen pasien celah bibir dan palatum (nilai p = 0.156). Kesimpulan: Sel stromal pulpa gigi sulung dan gigi permanen memiliki kemampuan diferensiasi osteogenik karena dapat mengekspresikan marker osteogenik ALP.

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Background: Cleft and lip palate is a condition where there is fusion disturbance or abnormal congenital cleft in the upper lip, alveolar, and palate area that can cause problems in patients such as aesthetic disorder and problem with talking. Autologous bone graft reconstruction treatment is the gold standard in treating cleft lip and palate patients, but this treatment has associated shortcomings so that alternative treatments such as tissue engineering techniques have been developed. The source of the mesenchymal stromal cells used can be derived from dental pulp tissue namely stem cells from human deciduous teeth and permanent dental pulp stromal cells. The osteogenic differentiation ability from dental pulp stromal cells of primary and permanent teeth in cleft lip and palate patients is one of the considerations for the use of autologous cells in the treatment of tissue engineering techniques, while the osteogenic differentiation ability of dental pulp stromal cells in cleft lip and palate patients has not been fully explored.

Objective: To compare the osteogenic differentiation capacity of primary and permanent dental pulp stromal cells in cleft lip and palate patients.

Methods: Samples isolated from primary and permanent dental pulp stromal cells in cleft lip and palate patients were cultured, RNA were extracted and tested by Real-Time Polymerase Chain Reaction (RT PCR) using alkaline phosphatase primers (ALP), and housekeeping gene in the form of 18s. Results: The relative expression of ALP in primary dental pulp stromal cells in cleft lip and palate patients was comparable to permanent dental pulp stromal cells in cleft lip and palate patients (p value = 0.156).

Conclusion: The primary and permanent dental pulp stromal cells have comparable ability to differentiate into osteogenic lineage and both cells tested can express the osteogenic gene of ALP."

"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."