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"Kitosan telah diketahui sebagai material scaffold poten pada rekayasa jaringan tulang. Penelitian ini bertujuan menganalisis potensi lain kitosan sebagai induktor diferensiasi sel punca pulpa gigi (SPPG) menjadi osteoblas dibandingkan dengan deksametason yang telah umum digunakan, melalui ekspresi mRNA Col1A1. SPPG dikultur pada medium yang mengandung kitosan, deksametason dan kombinasi keduanya selama 7 hari. Ekspresi mRNA Col1A1 diuji dengan metode comparative CT real-time PCR. Medium mineralizing yang ditambahkan 5 ng/ml kitosan dapat meningkatkan ekspresi mRNA Col1A1 pada SPPG dibandingkan dengan kontrol dan kelompok perlakuan lainnya (p<0,05). Kitosan dapat menginduksi diferensiasi tahap awal SPPG menjadi osteoblas bergantung pada konsentrasi yang diberikan.

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Chitosan has been proven as potential scaffold in bone tissue engineering. This research intends to analyze the other potency of chitosan as inductor in dental pulp stem cell (DPSC) differentiation into osteoblast compared to dexamethasone which is commonly used, by Col1A1 mRNA expression. DPSC were cultured in medium loaded with chitosan, dexamethasone and combination of both for 7 days. Col1A1 mRNA expression was analyzed by comparative CT method real time PCR. Mineralizing medium loaded with 5 ng/ml chitosan could enhance Col1A1 mRNA expression compared to control and another treated group on p<0,05. Chitosan could stimulate early stage of osteoblast differentiation. The effect was dose-dependent."

"Kitosan dan deksametason merupakan material yang digunakan dalam rekayasa jaringan tulang. Kitosan biasa digunakan sebagai scaffold, sedangkan deksametason sering digunakan sebagai sinyal. Salah satu penanda diferensiasi osteoblas adalah Alkaline phosphatase (ALP). Penelitian ini bertujuan menganalisis efek kitosan dibandingkan deksametason dalam menginduksi diferensiasi osteoblas melalui aktivitas ALP pada sel punca pulpa gigi (SPPG). Aktivitas ALP dianalisis dengan ALP assay. Terdapat 4 perlakuan yang memiliki aktivitas ALP diatas kontrol, yakni kitosan 5 ng/ml, deksametason 10 nM dan 100 nM, serta campuran kitosan 5 ng/ml dan deksametason 10 nM. Peningkatan konsentrasi deksametason meningkatkan aktivitas ALP. Peningkatan konsentrasi kitosan menurunkan aktivitas ALP.

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Chitosan and dexamethasone are materials that can be used in bone tissue engineering. Chitosan is often used as a scaffold, while dexamethasone is often used as signal. One of the markers of osteoblast differentiation is Alkaline phosphatase (ALP). This research objective is to analyse the effects of chitosan compared to dexamethasone in inducing osteoblast differentiation through ALP activity in Dental Pulp Stem Cells (DPSCs). ALP activity determined by ALP Assay. There were four treatments that have higher activity than the control, they are chitosan 5 ng/ml, 10 nM dexamethasone and 100 nM, and mixture of chitosan 5 ng/ml and 10 nM dexamethasone. The increased concentrations of dexamethasone increases ALP activity and the higher concentration of chitosan will decrease the ALP activity."

"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: 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: 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 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."

"Pendahuluan: Ekspresi berlebih suatu molekul bernama survivin dapat mencegah terjadinya apoptosis dengan menghambat caspase 9 dan mempertahankan keganasan pada sel glioblastoma. Penggunaan secretome sel punca tali pusat dapat mencegah perkembangan glioblastoma meskipun penggunaanya masih kontroversial. Untuk menguji efektifitas apoptosis secretome sel punca tali pusat pada glioblastoma, dapat dilakukan pengukuran survivin dan caspase 9 yang merupakan komponen penting pada jalur apoptosis intrinsik. Dengan demikian penelitian ini bertujuan untuk menganalisis pengaruh sekretom sel punca tali pusat terhadap ekspresi survivin dan caspase9 sebagai molekul yang mempengaruhi pertumbuhan glioblastoma Metode: Desain eksperimental in vitro dengan menggunakan galur sel glioblastoma T98G. Conditioned medium (CM) yang mengandung secretome sel punca tali pusat diperoleh dari medium sel punca  tali pusat yang dikultur selama 24 jam dengan  Î±MEM yang tidak mengandung serum. CM diencerkan 2 kali  dengan  medium tinggi glukosa Dulbecco's Modified Eagle Medium (DMEM) (50% konsentrasi). Perlakuan Sel glioblastoma T98G yang diberikan CM 50% dilakukan selama 24 jam, kemudian dilakukan ekstraksi RNA total dari sel T98G tersebut. RNA total digunakan untuk analisis ekspresi gen dengan menggunakan real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Metode Livak dilakukan untuk menghitung ekspresi relatif  caspase 9 dan survivin dengan 18srRNA sebagai gen referensi.

Hasil: Ekspresi mRNA survivin meningkat 3.5 kali (p=0.002) dan ekspresi mRNA caspase 9 meningkat 1.6 kali (p=0.017) pada sel T98G yang diberikan CM50% dibandingkan sampel kontrol. Kesimpulan : CM  sel punca pada tali pusat meningkatkan ekspresi survivin dan caspase 9 pada sel glioblastoma T98G. Penelitian selanjutnya diperlukan untuk mengetahui pengaruh peningkatan ekspresi gen tersebut terhadap proliferasi sel glioblastoma serta mekanisme molekulernya.

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Introduction: Aberrant expression of a molecule called survivin has been suspected to prevent apoptosis by indirectly inhibits a critical apoptosis enzyme called caspase 9, maintaining tumorigenicity of glioblastoma. Umbilical cord mesenchymal stem cells (UCMSC) has been discovered to inhibit glioblastoma growth but its usage is still controversial. To measure the apoptosis effectiveness of UCMSC-CM secretome against glioblastoma, measuring survivin and caspase 9, as essential components in intrinsic apoptosis pathway, are required. Therefore, this research aims to analyze the effect of UCMSC-CM against survivin and caspase 9 expression as the molecules that affect the growth of glioblastoma.

Method: In vitro experimental design by using glioblastoma cell line T98G. Conditioned medium (CM) which contain umbilical cord mesenchymal stem cell (UCMSC) secretome was obtained from UCMSC-CM that was cultured for 24 hours with αMEM that does not contain serum. CM was diluted twice with high glucose Dulbecco’s Modified Eagle Medium (DMEM) (50% concentration). The treatment of T98G glioblastoma cell, that had been exposed to 50% CM, was performed for 24 hours, then total RNA extraction was carried out from the T98G cells. Total RNA was used to analyze genetic expression by using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Livak’s method was used to calculate relative expression of Survivin and Caspase 9 expression with 18srRNA as reference gene.

Results: Survivin mRNA expression increased 3.5-fold (p=0.002) while caspase 9 mRNA expression increased 1.6-fold (p=0.017) in T98G cell that was given CM50% compared with control sample.

Conclusion: UCMSC-CM increases survivin and caspase 9 expression in glioblastoma T98G cells. Future researches are required to identify the effect of the elevated genetic expressions against glioblastoma cell proliferation as well as their molecular mechanisms."

"Latar Belakang: Rekayasa jaringan tulang memerlukan tiga komponen utama, yaitu sel punca, scaffold, dan faktor pertumbuhan. IGF-1 merupakan salah satu faktor pertumbuhan yang berperan dalam proliferasi dan diferensiasi sel osteoblast. IGF-1 akan berikatan dengan reseptornya, yaitu IGF-1R untuk mengaktivasi jalur hilir. Dalam sirkulasi tubuh manusia, IGF berikatan dengan IGFBP-3 yang dapat memperpanjang waktu paruh serta menghambat IGF-1 berikatan dengan IGF-1R. Pada penelitian sebelumnya, tercatat bahwa tidak ada perbedaan kemampuan proliferasi dan diferensiasi antara DPSC subjek normal dan subjek CLP, namun ada perbedaan signifikan dalam jumlah ekspresi IGF-1. OCT-4, SOX-2 dan NANOG merupakan faktor transkripsi utama pluripotensi yang telah diteliti dapat mengatur pluripotensi, pembaruan diri, proliferasi, serta diferensiasi DPSC. Penelitian terbaru mencatat peningkatan ekspresi ketiga gen tersebut pasca dilakukan penghambatan jalur GSK-3 dan m-TOR yang merupakan jalur hilir dari aksi IGF-1 pada sel DPSC. Namun, belum diketahui secara pasti ekspresi ketiga gen tersebut pada DPSC subjek normal dan CLP setelah dilakukannya penghambatan IGF-1 menggunakan anti IGF-1R dan IGFBP-3. Tujuan: Menganalisis pengaruh anti IGF-1 dan IGFBP-3 terhadap ekspresi gen OCT4, SOX2, dan NANOG pada DPSC subjek normal dan CLP. Metode: Sampel RNA DPSC subjek normal (n=4) dan DPSC subjek CLP (n=3), sebelum dan setelah diberikan perlakuan anti IGF-1R atau IGFBP-3, diperoleh dari bahan biologis tersimpan di Laboratorium Oral Biologi Fakultas Kedokteran Gigi Universitas Indonesia. Selanjutnya, ekspresi gen OCT4, SOX2, NANOG, dan housekeeping gene GAPDH diuji dengan two step Real-Time PCR (RT-PCR). Hasil: Tidak terdapat perbedaan ekspresi gen OCT4, SOX2, dan NANOG, baik antara DPSC subjek normal dan CLP sebelum dan setelah diberikan perlakuan anti IGF-1R dan IGFBP-3 (p³0,05). Kesimpulan: Perlakuan anti IGF-1R dan IGFBP-3 tidak memengaruhi tingkat ekspresi gen OCT4, SOX2, dan NANOG sel punca pulpa gigi permanen subjek normal dan subjek celah bibir dan palatum

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Background: Bone tissue engineering requires three main components, namely stem cells, scaffold, and growth factors. IGF-1 is a growth factor that plays role in osteoblast proliferation and differentiation. IGF-1 will bind to its receptor, namely IGF-1R, to activate the downstream pathway. In the human body circulation, IGF binds to IGFBP-3 which can inhibit IGF-1 from binding to IGF-1R. Previous studies noted that there were no differences in the ability to proliferate and differentiate between DPSC from normal subjects and CLP subjects, yet there were significant differences in the level of IGF-1 expression. OCT-4, SOX-2 and NANOG are core pluripotency factors which regulate pluripotency, self-renewal, proliferation and differentiation of DPSC. Recent study has noted an increase in the expression of these three genes after inhibition of GSK-3 and m-TOR pathways, which are the downstream pathways of IGF-1 on DPSC cells. However, the expression of these three genes in DPSC from normal and CLP subjects after inhibition of IGF-1 using anti IGF-1R and IGFBP-3 is still unknown. Objective: To analyze the effect of anti IGF-1 and IGFBP-3 on OCT4, SOX2, and NANOG gene expression in DPSC of normal and CLP subjects. Methods: RNA samples of DPSC from normal and CLP subjects, before and after being treated with anti-IGF-1R or IGFBP-3, were obtained from Laboratory of Oral Biology, Faculty of Dentistry, Universitas Indonesia. Furthermore, the expression of OCT4, SOX2, NANOG, and housekeeping gene GAPDH were tested using two step Real-Time PCR (RT-PCR). Results: There was no difference between the expression of the OCT4, SOX2, and NANOG in DPSC from normal and CLP subjects before and after anti IGF-1R and IGFBP-3 treatment (p≥0.05). Conclusion: Anti-IGF-1R and IGFBP-3 did not affect the expression level of OCT4, SOX2, and NANOG in dental pulp stem cells of normal subjects and cleft lip and palate subjects.

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"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."