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Andi Adytha Mutiah Ittie Rusiaty
"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.

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."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2012
S44749
UI - Skripsi Membership  Universitas Indonesia Library
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Nurul Ramadiani
"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.

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."
Depok: Universitas Indonesia, 2012
S44220
UI - Skripsi Membership  Universitas Indonesia Library
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Alya Hana Firdanisa
"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.

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.
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Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2021
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UI - Skripsi Membership  Universitas Indonesia Library
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Puput Wulandari
"Latar belakang: Celah bibir dan palatum atau cleft lip and palate (CLP) merupakan kelainan kongenital multifaktorial yang mengakibatkan pasien memiliki defek pada jaringan lunak dan keras di bagian bibir dan palatum. Pasien celah bibir dan palatum umumnya menderita gangguan estetik dan fungsi stomatognatik. Sehingga, untuk mengembalikan fungsinya maka harus dilakukan perawatan rekonstruksi tulang alveolar. Baku emas dalam perawataan ini ialah menggunakan autologous bone grafting. Namun, perawatan ini masih memiliki kekurangan sehingga dikembangkan perawatan yang baru dengan teknik rekayasa jaringan dengan sel punca mesenkim. Salah satu sumber sel punca mesenkim yaitu berasal dari jaringan pulpa gigi yaitu sel punca pulpa gigi permanen atau dental pulp stem cells (DPSCs) dan sel punca pulpa gigi sulung atau stem cells from human deciduous teeth (SHED). Kemampuan osteogenik dari sel punca merupakan salah satu faktor pertimbangan untuk pemakaian sel dalam rekayasa jaringan rekonstruksi tulang. Sementara kemampuan osteogenik dari SHED dan DPSCs CLP belum diketahui.
Tujuan: Mengetahui potensi kemampuan dan perbandingan potensi osteogenik dari sel punca gigi permanen dan sulung pasien celah bibir dan palatum dengan melihat ekspresi gen Alkaline phosphatase (ALP) dan Collagen Type I Alpha 1 (COL1A1).
Metode: Sampel RNA yang diperoleh dari ekstraksi RNA sel jaringan pulpa gigi sulung dan permanen pasien celah bibir dan palatum diuji dengan Real-Time Polymerase Chain Reaction (RT-PCR) menggunakan primers Alkaline Phosphatase (ALP), Collagen Type-I (COL1A1) dan Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) sebagai housekeeping gene.
Hasil: Ekspresi relatif gen ALP pada sel punca pulpa gigi sulung pasien celah bibir dan palatum mengalami penurunan dibandingkan dengan sel punca gigi permanen pasien celah bibir dan palatum. Sementara untuk ekspresi gen COL1A1 pada sel punca pulpa gigi sulung pasien celah bibir dan palatum tidak memiliki perbedaan dibandingkan dengan sel punca gigi permanen pasien celah bibir dan palatum.
Kesimpulan: Sel punca pulpa gigi sulung dan permanen pasien celah bibir dan palatum memiliki potensi kemampuan osteogenik dikarenakan keduanya mengekspresikan gen marker osteogenik seperti ALP dan COL1A1.

Background: Cleft lip and palate (CLP) is a multifactorial congenital disorder that results in patients having soft and hard tissue defects in the lips and palate. Patients with cleft lip and palate commonly suffer from aesthetic and stomatognathic function disorders. Therefore, to restore its function, alveolar bone reconstruction treatment must be done. The gold standard in this treatment is to perform autologous bone grafting. However, as autologous bone grafting still has associated shortcomings, new treatments using tissue engineering techniques with mesenchymal stem cells are being developed. One of the mesenchymal stem cells sources that can be used is derived from dental pulp tissue, namely permanent dental pulp stem cells (DPSCs) and primary dental pulp stem cells or stem cells from human deciduous teeth (SHED). Osteogenic ability of the stem cells is one of the factors considered for the use of cells in tissue engineering bone reconstruction. Osteogenic ability of SHED and DPSCs hasn’t been fully explored.
Objective: To determine the potential ability and to compare osteogenic potential of DPSCs and SHED from cleft lip and palate patients by looking at the expression of the Alkaline Phosphatase (ALP) and Collagen Type I Alpha 1 (COL1A1) genes.
Methods: RNA samples obtained from RNA cells extraction in deciduous and permanent dental pulp tissue of patients with cleft lip and palate were tested with Real-Time Polymerase Chain Reaction (RT-PCR) using Alkaline Phosphatase (ALP) primers, Collagen Type I Alpha 1 (COL1A1) primers and Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) primers as housekeeping gene.
Results: The relative expression of ALP genes in SHED from CLP patients decreased compared to DPSCs from patients with CLP. As for the expression of the COL1A1 gene, there was no difference in expression between SHED from patients with cleft lip and palate and DPSCs in patients with cleft lip and palate.
Conclusion: SHED and DPSCs CLP has osteogenic abilities.
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Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2019
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UI - Skripsi Membership  Universitas Indonesia Library
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Najmi Affifi
"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) dan
pulpa gigi permanen (DPSCs) yang dapat berdiferensiasi menjadi osteoblas. Pada penelitian
sebelumnya 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 melalui
ekspresi 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.

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.
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Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2020
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UI - Skripsi Membership  Universitas Indonesia Library
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Hawa Annisa Sudadiyo
"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."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2020
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UI - Skripsi Membership  Universitas Indonesia Library
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Rizqa Inayati
"Diferensiasi osteogenik dari Sel punca mesenkim (MSC) menjadi osteoblas memiliki signifikansi klinis yang sangat penting untuk mengobati cedera tulang. Berbagai penelitian telah dilakukan untuk meneliti faktor-faktor yang dapat meningkatkan diferensiasi osteogenik, termasuk pengembangan perancah untuk kultur MSC. Perancah Polivinil Alkohol (PVA)/ Fibroblast-derived Matrix (hFDM) asal manusia menjadi salah satu kandidat perancah yang diduga dapat mendukung diferensiasi osteogenik MSC. Keberadaan matriks ekstraseluler (ECM) pada perancah dapat meregulasi berbagai aktivitas seluler melalui komponen protein matriks yang terdapat pada ECM. Protein matriks berperan sebagai sekuesterasi berbagai faktor pertumbuhan. Faktor pertumbuhan seperti BMP2 dan chordin diketahui dapat meregulasi diferensiasi osteogenik. Proses terjadinya diferensiasi osteogenik dapat diamati melalui akumulasi mineral kalsium yang terdeposit pada matriks ekstraseluler. Tujuan dari penelitian ini adalah untuk mengetahui metode optimum dalam pembuatan perancah PVA/hFDM, danmengetahui peran perancah PVA/hFDM dalam mempengaruhi diferensiasi osteogenik MSC dengan mengukur ekspresi gen BMP2, dan chordin, serta ekspresi kadar kalsium relatif pada matriks ekstraseluler. Optimasi pembuatan perancah PVA hFDM dimulai dengan optimasi medium kultur, waktu kultur, preparasi, dan teknik deselulerisasi. hFDM dikarakterisasi menggunakan pewarnaan Hematoxylin, Masson Trichrome, dan Imunohistokimia untuk mengetahui keberadaan protein matriks. MSC dikultur pada perancah PVA/hFDM untuk uji diferensiasi osteogenik selama 21 hari. Sampel RNA diisolasi pada hari ke-7,14, dan 21. Ekspresi gen BMP2 dan chordin dianalisis menggunakan metode qRT-PCR. Adapun ekspresi kadar kalsium relatif dianalisis dengan uji kualitatif dan kuantitatif pewarnaan Alizarin Red. Hasil penelitian ini menunjukkan protokol pembuatan perancah PVA/hFDM telah dioptimasi, dan karakterisasi hFDM memperlihatkan keberadaan protein matriks berupa kolagen dan biglycan. Ekspresi gen BMP2 menurun pada kelompok MSC yang dikultur pada perancah PVA/hFDM baik di hari ke-7, 14, dan 21. Sedangkan ekspresi gen chordin meningkat pada kelompok MSC yang dikultur pada perancah PVA/hFDM di hari ke 7, dan 14, kembali menurun di hari ke-21. Ekspresi kadar kalsium relatif cenderung meningkat pada kelompok MSC yang dikultur pada perancah PVA/hFDM dengan gambaran mikroskopis berupa bercak merah pada permukaan perancah. Kesimpulan dari penelitian ini adalah perancah PVA/hFDM cenderung dapat mendukung diferensiasi osteogenik MSC. Hasil penelitian menunjukkan bahwa penggunaan perancah PVA/hFDM dapat menurunkan ekspresi gen BMP2, dan meningkatkan ekspresi gen chordin, serta cenderung meningkatkan ekspresi kadar kalsium relatif yang terdeposit pada matriks ekstraseluler.

Osteogenic differentiation from Mesenchymal Stem Cell (MSC) to osteoblasts has great clinical significance for treating bone injury. Various studies have been conducted to investigate factors that can enhance osteogenic differentiation, including scaffold development for MSC culture. Scaffold Polyvinyl Alcohol (PVA) / human Fibroblast-derived Matrix (hFDM) is a scaffold candidate assumed to support osteogenic differentiation of MSCs. The extracellular matrix (ECM) presence on the scaffold can regulate various cellular activities through the matrix protein components contained in the ECM. Matrix protein plays a role in sequestering multiple growth factors. Growth factors such as BMP-2 and chordin are to regulate osteogenic differentiation. The process of osteogenic differentiation can be observed by accumulating calcium minerals in the extracellular matrix. The purpose of this study was to determine the optimal method for making PVA / hFDM scaffold and to determine the role of the PVA / hFDM scaffold in affecting MSC osteogenic differentiation by measuring the expression of BMP2 and chordin genes, as well as the expression of relative calcium levels in the extracellular matrix. Optimization of making hFDM PVA scaffold begins with the optimization of culture medium, culture time, preparation, and decellularization techniques. hFDM was characterized using Hematoxylin, Masson Trichrome, and Immunohistochemical staining to determine matrix proteins' presence. MSCs were cultured on the PVA / hFDM scaffold for osteogenic differentiation assay for 21 days. RNA samples were isolated on day 7,14 and 21. Expression of BMP2 and chordin genes were analyzed using the qRT-PCR method. The expression of relative calcium levels was analyzed by qualitative and quantitative tests of Alizarin Red staining. The results of this study indicate that the PVA / hFDM scaffold preparation protocol has been optimized, and the hFDM characterization shows the presence of matrix proteins in the form of collagen and biglycan. BMP-2 gene expression decreased in the MSC group cultured on the PVA / hFDM scaffold on days 7, 14, and 21. In contrast, the chordin gene expression increased in the MSC group cultured on the PVA / hFDM scaffold on days 7, and 14, back down on day 21. The expression of relative calcium levels tended to increase in the MSC group cultured on the PVA / hFDM scaffold with a microscopic appearance of red spots on the scaffold surface. This study concludes that Scaffold PVA / hFDM tends to support osteogenic differentiation of MSCs. The results showed that the use of the PVA / hFDM scaffold could decrease the expression of the BMP2 gene, and increase the expression of the chordin gene, and tended to increase the expression of the relative calcium levels deposited in the extracellular matrix.
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Jakarta: Fakultas Kedokteran Universitas Indonesia, 2020
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UI - Tesis Membership  Universitas Indonesia Library
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Insyirah Nabil Nismara
"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.

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."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2021
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UI - Skripsi Membership  Universitas Indonesia Library
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Azra Nadhira
"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.

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."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2021
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UI - Skripsi Membership  Universitas Indonesia Library
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Zulkarnain Edward
"Ruang Lingkup dan Cara Penelitian: Enzim fosfatase alkali antara lain digunakan dalam teknik ?enzyme immunoassay?, untuk mengukur kadar sesuatu zat dalam cairan tubuh dalam jumlah yang sangat kecil. Dalam penelitian ini diusahakan isolasi dan pemurnian enzim fosfatase alkali dari E. coli. Identifikasi kuman dilakukan dengan agar Endo, agar darah, tes pewarnaan Gram, sifat-sifat biokimia, dan tes serologik. Untuk pemurnian enzim digunakan sonikator, kromatografi pertukaran ion dengan DEAE Biogel, dan kromatografi gel dengan Sephadex G-100. Kemurnian enzim diperiksa dengan elektroforesis pada membran selulosa asetat. Aktivitas enzim secara kuantitatif ditentukan dengan spektrofotometer, dan secara kuaLitatif dapat dilihat dengan agar substrat. Kadar protein diukur dengan metoda Lowry. Terhadap fraksi gel diteliti pengaruh suhu, pH, ion logam, dan jenis bufer atas aktivitas enzim. Demikian pula ditentukan nilai Km dan Vmax, serta reaksi hidrolisis tanpa dan dengan transfosforilasi.
Hasil dan Kesimpulan: Kuman diidentifikasi sebagai E. coli non-patogen. Enzim diperoleh setelah fraksi gel dengan ,pemurnian 242 kali dan hasil 59%. Pada eLektroforesis ditemukan kadar protein enzim 52,8%. Enzim memiliki pH optimum 8,0, dan tidak stabil bila diinkubasi selama 1 jam diluar pH optimum. Aktivitas enzimeningkat secara Linier sampai suhu 45° C, dengan koefisien suhu 1,49. Enzim stabil pada inkubasi selama 20 menit pada suhu 25 - 45° C. Aktivitas enzim tidak dipengaruhi penambahan ion Mg dan Zn (0,01 M). Aktivitas meningkat dengan meningkatnya molaritas bufer, Vmax terbesar didapatkan dalam buffer Tris dan Km terkeciL pada bufer AMP. Reaksi hidrolisis dan transfosforilasi berlangsung pada bufer Tris dan AMP, sedangkan pada bufer glisin hanya terjadi reaksi hidrolisis.

Scope and Method of Study: The enzyme alkaline phosphatase is used among others in enzyme immunoassay, to enable the quantitation of a small amount of substances in body fluids. In this study, an attempt was carried out for the isolation and purification of the enzyme from E. coli. The bacteria was identified through culture on Endo and blood agar, Gram staining, biochemical tests, and serology. The bacteria were disrupted by ultrasonication, and the enzyme purified by ion exchange chromatography on DEAE Biogel followed by gel chromatography on Sephadex G-100. Enzyme purity was examined by electrophoresis on cellulose acetate. Enzyme activity was determined by spectrophotometry, and protein concentration was measured by the method of Lowry. The gel fraction was tested for the effect of pH, temperature, metal ion, and type of buffer. The Km and Vmax was measured, for hydrolysis with and without transphosphorylation.
Findings and Conclusions: The bacteria was identified as non-pathogenic E. coli. After gel chromatography the enzyme was purified 242 fold, at 59%, yield. Electrophoresis revealed that the enzyme content was 52.8 %. The enzyme has a pH optimum of 8.0, and it was unstable on standing for 1 hour outside the pH optimum. Enzyme activity increased Linearly with temperature (to 45° C), with a temperature coefficient of 1.49. The enzyme is stable for 20 minutes at 5° - 45++ C, and the activity not influenced by Mg++ and Zn++ ions (0.01 M). The activity increased with increased molarity of the buffer, the highest Vmax was observed with Tris buffer, and the Lowest Km with AMP buffer. Hydrolysis and transphosphorylation occurred in Tris and AMP buffer, while in glycine buffer only hydrolysis was observed."
Jakarta: Fakultas Kedokteran Universitas Indonesia, 1986
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UI - Tesis Membership  Universitas Indonesia Library
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