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Zahra Savira
Kapasitas Proliferasi Sel Punca Pulpa Gigi Sulung dan Sel Punca Pulpa Gigi Permanen Pasien Celah Bibir dan Palatum = Proliferative Capacity of Stem Cells from Exfoliated Deciduous Teeth and Dental Pulp Stem Cells in Cleft Lip and Palate Patients
"Latar Belakang: Celah bibir dan palatum (CLP) merupakan salah satu kelainan kongenital yang menghasilkan defek jaringan lunak maupun jaringan keras dan membutuhkan perawatan rekonstruksi tulang alveolar dan palatum. Celah bibir dan palatum dianggap berasal dari anomali proliferasi sel akibat faktor genetika. Autologous bone graft adalah baku emas untuk memperbaiki defek tulang palatum pada pasien CLP. Namun demikian, perawatan tersebut membutuhkan prosedur yang invasif. Perawatan melalui rekayasa jaringan dapat menjadi alternatif perawatan. Rekonstruksi tulang alveolar melalui rekayasa jaringan membutuhkan jumlah sel yang banyak sehingga kapasitas proliferasi sel punca merupakan aspek penting dalam penerapan klinis. Sel punca pulpa gigi sulung (SHED) dan sel punca pulpa gigi permanen (DPSCs) dapat menjadi sumber sel yang ideal karena memiliki kapasitas proliferasi yang tinggi, kemampuan diferensiasi ke berbagai tipe sel, isolasi yang mudah, dan aksesibilitas yang baik. Namun, kapasitas proliferasi SHED dan DPSCs pasien CLP belum diketahui.
Tujuan: Penelitian ini bertujuan membandingkan kapasitas proliferasi SHED dan DPSCs pasien celah bibir dan palatum.
Metode: SHED dan DPSCs dari pasien CLP dikultur hingga mencapai 70%-80% confluent. Kapasitas proliferasi sel setelah dikultur selama 24 jam, 48 jam, dan 72 jam dianalisis melalui uji MTT.
Hasil: SHED setelah dikultur 24 jam menunjukkan nilai rata-rata optical density yang lebih tinggi secara signifikan (p<0,05). SHED dan DPSCs setelah dikultur 48 jam dan 72 jam tidak menunjukkan perbedaan nilai rata-rata optical density secara statistik (p>0,05).
Kesimpulan: SHED pasien CLP memiliki kapasitas proliferasi lebih tinggi secara signifikan hanya pada 24 jam pertama. Pada 48 jam dan 72 jam pertama, SHED dan DPSCs pasien CLP memiliki kesamaan kapasitas proliferasi.
Background: Cleft lip and palate (CLP) is one of orofacial congenital malformations that results in both soft tissue and hard tissue defect. It requires reconstruction of the maxillary alveolar cleft. Cleft lip and palate is thought to be came from anomalies of cell proliferation caused by genetic factors. Autologous bone graft have been the gold standard treatment to repair maxillary alveolar and palate clefts. However, such treatment needs an invasive procedure that may induce pain. To overcome those disadvantages, tissue engineering has received attention to be new alternative treatment.
Reconstruction of maxillary alveolar cleft requires huge number of stem cells so that proliferative capacity is important traits before clinical application. Stem Cells from Exfoliateed Deciduous Teeth (SHED) and Dental Pulp Stem Cells (DPSCs) can be ideal sources of stem cell since they are known to have high proliferative capacity, multilineage differentiation, ease of isolation, and well accesibility. However, proliferative capacity of SHED and DPSCs isolated from CLP patients have not yet known.
Objective: The aim of this study was to compare proliferative capacity between cultured stem cells from exfoliated deciduous teeth and dental pulp stem cells isolated from cleft lip and palate patients.
Methods: SHED and DPSCs isolated from cleft patient were cultured until it reached 70%-80% confluency. Proliferative capacity after culturing for 24 hours, 48 hours, and 72 hours were analyzed using MTT Assay.
Results: SHED after culturing for 24 hours showed higher optical density average value significantly (p<0,05). SHED and DPSCs after culturing for 48 hours and 72 hours has no difference optical density average value significantly (p>0,05).
Conclusions: SHED from cleft patients showed higher proliferative capacity significantly only on first 24 hours culturing. SHED and DPSCs have similar proliferative capacity on 48 hous and 72 hours culturing."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2019
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Athira Astari
Population doubling time sel punca pulpa gigi sulung dan sel punca pulpa gigi permanen pasien celah bibir dan palatum = Population doubling time of stem cells from exfoliated decidious teeth and dental pulpa stem cells in cleft lip and palate patients
"Latar Belakang: Celah bibir dan palatum merupakan salah satu kelainan kongenital yang paling sering terjadi. Kelainan ini dapat menyebabkan kendala dalam berbicara, abnormalitas telinga tengah, masalah psikologis, serta kelainan dental seperti anodontia parsial dan supernumerary teeth. Perawatan autologous alveolar bone grafting yang diambil dari tulang ilium pasien merupakan standar perawatan bagi pasien celah bibir dan palatum. Namun, pengambilan tulang tersebut bersifat invasif dan memiliki tingkat morbiditas dan mortalitas yang tinggi. Teknik rekayasa jaringan yang terdiri dari scaffold, faktor pertumbuhan, dan sel punca dapat menjadi solusi untuk masalah
tersebut. Sumber donor sel punca yang tidak invasif bisa didapatkan dari sel punca pulpa gigi sulung (SHED) dan sel punca pulpa gigi permanen (DPSCs). Salah satu syarat dapat digunakannya suatu sel punca adalah memiliki kapasitas proliferasi yang baik. Perbandingan antara kapasitas proliferasi SHED dan DPSCs pada pasien normal
telah diketahui, namun pada pasien celah palatum belum pernah diteliti. Tujuan: Penelitian ini bertujuan untuk mengetahui perbandingan kapasitas proliferasi sel punca pulpa gigi sulung dan sel punca pulpa gigi permanen pasien celah bibir dan palatum. Metode: SHED dan DPSCs pasien celah bibir dan palatum dikultur hinga tingkat
confluency 70-80%, setelah itu sel dipanen dan dilakukan Uji PDT pada sel yang telah dikultur selama 7 hari. Hasil: SHED pasien celah bibir dan palatum menunjukkan nilai PDT yang lebih tinggi dibandingkan dengan DPSCs, namun secara statistik perbedaan tersebut tidak berbeda bermakna. Kesimpulan: SHED dan DPSCs penderita celah bibir dan palatum memiliki kapasitas proliferasi yang sama baiknya.
Background: Cleft lip and palate is one of the most common congenital abnormalities. This disorder can cause speech impediments, middle ear abnormalities, psychological problems, and dental abnormalities such as partial anodontia and supernumerary teeth. Treatment of autologous alveolar bone grafting taken from the patient's ilium bone is the standard of care for cleft lip and palate patients. However, bone removal is invasive and carries a high rate of morbidity and mortality. Tissue engineering techniques consisting of scaffolds, growth factors, and stem cells can be a solution to the problem that. Sources of non-invasive stem cell donors can be obtained from primary dental pulp stem cells (SHED) and permanent dental pulp stem cells (DPSCs). One of the conditions for the use of a stem cell is to have a good proliferative capacity. Comparison between the proliferative capacity of SHED and DPSCs in normal patients known, but in cleft palate patients it has not been studied. Objective: This study was aimed to compare the proliferative capacity of pulp stem cells of primary teeth and pulp stem cells of permanent teeth in cleft lip and palate patients. Methods: SHED and DPSCs of cleft lip and palate patients were cultured to level
70-80% confluency, after that the cells were harvested and PDT test was performed on cells that had been cultured for 7 days. Results: SHED of cleft lip and palate patients showed a higher PDT value than DPSCs, but statistically the difference was not significantly different. Conclusion: SHED and DPSCs patients with cleft lip and palate have the same good proliferative capacity."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2019
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Nadya Firiza Salsabila
Identifikasi Karakteristik Sel Stromal Pulpa Gigi Sulung dan Permanen pada Subjek Normal dan Pasien Celah Bibir dan Palatum Melalui Ekspresi Gen CUX1 = Identification of Characteristics of Pulp Stromal Cells in Primary and Permanent Teeth in Normal Subjects and Cleft Lip and Palate Patients Through CUX1 Gene Expression
"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.
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."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2021
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Lulu Amanda Zatalini
Identifikasi Sel Punca Mesenkim pada Pulpa Gigi Sulung dan Gigi Permanen Pasien Celah Bibir dan Palatum = Identification of Mesenchymal Stem Cells in Dental Pulp from Cleft Lip and Palate Patients
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Latar Belakang: Celah bibir dan palatum merupakan kelainan kongenital yang paling sering terjadi pada regio orofasial. Pasien celah bibir dan palatum menunjukkan sejumlah permasalahan seperti defek tulang alveolar yang lebar, kehilangan gigi kongenital, supernumerary teeth, hipoplasia dan gigi impaks. Autologous alveolar bone grafting dianggap sebagai perawatan gold standard untuk rekonstruksi tulang alveolar dengan menggunakan tulang kanselus dari puncak ilium anterior. Namun, pengambilan tulang ilium bersifat invasif dan memiliki potensi terjadinya komplikasi. Mengingat hal tersebut, teknik rekayasa jaringan yang memanfaatkan scaffolds, faktor pertumbuhan, dan sel punca dipertimbangkan sebagai pilihan perawatan yang baru. Sel punca mesenkim bisa didapatkan dari jaringan pulpa, yang disebut dengan sel punca pulpa gigi sulung atau stem cells from exfoliated deciduous teeth (SHED) dan sel punca pulpa gigi permanen atau dental pulp stem cells (DPSC). Salah satu kriteria yang harus dimiliki sel punca mesenkim adalah mengekspresikan surface marker CD73, CD90, dan CD105. Pada pasien normal, penelitian yang membandingkan karakteristik antara SHED dan DPSC telah membuktikan bahwa keduanya merupakan sel punca mesenkim dengan mengekspresikan surface markersesuai dengan kriteria. Namun, pada pasien celah bibir dan palatum belum banyak diteliti. Tujuan: Menganalisis perbedaan persentase sel yang mengekspresikan surface marker (CD73, CD90, dan CD105) pada SHED dan DPSC pasien celah bibir dan palatum. Metode: Sel punca pulpa gigi sulung dan gigi permanen diisolasi dari jaringan pulpa pasien celah bibir dan palatum. Persentase sel yang mengekspresikan surface marker (CD73, CD90, dan CD105) dianalisis dengan uji flow cytometry. Hasil: Analisis flow cytometry menunjukkan bahwa baik SHED maupun DPSC mengekspresikan masing-masing surface marker dalam persentase yang tinggi (>90%). Setelah dilakukan uji Independent T-test untuk membandingkan ekspresi masing-masing surface markerpada kedua grup, didapatkan hasil >0,05. Kesimpulan:  Tidak terdapat perbedaan bermakna antara ekspresi masing-masing surface marker pada SHED dan DPSC pasien celah bibir dan palatum.

Background: Cleft lip and palate is the most common congenital anomaly in the orofacial region. Cleft lip and palate patients present with a number of complaints such as wide alveolar bone defects, congenitally missing teeth, supernumerary teeth, hypoplastic dan impacted teeth. Autologous bone grafting is considered to be the gold standard for alveolar bone reconstruction using the cancellous bone harvested from the anterior iliac crest. However, the procedure is invasive and carries a risk of complications. Bearing all that in mind, tissue engineering that utilizes scaffolds, growth factors, and stem cells arises as a new therapeutic option. Mesenchymal stem cells can be obtained from dental pulp, which are called stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC). One of the criterias to define mesenchymal stem cells is the expression of surface markers CD73, CD90, and CD105. In normal patients, both SHED and DPSC have been known to express those surface markers. However, the expression of CD73, CD90, and CD105 in SHED and DPSC from cleft lip and palate patients has not been fully explored. Objective: To analyze the difference in the percentage of cells that express CD73, CD90, and CD105 in SHED and DPSC from cleft lip and palate patients. Methods: SHED and DPSC were isolated from dental pulp. The expression of surface markers were analyzed with flow cytometry. Results: Flow cytometry analysis showed that SHED and DPSC from cleft lip and palate patients highly express (>90%) surface markers that are associated with mesenchymal stem cells such as CD73, CD90, and CD105. The Independent T-Test was then performed to see a comparison between the expression of each surface marker in both groups and the value was >0.05. Conclusions: There is no significant difference between the percentage of cells that express each surface marker in SHED and DPSC from cleft lip and palate patients.

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Jakarta: Fakultas Kedokteran Gigi Universitas Indonesia , 2019
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Alya Hana Firdanisa
Kemampuan diferensiasi osteogenik sel stromal pulpa gigi sulung dan gigi permanen pasien celah bibir dan palatum melalui ekspresi gen alkaline phosphatase (ALP) = Osteogenic differentiation capacity of stromal cell from human deciduous teeth and dental pulp stromal cells of cleft lip and palate patients through alkaline phosphatase (ALP) gene expression.
"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."
Depok: Fakultas Kedokteran Gigi Universitas Indonesia, 2021
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Azra Nadhira
Diferensiasi osteogenik sel stromal pulpa gigi pasien celah bibir dan palatum melalui deposisi kalsium = Osteogenic differentiation of dental stromal cells in cleft lip and palate patients by calcium deposition
"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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