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Title:	Morphology, viability and osteogenic differentation assessment of stem cells from exfoliated deciduous teeth (SHED) and Dental Pulp Stem Cell (DPSC) with and wihout hydroxyapatite scaffold
Authors:	Manal Nabil Abdullah Hagar (P98423)
Supervisor:	Rohaya Megat Abdul Wahab, Prof. Dr.
Keywords:	Dental Pulp Morphology Osteosarcoma Universiti Kebangsaan Malaysia -- Dissertations Dissertations, Academic -- Malaysia
Issue Date:	21-Jul-2020
Description:	Bone defects due to congenital abnormalities, diseases or trauma are problems that many people have suffered. Characterisation and osteogenic differentiation of dental pulp stem cells (DPSC) and stem cells from human exfoliated cells (SHED) are important in manipulating these cells for regenerative medicine purposes. This study was conducted to investigate the osteogenic differentiation of DPSC and SHED in granular hydroxyapatite scaffold (HA). DPSC and SHED were isolated from permanent and deciduous teeth. Preosteoblast cells (MC3T3-E1) were used as a control group. The experiment was done in two forms, 2D and 3D cultures. MC3T3-E1, DPSC, and SHED were analyzed in terms of morphology using cell B software. The proliferation rate of these cells was assessed by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay in 2D culture. The expression of stemness markers for DPSC and SHED was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR). Alkaline phosphatase (ALP) assay was used to compare the osteoblastic differentiation of these cells. After that, cells were seeded on granular hydroxyapatite scaffold and incubated for 21 days. Observation using Field Emission Scanning Electron Microscopy (FESEM) was carried out on day 7 and day 21 of the seeded cells and osteogenic differentiation was done using ALP assay. The results showed that the morphology of the cells was mononucleated, fibroblast-like shaped cells with extended cytoplasmic projection. The cell viability and differentiation potential were significantly increased in DPSC and SHED as compared to the control group (p < 0.01). Besides, ALP activity in 2D and 3D cultures showed a significant increase in the differentiating medium compared to the complete medium (p < 0.01). In RT-PCR study, DPSC and SHED expressed stemness markers such as CD73, CD105, and CD146 and negatively expressed CD34, CD45, and CD11b blood cell markers which indicate that these cells belong to mesenchymal cells. SEM image of SHED in the HA scaffold showed an abundance of mineralisation and larger nodules on day 7 as compared to DPSC and MC3T3-E1 . HA scaffold fulfills the criteria of an ideal scaffold as it promoted cell adhesion and induced osteogenesis in vitro. In conclusion, SHED had the highest osteogenic potential in both 2D and 3D cultures. Therefore, it serves as a potential candidate for bone tissue engineering using the HA scaffold as a model,Ijazah Sarjana Sains Pergigian
Pages:	46
Call Number:	WU230.M266m 2020 9 tesis
Publisher:	UKM, Kuala Lumpur
Appears in Collections:	Faculty of Dentistry / Fakulti Pergigian

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