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Sains Malaysiana 51(9)(2022):
http://doi.org/10.17576/jsm-2022-5109-21
Growth
Factor Cocktail to Facilitate Epithelial Differentiation of Exfoliated
Deciduous Teeth Stem Cells
NURUL
HAFIZAH MOHD NOR1,3, ZURAIRAH BERAHIM1,*, AHMAD AZLINA1 & THIRUMULU PONNURAJ KANNAN1,2
1School of Dental Sciences, Universiti
Sains Malaysia, 16150 Kubang Kerian, Kelantan Darul Naim, Malaysia
2Human Genome Centre, School of
Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan
Darul Naim, Malaysia
3Institut Islam Hadhari, Universiti
Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia
Diserahkan: 3 November 2021/ Diterima: 21 April 2022
Abstract
Epithelial
cells are important in the regeneration of oral mucosal tissue. The cell source
is commonly derived from tissue biopsy, which is obtained through surgery. Stem
cells from human exfoliated deciduous teeth (SHED) were demonstrated to
differentiate into multiple cell types. As it can be readily available from
exfoliated deciduous teeth, it can be induced and become a potential source of
epithelial cell for oral tissue study. This study aims to examine a mixture of
growth factors in the differentiation of epithelial-like cells obtained from human
exfoliated deciduous teeth (SHED) stem cells. This growth factor cocktail constitutes hepatocyte growth factor
(HGF), keratinocyte growth factor (KGF), insulin-like growth factor 2 (IGF-II),
and epidermal growth factor (EGF). After introducing the cocktail, the
treated SHED were assessed for epithelial characteristics and markers using cell proliferation test,
morphological transformation, protein and gene expression by immunofluorescence
staining and cytometry assessment, respectively. The proliferation rate was
analysed statistically using Analysis of Variance (ANOVA) with Repeated
Measures (p<0.05). SHED cells
demonstrated morphological changes on the 7th day after using the
cocktail. These transformations are in alignment with the identification of
genes associated with epithelial cells and positive stain outcomes for
pan-cytokeratin, E-cadherin, and p63. The cell proliferation test indicated
that proliferation of cells and growth factor introduction were significantly
correlated. The growth factor cocktail used for this research facilitated SHED
differentiation for epithelial-like cells. The outcomes validate the production
of epithelial cells using SHED; tissue production studies focus on these
aspects immensely.
Keywords: Epithelial characterization; epithelial growth factors;
epithelial markers; SHED differentiation
Abstrak
Sel epitelium adalah sel utama untuk
penjanaan semula tisu mukosa mulut. Sel ini selalunya diperoleh daripada biopsi tisu
melalui pembedahan. Sel stem gigi susu manusia terkelupas didapati boleh
diterbitkan kepada beberapa jenis sel. Oleh kerana ia boleh diperoleh dengan
mudah di dalam gigi susu yang terkelupas, maka ia boleh diinduksi dan
berpotensi menjadi sumber sel epitelium untuk kajian tisu mulut. Kajian ini bertujuan untuk
mengkaji campuran faktor pertumbuhan di dalam pembezaan sel epitelium yang
diperoleh daripada sel stem gigi susu manusia terkelupas (SHED). Koktel faktor pertumbuhan ini dirumus daripada faktor pertumbuhan
hepatosit (HGF), faktor pertumbuhan keratinosit (KGF), faktor pertumbuhan
seperti insulin -2 (IGF-II) dan faktor pertumbuhan epidermis (EGF). Selepas
dirawat dengan koktel faktor pertumbuhan, sel SHED dinilai untuk ciri dan
penanda epitelium menggunakan kelajuan percambahan sel, transformasi morfologi
dan pengekspresan protein dan gen menggunakan pewarnaan imunofluoresensi dan
penilaian sitometri. Kadar percambahan dianalisis secara statistik menggunakan
ANOVA sehala dengan pengukuran berulang (p<0.05). Sel SHED menunjukkan
perubahan morfologi pada hari ke-7 selepas menggunakan koktel. Transformasi ini
adalah sejajar dengan pengenalpastian gen yang dikaitkan dengan sel epitelium
dan hasil pewarnaan positif untuk pan-sitokeratin, E-cadherin dan p63
menggunakan pewarnaan imunofluoresensi dan penilaian sitometri. Analisis
statistik ujian menunjukkan bahawa percambahan sel dan pengenalan faktor
pertumbuhan berkorelasi dengan ketara. Koktel faktor pertumbuhan yang digunakan
untuk kajian ini didapati menggalakkan pembezaan SHED kepada sel seperti
epitelium. Hasil kajian mengesahkan penerbitan sel epitelium daripada sel SHED
setelah koktel digunakan; kajian penghasilan tisu meletakkan nilai yang tinggi
pada aspek ini.
Kata kunci: Faktor pertumbuhan epitelium; pembezaan SHED; penanda epitelium; pencirian epitelium
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*Pengarang untuk surat-menyurat; email: zurairah@usm.my
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