Sains Malaysiana 51(9)(2022): 3009-3018

http://doi.org/10.17576/jsm-2022-5109-21

Growth Factor Cocktail to Facilitate Epithelial Differentiation of Exfoliated Deciduous Teeth Stem Cells

(Koktel Faktor Pertumbuhan untuk Memudahkan Pembezaan Epitelium bagi Sel Stem Gigi Susu yang Terkelupas)

NURUL HAFIZAH MOHD NOR^1,3, ZURAIRAH BERAHIM^1,*, AHMAD AZLINA¹ & THIRUMULU PONNURAJ KANNAN^1,2

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

Received: 3 November 2021/ Accepted: 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 7^th 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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^*Corresponding author; email: zurairah@usm.my