Sains Malaysiana 51(5)(2022): 1449-1464

http://doi.org/10.17576/jsm-2022-5105-15

Higher Concentration of Ascorbic Acid as a Sole Induction Factor for Osteogenesis on MC3T3-E1 Cell Model

(Kepekatan Tinggi Asid Askorbik sebagai Faktor Aruhan Tunggal untuk Osteogenesis pada Model Sel MC3T3-E1)

FARINAWATI YAZID¹, WAN CHING NG², NUR ATMALIYA LUCHMAN^2, SHAHRUL HISHAM ZAINAL ARIFFIN³ & ROHAYA MEGAT ABDUL WAHAB^2,*

¹Discipline of Paediatric Dentistry, Department of Family Oral Health, Universiti Kebangsaan Malaysia, Jalan Raja Muda Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

²Discipline of Orthodontics, Department of Family Oral Health, Universiti Kebangsaan Malaysia, Jalan Raja Muda Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

³Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

Diserahkan: 31 Mac 2021/Diterima: 4 Oktober 2021

Abstract

This research aimed to investigate the potential of ascorbic acid (Asc) to act independently as an osteogenic induction factor in a murine pre-osteoblast (MC3T3-E1) model. MC3T3-E1 cells were seeded in culture wells and hydroxyapatite scaffold for two-dimensional and three-dimensional analyses respectively. Cell morphology, viability, osteoblast differentiation, and mineralisation potentials of MC3T3-E1 cells were compared between induction of standard (50 µg/mL) and doubled (100 µg/mL) Asc concentrations in growth media. Cells with fibroblast-like morphology became confluent earlier on day 6 in the standard group compared to the doubled group on day 9. Cell viability and differentiation potential were significantly increased in the doubled group (p < 0.01). Mineralisation occurred in the doubled group after 15 days of seeding but no mineralisation was seen in the standard group. Findings were similar in 3D analysis whereby mineralized nodules were seen only in the doubled group. The relative expression of collagen 1(α) and osteocalcin genes were increased in the doubled group than the standard group. Doubling Asc concentration in a growth medium to 100 µg/mL can induce viability, differentiation, and mineralisation of MC3T3-E1 cells. Thus, higher concentration of ascorbic acid can potentially be used as the sole induction factor in osteogenic medium.

Keywords: Ascorbic acid; growth media; MC3T3-E1 cells; osteogenic differentiation; sole induction

Abstrak

Penyelidikan ini bertujuan untuk mengkaji potensi asid askorbik (Asc) bagi tindak balas secara bebas sebagai faktor induksi osteogenik dalam model pra-osteoblas murine (MC3T3-E1). Sel MC3T3-E1 dibenihkan dalam telaga kultur dan perancah hidroksi apatit untuk analisa dua dimensi dan tiga dimensi. Morfologi sel, kebolehidupan sel, pembezaan osteoblas dan potensi pemineralan sel MC3T3-E1 dibandingkan antara induksi kepekatan piawai (50 µg/mL) dan kumpulan dua kali ganda (100 µg/mL) kepekatan asid askorbik di dalam media pertumbuhan. Sel dengan morfologi seperti fibroblas menjadi konfluensi pada hari ke 6 dalam kumpulan piawai berbanding kumpulan dua kali ganda pada hari ke 9. Kebolehidupan sel dan potensi pembezaan meningkat dengan ketara pada kumpulan dua kali ganda (p < 0.01). Pemineralan berlaku pada kumpulan yang berlipat ganda selepas 15 hari pengkulturan tetapi tiada pemineralan dilihat pada kumpulan piawai. Penemuan yang sama dilihat dalam analisis 3D manakala nodul mineral hanya dilihat pada kumpulan dua kali ganda. Ekspresi relatif gen kolagen 1(α) dan osteokalsin meningkat pada kumpulan dua kali ganda berbanding kumpulan piawai. Menggandakan kepekatan asid askorbik dalam medium pertumbuhan hingga 100 µg/mL boleh mengaruh keviabelan, pembezaan dan pemineralan sel MC3T3-E1. Oleh itu, kepekatan asid askorbik yang lebih tinggi berpotensi digunakan sebagai faktor induksi tunggal dalam medium osteogenik.

Kata kunci: Aruhan tunggal; asid askorbik; media pertumbuhan; sel MC3T3-E1; pembezaan osteogenik

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^*Pengarang untuk surat-menyurat; email: rohaya_megat@ukm.edu.my