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Sains Malaysiana 49(3)(2020): 593-602 http://dx.doi.org/10.17576/jsm-2020-4903-14
Cytotoxicity of L- and D-Ascorbic
Acid on Murine and Human Suspension Peripheral Blood Cells (Sitotoksisiti L- dan D-Asid
Askorbik ke atas Sel Ampaian Darah Periferi Mencit dan Manusia) INTAN
ZARINA ZAINOL ABIDIN1, THANALETCHUMI MANOGARAN2
& SHAHRUL HISHAM ZAINAL ARIFFIN2,3* 1Centre for Research
and Graduate Studies, University of Cyberjaya, 63000 Cyberjaya,
Selangor Darul Ehsan, Malaysia 2Centre for Biotechnology
and Functional Food, Faculty of Science and Technology, Universiti
Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia 3Malaysian Genome Institute
(MGI), National Institute of Biotechnology Malaysia (NIBM), Jalan
Bangi, 43600 Kajang, Selangor Darul Ehsan, Malaysia Received:
29 July 2019/Accepted: 18 November 2019 ABSTRACT Ascorbic acid has two isoforms,
i.e., L-ascorbic acid which exists naturally and D-ascorbic acid
which only can be found in synthetic form. This study aimed
to determine the cytotoxic effect of L- and D- ascorbic acid on
primary cell, i.e. murine and human suspension cells of peripheral
blood. Murine and human suspension blood cells were obtained through
density gradient centrifugation using Ficoll-Paque PLUS. Non-adherent
cells were identified after 7 days in culture and seeded at 1 ×
105 cells/mL. Both newly isolated primary cells were
analyzed for in vitro proliferation ability
for 7 days. The cells were treated with L- and D-ascorbic acid at
concentrations 30, 50, and 90 µg/mL followed by Trypan
blue viability assay at day 0, 3, 7, and 14. Cells cultured in the complete medium were represented
as control. Murine and human suspension blood cells showed round
morphology and significant increase of viable cells after 7 days
in complete medium. Both cells treated with L-ascorbic acid exhibited
low cytotoxic effect at 30, 50, and 90 µg/mL concentrations.
In addition, murine blood cells treated with D-ascorbic acid caused
total population cell death in all three concentrations at day 14, whereas human suspension
blood cells only exhibited total population cell death at higher
concentration, i.e. 90 µg/mL. In conclusion, L-ascorbic acid exhibited minimal
cytotoxic effect on both primary cell sources as compared to lethal
effect of D-ascorbic acid treatment. Keywords: Cytotoxicity; D-ascorbic acid; human suspension
blood cells; L-ascorbic acid; Mus musculus ABSTRAK Asid askorbik terdiri daripada
dua isoform iaitu L-asid askorbik yang wujud secara semula jadi dan D-asid askorbik yang hanya terdapat dalam bentuk sintetik. Kajian
ini adalah bertujuan untuk menentukan kesan sitotoksik L-
dan D-asid askorbik ke atas sel primari iaitu sel ampaian darah
periferi mencit dan manusia. Sel ampaian darah mencit dan manusia
diperoleh melalui kaedah pengemparan kecerunan ketumpatan menggunakan
Ficoll-Paque PLUS. Sel ampaian dikenal pasti selepas 7 hari pengkulturan dan dikulturkan pada ketumpatan
1 × 105 sel/mL. Kedua-dua sumber sel yang baru dikulturkan
dianalisis bagi menentukan
keupayaan proliferasi in vitro selama 7 hari. Sel seterusnya dirawat
dalam L- dan D-asid askorbik pada kepekatan 30, 50 dan 90 µg/mL
diikuti dengan pengasaian keviabelan tripan biru pada hari 0, 3,
7 dan 14. Sel yang dikulturkan dalam medium lengkap mewakili kawalan.
Sel ampaian darah mencit dan manusia menunjukkan morfologi bulat
dan sel viabel didapati meningkat secara signifikan selepas 7 hari
di dalam medium lengkap. Kedua-dua sel yang dirawat dengan L-asid
askorbik menunjukkan kesan sitotoksik yang rendah pada kepekatan
30, 50 dan 90 µg/mL. Walau bagaimanapun, sel yang dirawat dengan
D-asid askorbik menyebabkan kematian sel pada ketiga-tiga kepekatan pada hari ke-14, manakala sel
ampaian manusia hanya menunjukkan kematian keseluruhan sel pada
kepekatan tinggi iaitu 90 µg/mL.
Kesimpulannya, L-asid askorbik menunjukkan kesan sitotoksik
yang minima ke atas kedua-dua sumber sel primari berbanding kesan
kematian dalam rawatan D-asid askorbik. Kata kunci: D-asid askorbik; L-asid askorbik; Mus musculus; sel ampaian darah manusia;
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