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
Diserahkan: 29 Julai 2019/Diterima: 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; sitotoksisiti
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*Pengarang
untuk surat-menyurat;
email: hisham@ukm.edu.my
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