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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