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Sains Malaysiana
50(2)(2021): 461-473
http://dx.doi.org/10.17576/jsm-2021-5002-17
Aqueous
Polyphenol Semi-Purified Fractions from Galls of Quercus infectoria Increase ALP Activity and Mineralisation of hFOB 1.19 Cells
(Fraksi Separa
Tulen Polifenol Akueus daripada
Puru Quercus infectoria Meningkatkan Aktiviti ALP dan Pemineralan
Sel hFOB 1.19)
HERMIZI HAPIDIN1*,
FAIRUZA MUNIRAH MAZLAN1, HASMAH ABDULLAH1 & IMA NIRWANA SOELAIMAN2
1Biomedicine
Programme, School of Health Sciences, Universiti Sains Malaysia, Health Campus,
16150 Kubang Kerian, Kelantan Darul Naim, Malaysia
2Department
of Pharmacology, Universiti Kebangsaan
Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras,
Kuala Lumpur, Malaysia
Received: 14 April 2020/Accepted: 5 August
2020
ABSTRACT
The galls of Quercus infectoria (QI) have been reported to
possess numerous medicinal values and give a positive effect on bone
metabolism. This study investigated the effects of semi-purified fractions of
QI gall extract on the proliferation, alkaline phosphatase (ALP) activity, and
mineralisation of human foetal osteoblast cell line (hFOB 1.19). The
semi-purified fractions (fraction A and B) were prepared by column
chromatography methods. MTT assay was used to measure cell proliferation
activity to obtain half maximal concentration (EC50) of the cells
treated with fraction A (phenolic components and contained amide), fraction B (phenolic components
with the presence of alkene), and pamidronate (drug control). The most potent or lowest EC50 was further used to
measure ALP activity in the treated and untreated cells at day 1, 3, 7, 10, and
14 by ELISA. Cell mineralisation was determined by von Kossa staining for
phosphate depositions and Alizarin
Red S staining for calcium depositions. The EC50 values for
fraction A and B were 8.86 and 9.92 μg/mL, respectively, which showed a
greater effect compared to the pamidronate (15.27 μg/mL). The ALP activity
of both fractions in the treated cells were also greater compared to the two
control groups (cells treated with pamidronate and untreated cells), starting
from day 3 onwards. The calcium
depositions appeared as red spots, while phosphate depositions appeared as
black spots. Interestingly, the calcium depositions
of cells treated with both fractions were higher than those of the two control
groups. In conclusion, semi-purified fractions of QI gall extract enhanced
proliferation, improved mineralisation, and increased ALP activity of hFOB 1.19
cells.
Keywords: Alizarin Red S; alkaline phosphatase (ALP); hFOB
1.19 cell line; Quercus infectoria; von Kossa
ABSTRAK
Puru Quercus
infectoria (QI) telah dilaporkan mempunyai nilai perubatan yang banyak dan
memberi kesan positif pada metabolisme tulang. Penyelidikan ini dijalankan
untuk mengkaji kesan fraksi separa tulen QI pada proliferasi, aktiviti alkalin
fosfatase (ALP) dan pemineralan sel osteoblas fetal manusia (hFOB 1.19).
Fraksi separa tulen QI (fraksi A dan B) telah disediakan melalui kaedah
kromatografi kolum. Asai MTT digunakan untuk mengukur aktiviti proliferasi sel
bagi menentukan kepekatan separa maksimum (EC50)
sel yang dirawat dengan
fraksi A (komponen fenolik dan
mengandungi amida), fraksi B (komponen fenolik dengan kehadiran alkena) dan pamidronat. Nilai EC50 paling poten atau rendah yang diperoleh digunakan untuk mengukur aktiviti ALP dalam sel yang dirawat dan juga sel yang
tidak dirawat pada hari 1, 3, 7, 10 dan 14 melalui ujian ELISA. Pemineralan sel
ditentukan dengan pewarnaan von Kossa bagi pemendapan fosfat dan pewarnaan Alizarin Red S bagi pemendapan kalsium. Nilai EC50 bagi sel yang dirawat dengan fraksi
A fraksi B masing-masing adalah 8.86 dan 9.92
μg/mL yang menunjukkan kesan lebih hebat berbanding pamidronat (15.27
μg/mL). Kedua-dua fraksi juga menunjukkan aktiviti ALP yang lebih tinggi berbanding
kedua-dua kumpulan kawalan (sel yang dirawat dengan pamidronat dan sel yang
tidak dirawat), bermula pada hari ke-3 dan hari-hari berikutnya. Pemendapan
kalsium kelihatan sebagai tompokan merah manakala pemendapan fosfat kelihatan
tompokan hitam. Menariknya, pemendapan kalsium oleh sel yang dirawat dengan
kedua-dua fraksi adalah lebih tinggi berbanding dua kumpulan kawalan. Kesimpulannya,
fraksi separa tulen QI menambahkan proliferasi, memperbaiki pemineralan dan
meningkatkan aktiviti ALP sel hFOB 1.19.
Kata
kunci: Alizarin Red
S; alkalin fosfatase (ALP); Quercus infectoria; sel hFOB 1.19; von
Kossa
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*Corresponding author; email: hermizi@usm.my
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