Sains Malaysiana 40(6)(2011): 613–621
Kesan ß-gliserofosfat Terhadap Ampaian Sel Mononukleus
daripada Darah Periferi Manusia
(Effect of ß-glycerophosphate on Suspension Mononucleus
Cell from Human Peripheral Blood)
Nurul Atikah Ahmad1, Shahrul Hisham Zainal Ariffin* 1, Rohaya Megat Abdul Wahab2, Muhamad Abdul Razak3 & Sahidan Senafi1
1Pusat Pengajian Biosains dan
Bioteknologi, Fakulti Sains dan Teknologi
Universiti
Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor D.E., Malaysia
2Jabatan Ortodontik, Fakulti
Pergigian, Universiti Kebangsaan Malaysia
Jalan
Raja Muda Abdul Aziz, 53000 Kuala Lumpur, Malaysia
3Jabatan Ortopedik, Pusat
Perubatan Universiti
Kebangsaan Malaysia
Jalan
Yaacob Latif, Bandar Tun Razak, 56000, Cheras, Kuala Lumpur, Malaysia
Diserahkan:
15 Julai 2010 / Diterima: 11 Ogos 2010
ABSTRAK
Sel osteoblas merupakan sel mononukleus yang bertanggungjawab
untuk pembentukan tulang. Sel mononukleus telah terbukti mampu membeza kepada
sel osteoblas setelah diaruh oleh kombinasi asid askorbik dan ß-gliserofosfat
sebagai faktor pembezaan. Tujuan kajian ini adalah bagi melihat kesan aruhan
ß-gliserofosfat terhadap ampaian sel mononukleus daripada darah periferi
manusia secara in vitro. Sel mononukleus dipencilkan daripada darah periferi
manusia dengan menggunakan larutan Ficoll-Paque™ Plus melalui kaedah
pengemparan kecerunan ketumpatan. Sel mononukleus kemudian dikultur selama 7
hari di dalam medium proliferasi sebelum diaruh dengan ß-gliserofosfat pada
kepekatan 1 mM, 5 mM, 10 mM, 20 mM, dan 100 mM. Pada hari 0, 1, 3, 7, dan 14,
penentuan profil aktiviti enzim alkalin fosfatase (ALP)
dan analisis morfologi bagi sel osteoblas dilakukan dalam medium masing-masing.
Aktiviti enzim ALP dan analisis morfologi menunjukkan
peningkatan yang signifikan (p<0.05) antara sel yang diaruh dengan sel
kawalan negatif melalui statistik ujian-t berpasangan. Kesimpulannya, kehadiran
ß-gliserofosfat sahaja mampu untuk mengaruh pembezaan sel mononukleus kepada
sel osteoblas. Kesan ß-gliserofosfat terhadap pembezaan sel mononukleus kepada
sel osteoblas menunjukkan profil enzimologi (aktiviti enzim ALP)
dan morfologi yang hampir sama dengan kawalan positif (asid askorbik dan
ß-gliserofosfat). Berdasarkan kepada analisis enzimologi dan morfologi 1 mM
ß-gliserofosfat adalah kepekatan yang paling sesuai untuk pembezaan sel
osteoblas secara in vitro.
Kata kunci: ß-gliserofosfat; ampaian sel mononukleus
ABSTRACT
Osteoblast cell is a mononucleus cell that is responsible for
bone formation. Mononucleus cell have been proven to differentiate into
osteoblast cell after being induced with combinations of ascorbic acid and
ß-glycerophosphate as differentiation factors. This study aimed to observe the
in vitro effect of a single ß-glycerophosphate induction towards primary
mononucleated cells suspension that have been isolated from human peripheral
blood. Mononucleus cells were isolated from human peripheral blood using
density gradient centrifugation method in the present of Ficoll-Paque™ Plus.
Mononucleus cells were cultured for 7 days in proliferation medium before being
induced by 1 mM, 5 mM, 10 mM, 20 mM, and 100 mM of ß-glycerophosphate. Alkaline
phosphatase (ALP) activity profile and morphological analysis were
done at day 0, 1, 3, 7 and 14 in their respective medium. The activity of ALP enzyme
and morphology analysis showed a significant increase (p<0.05) between cells
that have been induced with the negative control cells by using paired t-test
statistical analysis. In conclusion, the presence of only ß-glycerophosphate is
able to induce mononucleated cell differentiation into osteoblast cells. The
effect of ß-glycerophosphate towards mononucleated cells differentiation showed
similar enzymological (ALP activities) and morphological
profile as positif control (ascorbic acid and ß-glycerophosphate). Based on
enzymological and morphological analyses, 1 mM of ß-glycerophosphate is the
best concentration for in vitro osteoblast differentiation.
Keywords:
ß-glycerophosphate; suspension mononucleus cell
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*Pengarang untuk
surat-menyurat; email: hisham@ukm.my
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