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

Received: 15 July 2010 / Accepted: 11 August 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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*Corresponding author; email: hisham@ukm.my