Sains Malaysiana 47(4)(2018): 691-698

http://dx.doi.org/10.17576/jsm-2018-4704-06

Proliferation and Osteoblast Differentiation Mice Dental Pulp Stem Cells between Enzyme Digestion and Outgrowth Method

(Proliferasi dan Pembezaan Osteoblas Sel Stem Pulpa Gigi Mencit antara Kaedah Pencernaan

Enzim dan Eksplan)

FARINAWATI YAZID^1*, NUR ATMALIYA LUCHMAN¹, ROHAYA MEGAT ABDUL WAHAB¹, SHAHRUL HISHAM ZAINAL ARIFFIN² & SAHIDAN SENAFI²

¹Faculty of Dentistry, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

²School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor Darul Ehsan, Malaysia

Diserahkan: 26 Julai 2017/Diterima: 6 November 2017

ABSTRACT

The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential of mice DPSCs into bone cells. Dental pulp was extracted from mice’s incisors and subjected to isolation either by collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblast- liked morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation. Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.

Keywords: Adherent cells; alkaline phosphatase; in vitro; mesenchymal stem cells

ABSTRAK

Kaedah pemencilan bagi sel stem pulpa gigi (DPSCs) masih kurang jelas terutamanya bagi mendapatkan persekitaran yang kondusif bagi DPSCs berproliferasi. Kaedah pencernaan enzim dan eksplan merupakan dua kaedah yang biasa digunakan untuk memencilkan DPSCs namun kurang dicirikan pada DPSCs mencit. Kajian ini bertujuan untuk membandingkan kaedah pemencilan dan potensi perbezaan DPSCs mencit kepada sel tulang. Pulpa gigi diekstrak daripada gigi kacip mencit dan pemencilan sel dilakukan sama ada menggunakan kolagenase 1A atau pengkulturan tisu pulpa pada medium lengkap alpha-modified eagle medium (αMEM). Kedua-dua sel yang dipencilkan dikulturkan sehingga pasaj 4 dan analisis proliferasi dan pembezaan secara in vitro dilakukan. Kedua-dua sel menunjukkan morfologi fibroblas namun sel yang diasingkan melalui pencernaan enzim berproliferasi lebih cepat berbanding dengan kaedah eksplan. Selepas 21 hari pembezaan kepada sel osteoblas, DPSCs yang dipencilkan melalui kaedah pencernaan enzim menunjukkan aktiviti alkali fosfatase (ALP) sedikit berbeza berbanding kaedah eksplan. Kesimpulannya, terdapat perbezaan yang signifikan daripada sel yang dipencilkan melalui kaedah percernaan enzim berbanding eksplan terutamanya daripada segi proliferasi dan pembezaan osteoblas. Oleh itu, adalah lebih baik untuk memencilkan sel melalui kaedah pencernaan enzim kerana ia adalah lebih cepat dan konsisten berbanding dengan kaedah eksplan.

Kata kunci: Alkali fosfatase; in vitro; sel melekat; sel stem mesenkima

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*Pengarang untuk surat-menyurat; email: drfarinawati@ukm.edu.my