Sains Malaysiana 47(10)(2018): 2349–2358

http://dx.doi.org/10.17576/jsm-2018-4710-11

 

Transforming Growth Factor Beta 3 Induced Human Adipose-Derived Stem Cells for Auricular Chondrogenesis

(Mengubah Faktor Pertumbuhan Beta 3 Aruhan Stem Asal Manusia Terbitan Adipos untuk Aurikul Kondrogenesis)

 

SITI NURHADIS CHE OMAR¹, BEE SEE GOH², MUHAMMAD AZHAN UBAIDAH², KHAIRUL ANUARKHAIROJI³, SHAMSUL SULAIMAN³, LOKMAN SAIM^2,4 & KIEN-HUI CHUA^1*

 

¹Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

²Department of Otorhinolaringology, Universiti Kebangsaan Malaysia Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

³Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

⁴School of Medicine, KPJ Healthcare University College, Lot PT 17010 Persiaran Seriemas, Kota Seriemas, 71800 Nilai, Negeri Sembilan Darul Khusus, Malaysia

 

Received: 22 March 2018/Accepted: 5 June 2018

 

ABSTRACT

The limitation of self-repair and proliferation capacity of chondrocytes in cartilage reconstruction lead to alternative search of cell source that can improve the auricular regeneration. Human adipose-derived stem cells (HADSC) are an alternative cell source that have unique characteristics to self-renew and differentiate into various tissues making it suitable for cell therapy and tissue engineering. This study aimed to examine the chondrogenic differentiation potential of (HADSC) in monolayer culture by the presence of different transforming growth factor beta’s, TFG-β1, -β2 and -β3. HADSC at passage 3 (1.5 × 105 cell/mL) were cultured in chondrogenic medium containing 5 ng/mL of different transforming growth factor beta’s, TFG-β1, -β2 and -β3 for 7, 14 and 21 days. Data analysis was evaluated based on the growth rate of cells, cells morphological changed, production of collagen type II and glycosaminoglycan sulphate (sGAG). The quantitative RT-PCR was carried out to determine the chondrogenic, fibrogenic and hypertrophic gene expression levels. Differentiation of HADSC into chondrocytes using TFG-β indicates the occurrence of the chondrogenesis process. The best chondrogenic differentiation was observed in HADSC induced by TFG-β3 through the chondrocytes-like cells morphology with cells aggregation and high production of proteoglycan matrices compared to other TGF-βs groups. Additionally, the expression of chondrocytes-specific genes such as Type II collagen, Aggrecan core protein, Elastin and Sox 9 was high. In conclusion, this study has showed that TGF-β3 is the potential growth factor in producing chondrogenic cells for auricular cartilage tissue engineering.

 

Keywords: Chondrocytes; chondrogenic differentiation; human adipose-derived stem; induction; tgf-β3

 

ABSTRAK

Kekurangan kapasiti pembaikan dan proliferasi kondrosit dalam pembentukan semula tisu rawan menyebabkan penyelidikan kini cenderung untuk mencari sumber sel baru yang dapat membantu memperbaiki tisu rawan aurikul dengan lebih baik. Sel stem adipos manusia (HADSC) dilihat sebagai sumber sel yang mempunyai ciri-ciri unik, boleh membahagi dengan sendiri dan membeza kepada pelbagai tisu, menyebabkan ia sesuai digunakan dalam terapi sel dan kejuruteraan tisu. Kajian ini bertujuan menilai tahap pembezaan kondrogeniknya secara monolapisan kultur dengan kehadiran faktor pertumbuhan transformasi beta yang berbeza iaitu TFG-β1,-β2 dan -β3. HADSC pada pasaj 3 (1.5 105 sel/mL) dikultur di dalam media kondrogenik yang mengandungi 5 ng/mL faktor pertumbuhan transformasi beta yang berbeza iaitu TFG-β1,-β2 dan -β3 selama 7, 14 dan 21 hari. Analisis data dinilai berdasarkan kepada kadar pertumbuhan sel, perubahan morfologi sel, pengeluaran kolagen jenis II dan glikoaminoglikan fosfat (sGAG). Kuantitatif RT-PCR dijalankan untuk menentukan kadar ekspresi gen kondrogenik, fibrogenik dan hipertrofik. Pembezaan HADSC kepada kondrosit menggunakan TFG-β menunjukkan berlakunya proses kondrogenesis. Pembezaan kondrogenik yang terbaik dilihat pada HADSC yang diaruhkan oleh TFG-β3 melalui perubahan morfologi dengan pembentukan sel agregasi dan penghasilan matrik proteoglikan yang tinggi apabila dibandingkan dengan kumpulan TFG-β yang lain. Tambahan pula, pengekspresan gen spesifik kondrogenik iaitu Kolagen jenis II, Elastin, Protien teras agrekan dan Sox 9 menunjukkan peningkatan. Kesimpulannya, kajian ini telah menunjukkan TFG-β3 berpotensi sebagai faktor pertumbuhan yang berupaya membantu penghasilan sel yang bercirikan kondrosit aurikular bagi strategi kejuruteraan tisu rawan aurikular.

 

Kata kunci: Aruhan; kondrosit; pembezaan kondrogenik; sel stem adipos manusia; tgf-β3

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*Corresponding author; email: ckienhui@gmail.com