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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
SAIM2,4 & KIEN-HUI
CHUA1* 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 Diserahkan: 22 Mac 2018/Diterima: 5 Jun 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.
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