Sains Malaysiana 47(10)(2018): 2325–2336

http://dx.doi.org/10.17576/jsm-2018-4710-09

 

Effects of PLGA Nanofibre on Osteoarthritic Chondrocytes

(Kesan Gentian Nano PLGA ke atas Sel Kondrosit Osteoartritis)

 

B.S. SHAMSUL¹, S.R. CHOWDHURY^1*, B.H.I. RUSZYMAH² & B.M.Y. NOR HAMDAN³

 

¹Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia, Clinical Block, Jalan Yaacob Latiff, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

²Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Pre-clinical Block, Jalan Yaacob Latif, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

 

³Department of Orthopedic & Traumatology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Clinical Block, Jalan Yaacob Latif, 56000 Cheras, Kuala Lumpur, Federal Territory

Malaysia

 

Received: 31 March 2018/Accepted: 5 June 2018

 

ABSTRACT

Chondrocytes obtained from osteoarthritis (OA) joints has been recognized as an abnormal cell; however, it’s proven to have potential in supporting cartilage regeneration. We have isolated chondrocytes from OA joints (OAC) and expanded chondrocytes growth medium (CGM). The growth kinetic, immunophenotyping and cell multilineage differentiation were analyzed to confirm the OAC stemness. The optimal condition to developed PLGA nanofiber with ratio 50:50 were 20% concentration of PLGA, flow rate with 0.3 mL/h, 10 kv voltage and 10 cm distance from nozzle to the collector. The toxicity level, scanning electron microscopy (SEM) and q-PCR analysis was performed in the present study. OAC fulfills the minimal criteria to be known to have stem cell as the cell easily adheres to the culture plate, shows high expression (≥95%) for CD13, CD29, CD44, CD73 and CD90 and less expression (≤2%) for CD45 and HLA-DR and potentially induced to mesodermal multilineage, which is osteocytes, adipocytes, and chondrocytes. Toxicity test showed no adverse effect of PLGA towards the cell. Based on the cell-PLGA nanofiber interaction, difference in fiber size will influence the proliferation of the cell. Nanofibers with 100 nm in size showed high proliferation of OAC and better gene and protein expression compared to monolayer culture. Thus, we concluded that OAC has the potential to be used in cartilage regeneration based on the presence of stem cell markers as similar to the human bone marrow. The cartilage regeneration will be more efficient if OAC cultured on 3D microenvironment as showed in the present study.

 

Keywords: Cytoskeleton; differentiation; electrospinning; osteoarthritis; tissue engineering of cartilage

 

ABSTRAK

Sel kondrosit yang diperoleh daripada sendi pesakit osteoartritis (OA) terbukti berpotensi untuk membantu pertumbuhan semula tulang rawan. Walau bagaimanapun, sel kondroit mempunyai kecenderungan untuk melalui proses pembezaan semasa pengkulturan, maka persekitaran tiga dimensi diperlukan untuk mengatasi masalah pembezaan tersebut. Oleh itu, sel kondrosit yang diisolasi daripada sendi OA (OAC) telah dikulturkan dalam media CGM dan proses analisis seperti pertumbuhan kinetik dan pencirian sel stem dilakukan sebelum disemai ke atas gentian nano yang dihasilkan. Gentian nano PLGA dipilih berdasarkan ciri-ciri versetilnya berserta kondisi optimum kepekatan cecair, kadar aliran, voltan dan jarak dari jarum picagari ke pengumpul yang digunakan. Tahap ketoksikan, pengimbasan mikroskop elektron (SEM) dan analisis q-PCR juga telah dilakukan dalam kajian ini. Keputusan kajian menunjukkan, OAC memenuhi kriteria minimum memiliki ciri sel stem dan ia mudah berpoliferasi di atas plat kultur, ekspresi penanda sel stem yang tinggi (≥95%) untuk CD13, CD29, CD44, CD73 dan CD90 (≤2%) CD45 dan HLA-DR serta berpotensi diinduksi kepada leluhur mesoderm. Gentian nano elektrospun tidak menunjukkan kesan toksik kepada sel malah mempengaruhi proliferasi sel semasa pengkulturan. Berdasarkan interaksi gentian nano PLGA dan OAC, perbezaan saiz gentian akan mempengaruhi proliferasi sel. Gentian nano bersaiz 100 nm telah menunjukkan berlakunya proliferasi OAC yang lebih tinggi dan ekspresi gen dan protein yang lebih baik berbanding dengan kultur monolapisan. Kesimpulannya, OAC berpotensi sebagai sumber sel untuk penjanaan semula tulang rawan berdasarkan kepada ekspresi penanda sel stem seperti yang diekspresikan oleh sumsum tulang manusia. Penjanaan tulang rawan akan lebih berkesan jika OAC dikulturkan di atas persekitaran mikro tiga dimensi seperti yang ditunjukkan di dalam kajian ini.

 

Kata kunci: Elektrospun; kejuruteraan tisu tulang rawan; perbezaan; osteoartritis; sitorangka

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*Corresponding author; email: shiplu@ppukm.ukm.edu.my