Sains Malaysiana 48(3)(2019): 637–644

http://dx.doi.org/10.17576/jsm-2019-4803-17

Exploring The Potential of Dermal Fibroblast Conditioned Medium on Skin Wound Healing and Anti-Ageing

(Meneroka Potensi Fibroblas Derma Medium Terkondisi untuk Penyembuhan Luka Pada Kulit dan Anti-Penuaan)

SHIPLU ROY CHOWDHURY*, LIM SHAUN JING, MUHAMMAD NOOR HAZIQ BIN ZOLKAFLI, NUR’ AQILAH BINTI MOHD ABU ZARIN, WAN ALYANI BINTI WAN ABDULLAH, NURUL AIDAH BINTI MD MOTHAR, MANIRA MAAROF & NUR ATIQAH HAIZUM ABDULLAH

Tissue Engineering Centre, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Received: 17 April 2018/Accepted: 29 November 2018

ABSTRACT

Skin ageing is associated with a decrease in collagen and delayed wound healing. The previous study has shown that supplementation of growth factors can enhance the production of extracellular matrix (ECM) and efficiency of wound healing. This study aimed to produce a Dermal Fibroblast Conditioned Medium (DFCM) and evaluate its potential on in vitro skin wound healing and anti-ageing properties. DFCM were obtained by culturing confluent human dermal fibroblasts (n=3) in keratinocyte-specific medium (DFCM-KM), fibroblast-specific medium (DFCM-FM) and fibroblast-specific medium with growth supplements (DFCM-GM). Protein concentration was determined by Bicinchoninic Acid (BCA) assay. To evaluate the wound healing and anti-ageing properties, human dermal fibroblasts were supplemented with three different DFCM. Fibroblasts cultured with F12:DMEM+10%FBS was used as a control. The growth rate of fibroblasts was evaluated by culturing cells until day 4. Migration rate was evaluated at 12 h after seeding. In vitro healing rate was evaluated via scratch assay. Anti-ageing potential of DFCM was evaluated via RT2 Profiler PCR Array. Concentration of total protein was found to be significantly higher in DFCM-GM (2914.79±150.67mg/mL) compared to DFCM-KM and DFCM-FM. Out of the tested 12 anti-ageing genes, fibroblasts supplemented with DFCM-GM demonstrated a significantly higher expression of WRN compared to other conditions. In conclusion, DFCM-GM contains more proteins and found superior in modulating wound healing and anti-ageing properties.

Keywords: Anti-ageing; conditioned medium; dermal fibroblast conditioned medium; fibroblasts; wound healing

ABSTRAK

Penuaan kulit dikaitkan dengan pengurangan kolagen dan penyembuhan luka yang lambat. Kajian terdahulu menunjukkan bahawa penambahan faktor pertumbuhan boleh meningkatkan penghasilan matriks ekstrasel (ECM) serta keberkesanan proses penyembuhan luka. Kajian ini bertujuan untuk menghasilkan medium terkondisi daripada fibroblas (DFCM) serta menilai kesannya terhadap proses penyembuhan luka dan anti-penuaan. DFCM diperoleh dengan mengkultur sel fibroblas derma manusia (n = 3) di dalam medium khusus keratinosit (DFCM-KM), medium khusus fibroblas (DFCM-FM) dan medium khusus fibroblas dengan penambahan faktor pertumbuhan (DFCM-GM). Kepekatan protein ditentukan dengan menggunakan asai acid Bicinchoninic (BCA). Untuk menilai proses penyembuhan luka dan anti-penuaan, kultur fibroblas ditambah dengan tiga DFCM yang berbeza. Fibroblas yang dikultur di dalam F12: DMEM + 10% FBS digunakan sebagai kumpulan kawalan. Kadar pertumbuhan fibroblas dinilai melalui pengkulturan sel sehingga hari ke-4 manakala kadar migrasi dinilai bermula dari 12 jam selepas pengkulturan. Kadar penyembuhan luka secara in vitro dinilai menggunakan teknik pencalaran. Potensi anti-penuaan DFCM dinilai melalui RT2 Profiler PCR Array. Kepekatan protein DFCM-GM (2914.79 ± 150.67 μg/mL) didapati jauh lebih tinggi berbanding dengan DFCM-KM dan DFCM-FM. Analisis pengekspresan 12 gen anti-penuaan menunjukkan bahawa ekspresi WRN pada fibroblas yang ditambah dengan DFCM-GM adalah jauh lebih tinggi berbanding kumpulan yang lain. Kesimpulannya, DFCM-GM mengandungi lebih banyak protein, boleh meningkatkan serta mengawal proses penyembuhan luka dan bersifat anti-penuaan.

Kata kunci: Anti-penuaan; fibroblas; medium terkondisi; medium terkondisi daripada fibroblas; penyembuhan luka

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