Sains Malaysiana 52(1)(2023): 223-231

http://doi.org/10.17576/jsm-2023-5201-18

Effect of Dermal Fibroblast Conditioned Medium on Keratinocytes Irrespective of Age Group

(Kesan Medium Terkondisi Fibroblas Kulit pada Keratinosit Tanpa mengira Kumpulan Umur)

NURUL ‘IZZAH ABDUL GHANI¹, MANIRA MAAROF^1,*, SHIPLU ROY CHOWDHURY¹, AMINUDDIN BIN SAIM², RUSZYMAH BINTI HAJI IDRUS^1,3 & FAUZI MH BUSRA¹

¹Centre for Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

²KPJ Ampang Puteri Specialist Hospital, 1 Jalan Mamanda 9, 68000 Ampang, Kuala Lumpur, Federal Territory, Malaysia

³Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Diserahkan: 16 Mac 2022/Diterima: 5 Oktober 2022

Abstract

Skin aging causes delayed re-epithelialisation and impaired wound healing. Thus, supplementation of wound healing mediators and extracellular matrix (ECM) components may be a potential treatment strategy for age-related impaired wound healing. Fibroblasts secrete wound-healing factors and can be collected from used medium, i.e., dermal fibroblast conditioned medium (DFCM). In this study, we elucidated the effect of DFCM on the in vitro wound healing of keratinocytes isolated from different age groups (≥18–35, 36–54, ≥55 years) via cell attachment, growth rate, and wound healing rate assays. The DFCM was prepared by culturing confluent fibroblasts with serum-free keratinocyte-specific (DFCM-KM) and fibroblast-specific (DFCM-FM) medium. The cell attachment efficiency decreased with the increase of age. However, keratinocyte attachment was enhanced in the DFCM-KM group, where it was 1.24, 1.27, and 1.32 times higher of cells concentration for the ≥18–35-, 36–54-, and ≥55-year age groups, respectively, as compared to the control group. The keratinocytes from each age group demonstrated a similar growth profile for all culture conditions, where the DFCM-KM group exhibited a comparable growth rate with the control group whilst the DFCM-FM group exhibited a significantly lower growth rate compared to the other groups. In contrast, the DFCM-FM group demonstrated a significantly higher healing rate in all age groups as compared to the DFCM-KM and control groups. However, there was no significant difference between the healing rates of the DFCM-KM and control groups. In conclusion, DFCM-KM enhanced keratinocyte attachment while DFCM-FM enhanced the keratinocyte healing rate irrespective of donor age, which indicated the potential application of DFCM in wound healing in aged skin.

Keywords: Aging; conditioned medium; re-epithelialisation; wound

Abstrak

Penuaan kulit menyebabkan kelewatan dalam pengepiteliuman semula dan penyembuhan luka. Oleh itu, penambahan faktor penyembuhan luka dan komponen matriks ekstrasel (ECM) merupakan strategi rawatan yang berpotensi bagi penyembuhan luka yang terjejas akibat faktor usia. Fibroblas merupakan sel yang merembeskan faktor pertumbuhan yang terlibat dalam proses penyembuhan luka dan boleh dikumpul daripada medium terpakai, iaitu medium terkondisi daripada fibroblas kulit (DFCM). Kajian ini bertujuan untuk mengkaji kesan DFCM pada penyembuhan luka secara in vitro bagi sel keratinosit daripada kumpulan umur yang berbeza (≥18-35, 36-54 dan ≥55 tahun) melalui pelekatan sel, kadar pertumbuhan dan penyembuhan luka. DFCM disediakan daripada pengkulturan fibroblas di dalam media tanpa serum khusus keratinosit (DFCM-KM) dan khusus fibroblas (DFCM-FM). Tahap pelekatan sel berkurangan dengan peningkatan umur. Walau bagaimanapun, pelekatan keratinosit menunjukkan peningkatan dengan tambahan DFCM-KM berbanding kumpulan kawalan yang masing-masing 1.24, 1.27 dan 1.32 kali lebih tinggi kepekatan sel bagi umur ≥18-35, 36-54 dan ≥55 tahun. Keratinosit daripada kumpulan umur yang berbeza menunjukkan profil pertumbuhan yang sama untuk semua keadaan kultur, dengan DFCM-KM menunjukkan kadar yang setanding dengan kumpulan kawalan manakala DFCM-FM adalah jauh lebih rendah berbanding kumpulan lain. Sebaliknya, kumpulan DFCM-FM menunjukkan kadar penyembuhan yang jauh lebih tinggi dalam semua kumpulan umur berbanding DFCM-KM dan kumpulan kawalan. Walau bagaimanapun, tiada perbezaan yang ketara dalam kadar penyembuhan luka untuk keratinosit dengan tambahan DFCM-KM dan kumpulan kawalan. Kesimpulannya, tanpa mengira umur penderma, DFCM-KM meningkatkan pelekatan keratinosit manakala DFCM-FM meningkatkan kadar penyembuhan keratinosit, menunjukkan potensi penggunaan DFCM dalam penyembuhan luka pada kulit pesakit yang sudah tua.

Kata kunci: Luka; medium terkondisi; pengepiteliuman semula; penuaan

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