Search for Author, Title, Keyword
Hydrolyzed Collagen from Salmon Skin Increases the Migration and Filopodia Formation of Skin Keratinocytes by Activation of FAK/Src Pathway
More details
Hide details
Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand
International Center of Excellence in Seafood Science and Innovation, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand
Department of Food Technology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla 90110, Thailand
Expert Centre of Innovative Health Food (InnoFood), Thailand Institute of Scientific and Technological Research (TISTR), Khlong Luang, Pathum Thani 12120, Thailand
Submission date: 2021-04-27
Final revision date: 2021-08-19
Acceptance date: 2021-08-23
Online publication date: 2021-09-03
Publication date: 2021-09-03
Corresponding author
Wanida Sukketsiri   

Division of Health and Applied Sciences, Faculty of Science, Prince of Songkla University, 90110, Songkhla, Thailand
Pol. J. Food Nutr. Sci. 2021;71(3):323-332
Previous studies reported hydrolyzed collagen increase cell proliferation and migration involved in the wound repair process. Nevertheless, the knowledge related with wound repair mechanism of hydrolyzed collagen from salmon skin (HCSS) has not been fully elucidated. Therefore, this study aimed to elucidate the effects of HCSS on the migration of keratinocyte HaCaT cells. Additionally, its molecular mechanism through cell division control protein 42 (Cdc42), Ras-related C3 botulinum toxin substrate 1 (Rac1), and Ras homolog family member A (RhoA) via focal adhesion kinase (FAK)-steroid receptor coactivator (Src) regulation and keratinocyte stem cells (KSCs) markers were also evaluated. After 24 h of incubation, keratinocyte proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and double stranded DNA (dsDNA) assays, and by determining the total cellular protein content. Keratinocyte migration and filopodia formation were measured by wound healing assay and phalloidin‐rhodamine staining, respectively. The migratory related proteins were evaluated by western blot analysis. HCSS had a high content of hydrophobic amino acids and imino acids. HaCaT cell proliferation and migration were significantly increased in response to HCSS at the concentration of 100-1000 μg/mL. The formation of filopodia was subsequently increased in response to HCSS at concentrations of 100-1000 μg/mL. Moreover, HCSS upregulated Cdc42, Rac1, and RhoA protein expression and activated the phosphorylation of FAK and Src pathway. HCSS at the concentration of 100-1000 μg/mL could trigger stemness by increased KSC markers, including keratin 19 and β-catenin expression. This study has demonstrated that HCSS induces proliferation and migration of keratinocytes, subsequently promotes the second phase of wound healing process by FAK-Src activation and also increases the KSC properties.
We thank Publication clinic, Prince of Songkla University for the assistance in manuscript language editing.
This research was funded by Participation in the Partnership Program in Production of Graduates in Master’s Degree between the Thailand Institute of Scientific and Technological Research (TISTR) and Educational Institutions (6110220098).
Abate, M., Citro, M., Pisanti, S., Caputo, M., Martinelli, R. (2021). Keratinocytes migration promotion, proliferation induction, and free radical injury prevention by 3-hydroxytirosol. International Journal of Molecular Sciences, 22(5), art. no. 2438.
Abbas, O., Richards, J.E., Yaar, R., Mahalingam, M. (2011). Stem cell markers (cytokeratin 15, cytokeratin 19 and p63) in in situ and invasive cutaneous epithelial lesions. Modern Pathology, 24, 90-97.
Abreu-Blanco, M.T., Watts, J.J., Verboon, J.M., Parkhurst, S.M. (2012). Cytoskeleton responses in wound repair. Cellular and Molecular Life Sciences, 69, 2469-2483.
Baroni, A., Buommino, E., De Gregorio, V., Ruocco, E., Ruocco, V., Wolf, R. (2012). Structure and function of the epidermis related to barrier properties. Clinics in Dermatology, 30(3), 257-262.
Benjakul, S., Karnjanapratum, S., Visessanguan, W. (2018a). Production and characterization of odorless antioxidative hydrolyzed collagen from seabass (Lates calcarifer) skin without descaling. Waste and Biomass Valorization, 9, 549-559.
Benjakul, S., Karnjanapratum, S., Visessanguan, W. (2018b). Hydrolysed collagen from Lates calcarifer skin: its acute toxicity and impact on cell proliferation and collagen production of fibroblasts. International Journal of Food Science & Technology, 53(8), 1871-1879.
Chalamaiah, M., Ulug, S.K., Hong, H., Wu, J.P. (2019). Regulatory requirements of bioactive peptides (protein hydrolysates) from food proteins. Journal of Functional Foods, 58, 123-129.
Chen, T., Hu, H., Fan, Y., Wang, S., Chen, Q., Si, L., Li, B. (2016). Protective effect of gelatin peptides from pacific cod skin against photoaging by inhibiting the expression of MMPs via MAPK signaling pathway. Journal of Photochemistry and Photobiology B: Biology, 165, 34-41.
Chen, J., Gao, K., Liu, S., Wang, S., Elango, J., Bao, B., Dong, J., Liu, N., Wu, W. (2019). Fish collagen surgical compress repairing characteristics on wound healing process in vivo. Marine Drugs, 17(1), art. no. 33.
Chiu, L.H., Lai, W.F., Chang, S.F., Wong, C.C., Fan, C.Y., Fang, C.L., Tsai, Y.H. (2014). The effect of type II collagen on MSC osteogenic differentiation and bone defect repair. Biomaterials, 35(9), 2680-2691.
Chotphruethipong, L., Aluko, R.E., Benjakul, S. (2019). Hydrolyzed collagen from porcine lipase‐defatted seabass skin: antioxidant, fibroblast cell proliferation, and collagen production activities. Journal of Food Biochemistry, 43(5), art. no. e12825.
Chotphruethipong, L., Sukketsiri, W., Aluko, R.E., Sae-leaw, T., Benjakul, S. (2021a). Effect of hydrolyzed collagen from defatted Asian sea bass (Lates calcarifer) skin on fibroblast proliferation, migration and antioxidant activities. Journal of Food Science and Technology, 58, 541-551.
Chotphruethipong, L., Sukketsiri, W., Battino, M., Benjakul, S. (2021b). Conjugate between hydrolyzed collagen from defatted seabass skin and epigallocatechin gallate (EGCG): characteristics, antioxidant activity and in vitro cellular bioactivity. RSC Advances, 11, 2175-2184.
Chotphruethipong, L., Binlateh, T., Hutamekalin, P., Sukketsiri, W., Aluko, R.E., Benjakul, S. (2021c). In vitro antioxidant and wound-healing activities of hydrolyzed collagen from defatted Asian sea bass skin as influenced by different enzyme types and hydrolysis processes. RSC Advances, 11, 18144-18151.
Chotphruethipong, L., Binlateh, T., Hutamekalin, P., Aluko, R.E., Tepaamorndech, S., Zhang, B., Benjakul, S. (2021d). Impact of hydrolyzed collagen from defatted sea bass skin on proliferation and differentiation of preosteoblast MC3T3-E1 cells. Foods, 10(7), art. no. 1476.
Crampton, S.P., Wu, B., Park, E.J., Kim, J.H., Solomon, C., Waterman, M.L., Hughes, C.C. (2009). Integration of the β-catenin-dependent Wnt pathway with integrin signaling through the adaptor molecule Grb2. PLoS One, 4, art. no. e7841.
Desgrosellier, J.S., Cheresh, D.A. (2010). Integrins in cancer: biological implications and therapeutic opportunities. Nature Reviews Cancer, 10, 9-22.
Edgar, S., Hopley, B., Genovese, L., Sibilla, S., Laight, D., Shute, J. (2018). Effects of collagen-derived bioactive peptides and natural antioxidant compounds on proliferation and matrix protein synthesis by cultured normal human dermal fibroblasts. Scientific Reports, 8, art. no. 10474.
Elango, J., Robinson, J., Zhang, J., Bao, B., Ma, N., de Val, J.E.M.S., Wu, W. (2019). Collagen peptide upregulates osteoblastogenesis from bone marrow mesenchymal stem cells through MAPK- Runx2. Cells, 8(5), art. no. 446.
Fuchs, E. (2008). Skin stem cells: rising to the surface. Journal of Cell Biology, 180(2), 273-284.
Horikoshi, Y., Kamizaki, K., Hanaki, T., Morimoto, M., Kitagawa, Y., Nakaso, K., Kusumoto, C., Matsura, T. (2018). α-Tocopherol promotes HaCaT keratinocyte wound repair through the regulation of polarity proteins leading to the polarized cell migration. BioFactors, 44(2), 180-191.
Hu, Z., Yang, P., Zhou, C., Li, S., Hong, P. (2017). Marine collagen peptides from the skin of nile tilapia (Oreochromis niloticus): characterization and wound healing evaluation. Marine Drugs, 15(4), art. no. 102.
Huang, R., Li, W., Lv, X., Lei, Z., Bian, Y., Deng, H., Wang, H., Li, J., Li, X. (2015). Biomimetic LBL structured nanofibrous matrices assembled by chitosan/collagen for promoting wound healing. Biomaterials, 53, 58-75.
Kawaguchi, T., Nanbu, P.N., Kurokawa, M. (2012). Distribution of prolylhydroxyproline and its metabolites after oral administration in rats. Biological and Pharmaceutical Bulletin, 35(3), 422-427.
Kimira, Y., Odaira, H., Nomura, K., Taniuchi, Y., Inoue, N., Nakatani, S., Shimizu, J., Wada, M., Mano, H. (2017). Collagen-derived dipeptide prolylhydroxyproline promotes osteogenic differentiation through Foxg1. Cellular & Molecular Biology Letters, 22, art. no. 27.
Kirkland, S.C. (2009). Type I collagen inhibits differentiation and promotes a stem cell-like phenotype in human colorectal carcinoma cells. British Journal of Cancer, 101, 320-326.
Leng, L., Ma, J., Lv, L., Wang, W., Gao, D., Zhu, Y., Wu, Z. (2020). Both Wnt signaling and epidermal stem cell-derived extracellular vesicles are involved in epidermal cell growth. Stem Cell Research & Therapy, 11, art. no. 415.
Lü, L., Zhang, L., Wai, M.S.M., Yew, D.T.W., Xu, J. (2012). Exocytosis of MTT formazan could exacerbate cell injury. Toxicology in Vitro, 26(4), 636-644.
Martin, P., Nunan, R. (2015). Cellular and molecular mechanisms of repair in acute and chronic wound healing. British Journal of Dermatology, 173(2), 370-378.
Masraksa, W., Tanasawet, S., Hutamekalin, P., Wongtawatchai, T., Sukketsiri, W. (2020). Luteolin attenuates migration and invasion of lung cancer cells via suppressing focal adhesion kinase and non-receptor tyrosine kinase signaling pathway. Nutrition Research and Practice, 14(2), 127-133.
Morgan, P.T., Breen, L. (2021). The role of protein hydrolysates for exercise-induced skeletal muscle recovery and adaptation: a current perspective. Nutrition & Metabolism, 18, art. no. 44.
Petpiroon, N., Suktap, C., Pongsamart, S., Chanvorachote, P., Sukrong, S. (2015). Kaempferol-3-O-rutinoside from Afgekia mahidoliae promotes keratinocyte migration through FAK and Rac 1 activation. Journal of Natural Medicines, 69, 340-348.
Pincelli, C., Marconi, A. (2010). Keratinocyte stem cells: friends and foes. Journal of Cellular Physiology, 225(2), 310-315.
Ritto, D., Tanasawet, S., Singkhorn, S., Klaypradit, W., Hutamekalin, P., Tipmanee, V., Sukketsiri, W. (2017). Astaxanthin induces migration in human skin keratinocytes via Rac1 activation and RhoA inhibition. Nutrition Research and Practice, 11(4), 275-280.
Sánchez, A., Vázquez, A. (2017). Bioactive peptides: A review. Food Quality and Safety, 1, 29-46.
Shigemura, Y., Iwai, K., Morimatsu, F., Iwamoto, T., Mori, T., Oda, C., Taira, T., Park, E.Y., Nakamura, Y., Sato, K. (2009). Effect of prolyl-hydroxyproline (Pro-Hyp), a food-derived collagen peptide in human blood, on growth of fibroblasts from mouse skin. Journal Agricultural and Food Chemistry, 57(2), 444-449.
Singkhorn, S., Tantisira, M.H., Tanasawet, S., Hutamekalin, P., Wongtawatchai, T., Sukketsiri, W. (2018). Induction of keratinocyte migration by ECa 233 is mediated through FAK/Akt, ERK, and p38 MAPK signaling. Phytotherapy Research, 32(7), 1397-1403.
Seo, G.Y., Hyun, C., Koh, D., Park, S., Lim, Y., Kim, Y.M., Cho, M. (2018). A novel synthetic material, BMM, accelerates wound repair by stimulating re-epithelialization and fibroblast activation. International Journal of Molecular Sciences, 19(4), art. no. 1164.
Sotiropoulou, P.A., Blanpain, C. (2012). Development and homeostasis of the skin epidermis. Cold Spring Harbor Perspectives in Biology, 4, art. no. a008383.
Thaweekitphathanaphakdee, S., Chanvorachote, P., Prateepchinda, S., Khongkow, M., Sucontphunt, A. (2019). Abalone collagen extracts potentiate stem cell properties of human epidermal keratinocytes. Marine Drugs, 17(7), art. no. 424.
Van Tonder, A., Joubert, A.M., Cromarty, A.D. (2015). Limitations of the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay when compared to three commonly used cell enumeration assays. BMC Research Notes, 8, art. no. 47.
Wickett, R.R., Visscher, M.O. (2006). Structure and function of the epidermal barrier. American Journal of Infection Control, 34(10), Suppl., S98-S110.
Wikramanayake, T.C., Stojadinovic, O., Tomic-Canic, M. (2014). Epidermal differentiation in barrier maintenance and wound healing. Advances in Wound Care, 3(3), 272-280.
Yang, R., Liu, F., Wang, J., Chen, X., Xie, J., Xiong, K. (2019a). Epidermal stem cells in wound healing and their clinical applications. Stem Cell Research & Therapy, 10, art. no. 229.
Yang, F., Jin, S., Tang, Y. (2019b). Marine collagen peptides promote cell proliferation of NIH-3T3 fibroblasts via NF-κB signaling pathway. Molecules, 24(22), art. no.4201.
Combined Effects of Defatted Hydrolyzed Collagen from Salmon Skin and Vitamin C on Proliferation and Migration of Human Fibroblast Cell
Lalita Chotphruethipong, Pilaiwanwadee Hutamekalin, Krisana Nilsuwan, Wanida Sukketsiri, Rotimi Aluko, Nazeer Abdul, Soottawat Benjakul
Utilization of feather keratin waste to antioxidant and migration-enhancer peptides by Bacillus licheniformis 8-4
Xiao-Dong Pei, Fan Li, Tian-Tian Gao, Lin-Ying Su, Fu-Tian Yu, Ping Shi, Chen-Xing Liu, Cheng-Hua Wang
Journal of Applied Microbiology
Food-derived collagen peptides: safety, metabolism, and anti-skin-aging effects
Lin Wang, Yue Zhang, Zhiling Zhu, Fuping Zheng, Ruichang Gao
Current Opinion in Food Science
M Gomathy, Krishnakumar Velayuthannair, Paul John
Advances in Wound Care
In vitro antioxidant and wound healing activity of Sargassum polycystum hydroethanolic extract in fibroblasts and keratinocytes
Wanwipha Woonnoi, Furoida Moolsap, Supita Tanasawet, Nattakanwadee Khumpirapang, Chakkapat Aenglong, Wanida Sukketsiri
Asian Pacific Journal of Tropical Biomedicine
Phytochemical analysis and impact of Citrus hystrix peel water extract on proliferation and migration of skin keratinocytes by activating FAK/Src/MAPK/Akt pathway
Furoida Moolsap, Supita Tanasawet, Wanwipha Woonnoi, Supawadee Daodee, Supawadee Parhira, Pennapa Chonpathompikunlert, Nareenath Muneerungsee, Tulaporn Wongtawatchai, Wanida Sukketsiri
Journal of Herbal Medicine
Antimetastatic effect of fucoidan against non-small cell lung cancer by suppressing non-receptor tyrosine kinase and extracellular signal-related kinase pathway
Nareenath Muneerungsee, Supita Tanasawet, Wanida Sukketsiri
Nutrition Research and Practice
The Combined Anti-Aging Effect of Hydrolyzed Collagen Oligopeptides and Exosomes Derived from Human Umbilical Cord Mesenchymal Stem Cells on Human Skin Fibroblasts
Huimin Zhu, Xin Guo, Yongqing Zhang, Ajab Khan, Yinuo Pang, Huifang Song, Hong Zhao, Zhizhen Liu, Hua Qiao, Jun Xie
Extraction of proteinaceous components and biominerals from cold water fish fileting side streams: a review
Sophie Kendler, Abhilash Sasidharan, Turid Rustad
Frontiers in Sustainable Food Systems
Journals System - logo
Scroll to top