Abalone Collagen Extracts Potentiate Stem Cell Properties of Human Epidermal Keratinocytes

Abstract

Stem cell activities in human tissues are critical for tissue integrity and function. Maintaining keratinocyte stem cells (KSCs) stemness helps sustain healthy skin by supporting keratinocyte renewal, involving the formation of epidermal barriers. In this study, abalone collagen (AC) extracts with molecular weights of 3 kDa (AC 1) and 300 kDa (AC 2) were compared to the epidermal growth factor (EGF) for their effects on cell proliferation, cell migration (wound healing), spheroid formation, and the expression level of stem cell markers on human keratinocytes (HaCaT cells). Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell proliferation was quantified by ATP and DNA content analysis and Sulforhodamine B (SRB) assays. Cell migration assay was determined using the scratch wound healing test. Spheroid formation was evaluated and the expression level of stem cell markers was investigated by western blot analysis. The results showed that AC 1 at the concentration of 100 µg/mL could stimulate HaCaT cell proliferation, migration, spheroid formation, and the expression level of stem cell markers (keratin 19, β-catenin, ALDH1A1) compared to the control. In conclusion, a smaller molecular weight of abalone collagen extract exhibits a better effect on keratinocytes proliferation, migration, and stemness, which could be a potential active ingredient in cosmeceutical products.

Hydrolyzed Collagen from Salmon Skin Increases the Migration and Filopodia Formation of Skin Keratinocytes by Activation of FAK/Src Pathway

“…The present study showed that HCSS treatment at the concentrations 100, 500, and 1000 μg/mL significantly enhanced keratin 19 and β-catenin protein expression in keratinocyte HaCaT cells when compared to the untreated control (Figure 6A and Figure 6B). Our results were similar to those from the previous studies reporting that the collagen extract from abalone caused an increase in KSCs marker expression, such as ALDH1A1, keratin 19, and β-catenin in keratinocyte HaCaT cells [Thaweekitphathanaphakdee et al, 2019]. In addition, collagen type I enhanced the properties of stem cell-like phenotype by the α 2 β 1 -integrin activation [Kirkland, 2009].…”

“…Research conducted by Yang et al [2019b] also showed that peptides from Nibea japonica skin collagen increased cell viability of NIH-3T3 fibroblasts. Additionally, abalone collagen was observed to increase keratinocyte viability [Thaweekitphathanaphakdee et al, 2019]. Based on the results, we selected the HCSS concentrations of 50, 100, 500, and 1000 μg/mL for further experiments.…”

“…Fish skins are a by-product of the food industry and a rich source of collagen. Marine collagen has been noted to promote cell proliferation and migration of skin cells by augmenting various mediators in the stimulation of skin wound repair process [Chotphruethipong et al, 2021a;Hu et al, 2017;Yang et al, 2019b] and KSCs function [Thaweekitphathanaphakdee et al, 2019]. To date, bioactive peptides released from marine origin proteins, for example fish collagen have been reported to exhibit many biological effects including anti-inflammatory, anti-oxidant, anti-aging, and wound repair activities [Chotphruethipong et al, 2021a;Edgar et al, 2018;Huang et al, 2015].…”

“…2018b;Chotphruethipong et al, 2021a;Yang et al, 2019b] and keratinocyte proliferation[Chotphruethipong et al, 2021b;Thaweekitphathanaphakdee et al, 2019] was promoted in response to peptides with hydrophobic amino acids (AAs) treatment Sánchez & Vázquez [2017]. reported that the size of peptides, their AAs composition and sequence affect their cell proliferation potential.…”

Recombinant expression a novel fibronectin—collage fusion peptide modulating stem cell stemness via integrin β3

“…FN10, a part of the fibronectin type III module, contains RGD sequences that specifically bind αvβ1 to guide the fate of the cells (Bharadwaj et al 2017 ; Hocking et al 1996 ). Moreover, collagen and fibronectin support the growth of human embryonic stem cells (hESC) without the use of a feeder layer under certain conditions (Lu et al 2006 ) and play a role in maintaining the stemness of stem cells (Akhir and Teoh 2020 ; Thaweekitphathanaphakdee et al 2019 ). Notably, as the main strain matrix element, collagen relies on fibronectin, fibronectin-bound, and collagen-bound integrins to complete its corresponding functions (Kadler et al 2008 ; Kubow et al 2015 ).…”