Peptide‐Functionalized Hydrogels Modulate Integrin Expression and Stemness in Adult Human Epidermal Keratinocytes. Issue 10 (23rd August 2019)

• Record Type: Journal Article
• Authors: Davis‐Hall, Duncan; Nguyen, Vy; D'Ovidio, Tyler J.; Tsai, Ethan; Bilousova, Ganna; Magin, Chelsea M.
• Is Part Of: Advanced biosystems. Volume 3:Issue 10(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-08-23
• Subjects: extracellular matrix -- hydrogels -- keratinocytes -- peptides -- poly(ethylene glycol); Biological systems -- Periodicals; Biotechnology -- Periodicals; Bioengineering -- Periodicals; Biomedical engineering -- Periodicals; Biological Science Disciplines; Periodicals; Periodicals; 660.6
• DOI: 10.1002/adbi.201900022
• Languages: English
• ISSNs: 2366-7478

Abstract

Abstract: The extracellular matrix (ECM) controls keratinocyte proliferation, migration, and differentiation through β‐integrin signaling. Wound‐healing research requires expanding cells in vitro while maintaining replicative capacity; however, early terminal differentiation under traditional culture conditions limits expansion. Here, a design of experiments approach identifies poly(ethylene glycol)‐based hydrogel formulations with mechanical properties (elastic modulus, E = 20.9 ± 0.56 kPa) and bioactive peptide sequences that mimic the epidermal ECM. These hydrogels enable systematic investigation of the influence of cell‐binding domains from fibronectin (RGDS), laminin (YIGSR), and collagen IV (HepIII) on keratinocyte stemness and β1 integrin expression. Quantification of 14‐day keratin protein expression shows four hydrogels improve stemness compared to standard techniques. Three hydrogels increase β1 integrin expression, demonstrating a positive linear relationship between stemness and β1 integrin expression. Multifactorial statistical analysis predicts an optimal peptide combination ([RGDS] = 0.67 mm, [YIGSR] = 0.13 mm, and [HepIII] = 0.02 mm ) for maintaining stemness in vitro. Best‐performing hydrogels exhibit no decrease in Ki‐67‐positive cells compared to standards (15% decrease, day 7 to 14; p < 0.05, Tukey Test). These data demonstrate that precisely designed hydrogel biomaterials direct integrin expression and promote proliferation, improving the regenerative capability of cultured keratinocytes for basic science and translational work. Abstract : Hydrogels that mimic the mechanical and biochemical properties of native human epidermis improve stemness and proliferation of adult human epidermal keratinocytes. Results show increases in expression of β1 integrins that maintain a stem‐like ratio of keratin 14 (K14 ) to keratin 1 (K1 ) compared to standard techniques over 14 days. These hydrogels could improve current techniques for expanding cells ex vivo.