title: Conjugation of extracellular matrix proteins to basal lamina analogs enhances keratinocyte attachment.
abstract: The dermal-epidermal junction of skin contains extracellular matrix proteins that are involved in initiating and controlling keratinocyte signaling events such as attachment, proliferation, and terminal differentiation. To characterize the relationship between extracellular matrix proteins and keratinocyte attachment, a biomimetic design approach was used to precisely tailor the surface of basal lamina analogs with biochemistries that emulate the native biochemical composition found at the dermal-epidermal junction. A high-throughput screening device was developed by our laboratory that allows for the simultaneous investigation of the conjugation of individual extracellular matrix proteins (e.g. collagen type I, collagen type IV, laminin, or fibronectin) as well as their effect on keratinocyte attachment, on the surface of an implantable collagen membrane. Fluorescence microscopy coupled with quantitative digital image analyses indicated that the extracellular matrix proteins adsorbed to the collagen-GAG membranes in a dose-dependent manner. To determine the relationship between extracellular matrix protein signaling cues and keratinocyte attachment, cells were seeded on protein-conjugated collagen-GAG membranes and a tetrazolium-based colorimetric assay was used to quantify viable keratinocyte attachment. Our results indicate that keratinocyte attachment was significantly enhanced on the surfaces of collagen membranes that were conjugated with fibronectin and type IV collagen. These findings define a set of design parameters that will enhance keratinocyte binding efficiency on the surface of collagen membranes and ultimately improve the rate of epithelialization for dermal equivalents.
authors.affiliation: Worcester Polytechnic Institute, Biomedical Engineering, Worcester, Massachusetts, USA.
authors.forename: Katie A
authors.initials: KA
authors.lastname: Bush
authors.forename: Brett R
authors.initials: BR
authors.lastname: Downing
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Walsh
authors.forename: George D
authors.initials: GD
authors.lastname: Pins
mesh_terms: D001485:Basement Membrane / Q000737:chemistry* / Q000378:metabolism; D040761:Biomimetic Materials / Q000737:chemistry*; D024141:Collagen Type IV / Q000737:chemistry; D020405:Dermis; D004817:Epidermis; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D019736:Prostheses and Implants; D011485:Protein Binding; D015398:Signal Transduction
chemical_list: D024141:Collagen Type IV; D016326:Extracellular Matrix Proteins; D005353:Fibronectins