title: Methods to promote Notch signaling at the biomaterial interface and evaluation in a rafted organ culture model.
abstract: The Notch signaling pathway is a promising target for controlling cell fate choices at the biomaterial-tissue interface. Building on our previous work in developing Notch-signaling biomaterials, we evaluated various immobilization schemes for Notch ligands, and their effect on human foreskin keratinocytes. A peptide sequence derived from the Jagged-1 DSL-region and immobilized to poly(2-hydroxyethyl methacrylate) (polyHEMA) showed no bioactivity in relation to the Notch-CSL pathway. The full-length Jagged-1 protein immobilized directly to the polyHEMA surface showed activity in signaling the Notch-CSL pathway. However, an indirect affinity immobilization approach yielded a stronger signal. Human keratinocytes plated on bound Jagged-1 showed upregulated involucrin, keratin 10, and loricrin protein expression, with this expression being cell density-dependent. Utilizing a human foreskin rafted organ culture model as a bridge between in vitro and in vivo studies, Jagged-1-modified or control polyHEMA rods were implanted in human foreskin and cultured at the air-medium interface. Keratinocyte proliferation was suppressed and intermediate-stage differentiation promoted in Jagged-1-modified rods compared with control rods. Thus, Notch-signaling biomaterials provide a robust approach to control keratinocyte differentiation and may find application to other progenitor and stem cells.
authors.affiliation: Department of Bioengineering, University of Washington, Seattle, WA 98195, USA.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Beckstead
authors.forename: Jason C
authors.initials: JC
authors.lastname: Tung
authors.forename: Katharine J
authors.initials: KJ
authors.lastname: Liang
authors.forename: Zarry
authors.initials: Z
authors.lastname: Tavakkol
authors.forename: Marcia L
authors.initials: ML
authors.lastname: Usui
authors.forename: John E
authors.initials: JE
authors.lastname: Olerud
authors.forename: Cecilia M
authors.initials: CM
authors.lastname: Giachelli
mesh_terms: D000595:Amino Acid Sequence; D001672:Biocompatible Materials / Q000737:chemistry / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D020007:Implants, Experimental; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D000072100:Jagged-1 Protein; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008689:Methacrylates / Q000737:chemistry; D008969:Molecular Sequence Data; D009924:Organ Culture Techniques / Q000379:methods*; D051880:Receptors, Notch / Q000378:metabolism*; D000072099:Serrate-Jagged Proteins; D015398:Signal Transduction; D012867:Skin / Q000166:cytology
chemical_list: D001672:Biocompatible Materials; D002135:Calcium-Binding Proteins; D055767:Immobilized Proteins; D036341:Intercellular Signaling Peptides and Proteins; C000606273:JAG1 protein, human; D000072100:Jagged-1 Protein; D008565:Membrane Proteins; D008689:Methacrylates; D051880:Receptors, Notch; D000072099:Serrate-Jagged Proteins; C005044:hydroxyethyl methacrylate