title: pH-responsive collagen fibrillogenesis in confined droplets induced by vapour diffusion.
abstract: A novel methodology for the assembly of collagen fibrils in microliter drops is proposed. It consists in the gradual increase of pH by means of vapour diffusion coming from the decomposition of NH4HCO3 solutions. The pH increase rate as well as the final steady pH of solutions containing collagen can be adjusted by varying the concentration of NH4HCO3. Both parameters are of predominant importance in collagen fibrillogenesis. The effect of these parameters on the kinetic of the fibrillogenesis process and on the fibrils morphology was studied. We found that both the kinetic and the morphology are mainly driven by electrostatic interactions. A gradual increase of pH slows down the formation of collagen fibres and favours the lateral interaction between fibrils producing broader fibres. On the other hand, a rapid increase of pH reduces the lateral electrostatic interactions favouring the formation of thinner fibres. The formation of the D-band periodicity is also a pH-dependent process that occurs after fibrillogenesis when the most stable state of fibres formation has been reached.
authors.affiliation: Laboratorio de Estudios Cristalográficos, Instituto Andaluz de Ciencias de la Tierra (IACT, CSIC-UGR), Avda. de las Palmeras 4, 18100, Armilla (Granada), Spain.
authors.forename: Gloria Belén
authors.initials: GB
authors.lastname: Ramírez-Rodríguez
authors.forename: Michele
authors.initials: M
authors.lastname: Iafisco
authors.forename: Anna
authors.initials: A
authors.lastname: Tampieri
authors.forename: Jaime
authors.initials: J
authors.lastname: Gómez-Morales
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Delgado-López
mesh_terms: D001639:Bicarbonates / Q000737:chemistry; D024042:Collagen Type I / Q000737:chemistry / Q000378:metabolism*; D004058:Diffusion; D024022:Fibrillar Collagens / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D057139:Nanofibers / Q000737:chemistry; D055503:Protein Multimerization; D012996:Solutions; D014835:Volatilization
chemical_list: D001639:Bicarbonates; D024042:Collagen Type I; D024022:Fibrillar Collagens; D012996:Solutions; C027043:ammonium bicarbonate