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.
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Acknowledgments
This work has been carried out within the frameworks of the projects from Spanish MINECO co-funded with FEDER (Projects MAT2011-28543 and Factoría de Cristalización, Consolider-Ingenio 2010), Junta de Andalucía (RMN 1344) and the Master in Crystallography and Crystallization (CSIC-UIMP). JMDL also acknowledges to the Spanish Servicio Público de Empleo Estatal (SEPE) for the funding during the writing of the manuscript. MI and AT would like to acknowledge the European projects SMILEY (FP7-NMP-2012-SMALL-6-310637) and OPHIS (FP7-NMP-2009-SMALL-3-246373).
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Ramírez-Rodríguez, G.B., Iafisco, M., Tampieri, A. et al. pH-responsive collagen fibrillogenesis in confined droplets induced by vapour diffusion. J Mater Sci: Mater Med 25, 2305–2312 (2014). https://doi.org/10.1007/s10856-014-5189-1
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DOI: https://doi.org/10.1007/s10856-014-5189-1