Verification of simple hydration/dehydration methods to characterize multiple water compartments on Tendon Type 1 Collagen

Ivan L. Cameron, Nicholas J. Short, Gary D. Fullerton

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

A molecular model of collagen hydration is used to validate centrifugal dehydration force (CDF) and re-hydration isotherm (RHI) methods to measure and characterize hydration compartments on bovine tendon. The CDF method assesses fluid flow rate from flexor and extensor tendons expressed in (g-water/g-dry mass-minute) and hydration capacity of compartments in (g-water/g-dry mass). Measured water compartment capacities agree with the molecular model of collagen hydration [Fullerton GD, Rahal A. Collagen structure: the molecular source of tendon magic angle effect. J Mag Reson Imag 2007;25:345-361; Fullerton GD, Amurao MR. Evidence that collagen and tendon have monolayer water coverage in the native state. Cell Biol Int 2006;30(1):56-65]. Native tendon hydration has monolayer coverage on collagen hm = 1.6 g/g which divides into primary hydration on polar surfaces hpp = 0.8 g/g and secondary hydration hs = 0.8 g/g bridging over hydrophobic surfaces. Primary hydration is hydrogen bonded to collagen polar side chains hpsc = 0.54 g/g with small free energy or to the protein main chain hydration hpmc = 0.26 g/g with greater free energy of binding. The CDF method replaces the more time consuming water proton NMR spin-lattice dehydration (NMR titration) method, confirms the presence of three non-bulk water compartments on collagen (hpmc = 0.26 g/g, hpp = 0.8 g/g and hm = 1.6 g/g). This CDF method provides the most reproducible experimental measure of total tissue non-bulk water (TNBW). The re-hydration isotherm method, on the other hand, provides the most accurate measure of the Ramachandran water-bridge capacity hRa = 0.0656 g/g. The only equipment needed are: microfilterfuge tubes, a microcentrifuge capable of 14,000 × g or 4 MPa, a vacuum drying oven, an accurate balance and curve fitting ability. The newly validated methods should be useful for characterizing multiple water compartments in biological and non-biological materials by allowing direct measurement of water compartment changes induced by pH, co-solute salt, glycation and protein cross-linking.

Original languageEnglish (US)
Pages (from-to)531-539
Number of pages9
JournalCell Biology International
Volume31
Issue number6
DOIs
StatePublished - Jun 2007

Keywords

  • Centrifugation
  • Collagen
  • Fluid flow
  • Glycation
  • Hydration
  • Hydration isotherm
  • Tendon
  • Water

ASJC Scopus subject areas

  • Cell Biology

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