Electrochemical Time-of-Flight in crosslinked collagen matrix solution: Implications of structural changes for drug delivery systems

Electrochemical Time-of-Flight (ETOF) method was used for the first time to measure the diffusion coefficient of 4-hydroxy-TEMPO, a molecular probe in collagen I matrix solution as a function of its concentration and the extent of crosslinking with glutaraldehyde (GA). The values of the diffusion coefficient were correlated with Circular Dichroism (CD) and viscosity data to assess the changes of the structure of a collagen matrix. The low value of probe diffusion coefficient indicates that the molecular collagen contributes to large diffusion hindrance of the medium. The combined Brinkman or effective medium model and the Carman–Kozeny model were used to estimate the average diameter of a matrix pore as a function of collagen solution composition. We show that 0.5% and 1% (w/w) collagen matrix crosslinked with the addition of GA above 0.1% (v/w) forms matrix with pores larger than in native collagen. This result suggests the existence of a micro-phase separation in the crosslinked collagen matrix. The implications of these findings for the design of small molecule drug delivery systems are discussed.