mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase:...

Mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase:

implications for load-bearing matrix growth and stability

by Amit P. Bhole, Brendan P. Flynn, Melody Liles, Nima Saeidi, Charles A. Dimarzio, and Jeffrey W. Ruberti

Philosophical Transactions AVolume 367(1902):3339-3362

September 13, 2009

©2009 by The Royal Society

(a) Schematic of the experimental setup used to carry out degradation of strained collagen under DIC microscopy.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

Contrast normalization and background noise removal.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

Time-series DIC and edge-detected ROI during both gelation and degradation of collagen.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

Representative plot showing quantification of edge loss against digestion time ΔT in control sample (no strain), where Istart is 95 per cent of (maximum edge detection intensity), Ifinish is 10

per cent of and ΔT is the time (s) between.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

DIC and edge-detected ROIs. (a) Intial strain on a heavily manipulated network of collagen fibrils.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

DIC and edge-detected ROIs. (a) Initial strain on a gently manipulated network of collagen fibrils.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

(a) Simulated fibril images (i) for diameters of 50, 150 and 250 nm and the result of the edge detection algorithm for each (ii).

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

Representative plot showing quantification of edge loss against digestion time ΔT, where Istart is 95 per cent of (maximum edge detection), Ifinish is 10 per cent of and ΔT is the time (s)

between.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

DIC images of the rapid degradation of the initially strained network shown in figure 2c.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

DIC and edge-detected ROIs in a similar experiment using MMP-8.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society

DIC image showing what appears to be a substantial accumulation of assembled collagen fibrils in the area of applied tensile strain on the network between the pipettes.

Amit P. Bhole et al. Phil. Trans. R. Soc. A 2009;367:3339-3362

©2009 by The Royal Society