Microfibrils.

<p>TEM micrographs showing the distribution of microfibrils in CDLs in standard (A) and stiff (B) states.</p

Schematic representation of the CDL dissection.

<p>Schematic representation of the CDL dissection.</p

Distance between collagen fibrils.

<p>Mean interfibrillar distance of CDLs in the three mechanical states. ***P<0.0001. Data are expressed as means ± SD.</p

Mean values ± S.D. of unnormalized complex modulus (n = 7).

<p>Mean values ± S.D. of unnormalized complex modulus (n = 7).</p

Arrangement of collagen fibrils.

<p>(A–C) FEG/ESEM and (D–F) TEM micrographs. (A, D) Compliant state. (B, E) Standard state. (C, F) Stiff state.</p

Mechanical properties of CDL.

<p>(A) Representative stress vs. strain curves of CDLs from one animal in the three mechanical states. (B) Effect of frequency on the complex modulus of CDLs from one animal in the three mechanical states tested at 13% strain.</p

MMPs of CDLs in compliant, standard and stiff conditions visualized by gelatin zymography.

<p>(A) Zymogram showing pro-enzyme and active MMP band profile of CDLs in the three mechanical states. (B) Zymogram comparing the band profiles of CDLs, compared with human cell line showing MMP-2 and MMP-9 activity; this zymogram was included because of its very good band separation. (C) Optical density of MMP activity in CDLs in the three mechanical states. (D) Comparative densitometric analysis of scanned gels of CDLs in the different mechanical states with and without 50 µM galardin. Data are expressed as means ± SD. The asterisk (*) represents statistically significant difference P<0.05. MMPs were detected in more than six animals for each of the three mechanical states. (E) Zymogram comparing standard CDLs with and without galardin treatment.</p

Effect of MMP inhibition on CDL viscoelasticity.

<p>(A) complex modulus, (B) storage modulus, (C) loss modulus and (D) tan delta of compliant, standard and stiff CDLs treated with 50 µM galardin in PPSW, SW, and AChSW respectively. The action of galardin was quantified by normalizing the maximum E* reached after galardin addition against the value just before the application of the chemical. The asterisk (*) represents statistically significant difference P<0.05 and the double asterisk (**) P<0.01.</p

Reversibility of MMP inhibition.

<p>(A–C) Examples of recordings showing the reversibility of the galardin effect on (A) standard, (B) stiff and (C) compliant CDLs. (D) Normalized results. Tissues in compliant, standard and stiff states were stimulated with 50 µM galardin, washed and treated again with galardin. Data are expressed as means ± SD.</p

JLCs in the different mechanical states.

<p>(A–C) TEM micrographs of compliant (A), standard (B) and stiff (C) CDLs. (D) Size of JLC granules in different mechanical states. *P<0.05. (E) Number of JLC granules in different mechanical states. **P<0.01; ***P<0.001. Data are expressed as means ± SD.</p