Fig. 6

HMF expression of myofibroblast markers and ECM proteins in mTG hydrogels HMFs were isolated from mTG cross-linked hydrogels and analyzed for expression of myofibroblast markers and ECM proteins. a Collagenase was used to isolate HMFs from stiff (e.g. 60 μg/ml) mTG hydrogels. Changes in the weight of the hydrogel were monitored during the incubation period wherein 1 h was shown to be a sufficient length of time for dissolution of the hydrogel. b Protein lysates from compliant, moderate and stiff hydrogels were subjected to western blot for analysis of myofibroblast and ECM proteins. α-SMA was most highly expressed in compliant hydrogels at day 5 and for moderate and stiff hydrogels at day 7. Collagen I and vimentin expression were upregulated in HMF in moderate and stiff hydrogels in comparison to HMFs in compliant hydrogels. Collagen IV was more abundant in HMFs encapsulated in the moderate and stiff hydrogels with expression greatest at days 3 and 5 for moderate and day 7 for stiff hydrogels. Fibronectin expression was highest overall at days 5 and 7 for HMFs in compliant and stiff hydrogels, respectively. Laminin was detectable, albeit negligibly, at days 5 and 7 for HMFs in compliant and stiff hydrogels, respectively. c Fibronectin expression was increased for stiff hydrogels at days 3 (significantly) and day 7 for compliant hydrogels. Collagen I was increased for moderate hydrogels at days 3 and 5 (significantly) and was similarly expressed for all hydrogel conditions at day 7. Collagen IV was increased for moderate hydrogels at days 3 and 5 (significantly) and for stiff hydrogels at day 7. Laminin was negligibly expressed. α-SMA was consistently increased for compliant hydrogels at each time point assessed. Vimentin was significantly increased at day 3 for compliant hydrogels