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Figure. Gelatin zymograms of MMPs from dentin extracts after sonication and ammonium sulphate treatments. MMP-2 and -9 forms present in whole blood and their relative molecular masses expressed in kDa are reported in Std lanes. (A) MMP forms detected in representative dentin extracts, after demineralizing/extraction with 0.87 M citric and 0.26 M acetic acids (lanes 1 and 2, respectively), after 0.5 M EGTA and 0.5M EDTA (lanes 3 and 4, respectively). (B) MMP forms detected in citric-acid-demineralized dentin extracts after activation with 2 mM APMA for 30, 60, and 90 min (lanes 1, 2, and 3, respectively). (C) Inhibition of MMPs in EDTA (lanes 1 and 4), acetic acid (lanes 2 and 4), and citric acid (lanes 3a and 3b) after pre-incubation with 2 mM 1,10-phenanthroline (lanes 1, 2, and 3a) and 0.5 M EDTA (lanes 3b, 4, and 5). (D) Western blot analysis of MMPs present in whole blood and citric-acid-demineralized dentin extracts (lanes Std and Dent), with the use of monoclonal anti-MMP-2 (72 kDa) and MMP-9 antibodies (92 kDa).
