title: Human macrophage-mediated biodegradation of polyurethanes: assessment of candidate enzyme activities.
abstract: A predominant cell type associated with explanted failed devices is the monocyte-derived macrophage (MDM). However, there is still very little known about the specific cellular enzyme activities involved in interactions with these devices. The current study investigates the nature of candidate enzymes that may be involved in the degradation of polymeric biomaterials through the use of specific enzyme inhibitor agents. When MDM were incubated with a polycarbonate-based polyurethane (PCNU) synthesized with 14C-labeled hexane diisocyanate (HDI), polycarbonate diol and butanediol (BD) (referred to as 14C-HDI431), the radiolabel release (RR) measured was inhibited by phenylmethylsulfonyl fluoride, diethyl-p-nitrophenyl phosphate (serine protease/esterase inhibitors), and sodium fluoride (NaF) (a carboxyl esterase (CXE) inhibitor). Sodium taurocholate (NaT) (a cholesterol esterase (CE) stimulator) had little effect on RR. The two candidate enzymes proposed were CE and CXE, based on the fact that both were identified by immunoblotting in the releasate of MDM following 48 h incubation with 14C-HDI431. The effect of the above reagents on the RR caused by purified CE and CXE, was measured and compared to changes in their activity with p-nitrophenylbutyrate (PNB). The effect of NaF on MDM was similar to that of purified CXE (inhibitory on both RR and lysate esterase activity), suggesting the involvement of CXE. However, NaT inhibited the PNB activity of purified CXE, but had no effect on MDM-mediated RR or PNB activity, implicating another esterase in the biomaterial degradation. Since NaT stimulated CE-mediated RR and PNB activity, it may also be involved in MDM-mediated biodegradation of PCNUs. The results of these studies point to both esterases as being candidates. However, the current methods were unable to determine the relative contribution of each one to the observed biodegradation.
authors.affiliation: Department of Biochemistry, Microbiology and Immunology, University of Ottawa Heart Institute, Ont, Canada. rlabow@ottawaheart.ca
authors.forename: Rosalind S
authors.initials: RS
authors.lastname: Labow
authors.forename: Erin
authors.initials: E
authors.lastname: Meek
authors.forename: Loren A
authors.initials: LA
authors.lastname: Matheson
authors.forename: J Paul
authors.initials: JP
authors.lastname: Santerre
mesh_terms: D001672:Biocompatible Materials / Q000378:metabolism; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015151:Immunoblotting; D008264:Macrophages / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism; D011140:Polyurethanes / Q000737:chemistry*; D011480:Protease Inhibitors / Q000494:pharmacology; D012969:Sodium Fluoride / Q000494:pharmacology; D002787:Sterol Esterase / Q000378:metabolism; D013656:Taurocholic Acid / Q000494:pharmacology; D013997:Time Factors; D014105:Tosyl Compounds / Q000494:pharmacology
chemical_list: D001672:Biocompatible Materials; D011140:Polyurethanes; D011480:Protease Inhibitors; D014105:Tosyl Compounds; C003614:4-toluenesulfonyl fluoride; D013656:Taurocholic Acid; D012969:Sodium Fluoride; D004247:DNA; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D002787:Sterol Esterase