Activity of enzyme immobilized on silanized Co-Cr-Mo.
The surface of an orthopedic biomaterial was modified by the covalent immobilization of biomolecules. Derivatization of Co-Cr-Mo samples with organic and aqueous solutions of gamma-aminopropyltriethoxysilane (APS) resulted in a concentration-dependent number of reactive NH2 groups on the surface available for coupling to protein. The enzyme trypsin was used as a model biomolecule to investigate the effect of immobilization on proteolytic activity. Trypsin was coupled to the silanized samples by formation of Schiff's base linkages via glutaraldehyde. The nature of the interaction between trypsin and biomaterial was then probed by treatment with concentrated guanidine hydrochloride (GuHCl) and urea. Residual activity (following treatment with chaotropic agents) of trypsin immobilized on silanized Co-Cr-Mo was dependent both on the nature of the silane solution and on the type of chaotropic agent. Organic silanization with APS required a minimum density of approximately 49 NH2 per nm2 of nominal surface area (> 0.021 M APS) for residual activity of immobilized trypsin. For aqueous silanization, approximately 5.4 NH2/nm2 (0.51 M APS) resulted in maximal residual trypsin activity. Treatment with GuHCl removed more trypsin activity from Co-Cr-Mo samples silanized with organic solutions of APS than did treatment with urea. On the contrary, with aqueous silanization the samples possessed greater residual activity following treatment with GuHCl than following urea. Compared to simple adsorption with protein onto Co-Cr-Mo, both methods of silanization with APS resulted in superior residual immobilized enzyme activity.
Puleo
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Center for Biomedical Engineering, University of Kentucky, Lexington 40506-0070, USA.
D000588:Amines / Q000737:chemistry; D002858:Chromium Alloys / Q000737:chemistry*; D004800:Enzymes, Immobilized; D005976:Glutaral; D008982:Molybdenum / Q000737:chemistry*; D011437:Propylamines; D012821:Silanes / Q000737:chemistry*; D013499:Surface Properties; D014357:Trypsin / Q000737:chemistry
D000588:Amines; D002858:Chromium Alloys; D004800:Enzymes, Immobilized; D011437:Propylamines; D012821:Silanes; D008982:Molybdenum; D014357:Trypsin; C477625:amino-propyl-triethoxysilane; D005976:Glutaral