title: The release kinetics, antimicrobial activity and cytocompatibility of differently prepared collagen/hydroxyapatite/vancomycin layers: Microstructure vs. nanostructure.
abstract: The aim of this study was to develop an osteo-inductive resorbable layer allowing the controlled elution of antibiotics to be used as a bone/implant bioactive interface particularly in the case of prosthetic joint infections, or as a preventative procedure with respect to primary joint replacement at a potentially infected site. An evaluation was performed of the vancomycin release kinetics, antimicrobial efficiency and cytocompatibility of collagen/hydroxyapatite layers containing vancomycin prepared employing different hydroxyapatite concentrations. Collagen layers with various levels of porosity and structure were prepared using three different methods: by means of the lyophilisation and electrospinning of dispersions with 0, 5 and 15wt% of hydroxyapatite and 10wt% of vancomycin, and by means of the electrospinning of dispersions with 0, 5 and 15wt% of hydroxyapatite followed by impregnation with 10wt% of vancomycin. The maximum concentration of the released active form of vancomycin characterised by means of HPLC was achieved via the vancomycin impregnation of the electrospun layers, whereas the lowest concentration was determined for those layers electrospun directly from a collagen solution containing vancomycin. Agar diffusion testing revealed that the electrospun impregnated layers exhibited the highest level of activity. It was determined that modification using hydroxyapatite exerts no strong effect on vancomycin evolution. All the tested samples exhibited sufficient cytocompatibility with no indication of cytotoxic effects using human osteoblastic cells in direct contact with the layers or in 24-hour infusions thereof. The results herein suggest that nano-structured collagen-hydroxyapatite layers impregnated with vancomycin following cross-linking provide suitable candidates for use as local drug delivery carriers.
authors.affiliation: Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, Prague 8, Czech Republic; Faculty of Mechanical Engineering, Czech Technical University in Prague, Prague 6, Czech Republic. Electronic address: suchyt@irsm.cas.cz.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Suchý
authors.affiliation: Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, Prague 8, Czech Republic.
authors.forename: Monika
authors.initials: M
authors.lastname: Šupová
authors.affiliation: Department of Medical Chemistry and Clinical Biochemistry, Charles University, 2nd Medical School and University Hospital Motol, Prague 5, Czech Republic.
authors.forename: Eva
authors.initials: E
authors.lastname: Klapková
authors.affiliation: Clinical Microbiology and ATB Centre, Institute of Medical Biochemistry and Laboratory Diagnostics, 1st Faculty of Medicine, Charles University in Prague and Motol University Hospital, Prague 2, Czech Republic.
authors.forename: Václava
authors.initials: V
authors.lastname: Adamková
authors.affiliation: Clinical Microbiology and ATB Centre, Institute of Medical Biochemistry and Laboratory Diagnostics, 1st Faculty of Medicine, Charles University in Prague and Motol University Hospital, Prague 2, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Závora
authors.affiliation: Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, Prague 8, Czech Republic.
authors.forename: Margit
authors.initials: M
authors.lastname: Žaloudková
authors.affiliation: Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, Prague 8, Czech Republic.
authors.forename: Šárka
authors.initials: Š
authors.lastname: Rýglová
authors.affiliation: 1st Department of Orthopaedics, 1st Faculty of Medicine, Charles University in Prague and Motol University Hospital, Prague 5, Czech Republic.
authors.forename: Rastislav
authors.initials: R
authors.lastname: Ballay
authors.affiliation: Department of Composites and Carbon Materials, Institute of Rock Structure and Mechanics, Academy of Sciences of the Czech Republic, Prague 8, Czech Republic; Faculty of Mechanical Engineering, Czech Technical University in Prague, Prague 6, Czech Republic.
authors.forename: František
authors.initials: F
authors.lastname: Denk
authors.affiliation: Contipro a.s., R&D Department, Dolni Dobrouc, Czech Republic.
authors.forename: Marek
authors.initials: M
authors.lastname: Pokorný
authors.affiliation: Institute of Inherited Metabolic Disorders, 1st Faculty of Medicine, Charles University in Prague, Ke Karlovu 2, Prague 2, Czech Republic; Biomedical Centre, Faculty of Medicine in Pilsen, Charles University in Prague, Alej Svobody 76, Pilsen, Czech Republic.
authors.forename: Pavla
authors.initials: P
authors.lastname: Sauerová
authors.affiliation: Institute of Inherited Metabolic Disorders, 1st Faculty of Medicine, Charles University in Prague, Ke Karlovu 2, Prague 2, Czech Republic; Biomedical Centre, Faculty of Medicine in Pilsen, Charles University in Prague, Alej Svobody 76, Pilsen, Czech Republic.
authors.forename: Marie
authors.initials: M
authors.lastname: Hubálek Kalbáčová
authors.affiliation: Faculty of Mechanical Engineering, Czech Technical University in Prague, Prague 6, Czech Republic.
authors.forename: Lukáš
authors.initials: L
authors.lastname: Horný
authors.affiliation: Faculty of Mechanical Engineering, Czech Technical University in Prague, Prague 6, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Veselý
authors.affiliation: Faculty of Mechanical Engineering, Czech Technical University in Prague, Prague 6, Czech Republic.
authors.forename: Tereza
authors.initials: T
authors.lastname: Voňavková
authors.affiliation: Department of Medical Chemistry and Clinical Biochemistry, Charles University, 2nd Medical School and University Hospital Motol, Prague 5, Czech Republic.
authors.forename: Richard
authors.initials: R
authors.lastname: Průša
mesh_terms: D000900:Anti-Bacterial Agents / Q000008:administration & dosage / Q000737:chemistry; D045744:Cell Line, Tumor; D003094:Collagen / Q000008:administration & dosage / Q000737:chemistry; D016503:Drug Delivery Systems; D017886:Durapatite / Q000008:administration & dosage / Q000737:chemistry; D005260:Female; D006801:Humans; D008297:Male; D049329:Nanostructures / Q000008:administration & dosage / Q000737:chemistry; D010006:Osteoblasts / Q000187:drug effects; D010949:Plasma / Q000737:chemistry; D013211:Staphylococcus aureus / Q000187:drug effects; D013212:Staphylococcus epidermidis / Q000187:drug effects; D014640:Vancomycin / Q000008:administration & dosage / Q000737:chemistry
chemical_list: D000900:Anti-Bacterial Agents; D014640:Vancomycin; D003094:Collagen; D017886:Durapatite
keywords: Coating; Controlled release; Drug delivery system; Nanoparticle; Pharmacokinetics; Polymeric drug delivery system