ABSTRACT
Marigold-like tyrosinase-entrenched nanostructures were developed by a facile method using a metal cofactor to overcome the limitations of conventional enzyme immobilization techniques. The protein-copper complex promotes the hierarchical self-assembly of nanopetals into marigold-like microstructures through a sequential germination process. Nanopetals, which originated from bead-like tiny projections, showed budding over the surface and promoted the anisotropic growth of copper phosphate nanocrystals upon co-ordination with the active functional groups in protein. This organic-inorganic hybrid showed excellent re-usability, comparable catalytic efficiency, faster reaction rate, improved storage, and thermal stability without affecting the enzyme activity.
Subject(s)
Coordination Complexes/chemistry , Copper/chemistry , Monophenol Monooxygenase/chemistry , Nanostructures/chemistry , Coordination Complexes/metabolism , Copper/metabolism , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Monophenol Monooxygenase/metabolism , Particle Size , Surface PropertiesABSTRACT
Advent of additive manufacturing in biomedical field has nurtured fabrication of complex, customizable and reproducible orthopaedic implants. Layer-by-layer deposition of biodegradable polymer employed in development of porous orthopaedic screws promises gradual dissolution and complete metabolic resorption thereby overcoming the limitations of conventional metallic screws. In the present study, screws with different pore sizes (916 × 918 µm to 254 × 146 µm) were 3D printed at 200 µm layer height by varying printing parameters such as print speed, fill density and travel speed to augment the bone ingrowth. Micro-CT analysis and scanning electron micrographs of screws with 45% fill density confirmed porous interconnections (40.1%) and optimal pore size (259 × 207 × 200 µm) without compromising the mechanical strength (24.58 ± 1.36 MPa). Due to the open pore structure, the 3D printed screws showed increased weight gain due to the deposition of calcium when incubated in simulated body fluid. Osteoblast-like cells attached on screw and infiltrated into the pores over 14 days of in vitro culture. Further, the screws also supported greater human mesenchymal stem cell adhesion, proliferation and mineralized matrix synthesis over a period of 21 days in vitro culture as compared to non-porous screws. These porous screws showed significantly increased vascularization in a rat subcutaneous implantation as compared to control screws. Porous screws produced by additive manufacturing may promote better osteointegration due to enhanced mineralization and vascularization.
