Bilayer nanostructure coated AZ31 magnesium alloy implants: in vivo reconstruction of critical-sized rabbit femoral segmental bone defect.

Healing or reconstruction of critical-sized bone defects is still challenging in orthopaedic practice. In this study, we developed a new approach to control the degradation and improve the bone regeneration of the AZ31 magnesium substrate, fabricated as mesh cage implants. Subsequently, bilayer nanocomposite coating was carried out using polycaprolactone (PCL) and nano-hydroxyapatite (nHA) by dip-coating and electrospinning. Lastly, the healing capacity of the implants was studied in New Zealand White (NZW) rabbit critical-sized femur bone defects. X-ray analysis showed the coated implant group bridged and healed the critical defects 100% during four weeks of post-implantation. Micro-computed tomography (Micro-CT) study showed higher total bone volume (21.10%), trabecular thickness (0.73), and total porosity (85.71%) with bilayer coated implants than uncoated. Our results showed that nanocomposite coated implants controlled the in vivo degradation and improved bioactivity. Hence, the coated implants can be used as a promising bioresorbable implant for critical segmental bone defect repair applications.

Synthesis of magnesium phosphate nanoflakes and its PCL composite electrospun nanofiber scaffolds for bone tissue regeneration.

This study describes the preparation of nano-magnesium phosphate (nMP) flakes by one step microwave irradiation method. The synthesized nMP was incorporated with polycaprolactone (PCL), hyperbranched polyglycerol (HPG) and nano-hydroxyapatite (nHA) to fabricate as composite electrospun nanofibrous scaffold for bone tissue engineering applications. The electrospun nanofibers were analyzed by scanning electron microscope, XRD, FTIR, DSC, TGA, and wettability measurement. The nanofibers were smooth, randomly oriented, and surface decorated with nMP. The water contact angle was 32 ± 1° (initial contact angle), which reduces to 0° after 1 min for HPG and nMP containing nanocomposites, while the contact angle of PCL is 104 ± 5°. The nanocomposite scaffolds exhibited higher swelling, biomineralization, and breakages during PBS immersion. The scaffolds were non-cytotoxic to MG63 osteosarcoma cells and hMSCs with higher viability after 72 h. They allowed good adhesion and spreading of these cells when compared to PCL and PCL/nHA electrospun nanofibers. These results indicated that HPG with surface decorated nMP electrospun nanocomposite scaffold can be a promising material for bone tissue engineering applications.

Influence of magnesium particles and Pluronic F127 on compressive strength and cytocompatibility of nanocomposite injectable and moldable beads for bone regeneration.

A novel one-step preparation of magnesium particles and Pluronic F127 incorporated with calcium sulfate hemihydrate (CSH) and nano-hydroxyapatite (nHA) ready to use injectable or moldable beads was developed for bone tissue regeneration applications. The nanocomposite showed setting time less than 15 min, very good injectability (75-85%) and good mechanical strength (52-80 MPa). Samples immersed in SBF showed controlled degradation (40-45% reduction in weight) in 28 days. The nanocomposite bone graft was cytocompatible against MG63 osteosarcoma cells and increased the osteogenic gene expression by 2-3 folds. These results indicate that it can be a potential defect filling biomaterial for bone tissue regeneration at the fracture site.

Differential Adhesive and Bioactive Properties of the Polymeric Surface Coated with Graphene Oxide Thin Film.

Surface engineering of implantable devices involving polymeric biomaterials has become an essential aspect for medical implants. A surface enhancement technique can provide an array of unique surface properties that improve its biocompatibility and functionality as an implant. Polyurethane-based implants that have found extensively acclaimed usage as an implant in biomedical applications, especially in the area of cardiovascular devices, still lack any mechanism to ward off bacterial or platelet adhesion. To bring out such a defense mechanism we are proposing a surface modification technique. Graphene oxide (GO) in very thin film form was wrapped onto the electrospun fibroporous polycarbonate urethane (PCU) membrane (GOPCU) by a simple method of electrospraying. In the present study, we have developed a simple single-step method for coating a polymeric substrate with a thin GO film and evaluated the novel antiadhesive activity of these films. SEM micrographs after coating showed the presence of very thin GO films over the PCU membrane. On the GOPCU surface, the contact angle was shifted by ∼30°, making the hydrophobic PCU surface slightly hydrophilic, while Raman spectral characterization and mapping showed the presence and distribution of GO over 75% of the membrane. A reduced platelet adhesion on the GOPCU surface was observed; meanwhile, bacterial adhesion also got reduced by 85% for Staphylococcus aureus (Gram positive, cocci) and 64% for Pseudomonas aeruginosa (Gram negative, bacilli). A cell adhesion study conducted using mammalian fibroblast cells projected its proliferation percentage in a MTT assay, with 82% cell survival on PCU and 86% on GOPCU after 24 h culture, while a study for an extended period of 72 h showed 87% of survival on PCU and 88% on GOPCU. This plethora of functionalities by a simple modification technique makes thin GO films a self-sufficient surface engineering material for future biomedical applications.

Effect of tobacco extract on surfactant synthesis and its reversal by retinoic acid-role of cell-cell interactions in vitro.

Tobacco induces oxidative stress in the alveolar epithelium and causes its damage. Retinoic acid (RA) has a cardinal role in alveolar cell growth, differentiation, and maturation. The aim of the study was to investigate the role of cell-cell interactions and whether RA could reverse the effect of tobacco extract on epithelial function as expressed by surfactant synthesis. For this, an in vitro model, which provides multiple cell type interactions, as seen in vivo, was used. We had used the major lung cell types, alveolar epithelial and mesenchymal cells represented by the cell lines A549 (human lung adenocarcinoma cell line), and human fetal lung fibroblast-1 (HFL-1) for developing the monoculture and co-culture systems and studied the effect of tobacco extract and retinoic acid. The effect of tobacco and retinoic acid both singly and in combination on proliferation and surfactant synthesis was analyzed. Retinoic acid induced proliferation and upregulated surfactant synthesis in monocultures and co-cultures. Tobacco extract at 100 µg/ml concentration decreased A549 proliferation and upregulated surfactant protein mRNA expression. In co-cultures treated with tobacco extract (100 µg/ml), retinoic acid (1 µM), regulated cell proliferation, and surfactant protein mRNA expression vis-à-vis the monoculture system. This clearly points to the fact that cell-cell interactions modulate the effect of additives or stimulants and help in assessing the in vivo combinatorial responses in vitro and that the retinoic acid effect is regenerative.