ZrN-ZrOxNy vs ZrO2-ZrOxNy coatings deposited via unbalanced DC magnetron sputtering

ZrN-ZrO$${ }_{x}$$ x N$${ }_{y}$$ y and ZrO$${ }_{2}$$ 2 -ZrO$${ }_{x}$$ x N$${ }_{y}$$ y coatings were deposited on 316L stainless steel substrates via the unbalanced DC magnetron sputtering technique in order to improve their corrosion resistance and evaluate their possible use as a coating biocompatible with bone cells. The composition, structure, morphology, and corrosion resistance were studied by sum means of x-ray photoelectron spectroscopy (XPS), x-Ray diffraction (XRD), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The corrosion resistance was evaluated in 3.5 wt.% NaCl using potentiodynamic polarization (PL) and electrochemical impedance techniques (EIS). The ZrN-ZrO$${ }_{x}$$ x N$${ }_{y}$$ y and ZrO$${ }_{2}$$ 2 -ZrO$${ }_{x}$$ x N$${ }_{y}$$ y coatings exhibited barrier-type protection of the substrate against corrosion. The growth of mouse osteoblast cells was evaluated in the coating that exhibited the greatest resistance to corrosion, ZrO$${ }_{2}$$ 2 -ZrO$${ }_{x}$$ x N$${ }_{y}$$ y , finding that the cell viability was maintained, so this material can be considered to be a candidate for use in osteosynthesis processes.

Fabrication and biological evaluation of porous β-TCP bioceramics produced using digital light processing

Beta-tricalcium phosphate ( β-TCP) refers to one ideal bone repair substance with good biocompatibility and osteogenicity. A digital light processing (DLP)-system used in this study creates bioceramic green part by stacking up layers of photocurable tricalcium phosphate-filled slurry with various β-TCP weight fractions. Results show that the sintering shrinkage is anisotropic and the shrinkage vertically reaches over that horizontally. The obtained porous β-TCP parts have both macroporous outer structure and microporous inner structure, the macropore size is 400–600 μm and the micropore size is 500–1500 nm. The mechanical tests show that the porous β-TCP bioceramic’s compressive strength reaches 16.53 MPa. The cell culture confirmed that the porous β-TCP bioceramic is capable of achieving the effective attaching, growing, and proliferating pertained to mouse osteoblast cells. This study identified considerable blood vessels and significant ectopic bone forming obviously based on the histologically-related assessment when implanting to rabbit femoral condyle deficiency for 3 months. Thus, under high bioactive property and osteoinductivity, and large precision and mechanical strength that can be adjusted, the DLP printed porous β-TCP ceramics is capable of being promising for special uses of bones repairing.

Osteoblast-like Cell Proliferation, ALP Activity and Photocatalytic Activity on Sintered Anatase and Rutile Titanium Dioxide

This study aimed to create a biomaterial from titanium dioxide (TiO2), which has been known to have photocatalytic and bone formation promoting effects. I expected that anatase titanium dioxide-based implants could promote bone augmentation and induce bone formation. Powdery anatase TiO2 was compression molded and sintered at 700, 800, 900, and 1000 °C to prepare sintered compact samples. X-ray diffraction and scanning electron microscopy were used to observe the surface of these samples. Furthermore, mouse osteoblast-like cells (MC3T3-E1 cell line) were seeded on the samples sintered at different temperatures, and cell proliferation was observed to evaluate the cell proliferation of the samples. The sample sintered at 700 °C was composed of anatase TiO2. The samples sintered at 800 °C and 900 °C were confirmed to consist of a mixture of anatase and rutile TiO2 crystalline phases. Moreover, the sample sintered at 700 and 800 °C, which contained anatase TiO2, showed remarkable photocatalytic activity. Those samples sintered at 1000 °C were transformed to the rutile TiO2. The cell proliferation after 7–14-days culturing revealed that cells cultured on the 700 °C sample decreased in number immediately after initiation of culturing. The cells cultured on TiO2 sintered at 900 °C markedly proliferated over time with an increase in the alkaline phosphatase activity, showing good MC3T3-E1 cell compatibility of the samples. The sample sintered at 1000 °C, which is rutile TiO2, showed the highest increase.

Delivery of Bioactive Gene Particles via Gelatin-Collagen-PEG-Based Electrospun Matrices

The fabrication of fiber mats via electrospinning has been adopted in the last decades to produce high quality scaffolds for tissue engineering. However, an effective combination of electrospinning methods with gene delivery therapies remains a challenge. In this study, we describe how the delivery of gene complexes via electrospun mats that contain different volumes of gelatin (Gel), collagen (Col), and polyethylene glycol (PEG) can affect gene expression by transfected cells. Non-viral complexes were formulated by using lipid modified polyethylenimine (PEI) polymer and plasmid DNAs (pDNA) like the reporter Green Fluorescent Protein (GFP) and the therapeutically relevant Bone Morphogenetic Protein-2 (BMP-2) and electrospuned after being mixed with different volumes of Gel-Col-PEG mats and delivered to human myoblast (C2C12) and mouse osteoblast cells (MC3T3). The entrapment of GFP complexes via different homogeneous electrospun fiber mats revealed that a high fraction of collagen in the mats affected the quality of the fibers and led to reduced transfection efficiency on target cells. On the other hand, the fabrication of double-layered mats that contained collagen without complexes as a first layer and gelatin-collagen-PEG with complexes as a second layer successfully induced GFP expression and ALP activity in C2C12 cells. We conclude that this study has established the advantage of formulating multilayered bioactive collagen-based mats for gene delivery applications.

Uniaxially fixed mechanical boundary condition elicits cellular alignment in collagen matrix with induction of osteogenesis

Osteocytes differentiated from osteoblasts play significant roles as mechanosensors in modulating the bone remodeling process. While the well-aligned osteocyte network along the trabeculae with slender cell processes perpendicular to the trabeculae surface is known to facilitate the sensing of mechanical stimuli by cells and the intracellular communication in the bone matrix, the mechanisms underlying osteocyte network formation remains unclear. Here, we developed a novel in vitro collagen matrix system exerting a uniaxially-fixed mechanical boundary condition on which mouse osteoblast-like MC3T3-E1 cells were subcultured, evoking cellular alignment along the uniaxial boundary condition. Using a myosin II inhibitor, blebbistatin, we showed that the intracellular tension via contraction of actin fibers contributed to the cellular alignment under the influence of isometric matrix condition along the uniaxially-fixed mechanical boundary condition. Furthermore, the cells actively migrated inside the collagen matrix and promoted the expression of osteoblast and osteocyte genes with their orientations aligned along the uniaxially-fixed boundary condition. Collectively, our results suggest that the intracellular tension of osteoblasts under a uniaxially-fixed mechanical boundary condition is one of the factors that determines the osteocyte alignment inside the bone matrix.

The Anatase Phase of Nanotopography Titania with Higher Roughness Has Better Biocompatibility in Osteoblast Cell Morphology and Proliferation

Previous studies have concluded that surface-modified titanium oxide (titania, TiO2) surface properties promote osteoblast cell morphology and proliferation. To screen a suitable structured titania coating with the best biocompatibility to be used in dental implants, five titania films (two amorphous, one rutile, and two anatases) with different surfaces were successfully synthesized on polished titanium by radio frequency (RF) magnetron sputtering. We applied atomic force microscopy (AFM) and X-ray diffraction (XRD) to depict the formulations. Furthermore, MC3T3-E1, the mouse osteoblast precursor cell, was used to assess cell proliferation and observe morphologic changes at the film surface. The data indicated that the overall number of MC3T3-E1 cells on anatase films was significantly higher as compared with cells on rutile and amorphous films. Meanwhile, the actin filaments of the cells grown on the anatase phase films were well defined and fully spread. In addition, the film with higher roughness had enhanced biocompatibility than that with lower roughness. The results showed that the crystal phase and titania coated roughness had a greater influence on the biocompatibility of nanostructured titania film. The higher the roughness of the anatase phase was, the better bioactivity for the morphology and proliferation of osteoblast. This is a good surface-modified biological material and may have a good application prospect in dental implants.