Comparing the bone regeneration potential between a trabecular bone and a porous scaffold through osteoblast migration and differentiation: A multiscale approach.

Both cell migration and osteogenic differentiation are critical for successful bone regeneration. Therefore, understanding the mechanobiological aspects that govern these two processes is essential in designing effective scaffolds that promote faster bone regeneration. Studying these two factors at different locations is necessary to manage bone regeneration in various sections of a scaffold. Hence, a multiscale computational model was used to observe the mechanical responses of osteoblasts placed in different positions of the trabecular bone and gyroid scaffold. Fluid shear stresses in scaffolds at cell seeded locations (representing osteogenic differentiation) and strain energy densities in cells at cell substrate interface (representing cell migration) were observed as mechanical response parameters in this study. Comparison of these responses, as two critical factors for bone regeneration, between the trabecular bone and gyroid scaffold at different locations, is the overall goal of the study. This study reveals that the gyroid scaffold exhibits higher osteogenic differentiation and cell migration potential compared to the trabecular bone. However, the responses in the gyroid only mimic the trabecular bone in two out of nine positions. These findings can guide us in predicting the ideal cell seeded sites within a scaffold for better bone regeneration and in replicating a replaced bone condition by altering the physical parameters of a scaffold.

Biocompatibility and corrosion evaluation of niobium oxide coated AZ31B alloy for biodegradable implants.

Biodegradable magnesium (Mg) based implants have considerable interest in the biomedical field as their use nullifies the necessity for implant removal surgery and avoids the long-standing adverse reaction of permanent bioimplants. The degradation resistance and biocompatibility of the Mg alloys can be improved by coating them with a suitable thin film. Here, thin films of niobium and niobium oxide were developed on the AZ31B Mg alloy by sputtering technique and their biocompatibility and corrosion resistance was examined. X-ray diffraction (XRD) and Transmission electron microscope (TEM) techniques confirmed the crystallinity of the thin films. Subsequently, scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) techniques were employed to evaluate the morphology and chemical composition of the thin film surfaces, respectively. Thin-film coated Mg alloys revealed good corrosion resistance compared to their uncoated bare counterparts in simulated body fluid (SBF). The contact angle study was performed on the coated specimens to investigate their wettability which revealed their hydrophobic character. The cell viability studies on thin-film coated specimens exhibited significant cell proliferation, and cell morphological studies showed good cell attachment and growth. The in vitro MTT assay on mouse osteoblast precursor cells (MC3T3-E1) indicated that the Nb-based coatings are cytocompatible and promote cell proliferation.

In vitro and in vivo degradation assessment and preventive measures of biodegradable Mg alloys for biomedical applications.

Magnesium (Mg) and its alloys have been widely explored as a potential biodegradable implant material. However, the fast degradation of Mg-based alloys under physiological environment has hindered their widespread use for implant applications till date. The present review focuses on in vitro and in vivo degradation of biodegradable Mg alloys, and preventive measures for biomedical applications. Initially, the corrosion assessment approaches to predict the degradation behavior of Mg alloys are discussed along with the measures to control rapid corrosion. Furthermore, this review attempts to explore the correlation between in vitro and in vivo corrosion behavior of different Mg alloys. It was found that the corrosion depends on experimental conditions, materials and the results of different assessment procedures hardly matches with each other. It has been demonstrated the corrosion rate of magnesium can be tailored by alloying elements, surface treatments and heat treatments. Various researches also studied different biocompatible coatings such as dicalcium phosphate dihydrate (DCPD), ß-tricalcium phosphate (ß-TCP), hydroxyapatite (HA), polycaprolactone (PCL), polylactic acid (PLA), and so on, on Mg alloys to suppress rapid degradation and examine their influence on new bone regeneration as well. This review shows the need for a standard method of corrosion assessment to predict the in vivo corrosion rate based on in vitro data, and thus reducing the in vivo experimentation.

Biological performance of metal metalloid (TiCuZrPd:B) TFMG fabricated by pulsed laser deposition.

The aim of our study is to investigate the effect of boron with different ratios in Ti-Cu-Pd-Zr metallic glass (MG) matrix (Ti-Cu-Pd-Zr:B) fabricated by Pulsed Laser Deposition (PLD) for biomedical implants. The Ti based Thin Film Metallic Glasses (TFMGs) in combination with boron (in different atomic %) was assessed in attaining the combined properties, like outstanding corrosion resistant properties and good biocompatibility in this work. The disordered structure and amorphous nature of the Ti-Cu-Pd-Zr:B thin films systems were achieved by the PLD process and affirmed by XRD and transmission electron microscopy. The boron incorporation in the TFMG has been elucidated by XPS analysis. The boron containing films displays distribution of boron protuberances interleaved in the amorphous matrix was stated from SEM analysis. It is found that increase in atomic percentage of boron contents in TFMG results in the improvement in glass transition temperatures. The electrochemical parameters suggest better corrosion resistance and capabilities of passivity when boron percentage was increased in the film thereby preventing adverse biological reactions. TFMGs exhibited excellent hemocompatibility by preventing the platelet activation. MTT assay manifests increase in cell concentration with culture period on the TFMGs for the MC3T3-E1 preosteoblasts cells. Cell morphology was also studied which confirmed the viable state of the cells on the TFMG surfaces. The combination of such distinctive properties marks these TFMG systems as prospective aspirants for biomedical implants.

Degradation, wettability and surface characteristics of laser surface modified Mg-Zn-Gd-Nd alloy.

This work evaluates the effects of laser surface modification on Mg-Zn-Gd-Nd alloy which is a potential biodegradable material for temporary bone implant applications. The laser surface melted (LSM) samples were investigated for microstructure, wettability, surface hardness and in vitro degradation. The microstructural study was carried out using scanning and transmission electron microscopes (SEM, TEM) and the phases present were analyzed using X-ray diffraction. The in vitro degradation behaviour was assessed in hank's balanced salt solution (HBSS) by immersion corrosion technique and the effect of LSM process parameters on the wettability was analyzed through contact angle measurements. The microstructural examination showed remarkable grain refinement as well as uniform redistribution of intermetallic phases throughout the matrix after LSM. These microstructural changes increased the hardness of LSM samples with an increase in energy density. The wetting behaviour of processed samples showed hydrophilic nature when processed at lower (12.5 and 17.5 J/mm2) and intermediate energy density (22.5 and 25 J/mm2), which can potentially improve cell-materials interaction. The corrosion rate of as cast Mg-Zn-Gd-Nd alloy decreased by ~83% due to LSM.

Synthesis, characterization and in vitro biocompatibility study of strontium titanate ceramic: A potential biomaterial.

Strontium (Sr), a mineral element present in trace in the human body, has significant effect on bone remodelling. Sr containing ceramics have huge potential to heal bone defects and improve osseointegration of implants. In this study, perovskite oxide - strontium titanate (SrTiO3) was synthesized and explored its potential for biomedical applications. The phase pure SrTiO3 powder was synthesized from solid state reaction of strontium carbonate (SrCO3) and titanium dioxide (TiO2) at 1200 °C for 2 h. The as synthesized SrTiO3 powder, pure hydroxyapatite (HAp) and SrTiO3-50 wt% HAp (SH50) premixed powders were sintered at different temperatures varies from 1100 to 1400 °C in air. The sintered samples were characterized using X-ray diffraction (XRD) for phases and scanning electron microscopy (SEM) for microstructure analysis. XRD results revealed no dissociation of HAp or reaction with SrTiO3 during sintering. The sintered samples were studied for mechanical properties, wettability, and biocompatibility. The relative density of the sintered SrTiO3 increases with increasing sintering temperature. The relative density of SrTiO3 was increased from 77% to 98% with increase in sintering temperature from 1250 to 1400 °C. The substantial improvement of hardness and compressive strength was observed for sintered SrTiO3 compared to HAp of similar porosity level. The hardness and compressive strength of SrTiO3 sintered at 1250 °C found ~6 and ~3.5 times higher than sintered HAp. In vitro dissolution study carried out in phosphate buffer solution at 37 °C, confirmed the release of Sr2+ ion from the bulk SrTiO3 sintered at 1250 °C. The in vitro cell materials interaction showed cytocompatibility of sintered SrTiO3 and SrTiO3-HAp composite. In summary, excellent biocompatibility of SrTiO3 with superior mechanical properties confirmed its potential as novel biomaterial for use in the repair of infected or aseptic bone defects.

Effect of fluorine substitution on sintering behaviour, mechanical and bioactivity of hydroxyapatite.

Fluorine substituted hydroxyapatite (FAp) with different degree of fluorine (F) substitution, has been synthesized using hydrothermal synthesis method. In the present work, as synthesized powders were consolidated by sintering at 1200 °C in air for 1 h. The sintered specimens were characterized using Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) for phase analysis. Further, fluorine intake in the sintered specimens was evaluated using ion chromatography (IC). XRD peaks clearly showed biphasic nature of the sintered specimen. However, the sintered samples containing more than ∼60% fluorine substitution showed no ß-tricalcium phosphate (ß-TCP) phase formation. The IC results revealed that the degree of fluoridation decreased significantly in the sintered specimen compare to the respective as synthesized powders. The effect of actual fluorine content in the sintered specimens was further evaluated in terms of sinterability, surface energy, mechanical properties and in vitro cytocompatibility study. The surface energy of the sintered specimen decreased from 51.8 mN/m to 42.5 mN/m, in which degree of fluoridation varies from 0% to 110%. The in vitro cytocompatibility of the sintered specimen were carried out against mouse osteoblast cell line (MC3T3-E1). In vitro study showed that all the samples were nontoxic but cell proliferation for the samples containing more than 40% fluorine substitution became significantly low.

Surface engineering of LENS-Ti-6Al-4V to obtain nano- and micro-surface topography for orthopedic application.

Two distinct surface topographies consisting of micro- and nano-surface were developed using laser texturing (LT) and anodization process respectively and their effect on the surface-related properties of Ti-6Al-4V fabricated using Laser Engineered Net Shaping (LENS) were determined. The topographies developed using laser texturing (25, 50 and 75% overlap) were examined using 3D profilometer, whereas, Field Emission Scanning Electron Microscopy (FE-SEM) was used to analyze Titania NanoTubes (TNT) formed using anodization. Though all the surface modified specimens exhibited hydrophilic behavior, least contact angle values were observed for the specimen surface modified with TNT. 25LT and 50LT specimens offered about 8 fold higher corrosion resistance than TNT specimens. All the surface modified samples exhibited non-toxicity to blood cells as well as to the mesenchymal stem cells (hMSCs) with a higher rate of proliferation and differentiation hMSCs observed on 75LT specimens and TNT specimen.

Strontium doped hydroxyapatite from Mercenaria clam shells: Synthesis, mechanical and bioactivity study.

Synthesis of strontium-doped hydroxyapatite from Mercenaria clam shells has been carried out by hydrothermal method. The doping of bioceramic, processed from biogenic resources is mostly unexplored. The objective is to understand the effect of strontium (Sr) incorporation on phase stability, sintering behaviour, mechanical properties and cytotoxicity of hydroxyapatite (HAp) derived from clam shells. The different molar concentrations of Sr, varies from 10, 30, 50, 70% of Ca, were substituted into the HAp. The synthesized powders were sintered at 1200 °C in air. The as synthesized powders and sintered specimens were characterized using X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and high resolution transmission electron microscopy. The crystallite size and cell parameters of sintered specimens were analyzed from XRD. The XRD of hydrothermally synthesized powders mostly matched with HAp with slight shifting due to Sr doping. However, some distinct Sr based compounds were also observed where Sr substitution is more that 50% of Ca. The XRD of sintered specimen showed increasing ß-tricalcium phosphate (ß-TCP) phase with Sr substitution. The sintered density of solid samples gradually increased from 3.04 g/cc to 3.50 g/cc and surface energy decreased with increasing Sr substitution. Similarly, microhardness, fracture toughness and nanohardness of solid samples found to be enhanced with Sr substitution. The elastic modulus gradually increased from 130 to 137 GPa for HAp and Sr substituted HAp (70% of Ca). The in vitro cytotoxicity of sintered specimen against mouse osteoblast cell line showed that all the samples were nontoxic. However cell proliferation found low for the solid samples containing more than 50% Sr substitution.

Phase pure, high hardness, biocompatible calcium silicates with excellent anti-bacterial and biofilm inhibition efficacies for endodontic and orthopaedic applications.

Here we report for the very first time the synthesis of 100% phase pure calcium silicate nanoparticles (CSNPs) of the α-wollastonite phase without using any surfactant or peptizer at the lowest ever reported calcination temperature of 850 °C. Further, the phase purity is confirmed by quantitative phase analysis. The nano-network like microstructure of the CSNPs is characterized by FTIR, Raman, XRD, FESEM, TEM, TGA, DSC etc. techniques to derive the structure property correlations. The performance efficacies of the CSNPs against gram-positive e.g., S. pyogenes and S. aureus (NCIM2127) and gram-negative e.g., E. coli (NCIM2065) bacterial strains are studied. The biocompatibility of the CSNPs is established by using the conventional mouse embryonic osteoblast cell line (MC3T3). In addition, the biofilm inhibition efficacies of two varieties of CSNPs e.g., CSNPs(W) and CSNPs(WC) are investigated. Further, the interconnection between ROS e.g., superoxide (O2.-) and hydroxyl radical (.OH) generation capabilities of CSNPs and their biofilm inhibition efficacies is clearly established for the very first time. Finally, the mechanical responses of the CSNPs at the microstructural length scale are investigated by nanoindentation. The results confirm that the α-wollastonite phases present in CSNPs(W) and CSNPs(WC) possess extraordinarily high nanohardness and Young's moduli values. Therefore, these materials are well suited for orthopaedic and endodontic applications.

Nano- and micro-tribological behaviours of plasma nitrided Ti6Al4V alloys.

Plasma nitriding of the Ti-6Al-4V alloy (TA) sample was carried out in a plasma reactor with a hot wall vacuum chamber. For ease of comparison these plasma nitrided samples were termed as TAPN. The TA and TAPN samples were characterized by XRD, Optical microscopy, FESEM, TEM, EDX, AFM, nanoindentation, micro scratch, nanotribology, sliding wear resistance evaluation and in vitro cytotoxicity evaluation techniques. The experimental results confirmed that the nanohardness, Young's modulus, micro scratch wear resistance, nanowear resistance, sliding wear resistance of the TAPN samples were much better than those of the TA samples. Further, when the data are normalized with respect to those of the TA alloy, the TAPN sample showed cell viability about 11% higher than that of the TA alloy used in the present work. This happened due to the formation of a surface hardened embedded nitrided metallic alloy layer zone (ENMALZ) having a finer microstructure characterized by presence of hard ceramic Ti2N, TiN etc. phases in the TAPN samples, which could find enhanced application as a bioimplant material.

Laser surface modification of 316L stainless steel.

Medical grade 316L stainless steel was laser surface melted (LSM) using continuous wave Nd-YAG laser in argon atmosphere at 1 and 5 mm/s. The treated surfaces were characterized using electron backscatter diffraction to study the influence of top surface crystallographic orientation and type of grain boundaries on corrosion resistance, wettability, and biocompatibility. The laser scan velocity was found to have a marginal influence on the surface roughness and the type of grain boundaries. However, the crystal orientation density was found to be relatively high in 1 mm/s samples. The LSM samples showed a higher concentration of {101} and {123} planes parallel to the sample surface as well as a higher fraction of low-angle grain boundaries. The LSM samples were found to exhibit better surface wettability and enhanced the viability and proliferation of human fetal osteoblast cells in vitro when compared to the untreated samples. Further, the corrosion protection efficiency of 316L stainless steel was improved up to 70% by LSM in as-processed condition. The increased concentration of {101} and {123} planes on surfaces of LSM samples increases their surface energy, which is believed to be responsible for the improved in vitro cell proliferation. Further, the increased lattice spacing of these planes and high concentration of low-energy grain boundaries in LSM samples would have contributed to the better in vitro corrosion resistance than untreated 316L stainless steel. Our results indicate that LSM can be a potential treatment option for 316L stainless steel-based biomedical devices to improve biocompatibility and corrosion resistance. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 569-577, 2018.

Nanotribological response of a plasma nitrided bio-steel.

AISI 316L is a well known biocompatible, austenitic stainless steel (SS). It is thus a bio-steel. Considering its importance as a bio-prosthesis material here we report the plasma nitriding of AISI 316L (SS) followed by its microstructural and nanotribological characterization. Plasma nitriding of the SS samples was carried out in a plasma reactor with a hot wall vacuum chamber. For ease of comparison these plasma nitrided samples were termed as SSPN. The experimental results confirmed the formations of an embedded nitrided metal layer zone (ENMLZ) and an interface zone (IZ) between the ENMLZ and the unnitrided bulk metallic layer zone (BMLZ) in the SSPN sample. These ENMLZ and IZ in the SSPN sample were richer in iron nitride (FeN) chromium nitride (CrN) along with the austenite phase. The results from nanoindentation, microscratch, nanoscratch and sliding wear studies confirmed that the static contact deformation resistance, the microwear, nanowear and sliding wear resistance of the SSPN samples were much better than those of the SS samples. These results were explained in terms of structure-property correlations.

Understanding compressive deformation behavior of porous Ti using finite element analysis.

In the present study, porous commercially pure (CP) Ti samples with different volume fraction of porosities were fabricated using a commercial additive manufacturing technique namely laser engineered net shaping (LENS™). Mechanical behavior of solid and porous samples was evaluated at room temperature under quasi-static compressive loading. Fracture surfaces of the failed samples were analyzed to determine the failure modes. Finite Element (FE) analysis using representative volume element (RVE) model and micro-computed tomography (CT) based model have been performed to understand the deformation behavior of laser deposited solid and porous CP-Ti samples. In vitro cell culture on laser processed porous CP-Ti surfaces showed normal cell proliferation with time, and confirmed non-toxic nature of these samples.

Microstructure and corrosion behavior of laser processed NiTi alloy.

Laser Engineered Net Shaping (LENS™), a commercially available additive manufacturing technology, has been used to fabricate dense equiatomic NiTi alloy components. The primary aim of this work is to study the effect of laser power and scan speed on microstructure, phase constituents, hardness and corrosion behavior of laser processed NiTi alloy. The results showed retention of large amount of high-temperature austenite phase at room temperature due to high cooling rates associated with laser processing. The high amount of austenite in these samples increased the hardness. The grain size and corrosion resistance were found to increase with laser power. The surface energy of NiTi alloy, calculated using contact angles, decreased from 61 mN/m to 56 mN/m with increase in laser energy density from 20 J/mm(2) to 80 J/mm(2). The decrease in surface energy shifted the corrosion potentials to nobler direction and decreased the corrosion current. Under present experimental conditions the laser power found to have strong influence on microstructure, phase constituents and corrosion resistance of NiTi alloy.

Effect of hydroxyapatite particle size, morphology and crystallinity on proliferation of colon cancer HCT116 cells.

The aim of the present work is to chemically and physically characterize the synthesized Hydroxyapatite (HAp) micro and nanoparticles and to explore the inhibitory effect of nano-HAps on the in vitro growth of human colon cancerous cells HCT116. HAp powder was synthesized using three different routes to achieve micro and nanosized powders, with different morphologies and crystallinity. The synthesized powders were characterized using X-ray diffraction, FTIR spectroscopy and scanning electron microscope. The results showed that the average crystallite size of HAp powder varies from 11nm to 177nm and respective crystallinity of powder found to be in the range of 0.12 and 0.92. The effect of these physico-chemical properties of HAp powders on human colon cancer HCT116 cells inhibition was determined in vitro. It was found that decreasing the HAp powder crystallite size between 11nm and 22nm significantly increases the HCT116 cell inhibition. Our results demonstrate that apart from HAp powder size their crystallinity and morphology also play an important role in cellular inhibition of human colon cancer cells.

In situ synthesized TiB-TiN reinforced Ti6Al4V alloy composite coatings: microstructure, tribological and in-vitro biocompatibility.

Wear resistant TiB-TiN reinforced Ti6Al4V alloy composite coatings were deposited on Ti substrate using laser based additive manufacturing technology. Ti6Al4V alloy powder premixed with 5wt% and 15wt% of boron nitride (BN) powder was used to synthesize TiB-TiN reinforcements in situ during laser deposition. Influences of laser power, scanning speed and concentration of BN on the microstructure, mechanical, in vitro tribological and biological properties of the coatings were investigated. Microstructural analysis of the composite coatings showed that the high temperature generated due to laser interaction with Ti6Al4V alloy and BN results in situ formation of TiB and TiN phases. With increasing BN concentration, from 5wt% to 15wt%, the Young's modulus of the composite coatings, measured by nanoindentation, increased from 170±5GPa to 204±14GPa. In vitro tribological tests showed significant increase in the wear resistance with increasing BN concentration. Under identical test conditions TiB-TiN composite coatings with 15wt% BN exhibited an order of magnitude less wear rate than CoCrMo alloy-a common material for articulating surfaces of orthopedic implants. Average top surface hardness of the composite coatings increased from 543±21HV to 877±75HV with increase in the BN concentration. In vitro biocompatibility and flow cytometry study showed that these composite coatings were non-toxic, exhibit similar cell-materials interactions and biocompatibility as that of commercially pure titanium (CP-Ti) samples. In summary, excellent in vitro wear resistance, high stiffness and suitable biocompatibility make these composite coatings as a potential material for load-bearing articulating surfaces towards orthopaedic implants.

Laser surface modification of 316 L stainless steel with bioactive hydroxyapatite.

Laser-engineered net shaping (LENS™), a commercial additive manufacturing process, was used to modify the surfaces of 316 L stainless steel with bioactive hydroxyapatite (HAP). The modified surfaces were characterized in terms of their microstructure, hardness and apatite forming ability. The results showed that with increase in laser energy input from 32 J/mm(2) to 59 J/mm(2) the thickness of the modified surface increased from 222±12 µm to 355±6 µm, while the average surface hardness decreased marginally from 403±18 HV0.3 to 372±8 HV0.3. Microstructural studies showed that the modified surface consisted of austenite dendrites with HAP and some reaction products primarily occurring in the inter-dendritic regions. Finally, the surface-modified 316 L samples immersed in simulated body fluids showed significantly higher apatite precipitation compared to unmodified 316 L samples.