Characterization Of Biomaterials Research Articles

Formulative Study and Characterization of Novel Biomaterials Based on Chitosan/Hydrolyzed Collagen Films.

To date, the need for biomaterials capable of improving the treatment of chronic skin wounds remains a clinical challenge. The aim of the present work is to formulate and characterize chitosan (Cs)/hydrolyzed collagen (HC) films as potential biomaterials with improved mechanical and hydration performances compared to single component formulations. Films were made by the solvent casting method, with or without glycerin and/or PEG1500 as plasticizers, resulting in a total of eight formulations. All films were characterized by their physico-chemical characteristics and their mechanical and hydration features. A full factorial design was also used to statistically assess the effect of HC concentration, type and concentration of plasticizers and their possible interactions on mechanical and swelling behaviors. Solid state characterization confirmed the hybrid nature of the films, with suggested electrostatic interactions between Cs and HC. Mechanical and swelling properties, along with the analysis of the experimental design, allowed the identification of formulations containing high HC concentration (2% w/v) and glycerin or glycerin/PEG1500 as more suitable candidates for skin wound treatment. Finally, viability assay of immortalized human keratinocytes (HaCaT) showed no statistical differences in cell survival compared to the complete culture medium, suggesting their potential as a promising tool for biomedical applications.

Beyond 2D: Novel biomaterial approaches for modeling the placenta

This review considers fully three-dimensional biomaterial environments of varying complexity as these pertain to research on the placenta. The developments in placental cell sources are first considered, along with the corresponding maternal cells with which the trophoblast interact. We consider biomaterial sources, including hybrid and composite biomaterials. Properties and characterization of biomaterials are discussed in the context of material design for specific placental applications. The development of increasingly complicated three-dimensional structures includes examples of advanced fabrication methods such as microfluidic device fabrication and 3D bioprinting, as utilized in a placenta context. The review finishes with a discussion of the potential for in vitro, three-dimensional placenta research to address health disparities and sexual dimorphism, especially in light of the exciting recent changes in the regulatory environment for in vitro devices.

Synthesis and mechanical characterization of hydroxyapatite-based biomaterials reinforced with nickel-titanium-magnesium alloys for use in cortical bone implants

In this work, biomaterials consisting of bioactive hydroxyapatite-based ceramic and nickel-titanium-magnesium alloys were made. The biomaterials were obtained through powder techniques involving mechanical milling, uniaxial pressing and pressureless sintering. The purpose of the study was to evaluate the effect of alloys on the mechanical properties of hydroxyapatite, the alloy additions were 0, 20, 30, 40, 40, 50 and 60 wt.%. The results were compared with cortical bone samples of bovine origin. From the results, it is possible to comment that, with increasing alloy contents in the hydroxyapatite, the mechanical properties increase considerably. The behavior of the composites reinforced with 50 and 60 wt.% NiTiMg alloy is similar to that of bovine cortical bone. The microstructures of the biomaterials are fine, homogeneous, and similar to those of bovine cortical bone. From the study it can be commented that given the mechanical characteristics of the biomaterials obtained, they could be used as possible bone implants.

Hybrid Polymer-Inorganic Materials with Hyaluronic Acid as Controlled Antibiotic Release Systems.

In recent years, significant developments have taken place in scientific fields such as tissue and materials engineering, which allow for the development of new, intelligent biomaterials. An example of such biomaterials is drug delivery systems that release the active substance directly at the site where the therapeutic effect is required. In this research, polymeric materials and ceramic-polymer composites were developed as carriers for the antibiotic clindamycin. The preparation and characterization of biomaterials based on hyaluronic acid, collagen, and nano brushite obtained using the photocrosslinking technique under UV (ultraviolet) light are described. Physical and chemical analyses of the materials obtained were carried out using Fourier transform infrared spectroscopy (FT-IR) and optical microscopy. The sorption capacities were determined and subjected to in vitro incubation in simulated biological environments such as Ringer's solution, simulated body fluid (SBF), phosphate-buffered saline (PBS), and distilled water. The antibiotic release rate was also measured. The study confirmed higher swelling capacity for materials with no addition of a ceramic phase, thus it can be concluded that brushite inhibits the penetration of the liquid medium into the interior of the samples, leading to faster absorption of the liquid medium. In addition, incubation tests confirmed preliminary biocompatibility. No drastic changes in pH values were observed, which suggests that the materials are stable under these conditions. The release rate of the antibiotic from the biomaterial into the incubation medium was determined using high-pressure liquid chromatography (HPLC). The concentration of the antibiotic in the incubation fluid increased steadily following a 14-day incubation in PBS, indicating continuous antibiotic release. Based on the results, it can be concluded that the developed polymeric material demonstrates potential for use as a carrier for the active substance.

Fundamentals and Applications of Raman-Based Techniques for the Design and Development of Active Biomedical Materials.

Raman spectroscopy is an analytical method based on light-matter interactions that can interrogate the vibrational modes of matter and provide representative molecular fingerprints. Mediated by its label-free, non-invasive nature, and high molecular specificity, Raman-based techniques have become ubiquitous tools for in situ characterization of materials. This review comprehensively describes the theoretical and practical background of Raman spectroscopy and its advanced variants. The numerous facets of material characterization that Raman scattering can reveal, including biomolecular identification, solid-to-solid phase transitions, and spatial mapping of biomolecular species in bioactive materials, are highlighted. The review illustrates the potential of these techniques in the context of active biomedical material design and development by highlighting representative studies from the literature. These studies cover the use of Raman spectroscopy for the characterization of both natural and synthetic biomaterials, including engineered tissue constructs, biopolymer systems, ceramics, and nanoparticle formulations, among others. To increase the accessibility and adoption of these techniques, the present review also provides the reader with practical recommendations on the integration of Raman techniques into the experimental laboratory toolbox. Finally, perspectives on how recent developments in plasmon- and coherently-enhanced Raman spectroscopy can propel Raman from underutilized to critical for biomaterial development areprovided.

Electrochemical corrosion study of biomaterials: A bibliometric study based on co-word analysis

Demand for better biomaterials has been increasing. To meet this demand, testing is necessary to screen potential options. Corrosion testing is essential as it relates to durability and biocompatibility. Usually, electrochemical tests are employed in screening candidates for in vivo testing. Researching in the biomaterials field, or any field for that matter, requires continuous consultation of the literature. When conducting a literature search, the amount of information available is often overwhelming. Identifying relevant research trends that can guide decisions about future projects becomes challenging. This study employs bibliometric analysis to identify research trends in electrochemical corrosion studies of biomaterials. By doing this, researchers interested in the subject are provided with relevant information on which to base decisions about future research projects. The method used is called co-word analysis. Keywords have been downloaded from Web of Science using an appropriate search query. The analysis period includes the months from January 2013 to December 2021. This period is divided into three subperiods of three years each: 2013–15, 2016–18, and 2019–21. Evolution analysis identified 5 areas of interest that remained active throughout the entire period: biodegradable magnesium alloys, titanium and its alloys, hydroxyapatite coatings, general biomaterial characterization, and the use of electrochemical impedance spectroscopy. All of them are likely to remain of interest in the following years. One area related to titanium anodization appears to be an active research area with few publications, leaving room for new research. Methodological limitations are properly addressed.

Noninvasive material characterization of biomaterials: Measuring viscosity and elasticity using ultrasound

The advent of tissue engineering has led to increased interest in the viscoelastic characterization of biomaterials. Gelatin Methacrylate (GelMA) is a particularly promising biomaterial, largely due to its tunability, yet the impact of different preparation parameters on the material’s viscoelasticity is not well understood. We characterized an array of GelMA scaffolds, fabricated by varying both GelMA concentration and ultraviolet (UV) lightexposure time. Pulse-echo ultrasound techniques were used to non-invasively determine the sound speed and attenuation of the scaffolds, revealing significant dependence on GelMA concentration. Steady shear rate tests in addition to strain- and frequency-controlled oscillatory shear tests were performed using a rotational rheometer (Model: DHR-2, TA Instruments) to ascertain the levels of shear-thinning and viscoelasticity at a wide range of strain rates, oscillation frequencies, and amplitudes. The rheological tests show moduli dependence on both GelMA concentration and curing time. Together, this acoustic and rheological characterization can be used to inform the selection of GelMA scaffolds in tissue engineering applications, and this method can be used as a guide for characterizing other polymeric hydrogels.

A Novel Injectable Piezoelectric Hydrogel for Periodontal Disease Treatment.

Periodontal disease is a multifactorial, bacterially induced inflammatory condition characterized by the progressive destruction of periodontal tissues. The successful nonsurgical treatment of periodontitis requires multifunctional technologies offering antibacterial therapies and promotion of bone regeneration simultaneously. For the first time, in this study, an injectable piezoelectric hydrogel (PiezoGEL) was developed after combining gelatin methacryloyl (GelMA) with biocompatible piezoelectric fillers of barium titanate (BTO) that produce electrical charges when stimulated by biomechanical vibrations (e.g., mastication, movements). We harnessed the benefits of hydrogels (injectable, light curable, conforms to pocket spaces, biocompatible) with the bioactive effects of piezoelectric charges. A thorough biomaterial characterization confirmed piezoelectric fillers' successful integration with the hydrogel, photopolymerizability, injectability for clinical use, and electrical charge generation to enable bioactive effects (antibacterial and bone tissue regeneration). PiezoGEL showed significant reductions in pathogenic biofilm biomass (∼41%), metabolic activity (∼75%), and the number of viable cells (∼2-3 log) compared to hydrogels without BTO fillers in vitro. Molecular analysis related the antibacterial effects to be associated with reduced cell adhesion (downregulation of porP and fimA) and increased oxidative stress (upregulation of oxyR) genes. Moreover, PiezoGEL significantly enhanced bone marrow stem cell (BMSC) viability and osteogenic differentiation by upregulating RUNX2, COL1A1, and ALP. In vivo, PiezoGEL effectively reduced periodontal inflammation and increased bone tissue regeneration compared to control groups in a mice model. Findings from this study suggest PiezoGEL to be a promising and novel therapeutic candidate for the treatment of periodontal disease nonsurgically.

Osmolytes Modulate Photoactivation of Phytochrome: Probing Protein Hydration.

Phytochromes are bistable red/far-red light-responsive photoreceptor proteins found in plants, fungi, and bacteria. Light-activation of the prototypical phytochrome Cph1 from the cyanobacterium Synechocystis sp. PCC 6803 allows photoisomerization of the bilin chromophore in the photosensory module and a subsequent series of intermediate states leading from the red absorbing Pr to the far-red-absorbing Pfr state. We show here via osmotic and hydrostatic pressure-based measurements that hydration of the photoreceptor modulates the photoconversion kinetics in a controlled manner. While small osmolytes like sucrose accelerate Pfr formation, large polymer osmolytes like PEG 4000 delay the formation of Pfr. Thus, we hypothesize that an influx of mobile water into the photosensory domain is necessary for proceeding to the Pfr state. We suggest that protein hydration changes are a molecular event that occurs during photoconversion to Pfr, in addition to light activation, ultrafast electric field changes, photoisomerization, proton release and uptake, and the major conformational change leading to signal transmission, or simultaneously with one of these events. Moreover, we discuss this finding in light of the use of Cph1-PGP as a hydration sensor, e.g., for the characterization of novel hydrogel biomaterials.

Mechanical characterization and design of biomaterials for nucleus pulposus replacement and regeneration.

Biomaterials for nucleus pulposus (NP) replacement and regeneration have great potential to restore normal biomechanics in degenerated intervertebral discs following nucleotomy. Mechanical characterizations are essential for assessing the efficacy of biomaterial implants for clinical applications. While traditional compression tests are crucial to quantify various modulus values, relaxation behaviors and fatigue resistance, rheological measurements should also be conducted to investigate the viscoelastic properties, injectability, and overall stability upon deformation. To recapitulate the physiological in vivo environment, the use of spinal models is necessary to evaluate the risk of implant extrusion and the restoration of biomechanics under different loading conditions. When designing devices for NP replacement, injectable materials are ideal to fully fill the nucleus cavity and prevent implant migration. In addition to achieving biocompatibility and desirable mechanical characteristics, biomaterial implants should be optimized to avoid implant extrusion or re-herniation post-operatively. This review discusses the most commonly used testing protocols for assessing mechanical properties of biomaterial implants and serves as reference material for enabling researchers to characterize NP implants through a unified approach whereby newly developed biomaterials may be compared and contrasted to existing devices.

Challenges and solutions of environmental scanning electron microscopy characterisation of biomaterials: Application to hygro‐expansion of paper

AbstractMost methodologies to measure the moisture‐induced deformation (hygro‐expansion) of paper microconstituents, including fibres and interfibre bonds, are low resolution or time‐consuming. Hence, here, a novel method is proposed and validated to measure high‐resolution full‐field strain maps of paper microconstituents during hygro‐expansion, based on environmental scanning electron microscopy (ESEM). To this end, a novel climate stage enables accurate control of the relative humidity (RH) near the specimen in the ESEM from 0%–100%. The fibre surface, which is decorated a priori with a microparticle pattern, is captured during RH change. Subsequently, correlating the fibre surface using a dedicated global digital image correlation algorithm enables high‐resolution hygro‐expansion strain maps. Method optimisation involved performing contrast enhancement, scan‐correction to reduce ESEM artefacts and a background correction, resulting in a strain resolution of . Method validation revealed that the fibres' crystallinity is affected by the electron beam, even for minimal invasive electron beam settings. Interestingly, however, the fibres consistently exhibit conventional hygro‐expansion behaviour during the drying slopes. Using the optimised procedure, hygro‐expansion characterisation of two interfibre bonds and four interfibre bond cross‐sections revealed the competition between the low longitudinal and large transverse fibre hygro‐expansion in the bonded area.

Proteomic Analysis of Mesenchymal Stem Cells and Monocyte Co-Cultures Exposed to a Bioactive Silica-Based Sol-Gel Coating.

New methodologies capable of extensively analyzing the cell-material interactions are necessary to improve current in vitro characterization methods, and proteomics is a viable alternative. Also, many studies are focused on monocultures, even though co-cultures model better the natural tissue. For instance, human mesenchymal stem cells (MSCs) modulate immune responses and promote bone repair through interaction with other cell types. Here, label-free liquid chromatography tandem mass spectroscopy proteomic methods were applied for the first time to characterize HUCPV (MSC) and CD14+ monocytes co-cultures exposed to a bioactive sol-gel coating (MT). PANTHER, DAVID, and STRING were employed for data integration. Fluorescence microscopy, enzyme-linked immunosorbent assay, and ALP activity were measured for further characterization. Regarding the HUCPV response, MT mainly affected cell adhesion by decreasing integrins, RHOC, and CAD13 expression. In contrast, MT augmented CD14+ cell areas and integrins, Rho family GTPases, actins, myosins, and 14-3-3 expression. Also, anti-inflammatory (APOE, LEG9, LEG3, and LEG1) and antioxidant (peroxiredoxins, GSTO1, GPX1, GSHR, CATA, and SODM) proteins were overexpressed. On co-cultures, collagens (CO5A1, CO3A1, CO6A1, CO6A2, CO1A2, CO1A1, and CO6A3), cell adhesion, and pro-inflammatory proteins were downregulated. Thus, cell adhesion appears to be mainly regulated by the material, while inflammation is impacted by both cellular cross-talk and the material. Altogether, we conclude that applied proteomic approaches show its potential in biomaterial characterization, even in complex systems.

Fast extraction of three-dimensional nanofiber orientation from WAXD patterns using machine learning.

Structural disclosure of biological materials can help our understanding of design disciplines in nature and inspire research for artificial materials. Synchrotron microfocus X-ray diffraction is one of the main techniques for characterizing hierarchically structured biological materials, especially the 3D orientation distribution of their interpenetrating nanofiber networks. However, extraction of 3D fiber orientation from X-ray patterns is still carried out by iterative parametric fitting, with disadvantages of time consumption and demand for expertise and initial parameter estimates. When faced with high-throughput experiments, existing analysis methods cannot meet the real time analysis challenges. In this work, using the assumption that the X-ray illuminated volume is dominated by two groups of nanofibers in a gradient biological composite, a machine-learning based method is proposed for fast and automatic fiber orientation metrics prediction from synchrotron X-ray micro-focused diffraction data. The simulated data were corrupted in the training procedure to guarantee the prediction ability of the trained machine-learning algorithm in real-world experimental data predictions. Label transformation was used to resolve the jump discontinuity problem when predicting angle parameters. The proposed method shows promise for application in the automatic data-processing pipeline for fast analysis of the vast data generated from multiscale diffraction-based tomography characterization of textured biomaterials.

Characterization of biomaterials with reference to biocompatibility dedicated for patient-specific finger implants

Purpose The research was focused on determining basic mechanical properties, surface and phase structure taking into consideration basic cytotoxicity analysis towards human cells. Methods Biological tests were performed on human C-12302 fibroblasts cells using 3D-printed Ti6Al4V alloy (Ti64), produced by laser-based powder bed fusion (LB-PBF) and Alumina Toughened Zirconia 20 (ATZ20), produced by lithography-based ceramic manufacturing (LCM). Surface modifications included electropolishing, and hydroxyapatite or hydroxyapatite/zinc coating. Structure analysis was carried out using a variety of techniques such as X-Ray diffraction, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and confocal laser scanning microscopy (CLSM), followed by mechanical properties evaluation using nanoindentation testing. Results Samples subjected to surface modifications showed diversity among surface and phase structure and mechanical properties. However, the cytotoxicity towards tested cells was not significantly higher than the control. Though, a trend was noted among the materials analysed, indicating that HAp/Zn coating on Ti64 and ATZ20 resulted in the best biological performance Conclusions Hydroxyapatite coating on Ti64 and ATZ20 resulted in the best biological properties. Tested materials are suitable for in vivo toxicity testing.

Elasticity imaging using physics-informed neural networks: Spatial discovery of elastic modulus and Poisson's ratio.

Elasticity imaging is a technique that discovers the spatial distribution of mechanical properties of tissue using deformation and force measurements under various loading conditions. Given the complexity of this discovery, most existing methods approximate only one material parameter while assuming homogeneous distributions for the others. We employ physics-informed neural networks (PINN) in linear elasticity problems to discover the space-dependent distribution of both elastic modulus (E) and Poisson's ratio (ν) simultaneously, using strain data, normal stress boundary conditions, and the governing physics. We validated our model on three examples. First, we experimentally loaded hydrogel samples with embedded stiff inclusions, representing tumorous tissue, and compared the approximations against ground truth determined through tensile tests. Next, using data from finite element simulation of a rectangular domain containing a stiff circular inclusion, the PINN model accurately localized the inclusion and estimated both E and ν. We observed that in a heterogeneous domain, assuming a homogeneous ν distribution increases estimation error for stiffness as well as the area of the stiff inclusion, which could have clinical importance when determining size and stiffness of tumorous tissue. Finally, our model accurately captured spatial distribution of mechanical properties and the tissue interfaces on data from another computational model, simulating uniaxial loading of a rectangular hydrogel sample containing a human brain slice with distinct gray matter and white matter regions and complex geometrical features. This elasticity imaging implementation has the potential to be used in clinical imaging scenarios to reliably discover the spatial distribution of mechanical parameters and identify material interfaces such as tumors. STATEMENT OF SIGNIFICANCE: Our work is the first implementation of physics-informed neural networks to reconstruct both material parameters - Young's modulus and Poisson's ratio - and stress distributions for isotropic linear elastic materials by having deformation and force measurements. We comprehensively validate our model using experimental measurements and synthetic data generated using finite element modeling. Our method can be implemented in clinical elasticity imaging scenarios to improve diagnosis of tumors and for mechanical characterization of biomaterials and biological tissues in a minimally invasive manner.

Development and biomaterial characterization of Mg-Li-Zn-Ca alloys

Development and biomaterial characterization of Mg-Li-Zn-Ca alloys

Characterization of porous biomaterials of 316L stainless steel made from polyethylene waxes as pore-forming agents

For biomedical purposes, the 316L stainless steel has been used due to its high mechanical strength and its biotolerability by the human body. Its porosity allows the growth of living tissue within the implant, which improves fixation to hard tissues. Porous 316L stainless steel specimens were produced by powder metallurgy with controlled porosity by the addition of different polyethylene waxes granulometries (50 and 100 mm) and concentrations (0.0, 1.0, 2.0, 3.0 and 4.0%). The elastic modulus, porous morphologies and densities of the steel samples were determined. Thus, the densities and the elastic modulus were evaluated statistically through Tukey’s test. The results showed that the porosity increases proportionally to polyethylene wax concentration, but the elastic modulus decreases. The Tukey’s Test showed that there are no significant differences in most samples using the two waxes at most concentrations.   Range values of 1.89 to 1.51 GPa using wax with 50 mm, and 1.72 to 1.19 GPa were respectively obtained using 100 mm particle size. The highest pore size (45 mm) and the lowest elastic modulus (1.19 GPa) were obtained by use of 4.0% of wax.

Translating Material Science into Bone Regenerative Medicine Applications: State-of-The Art Methods and Protocols.

In the last 20 years, bone regenerative research has experienced exponential growth thanks to the discovery of new nanomaterials and improved manufacturing technologies that have emerged in the biomedical field. This revolution demands standardization of methods employed for biomaterials characterization in order to achieve comparable, interoperable, and reproducible results. The exploited methods for characterization span from biophysics and biochemical techniques, including microscopy and spectroscopy, functional assays for biological properties, and molecular profiling. This review aims to provide scholars with a rapid handbook collecting multidisciplinary methods for bone substitute R&D and validation, getting sources from an up-to-date and comprehensive examination of the scientific landscape.

Visualization of the distribution of covalently cross-linked hydrogels in CLARITY brain-polymer hybrids for different monomer concentrations

CLARITY is a tissue preservation and optical clearing technique whereby a hydrogel is formed directly within the architectural confines of ex vivo brain tissue. In this work, the extent of polymer gel formation and crosslinking within tissue was assessed using Raman spectroscopy and rheology on CLARITY samples prepared with a range of acrylamide monomer (AAm) concentrations (1%, 4%, 8%, 12% w/v). Raman spectroscopy of individual neurons within hybrids revealed the chemical presence and distribution of polyacrylamide within the mouse hippocampus. Consistent with rheological measurements, lower %AAm concentration decreased shear elastic modulus G’, providing a practical correlation with sample permeability and protein retention. Permeability of F(ab)’2 secondary fluorescent antibody changes from 9.3 to 1.4 µm2 s−1 going from 1 to 12%. Notably, protein retention increased linearly relative to standard PFA-fixed tissue from 96.6% when AAm concentration exceeded 1%, with 12% AAm samples retaining up to ~ 99.3% native protein. This suggests that though 1% AAm offers high permeability, additional %AAm may be required to enhance protein. Our quantitative results on polymer distribution, stability, protein retention, and macromolecule permeability can be used to guide the design of future CLARITY-based tissue-clearing solutions, and establish protocols for characterization of novel tissue-polymer hybrid biomaterials using chemical spectroscopy and rheology.

Characterizing cell recruitment into isotropic and anisotropic biomaterials by quantification of spatial density gradients in vitro.

The success of cell-free in situ tissue engineering approaches depends on an appropriate recruitment of autologous cells from neighboring tissues. This identifies cellular migration as a critical parameter for the pre-clinical characterization of biomaterials. Here, we present a new method to quantify both the extent and the spatial anisotropy of cell migration in vitro. For this purpose, a cell spheroid is used as a cell source to provide a high number of cells for cellular invasion and, at the same time, to guarantee a controlled and spatially localized contact to the material. Therefore, current limitations of assays based on 2D cell sources can be overcome. We tested the method on three biomaterials that are in clinical use for soft tissue augmentation in maxilla-facial surgery and a substrate used for 3D in vitro cell culture. The selected biomaterials were all collagen-derived, but differed in their internal architecture. The analysis of cellular isodensity profiles within the biomaterials allowed the identification of the extent and the preferential directions of migration, as well as their relation to the biomaterials and their specific pore morphologies. The higher cell density within the biomaterials resulting from the here-introduced cell spheroid assay compared to established 2D cell layer assays suggests a better representation of the in vivo situation. Consequently, the presented method is proposed to advance the pre-clinical evaluation of cell recruitment into biomaterials, possibly leading to an improved prediction of the regeneration outcome.