Antibacterial and anti-inflammatory properties of host defense peptides against Staphylococcus aureus

Cationic host defense peptides (HDPs) are a promising alternative to antibiotics in the fight against Staphylococcus aureus infections. In this study, we investigated the antibacterial and immunomodulatory properties of three HDPs namely IDR-1018, CATH-2, and LL-37. Although all three HDPs significantly inhibited LPS-induced activation of human macrophages, only CATH-2 prevented S. aureus growth. When applied to different infection models focused on intracellularly surviving bacteria, only IDR-1018 showed a consistent reduction in macrophage bacterial uptake. However, this observation did not correlate with an increase in killing the efficiency of intracellular S. aureus. Here, we conclude that despite the promising antibacterial and anti-inflammatory properties of the selected HDPs, macrophages' intrinsic antibacterial functions were not improved. Future studies should either focus on combining different HDPs or using them synergistically with other antibacterial agents to improve immune cells' efficacy against S. aureus pathogenesis

Selective laser melting-produced porous titanium scaffolds regenerate bone in critical size cortical bone defects

Porous titanium scaffolds have good mechanical properties that make them an interesting bone substitute material for large bone defects. These scaffolds can be produced with selective laser melting, which has the advantage of tailoring the structure's architecture. Reducing the strut size reduces the stiffness of the structure and may have a positive effect on bone formation. Two scaffolds with struts of 120-μm (titanium-120) or 230-μm (titanium-230) were studied in a load-bearing critical femoral bone defect in rats. The defect was stabilized with an internal plate and treated with titanium-120, titanium-230, or left empty. In vivo micro-CT scans at 4, 8, and 12 weeks showed more bone in the defects treated with scaffolds. Finally, 18.4 ± 7.1 mm3(titanium-120, p = 0.015) and 18.7 ± 8.0 mm3(titanium-230, p = 0.012) of bone was formed in those defects, significantly more than in the empty defects (5.8 ± 5.1 mm3). Bending tests on the excised femurs after 12 weeks showed that the fusion strength reached 62% (titanium-120) and 45% (titanium-230) of the intact contralateral femurs, but there was no significant difference between the two scaffolds. This study showed that in addition to adequate mechanical support, porous titanium scaffolds facilitate bone formation, which results in high mechanical integrity of the treated large bone defects. Copyrigh

Additively manufactured Bi-functionalized bioceramics for reconstruction of bone tumor defects

Bone tissue exhibits critical factors for metastatic cancer cells and represents an extremely pleasant spot for further growth of tumors. The number of metastatic bone lesions and primary tumors that arise directly from cells comprised in the bone milieu is constantly increasing. Bioceramics have recently received significant attention in bone tissue engineering and local drug delivery applications. Additionally, additive manufacturing of bioceramics offers unprecedented advantages including the possibilities to fill irregular voids after the resection and fabricate patient-specific implants. Herein, we investigated the recent advances in additively manufactured bioceramics and ceramic-based composites that were used in the local bone tumor treatment and reconstruction of bone tumor defects. Furthermore, it has been extensively explained how to bi-functionalize ceramics-based biomaterials and what current limitations impede their clinical application. We have also discussed the importance of further development into ceramic-based biomaterials and molecular biology of bone tumors to: (1) discover new potential therapeutic targets to enhance conventional therapies, (2) local delivering of bio-molecular agents in a customized and “smart” way, and (3) accomplish a complete elimination of tumor cells in order to prevent tumor recurrence formation. We emphasized that by developing the research focus on the introduction of novel 3D-printed bioceramics with unique properties such as stimuli responsiveness, it will be possible to fabricate smart bioceramics that promote bone regeneration while minimizing the side-effects and effectively eradicate bone tumors while promoting bone regeneration. In fact, by combining all these therapeutic strategies and additive manufacturing, it is likely to provide personalized tumor-targeting therapies for cancer patients in the foreseeable future. Statement of significance: To increase the survival rates of cancer patients, different strategies such as surgery, reconstruction, chemotherapy, radiotherapy, etc have proven to be essential. Nonetheless, these therapeutic protocols have reached a plateau in their effectiveness due to limitations including drug resistance, tumor recurrence after surgery, toxic side-effects, and impaired bone regeneration following tumor resection. Hence, novel approaches to specifically and locally attack cancer cells, while also regenerating the damaged bony tissue, have being developed in the past years. This review sheds light to the novel approaches that enhance local bone tumor therapy and reconstruction procedures by combining additive manufacturing of ceramic biomaterials and other polymers, bioactive molecules, nanoparticles to affect bone tumor functions, metabolism, and microenvironment

Stimuli-responsive piezoelectricity in electrospun polycaprolactone (PCL)/Polyvinylidene fluoride (PVDF) fibrous scaffolds for bone regeneration

Polymeric scaffolds are a determinant part of modern tissue engineering owing to their great diversity, adaptability, and processability. Interestingly, the physical properties of these scaffolds, e.g., porosity, mechanical properties, and biocompatibility, can be tuned to make them smart and stimuli-responsive. In this regard, piezoelectric materials can be applied to stimulate bone regeneration by converting mechanical impulses to electrical signals. In the present research, fibers made of various blend ratios of polyvinylidene fluoride (PVDF)/polycaprolactone (PCL) were fabricated, investigated and optimized to promote bone regeneration. Uniform fibers containing β-phase PVDF were obtained due to the simultaneous stretching and high voltage applied during electrospinning. Furthermore, components interaction, crystallinity, and piezoelectric behavior were estimated through fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and piezometery, respectively. The samples showed improved wettability and controlled biodegradability, and the piezoelectric charge output reached up to 7.5 pC/N in the sample containing 70 wt% PVDF. At the same time, these scaffolds could provide mechanical properties close to the native bone tissue relying on the PVDF component. In vitro assessments demonstrated that the composite scaffolds were biocompatible and could support cell attachment and proliferation. Moreover, their piezoelectric behavior promoted stem cell differentiation into osteoblasts. Considering the obtained results, the potential of piezoelectric PVDF/PCL blend fibers for bone scaffolds is indisputable

Effect of Alkali-Acid-Heat Chemical Surface Treatment on Electron Beam Melted Porous Titanium and Its Apatite Forming Ability

Advanced additive manufacturing techniques such as electron beam melting (EBM), can produce highly porous structures that resemble the mechanical properties and structure of native bone. However, for orthopaedic applications, such as joint prostheses or bone substitution, the surface must also be bio-functionalized to promote bone growth. In the current work, EBM porous Ti6Al4V alloy was exposed to an alkali acid heat (AlAcH) treatment to bio-functionalize the surface of the porous structure. Various molar concentrations (3, 5, 10M) and immersion times (6, 24 h) of the alkali treatment were used to determine optimal parameters. The apatite forming ability of the samples was evaluated using simulated body fluid (SBF) immersion testing. The micro-topography and surface chemistry of AlAcH treated samples were evaluated before and after SBF testing using scanning electron microscopy and energy dispersive X-ray spectroscopy. The AlAcH treatment successfully modified the topographical and chemical characteristics of EBM porous titanium surface creating nano-topographical features ranging from 200–300 nm in size with a titania layer ideal for apatite formation. After 1 and 3 week immersion in SBF, there was no Ca or P present on the surface of as manufactured porous titanium while both elements were present on all AlAcH treated samples except those exposed to 3M, 6 h alkali treatment. An increase in molar concentration and/or immersion time of alkali treatment resulted in an increase in the number of nano-topographical features per unit area as well as the amount of titania on the surface

Crystal structure and nanotopographical features on the surface of heat-treaded and anodized porous titanium biomaterials produced using selective laser melting

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Repeatability of digital image correlation for measurement of surface strains in composite long bones

Digital image correlation (DIC) can measure full-field surface strains during mechanical testing of hard and soft tissues. When compared to traditional methods, such as strain gauges, DIC offers larger validation data (similar to 50,000 points) for, e.g., finite element models. Our main aim was to evaluate the repeatability of surface strain measurements with DIC during compressive testing of composite femurs mimicking human bones. We also studied the similarity of the composite femur samples using CT. Composite femurs were chosen as test material to minimize the uncertainties associated with the use of cadaveric tissues and to understand the variability of the DIC measurement itself. Six medium-sized fourth generation composite human proximal femora (Sawbones) were CT imaged and mechanically tested in stance configuration. The force-displacement curves were recorded and the 3D surface strains were measured with DIC on the anterior surface of the femurs. Five femurs fractured at the neck-trochanter junction and one at the site below the minor trochanter. CT image of this bone showed an air cavity at the initial fracture site. All femurs fractured through a sudden brittle crack. The fracture force for the composite bones was 5751 +/- 650 N (mean +/- SD). The maximum von Mises strain during the fractures was 2.4 +/- 0.8%. Noise in one experiment was 5-30 mu epsilon. When applied loads were equalized the variation in strains between the bones was 20-25%, and when the maximum strains were equalized, variation in the other regions was 5-10%. DIC showed that the ability of nominally identical composite bones to bear high strains and loads before fracturing may vary between the samples. (C) 2013 Elsevier Ltd. All rights reserved

Effects of anodizing parameters and heat treatment on nanotopographical features, bioactivity, and cell culture response of additively manufactured porous titanium

Anodizing could be used for bio-functionalization of the surfaces of titanium alloys. In this study, we use anodizing for creating nanotubes on the surface of porous titanium alloy bone substitutes manufactured using selective laser melting. Different sets of anodizing parameters (voltage: 10 or 20 V anodizing time: 30 min to 3 h) are used for anodizing porous titanium structures that were later heat treated at 500 °C. The nanotopographical features are examined using electron microscopy while the bioactivity of anodized surfaces is measured using immersion tests in the simulated body fluid (SBF). Moreover, the effects of anodizing and heat treatment on the performance of one representative anodized porous titanium structures are evaluated using in vitro cell culture assays using human periosteum-derived cells (hPDCs). It has been shown that while anodizing with different anodizing parameters results in very different nanotopographical features, i.e. nanotubes in the range of 20 to 55 nm, anodized surfaces have limited apatite-forming ability regardless of the applied anodizing parameters. The results of in vitro cell culture show that both anodizing, and thus generation of regular nanotopographical feature, and heat treatment improve the cell culture response of porous titanium. In particular, cell proliferation measured using metabolic activity and DNA content was improved for anodized and heat treated as well as for anodized but not heat-treated specimens. Heat treatment additionally improved the cell attachment of porous titanium surfaces and upregulated expression of osteogenic markers. Anodized but not heat-treated specimens showed some limited signs of upregulated expression of osteogenic markers. In conclusion, while varying the anodizing parameters creates different nanotube structure, it does not improve apatite-forming ability of porous titanium. However, both anodizing and heat treatment at 500 °C improve the cell culture response of porous titanium.status: publishe

Experimental Validation Of Finite Element Model For Proximal Composite Femur Using Optical Measurements

Patient-specific finite element models have been used to predict femur strength and fracture risk in individuals. Validation of the adopted finite element modelling procedure against mechanical testing data is a crucial step when aiming for clinical applications. The majority of the works available in literature used data from strain gages to validate the model, thus having up to 15 experimental measurements. Optical techniques, such as Digital Image Correlation, can help to improve the models by providing a continuous field of deformation data over a femoral surface. The main objective of this study was to validate finite element models of six composite femora against strain data from digital image correlation, obtained during fracture tests performed in quasi-axial loading configuration. The finite element models were obtained from CT scans, by means of a semi-automatic segmentation. The principal strains both during the elastic phase and close to the fracture were compared, and showed a correlation coefficient close to 0.9. In the linear region, the slope and intercept were close to zero and unity, while for the case when fracture load was simulated, the slope decreased somewhat. The accuracy of the obtained results is comparable with the state-of-the-art literature, with the significant improvement of having around 50000 data points for each femur. This large number of measurements allows a more comprehensive validation of the predictions by the finite element models, since thousand of points are tracked along the femoral neck and trochanter region, i.e., the sites that are most critical for femur fracture. Moreover, strain measurement biases due to the strain gage reinforcement effect, were avoided. The combined experimental-numerical approach proved to be ready for application to in-vitro tests of human cadaver femurs, thus helping to develop a suitable mechanistic fracture risk criterion

Full-field Strain Measurement During Mechanical Testing of the Human Femur at Physiologically Relevant Strain Rates

Understanding the mechanical properties of human femora is of great importance for the development of a reliable fracture criterion aimed at assessing fracture risk. Earlier ex vivo studies have been conducted by measuring strains on a limited set of locations using strain gauges. Digital Image Correlation (DIC) could instead be used to reconstruct the full-field strain pattern over the surface of the femur. The objective of this study was to measure the full-field strain response of cadaver femora tested at a physiological strain rate up to fracture in a configuration resembling single stance. The three cadaver femora were cleaned from soft tissues, and a white background paint was applied with a random black speckle pattern over the anterior surface. The mechanical tests were conducted up to fracture at a constant displacement rate of 15 mm/s, and two cameras recorded the event at 3000 frames per second. DIC was performed to retrieve the full-field displacement map, from which strains were derived. A low-pass filter was applied over the measured displacements before the crack opened in order to reduce the noise level. The noise levels were assessed using a dedicated control plate. Conversely, no filtering was applied at the frames close to fracture to get the maximum resolution. The specimens showed a linear behavior of the principal strains with respect to the applied force up to fracture. The strain rate was comparable to the values available in literature from in-vivo measurements during daily activities. The cracks opened and fully propagated in less than 1 ms, and small regions with high values of the major principal strains could be spotted just a few frames before the crack opened. This corroborates the hypothesis of a strain-driven fracture mechanism in human bone. The data represents a comprehensive collection of full-field strains, both at physiological load levels and up to fracture. About 10000 measurements were collected for each bone, providing superior spatial resolution compared to ~15 measurements typically collected using strain gauges. These experimental data collection can be further used for validation of numerical models, and for experimental verification of bone constitutive laws and fracture criteria