Three-dimensional force microscopy of cells in biopolymer networks.

We describe a technique for the quantitative measurement of cell-generated forces in highly nonlinear three-dimensional biopolymer networks that mimic the physiological situation of living cells. We computed forces of MDA-MB-231 breast carcinoma cells from the measured network deformations around the cells using a finite-element approach based on a constitutive equation that captures the complex mechanical properties of diverse biopolymers such as collagen gels, fibrin gels and Matrigel. Our measurements show that breast carcinoma cells cultured in collagen gels generated nearly constant forces regardless of the collagen concentration and matrix stiffness. Furthermore, time-lapse force measurements showed that these cells migrated in a gliding motion with alternating phases of high and low contractility, elongation, migratory speed and persistence.

Mechanical plasticity of cells.

Under mechanical loading, most living cells show a viscoelastic deformation that follows a power law in time. After removal of the mechanical load, the cell shape recovers only incompletely to its original undeformed configuration. Here, we show that incomplete shape recovery is due to an additive plastic deformation that displays the same power-law dynamics as the fully reversible viscoelastic deformation response. Moreover, the plastic deformation is a constant fraction of the total cell deformation and originates from bond ruptures within the cytoskeleton. A simple extension of the prevailing viscoelastic power-law response theory with a plastic element correctly predicts the cell behaviour under cyclic loading. Our findings show that plastic energy dissipation during cell deformation is tightly linked to elastic cytoskeletal stresses, which suggests the existence of an adaptive mechanism that protects the cell against mechanical damage.

Labeling of cancer cells with magnetic nanoparticles for magnetic resonance imaging.

PURPOSE: The process of invasion and metastasis formation of tumor cells can be studied by following the migration of labeled cells over prolonged time periods. This report investigates the applicability of iron oxide nanoparticles as a magnetic resonance imaging (MRI) contrast agent for cell labeling. METHODS: γFe2 O3 nanoparticles prepared with direct flame spray pyrolysis are biofunctionalized with poly-l-lysine (PLL). The nanoparticles within the cells were observed with transmission electron microscopy, bright-field microscopy, and magnetorelaxometry. MRI of labeled cells suspended in agarose was used to estimate the detection limit. RESULTS: PLL-coated particles are readily taken up, stored in intracellular clusters, and gradually degraded by the cells. During cell division, the nanoparticle clusters are divided and split between daughter cells. The MRI detection limit was found to be 25 cells/mm(3) for R2*, and 70 cells/mm(3) for R2. The iron specificity, however, was higher for R2 images. Due to the degradation of intracellular γFe2 O3 to paramagnetic iron ions within 13 days, the R1, R2, and R2* contrast gradually decreased over this time period to approximately 50% of its initial value. CONCLUSIONS: These results suggest that PLL-coated γFe2 O3 nanoparticles can be used as an MRI contrast agent for long-term studies of cell migration. Magn Reson Med 71:1896-1905, 2014. © 2013 Wiley Periodicals, Inc.

Estimating the 3D pore size distribution of biopolymer networks from directionally biased data.

The pore size of biopolymer networks governs their mechanical properties and strongly impacts the behavior of embedded cells. Confocal reflection microscopy and second harmonic generation microscopy are widely used to image biopolymer networks; however, both techniques fail to resolve vertically oriented fibers. Here, we describe how such directionally biased data can be used to estimate the network pore size. We first determine the distribution of distances from random points in the fluid phase to the nearest fiber. This distribution follows a Rayleigh distribution, regardless of isotropy and data bias, and is fully described by a single parameter--the characteristic pore size of the network. The bias of the pore size estimate due to the missing fibers can be corrected by multiplication with the square root of the visible network fraction. We experimentally verify the validity of this approach by comparing our estimates with data obtained using confocal fluorescence microscopy, which represents the full structure of the network. As an important application, we investigate the pore size dependence of collagen and fibrin networks on protein concentration. We find that the pore size decreases with the square root of the concentration, consistent with a total fiber length that scales linearly with concentration.

A novel polysaccharide/zein conjugate as an alternative green plastic.

The flax seed cake is a waste product from flax oil extraction. Adding value to this wasted material aligns with the concept of circularity. In this study, we explored zein protein conjugation with flax mucilage for packaging material development. Although both flax mucilage and zein have excellent film-forming properties, they lack the required mechanical properties for industrial processing and are sensitive to high humidity. We present a simple and non-toxic one-pot method for developing the novel flax mucilage/zein conjugate. Where the flax mucilage undergoes oxidation to form aldehyde groups, which then react with zein's amino groups in a glycation process. The conjugates were analyzed using different techniques. The flax mucilage conjugate had a water-holding capacity of 87-62%. Increasing the zein content improved the surface smoothness of the films. On the other hand, higher levels of zein led to a significant decrease in film solubility (p < 0.05). The flax mucilage conjugate exhibited thermoplastic and elastic properties; revealing Young's modulus of 1-3 GPa, glass transition temperature between 49 °C and 103 °C and excellent processability with various industrial techniques. Showing its potential as a sustainable alternative to traditional plastics.

Capacity fade in Sn-C nanopowder anodes due to fracture.

Sn based anodes allow for high initial capacities, which however cannot be retained due to the severe mechanical damage that occurs during Li-insertion and de-insertion. To better understand the fracture process during electrochemical cycling three different nanopowders comprised of Sn particles attached on artificial graphite, natural graphite or micro-carbon microbeads were examined. Although an initial capacity of 700 mAh g(-1) was obtained for all Sn-C nanopowders, a significant capacity fade took place with continuous electrochemical cycling. The microstructural changes in the electrodes corresponding to the changes in electrochemical behavior were studied by transmission and scanning electron microscopy. The fragmentation of Sn observed by microscopy correlates with the capacity fade, but this fragmentation and capacity fade can be controlled by controlling the initial microstructure. It was found that there is a dependence of the capacity fade on the Sn particle volume and surface area fraction of Sn on carbon.

Transverse mechanical properties of collagen fibers from nanoindentation.

The mechanical properties of collagenous tissues, such as tendon and ligaments, are of particular interest as they are found extensively in the human body. In the present study the transverse mechanical properties of collagen fibers are reported for the first time. The elastic modulus was found to be 63 ± 4 MPa, while the viscosity was estimated to be 14 GPa ≤ η ≤ 56 GPa s. Comparison with similar data in the literature, for bulk tendon and collagen fibrils, suggests that the apparent modulus of a network of interconnected building blocks is reduced as compared to the modulus of the individual building blocks; in particular E (tendon) < E (fiber) < E (fibril); this is due to the fact that as the scale of the microstructure increases (i) slippage and sliding between the respective building blocks (fibrils or fibers) increases, (ii) the volume fraction of the stiff collagen proteins decreases.

A First Molecular Dynamics Study for Modeling the Microstructure and Mechanical Behavior of Si Nanopillars during Lithiation.

This is the first study that employs large-scale atomistic simulations to examine the stress generation and deformation mechanisms of various Si nanopillars (SiNPs) during Li-ion insertion. First, a new robust and effective minimization approach is proposed to relax a lithiated amorphous SiNP (a-SiNP), which outperforms the known methods. Using this new method, our simulations are able to successfully capture the experimental morphological changes and volume expansions that SiNPs, hollow a-SiNPs, and solid crystalline SiNPs (c-SiNPs) experience upon maximum lithiation. These simulations enable us to selectively track the displacement of Si atoms and their atomic shear strain in the Li3.75Si alloy region, allowing us to observe the plastic flow and illustrate the atomistic mechanism of lithiation-induced deformation for various SiNPs for the first time. Based on the simulation results, a simple fracture mechanistic model is used to determine the fracture resistance of SiNPs, showing that the hollow a-SiNP is the optimal form of Si as an anode because it has the highest fracture resistance. The crack propagation simulation suggests that the preexisting dislocations in pristine c-Si can contribute toward the fracture of c-SiNPs during lithiation. These findings can guide the design of new Si-based anode geometries for the next-generation Li-ion batteries.

Strengthening in Metal/Graphene Composites: Capturing the Transition from Interface to Precipitate Hardening.

A promising materials engineering method for improving the strength of crystalline materials is to add obstacles to dislocation motion that induce interface hardening (IH) or precipitate hardening (PH). In this study, molecular dynamics simulations are performed for Ni/graphene composites, revealing for the first time that graphene can strengthen the Ni matrix not only strictly via IH or PH but also through a continuous transition between the two. When graphene behaves like an interface, dislocation pileups form, whereas when it behaves as a precipitate, complex Orowan looping occurs by dislocation cross-slip. IH transitions to PH when the integrity of the graphene-dislocation configuration (GDC) deteriorates, leading to a reduced strengthening effect. Furthermore, the deformation of graphene is found to be an effective signature to indicate the real-time strengthening. This observation relates the graphene strengthening effect on metals to a combination of parameters, such as the GDC integrity, graphene deformation, and dislocation evolution, opening an avenue to tune the mechanical properties by controlling the dislocation movements and manipulating the dislocation-obstacle interaction mechanisms.

The effect of carbon nanotubes on osteogenic functions of adipose-derived mesenchymal stem cells in vitro and bone formation in vivo compared with that of nano-hydroxyapatite and the possible mechanism.

It has been well recognized that the development and use of artificial materials with high osteogenic ability is one of the most promising means to replace bone grafting that has exhibited various negative effects. The biomimetic features and unique physiochemical properties of nanomaterials play important roles in stimulating cellular functions and guiding tissue regeneration. But efficacy degree of some nanomaterials to promote specific tissue formation is still not clear. We hereby comparatively studied the osteogenic ability of our treated multi-walled carbon nanotubes (MCNTs) and the main inorganic mineral component of natural bone, nano-hydroxyapatite (nHA) in the same system, and tried to tell the related mechanism. In vitro culture of human adipose-derived mesenchymal stem cells (HASCs) on the MCNTs and nHA demonstrated that although there was no significant difference in the cell adhesion amount between on the MCNTs and nHA, the cell attachment strength and proliferation on the MCNTs were better. Most importantly, the MCNTs could induce osteogenic differentiation of the HASCs better than the nHA, the possible mechanism of which was found to be that the MCNTs could activate Notch involved signaling pathways by concentrating more proteins, including specific bone-inducing ones. Moreover, the MCNTs could induce ectopic bone formation in vivo while the nHA could not, which might be because MCNTs could stimulate inducible cells in tissues to form inductive bone better than nHA by concentrating more proteins including specific bone-inducing ones secreted from M2 macrophages. Therefore, MCNTs might be more effective materials for accelerating bone formation even than nHA.

Conductive nanostructured Si biomaterials enhance osteogeneration through electrical stimulation.

It is well known that the differentiation of stem cells is affected by the cell culture medium, the scaffold surface and electrochemical signals. However, stimulation of patterned biomaterials seeded with stem cell cultures has not been explored. Herein the effect of electrical stimulation on osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) cultured on solid and nanoporous micropyramid patterned Si surfaces was evaluated. It was found that both stimulation and scaffold patterning significantly enhanced osteo-differentiation. The stimulated nanoporous micropyramid scaffolds were more promising compared to the stimulated solid micropyramid surfaces, as they significantly promoted the osteogenic differentiation of rBMSCs via BMP/Smad signaling pathway. Particularly, as compared to the unstimulated patterned biomaterials, the stimulated patterned scaffolds allowed for a significant increase in core binding factor alpha l, alkaline phosphatase, the alpha l chain of type I Col, osteocalcin, and osteonectin, all of which are characteristic for osteo-differentiation. The proposed combination of electrical stimulation with scaffold patterning may provide novel promising strategies for bone tissue engineering and regenerative medicine.

Relating the blood-thinning effect of pentoxifylline to the reduction in the elastic modulus of human red blood cells: an in vivo study.

The blood thinning properties of pentoxifylline have been attributed to its ability to increase the deformability of red blood cells and improve their rheological properties. To interpret and substantiate these observations a novel approach is taken by measuring the stiffness of individual red blood cells from healthy humans before and after subscription to pentoxifylline for nine days. Atomic force microscopy nanoindentation experiments reveal that the elastic modulus of the red blood cells decreased by 30%-40%, after pentoxifylline subscription. This decrease in elastic modulus is related to the ability of pentoxifylline to increase the production of ATP and lower Ca2+ concentrations in red blood cells. The present in vivo experiments provide a deeper understanding of the mode of action of pentoxifylline, and pave the way to using indentation in medicine. A further unique advantage of this study is that it was performed on healthy volunteers, rather than requiring in vitro incubation.

Optomechanical measurement of the role of lamins in whole cell deformability.

There is mounting evidence that the nuclear envelope, and particularly the lamina, plays a critical role in the mechanical and regulation properties of the cell and changes to the lamina can have implications for the physical properties of the whole cell. In this study we demonstrate that the optical stretcher can measure changes in the time-dependent mechanical properties of living cells with different levels of A-type lamin expression. Results from the optical stretcher shows a decrease in the deformability of cells as the levels of lamin A increases, for cells which grow both adherently and in suspension. Further detail can be probed by combining the optical stretcher with fluorescence microscopy to investigate the nuclear mechanical properties which show a larger decrease in deformability than for the whole cell.

Current investigations into magnetic nanoparticles for biomedical applications.

It is generally recognized that nanoparticles possess unique physicochemical properties that are largely different from those of conventional materials, specifically the electromagnetic properties of magnetic nanoparticles (MNPs). These properties have attracted many researchers to launch investigations into their potential biomedical applications, which have been reviewed in this article. First, common types of MNPs were briefly introduced. Then, the biomedical applications of MNPs were reviewed in seven parts: magnetic resonance imaging (MRI), cancer therapy, the delivery of drugs and genes, bone and dental repair, tissue engineering, biosensors, and in other aspects, which indicated that MNPs possess great potentials for many kinds of biomedical applications due to their unique properties. Although lots of achievements have been obtained, there is still a lot of work to do. New synthesis techniques and methods are still needed to develop the MNPs with satisfactory biocompatibility. More effective methods need to be exploited to prepare MNPs-based composites with fine microstructures and high biomedical performances. Other promising research points include the development of more appropriate techniques of experiments both in vitro and in vivo to detect and analyze the biocompatibility and cytotoxicity of MNPs and understand the possible influencing mechanism of the two properties. More comprehensive investigations into the diagnostic and therapeutic applications of composites containing MNPs with "core-shell" structure and deeper understanding and further study into the properties of MNPs to reveal their new biomedical applications, are also described in the conclusion and perspectives part.

The applications of conductive nanomaterials in the biomedical field.

As their name suggests, conductive nanomaterials (CNMs) are a type of functional materials, which not only have a high surface area to volume ratio, but also possess excellent conductivity. Thus far, CNMs have been widely used in biomedical applications, such as effectively transferring electrical signals, and providing a large surface area to adsorb proteins and induce cellular functions. Recent works propose further applications of CNMs in biosensors, tissue engineering, neural probes, and drug delivery. This review focuses on common types of CNMs and elaborates on their unique properties, which indicate that such CNMs have a potential to develop into a class of indispensable biomaterials for the diagnosis and therapy of human diseases.

Effect of substrate mechanics on cardiomyocyte maturation and growth.

Cardiac tissue engineering constructs are a promising therapeutic treatment for myocardial infarction, which is one of the leading causes of death. In order to further advance the development and regeneration of engineered cardiac tissues using biomaterial platforms, it is important to have a complete overview of the effects that substrates have on cardiomyocyte (CM) morphology and function. This article summarizes recent studies that investigate the effect of mechanical cues on the CM differentiation, maturation, and growth. In these studies, CMs derived from embryos, neonates, and mesenchymal stem cells were seeded on different substrates of various elastic modulus. Measuring the contractile function by force production, work output, and calcium handling, it was seen that cell behavior on substrates was optimized when the substrate stiffness mimicked that of the native tissue. The contractile function reflected changes in the sarcomeric protein confirmation and organization that promoted the contractile ability. The analysis of the literature also revealed that, in addition to matrix stiffness, mechanical stimulation, such as stretching the substrate during cell seeding, also played an important role during cell maturation and tissue development.

Biphasic response of cell invasion to matrix stiffness in three-dimensional biopolymer networks.

When cells come in contact with an adhesive matrix, they begin to spread and migrate with a speed that depends on the stiffness of the extracellular matrix. On a flat surface, migration speed decreases with matrix stiffness mainly due to an increased stability of focal adhesions. In a three-dimensional (3-D) environment, cell migration is thought to be additionally impaired by the steric hindrance imposed by the surrounding matrix. For porous 3-D biopolymer networks such as collagen gels, however, the effect of matrix stiffness on cell migration is difficult to separate from effects of matrix pore size and adhesive ligand density, and is therefore unknown. Here we used glutaraldehyde as a crosslinker to increase the stiffness of self-assembled collagen biopolymer networks independently of collagen concentration or pore size. Breast carcinoma cells were seeded onto the surface of 3-D collagen gels, and the invasion depth was measured after 3 days of culture. Cell invasion in gels with pore sizes >5 µm increased with higher gel stiffness, whereas invasion in gels with smaller pores decreased with higher gel stiffness. These data show that 3-D cell invasion is enhanced by higher matrix stiffness, opposite to cell behavior in two dimensions, as long as the pore size does not fall below a critical value where it causes excessive steric hindrance. These findings may be important for optimizing the recellularization of soft tissue implants or for the design of 3-D invasion models in cancer research.

Poly(glycerol sebacate)/poly(butylene succinate-butylene dilinoleate) fibrous scaffolds for cardiac tissue engineering.

The present article investigates the use of a novel electrospun fibrous blend of poly(glycerol sebacate) (PGS) and poly(butylene succinate-butylene dilinoleate) (PBS-DLA) as a candidate for cardiac tissue engineering. Random electrospun fibers with various PGS/PBS-DLA compositions (70/30, 60/40, 50/50, and 0/100) were fabricated. To examine the suitability of these fiber blends for heart patches, their morphology, as well as their physical, chemical, and mechanical properties were measured before examining their biocompatibility through cell adhesion. The fabricated fibers were bead-free and exhibited a relatively narrow diameter distribution. The addition of PBS-DLA to PGS resulted in an increase of the average fiber diameter, whereas increasing the amount of PBS-DLA decreased the hydrophilicity and the water uptake of the nanofibrous scaffolds to values that approached those of neat PBS-DLA nanofibers. Moreover, the addition of PBS-DLA significantly increased the elastic modulus. Initial toxicity studies with C2C12 myoblast cells up to 72 h confirmed nontoxic behavior of the blends. Immunofluorescence analyses and scanning electron microscopy analyses confirmed that C2C12 cells showed better cell attachment and proliferation on electrospun mats with higher PBS-DLA content. However, immunofluorescence analyses of the 3-day-old rat cardiomyocytes cultured for 2 and 5 days demonstrated better attachment on the 70/30 fibers containing well-aligned sarcomeres and expressing high amounts of connexin 43 in cellular junctions indicating efficient cell-to-cell communication. It can be concluded, therefore, that fibrous PGS/PBS-DLA scaffolds exhibit promising characteristics as a biomaterial for cardiac patch applications.

The application of nanomaterials in controlled drug delivery for bone regeneration.

Bone regeneration is a complicated process that involves a series of biological events, such as cellular recruitment, proliferation and differentiation, and so forth, which have been found to be significantly affected by controlled drug delivery. Recently, a lot of research studies have been launched on the application of nanomaterials in controlled drug delivery for bone regeneration. In this article, the latest research progress in this area regarding the use of bioceramics-based, polymer-based, metallic oxide-based and other types of nanomaterials in controlled drug delivery for bone regeneration are reviewed and discussed, which indicates that the controlling drug delivery with nanomaterials should be a very promising treatment in orthopedics. Furthermore, some new challenges about the future research on the application of nanomaterials in controlled drug delivery for bone regeneration are described in the conclusion and perspectives part.

Effects of physicochemical properties of nanomaterials on their toxicity.

Due to their unique size and properties, nanomaterials have numerous applications, which range from electronics, cosmetics, household appliances, energy storage, and semiconductor devices, to medical products such as biological sensors, drug carriers, bioprobes, and implants. Many of the promising properties of nanomaterials arise from their large surface to volume ratio and, therefore, nanobiomaterials that are implantable have a large contact area with the human body. Before, therefore, we can fully exploit nanomaterials, in medicine and bioengineering; it is necessary to understand how they can affect the human body. As a step in this direction, this review paper provides a comprehensive summary of the effects that the physicochemical properties of commonly used nanobiomaterials have on their toxicity. Furthermore, the possible mechanisms of toxicity are described with the aim to provide guidance concerning the design of the nanobiomaterials with desirable properties.