Bioreactor mimicking knee-joint movement for the regeneration of tissue-engineered cartilage

Efforts to minimize the sacrifice of laboratory animals have become a recent worldwide trend. This trend has triggered a number of studies toward developing effective methods to replace the animal experiments. In this study, we developed a biomimetic bioreactor system that simulates the movements of the human knee joint. The system consists of a knee-joint drive and a unit capable of culturing cells at the joint surface. The knee-joint drive is designed to apply dynamic stimulation similar to the real bending motion of the knee joint. We employed a commercial incubator for comparative evaluation and validation of our laboratory-made cell-culture unit mounted in a bioreactor. The results revealed that the ability of the proposed system in culturing cells was similar to that of the commercial incubator. The cell culture was evaluated by dividing the knee joint into zones according to the size of the stimulus. The results confirmed that the cell assessment stimulated by the knee-joint movement was two times higher than that having no stimulation. Overall, the study helped establish that the cell characteristics is more effective when an appropriate external stimulus is applied according to the target tissue. The study is also expected to form the basis for implementing an ex-vivo environment that can potentially replace animal and cadaver experiments in the future. In the future, follow-up studies will be conducted on the in-vivo environment and the characteristics of each organ and tissue for effective tissue regeneration.

     

Immune response by host after allogeneic chondrocyte transplant to the cartilage

We discovered adrenomedullin (AM) from human pheochromocytoma tissue by monitoring the elevating activity of intracellular cyclic AMP (cAMP) in rat platelets in 1993. Since the discovery of AM, it has attracted intense interest from cardiovascular researchers because AM elicits multiple biological activities, including a potent and powerful hypotensive activity caused by dilatation of resistance vessels. AM is biosynthesized and secreted from tissues, including cardiovascular organs. In addition to AM, "proadrenomedullin N-terminal 20 peptide (PAMP)," another biologically active peptide, was found to be processed from the AM precursor. Plasma AM levels are increased in various cardiovascular and renal diseases. AM, therefore, seems to function as a novel system that controls circulation and body fluid, and may be involved in pathophysiological changes in cardiovascular diseases. Therefore, in this review we will focus on the structure of AM and its gene, distribution, receptor, and the physiological and pathological roles of AM in cardiovascular disease.     

Anisotropic properties of bovine nasal cartilage

To investigate the structural anisotropy in bovine septal cartilage, quantitative procedures in microscopic magnetic resonance imaging (μMRI), polarized light microscopy (PLM), and mechanical indentation were used to measure the tissue in three orthogonal planes: vertical, medial, and caudocephalic. The quantitative T2 imaging experiments in μMRI found strong anisotropy in the images of both vertical and caudocephalic planes but little anisotropy in the images from the medial plane. The PLM birefringent experiments found that the retardation values in the medial section were only about 10% of these in the vertical and caudocephalic sections and that the angle values in all three sections followed the rotation of the tissue section in the microscope stage. The stress relaxation experiments in mechanical indentation showed reduced stiffness in the medial plane compared to stiffness in either the vertical or caudocephalic planes. Collectively, the results in this project coherently indicate a marked structural anisotropy in cartilage from the nasal septum, where the long axis of the collagen fibrils is oriented in parallel with the medial axis.     

Nanomechanical characterization of multilayered thin film structures for digital micromirror devices

The digital micromirror device (DMD), used for digital projection displays, comprises a surface-micromachined array of up to 2.07 million aluminum micromirrors (14 μm square and 15 μm pitch), which switch forward and backward thousands of times per second using electrostatic attraction. The nanomechanical properties of the thin-film structures used are important to the performance of the DMD. In this paper, the nanomechanical characterization of the single and multilayered thin film structures, which are of interest in DMDs, is carried out. The hardness, Young's modulus and scratch resistance of TiN/Si, SiO₂/Si, Al alloy/Si, TiN/Al alloy/Si and SiO₂/TiN/Al alloy/Si thin-film structures were measured using nanoindentation and nanoscratch techniques, respectively. The residual (internal) stresses developed during the thin film growth were estimated by measuring the radius of curvature of the sample before and after deposition. To better understand the nanomechanical properties of these thin film materials, the surface and interface analysis of the samples were conducted using X-ray photoelectron spectroscopy. The nanomechanical properties of these materials are analyzed and the impact of these properties on micromirror performance is discussed.     

Nanomechanical characterization of skin and skin cream

The mechanical properties of skin are an important characteristic of its resistance to damage and important indicators of pathological situations. Skin care products are the most common method to improve skin health and create a smooth, soft, and elastic perception by altering the mechanical properties of skin. It is of interest to study how skin cream affects the mechanical properties of skin. It also can help to quantify the effectiveness of cosmetic products. In this study, we present a systematic study of the mechanical properties of virgin skin and cream-treated skin. In nanoscratch measurements, the scratch wear tracks were generated at various loads using an atomic force microscope. Hardness and elastic moduli were measured using a nanoindenter. The in situ tensile properties of virgin skin and cream-treated skin were measured using a custom-built tensile stage that attaches to the atomic force microscope. Compared with virgin skin, cream-treated skin exhibits better scratch resistance up to a normal load of 15 μN. The indentation hardness and elastic modulus of cream-treated skin are lower than that of virgin skin, indicating that the skin cream moistens and softens the skin surface. In the stretching experiments, the elastic modulus is lower and ultimate strain is higher than that of virgin skin, indicating skin cream can improve the tensile response of skin. Mechanisms for the observed trends are discussed.     

摩擦表面膜纳米力学性能评定新方法

介绍了新型纳米压痕技术的基本测量原理，该技术利用加载-卸载过程中压痕对载荷和压入深度的敏感关系，测试材料的硬度和弹性模量等力学性能。由于该技术纳米级的压头位移和纳牛顿级的作用力，使之成为研究摩擦表面膜的有力工具，应用纳米压痕法对纳米铜摩擦表面膜进行了硬度和弹性模量的测试和分析。     

Nanomechanical characterization of human hair using nanoindentation and SEM

Human hair is a nanocomposite biological fiber with well-characterized microstructures. Nanomechanical characterization of human hair can help to evaluate the effect of cosmetic products on hair surface, can provide a better understanding of the physicochemical properties of a wide variety of composite biological systems, and can provide the dermatologists with some useful markers for the diagnosis of hair disorders. The paper presents systematic studies of nanomechanical properties of human hair including hardness, elastic modulus and creep, using the nanoindentation technique. The samples include Caucasian, Asian and African hair at virgin, chemo-mechanically damaged and treated conditions. Hair morphology was studied using scanning electron microscopy (SEM). Indentation experiments were performed on both the surface and cross-section of the hair, and the indents were studied using SEM. The nanomechanical properties of hair as a function of hair composition, microstructure, ethnicity, damage and treatment are discussed.     

Effects of ageing on the biomechanical properties of rat articular cartilage

Previous studies have demonstrated that male Sprague Dawley (SD) rats experience age-related bone loss with the same characteristics as that in ageing men. As articular cartilage, like bone, is a critical component of the health and function of the musculoskeletal system, the authors hypothesized that articular cartilage in the untreated male SD rats could be a suitable model for studying the age-related deterioration of articular cartilage in men. To test this hypothesis, male SD rats were killed at between 6 and 27 months. The right femur of each rat was removed. The effects of ageing on the structural integrity of the distal femoral articular cartilage were studied by biomechanical testing with a creep indentation apparatus. The aggregate modulus, Poisson's ratio, permeability, thickness, and percentage recovery of articular cartilage were determined using finite element/non-linear optimization modelling. No significant differences were observed in these biomechanical properties of the distal femoral articular cartilage as a function of age. Therefore, untreated male SD rats appear to be unsuitable for studying the age-related changes of articular cartilage as they occur in men. However, and more intriguingly, it is also possible that ageing does not affect the biomechanical properties of articular cartilage in the absence of cartilage pathology.     

Influence of protein conformation on frictional properties of poly (vinyl alcohol) hydrogel for artificial cartilage

Poly (vinyl alcohol) (PVA) hydrogel is one of the anticipated materials for artificial cartilage. In our previous studies, wear of PVA hydrogel depended on content of proteins in lubricants. The secondary structures of bovine serum albumin (BSA) and human gamma globulin (HGG) were investigated in circular dichroism spectroscopy to clarify the influence of the proteins on frictional properties. BSA and HGG were mainly composed of the α-helix and the β-sheet, respectively. BSA containing the α-helix structure showed low friction compared to HGG composed of the β-sheet structure in mixed or boundary lubrication mode. The α-helix structure forms low shear layer because the α-helix structure is easily released from surfaces and low cohesive strength. HGG forms uniform adsorption layer, but showed higher friction than BSA in the rubbing with single protein. In the repeated rubbing with changing of lubricants from HGG to BSA, however, the final friction was reduced, because an optimum layered structure of proteins was formed. Hence, layered structure of proteins appears to play an important role to protect rubbing surfaces and to reduce friction. In heat treatment tests, heat-induced BSA showed very low friction because of reduction of the α-helix structure. Heat-induced HGG did not show large differences from native HGG, but could not bring low friction with heat-induced BSA. Thus it was shown that the protein conformation has effective influences on friction.     

Substrate Effects on the Micro/Nanomechanical Properties of TiN Coatings

Nanoindentation and nanoscratch tests were performed for titanium nitride (TiN) coatings on different tool steel substrates to investigate the indentation/scratch induced deformation behavior of the coatings and the adhesion of the coating–substrate interfaces and their tribological property. In this work, TiN coatings with a thickness of about 500 nm were grown on GT35, 9Cr18 and 40CrNiMo steels using vacuum magnetic-filtering arc plasma deposition. In the nanoindentation tests, the hardness and modulus curves for TiN/GT35 reduced the slowest around the film thickness 500 nm with the increase of indentation depth, followed by TiN/9Cr18 and TiN/40CrNiMo. Improving adhesion properties of coating and substrate can decrease the differences of internal stress field. The scratch tests showed that the scratch response was controlled by plastic deformation in the substrate. The substrate plays an important role in determining the mechanical properties and wear resistance of such coatings. TiN/GT35 exhibited the best load-carrying capacity and scratch/wear resistance. As a consequence, GT35 is the best substrate for TiN coatings of the substrate materials tested.     

Gene therapy for articular cartilage repair

Articular cartilage serves as the gliding surface of joints. It is susceptible to damage from trauma and from degenerative diseases. Restoration of damaged articular cartilage may be achievable through the use of cell-regulatory molecules that augment the reparative activities of the cells, inhibit the cells' degradative activities, or both. A variety of such molecules have been identified. These include insulin-like growth factor I, fibroblast growth factor 2, bone morphogenetic proteins 2, 4, and 7, and interleukin-1 receptor antagonist. It is now possible to transfer the genes encoding such molecules into articular cartilage and synovial lining cells. Although preliminary, data from in-vitro and in-vivo studies suggest that gene therapy can deliver such potentially therapeutic agents to protect existing cartilage and to build new cartilage.     

Preparation of biomolecule gel matrices for electron microscopy

We report a new sample preparation method that allows the direct transmission electron microscopy evaluation of the architectural characteristics of biomolecules entrapped in gel matrices. We demonstrate that this sample preparation technique can be used for the identification and ultrastructural characterization of liposomes, collagen I and collagen III embedded in gel matrices, and has the potential to be useful for transmission electron microscopy (TEM) characterization of other biomolecule-gel matrix systems.     

Stem cells for tissue engineering of articular cartilage

Articular cartilage injuries are one of the most common disorders in the musculo-skeletal system. Injured cartilage tissue cannot spontaneously heal and, if not treated, can lead to osteoarthritis of the affected joints. Although a variety of procedures are being employed to repair cartilage damage, methods that result in consistent durable repair tissue are not yet available. Tissue engineering is a recently developed science that merges the fields of cell biology, engineering, material science, and surgery to regenerate new functional tissue. Three critical components in tissue engineering of cartilage are as follows: first, sufficient cell numbers within the defect, such as chondrocytes or multipotent stem cells capable of differentiating into chondrocytes; second, access to growth and differentiation factors that modulate these cells to differentiate through the chondrogenic lineage; third, a cell carrier or matrix that fills the defect, delivers the appropriate cells, and supports cell proliferation and differentiation. Stem cells that exist in the embyro or in adult somatic tissues are able to renew themselves through cell division without changing their phenotype and are able to differentiate into multiple lineages including the chondrogenic lineage under certain physiological or experimental conditions. Here the application of stem cells as a cell source for cartilage tissue engineering is reviewed.     

Mechanical and tribological properties of poly(hydroxyethyl methacrylate) hydrogels as articular cartilage substitutes

Poly(hydroxyethyl methacrylate) (p(HEMA)) hydrogels have been proposed as promising biomaterials to replace damaged articular cartilage. A major obstacle to their use as replacement bearing tissue is their poor mechanical properties in comparison with healthy articular cartilage. The purpose of this study was to obtain p(HEMA) hydrogels with physicochemical and mechanical properties close to healthy articular cartilage, by introducing a hydrophilic monomer, namely acrylic acid (AA). Formulations of hydrogels with different amounts of hydrophilic monomer (acrylic acid, AA) were synthesized and tested: p(HEMA), p(HEMA-co-5%AA), p(HEMA-co-25%AA). The macro-mechanical tests were reproduced at nanoscale in order to verify if the superficial properties of the hydrogels are similar to the bulk ones.     

Nanomechanical Probes of Single Corneal Epithelial Cells: Shear Stress and Elastic Modulus

Living human corneal epithelial cells have been probed in vitro via atomic force microscopy, revealing the frictional characteristics of single cells. Under cell media, measured shear stresses of 0.40 kPa demonstrate the high lubricity of epithelial cell surfaces in contact with a microsphere probe. The mechanical properties of individual epithelial cells have been further probed through nanometer scale indentation measurements. A simple elastic foundation model, based on experimentally verifiable parameters, is used to fit the indentation data, producing an effective elastic modulus of 16.5 kPa and highlighting the highly compliant nature of the cell surface. The elastic foundation model is found to more accurately fit the experimental data, to avoid unverifiable assumptions, and to produce a modulus significantly higher than that of the widely used Hertz–Sneddon model.     

Friction properties of nano-hydroxyapatite reinforced poly(vinyl alcohol) gel composites as an articular cartilage

Nano-hydroxyapatite reinforced poly(vinyl alcohol) gel (nano-HA/PVA gel) composite has been proposed as a promising biomaterial, especially used as an articular cartilage repair biomaterial. In this paper, nano-HA/PVA gel composite was prepared by in situ synthesis method and incorporation with freeze–thaw cycle process. The effects of various factors on the friction coefficient of the gel composites and stainless steel ball counterpart were investigated by a ball–plate friction and wear tester. The variations of the friction coefficient with influence factors were explored by Hertzian contact theory and elastohydrodynamic lubrication theory. The results show that the friction coefficient of the gel composites is positively to the normal load and inversely proportional to the sliding speed and the diameter of stainless steel ball. Furthermore, the friction coefficient of the gel composites is obviously lower than that in distilled water or in physiological saline, whereas, friction coefficient has little difference between in distilled water and in physiological saline.     

Techniques for assessment of wear between human cartilage surfaces

Osteoarthritis (OA) is a disease of joints, affecting a large number of people worldwide. One of the symptoms of OA is wear of articular cartilage; it is thought that among other factors this may be due to failure of lubrication. Injection of bio-lubricants into a joint may remedy this problem. Wear of cartilage and its prevention is a focus of much interest. The present paper describes wear tests performed using human cartilage on cartilage under various working conditions. Several techniques assessing wear are described, such as changes in surface morphology using optical profilometry and variation in the content of collagen and proteoglycans (PG) in the lubricating solution. Of all these techniques the PG content analysis was found to be the most efficient one.     

Tribology-optimised silk protein hydrogels for articular cartilage repair

Porous hydrogels were made from silk fibre as potential materials for cartilage repair. The aim was to develop materials which mimicked the tribological behaviour of cartilage, with controlled pore-sizes and optimised mechanical properties. Mechanical tests showed hydrogels had a comparable compressive modulus to cartilage, with stiffness improved by decreasing pore size. Under static loading and during shear hydrogels demonstrated significant interstitial fluid support. Friction testing showed the hydrogels had a cartilage-like frictional response, dominated by this interstitial fluid support. Silk hydrogels showed little wear, early signs of which were changes in surface morphology that did not correlate with the equilibrium friction coefficient. Consequently both wear and friction should be monitored when assessing the tribological performance of hydrogels.