Studying the rate of cell migration provides insight into fundamental cell biology as well as a tool to assess the functionality of synthetic surfaces and soluble environments used in tissue engineering. The traditional tools used to study cell migration include the fence and wound healing assays. In this paper we describe the development of a microchannel based device for the study of cell migration on defined surfaces. We demonstrate that this device provides a superior tool, relative to the previously mentioned assays, for assessing the propagation rate of cell wave fronts. The significant advantage provided by this technology is the ability to maintain a virgin surface prior to the commencement of the cell migration assay. Here, the device is used to assess rates of mouse fibroblasts (NIH 3T3) and human osteosarcoma (SaOS2) cell migration on surfaces functionalized with various extracellular matrix proteins as a demonstration that confining cell migration within a microchannel produces consistent and robust data. The device design enables rapid and simplistic assessment of multiple repeats on a single chip, where surfaces have not been previously exposed to cells or cellular secretions.
Cell attachment, expansion, and migration in three-dimensional biomaterials are crucial steps for effective delivery of osteogenic cells into bone defects. Complexes composed of vitronectin (VN), insulin-like growth factors (IGFs), and insulin growth factor-binding proteins (IGFBPs) have been reported to enhance cell attachment, proliferation, and migration in a variety of cell lines in vitro. The aim of this study was to examine whether prebound complexes of VN and IGFs +/- IGFBPs could facilitate human osteoblast serum-free expansion in vitro and enhance cell attachment, proliferation, and migration in three-dimensional biomaterial constructs. Human osteoblasts derived from alveolar bone chips and the established human osteoblast cell line Saos-2 were used. These cells were seeded on tissue culture plates and porous scaffolds of type I collagen sponges and polyglycolic acid (PGA), which had been coated with VN +/- IGFBP-5 +/- IGF-I. Cell attachment, proliferation, and migration were evaluated by cell counting, confocal microscopy, and scanning electron microscopy. The number of attached human osteoblasts was significantly higher in VN-coated polystyrene culture dishes. Furthermore, significant increases in cell proliferation were observed when growth factors were bound to these surfaces in the presence of VN. In the two scaffold materials examined, greater cell attachment was found in type I collagen sponges compared with PGA scaffolds. However, coating the scaffolds with complexes composed of VN + IGF-I or VN + IGFBP-5 + IGF-I enhanced cell attachment on PGA. Moreover, the presence of VN + IGFBP-5 + IGF-I resulted in significantly greater osteoblast migration into deep pore areas as compared with untreated scaffolds or scaffolds treated with fetal calf serum. These results demonstrated that complexes of VN + IGFBP-5 + IGF-I can be used to expand osteoblasts in vitro under serum-free conditions and enhance the attachment and migration of human osteoblasts in three-dimensional culture. This in turn suggests a potential application in surface modification of biomaterials for tissue reconstruction.
During the distribution phase, packaged consignments are exposed to a variety of environmental hazards (such as vibrations) that, if excessively severe, may cause damage to or even destroy the product. Structural deterioration can be tracked by monitoring variations in the packaging system's modal parameters, particularly its natural frequency (stiffness). Natural frequency estimates are often extracted using a least‐squares regression curve fit, applied to an estimate of the system's frequency response function (FRF). FRF estimates are generally obtained using the Fourier transform with a single input and single output (SISO). This approach is suitable for many applications; however, as the non‐linearity of the system under analysis increases, the ability of this technique to accurately monitor changes in the system will decrease. In addition, when the excitation to the non‐linear system is varied (increased or decreased amplitude), a SISO‐based approach may indicate a shift in natural frequency (as a result of the varied input) even though no change in the condition of the system has occurred. This paper discusses an approach that is designed to separate the linear component of the system's FRF using a reverse multiple input/single output (RMISO) algorithm. Such separation will allow traditional modal parameter extraction (curve fitting) techniques to be used to monitor the condition of non‐linear systems. The paper presents the results of experiments in which expanded polystyrene samples were subjected to broad‐band random base excitation with a free‐moving load placed atop the cushion sample. Continuous acceleration measurements of the vibration table and the free‐moving load were used to compute the FRFs of the cushions, and the differences between a conventional (SISO) approach and the proposed RMISO‐based parameter extraction technique were evaluated. Copyright © 2014 John Wiley & Sons, Ltd.
The Reverse Multiple Input–Single Output (R‐MISO) identification technique was implemented to determine vibration transmissibility frequency response functions (FRFs) of a cushion‐product system that accounts for nonlinearities known to exist in such systems. The signed quadratic x|x| nonlinear term was used. The results were compared with the classical linear approximation FRFs calculated with the single input–single output identification method. Comparisons were made for five types of expanded polystyrene (EPS) cushions with varying density and two levels of band‐limited random excitation. It was shown that vibration transmissibility FRFs obtained with the R‐MISO method and with the inclusion of a signed quadratic x|x| nonlinear term significantly (up to ~70%) improved the total coherence of the nonlinear vibration transmissibility FRFs for the EPS cushions subjected to the static load selected to coincide with the optimum of their respective cushion curves. The R‐MISO technique was successful in accounting for some of the nonlinearities in the EPS cushion‐product system. However, its ability to deal with nonlinearities is not specific to the EPS but rather common to cushion‐product systems subjected to vertical vibrations. Copyright © 2012 John Wiley & Sons, Ltd.
Retail grocery price variations in Dublin are examined with a view to confirming that systematic structural and locational variations exist at different time periods. The relevance of the price component to central-place theory, consumer shopping behaviour, and low-income consumers is briefly indicated prior to a detailed review of previous price-variation studies. Two surveys in Dublin indicate temporal consistency in certain hypotheses for different ranges of products, and more detailed analyses indicate the importance of organisational structure in accounting for retail price variations. A stepwise multiple-regression analysis is developed to determine the best predictors of price variations. The data are also examined to determine whether low-income consumers pay significantly more for their groceries than the more affluent and mobile socioeconomic groups.
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