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The clinical benefits of electromagnetic field (EMF) therapy in enhancing osteogenesis have been acknowledged for decades, but agreement regarding the underlying mechanisms continues to be sought. Studies have shown EMFs to promote osteoblast-like cell proliferation, or contrarily, to induce differentiation and enhance mineralization. Typically these disparities have been attributed to methodological differences. The present paper argues the possibility that the chosen osteoblast model impacts stimulation outcome. Phenotypically immature cells, particularly at low seeding densities, appear to be prone to EMF-amplified proliferation. Conversely, mature cells at higher densities seem to be predisposed to earlier onset differentiation and mineralization. This suggests that EMFs augment ongoing processes in cell populations. To test this hypothesis, mature SaOS-2 cells and immature MC3T3-E1 cells at various densities, with or without osteo-induction, were exposed to sinusoidal 50 Hz EMF. The exposure stimulated the proliferation of MC3T3-E1 and inhibited the proliferation of SaOS-2 cells. Baseline alkaline phosphatase (ALP) expression of SaOS-2 cells was high and rapidly further increased with EMF exposure, whereas ALP effects in MC3T3-E1 cells were not seen until the second week. Thus both cell types responded differently to EMF stimulation, corroborating the hypothesis that the phenotypic maturity and culture stage of cells influence stimulation outcome. [ABSTRACT FROM AUTHOR]

Background Recurrent aphthous ulcer ( RAU) is an ulcerative disease of non-keratinized oral mucosa. Colon and bronchial epithelial cells produce interleukin-17 C ( IL-17 C) upon stimulation of Toll-like receptor 2 ( TLR2), TLR3 and TLR5, which are highly expressed in epithelial cells in RAU lesions. We therefore investigated the eventual presence and function of IL-17 C in cultured human oral keratinocytes ( HOK) and control biopsies compared to RAU lesions. Methods Expression of IL-17 A, IL-17 C, IL-17 RA and IL-17 RE was analysed in cultured HOK cells using quantitative real-time polymerase chain reaction (q RT- PCR). HOK cells were stimulated with IL-17 C and analysed for IL-8 and tumour necrosis factor-α ( TNF-α) using q RT- PCR. Control mucosa ( n = 5) was immunostained for IL-17 A, IL-17 C, IL-8, TNF-α and mast cell tryptase and compared with RAU lesions ( n = 5) using the mean grey scale value. Results IL-17 C, but no IL-17 A, m RNA was found in cultured HOK cells. Components of the heterodimeric IL-17 RA/ IL-17 RE receptor for IL-17 C were also highly expressed. Stimulation of HOK with IL-17 C increased TNF-α m RNA ( P = 0.03; IL-8 increase was not statistically significant). HOK in RAU lesions stained intensively for IL-17 C compared to controls ( P = 0.006). This was associated with increased epithelial immunostaining of TNF-α ( P = 0.04) and IL-8 ( P = 0.02). Most of the inflammatory cells which stained for IL-17 A in control mucosa and RAU lesions were also mast cell tryptase positive. Conclusion IL-17 C is highly expressed in epithelial cells in RAU lesions, where it seems to stimulate oral keratinocytes via IL-17 RA/ IL-17 RE to produce pro-inflammatory cytokines. Human oral epithelial cells are probably important inflammatory cells in RAU. [ABSTRACT FROM AUTHOR]

Osteogenic responses of human mesenchymal stromal cells (hMSCs) were compared on square-patterned, inverse square-patterned, and planar titanium, chromium, diamond-like carbon (DLC), and tantalum; hypothesis was that both the materials and patterns affect osteogenesis. Samples were produced using photolithography and physical vapor deposition. Early-marker alkaline phosphatase (ALP) and mid-markers, small body size and mothers against decapentaplegic-related protein-1 (SMAD1), runt-related transcription factor-2 (RUNX2), and osteopontin were studied using quantitative real-time polymerase chain reaction. ALP and hydroxyapatite, were colorimetrically studied. ALP reached highest values on both patterned titanium samples, but mid-markers disclosed that it was already lagging behind planar and inverse patterned tantalum. Hydroxyapatite formation disclosed that osteo-induced hMSCs passed all the differentiation stages (except on planar chromium). Presence of hydroxyapatite disclosed that both types of patterning promoted (p < 0.001) osteogenesis compared to planar samples. Results suggest that the osseocompatibility/integration of implants could be improved by changing the monotonous and featureless implant–host interface into micro-patterned interface to provide physical differentiation cues. [ABSTRACT FROM AUTHOR]

We hypothesized that when compared with conventional two-dimensional (2D) cultures, substrates containing 3D micropillars would allow cells to grow at levels, activating their cytoskeleton to promote osteogenesis. Fibroblasts, osteoblast-like cells, and mesenchymal stem cells (MSCs) were studied. Planar substrates were compared with 200-nm-, 5-μm-, and 20-μm-high pillars of Ormocomp®, Si, diamond-like carbon, or TiO2. Scanning electron microscopy and staining of actin cytoskeleton showed 7.5-h adhesion to pillar edges and 5-day stretching between adhesion contacts > 100-μm distances of fibroblast and MSC in 3D networks, whereas SaOS-2 cells adhered flatly and individually on horizontal and vertical surfaces. ERK and ROCK immunostaining at 14 and 21 days confirmed activation of the cytoskeleton. In contrast to expectations, success to induce osteogenesis was dominated by the cytocompatibility of the substrate over the 3D structure. This was shown using early alkaline phosphatase, intermediate osteopontin, and late mineralization markers, together with bone nodule formation, which were seen in planar substrates and low-profile TiO2 pillars, but were poor in the 20-μm landscape. The lack of intercellular contacts seems to halt the osteogenesis-promoting effects of cytoskeletal organization and tension described earlier. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 101A: 842-852, 2013. [ABSTRACT FROM AUTHOR]

Regenerative medicine is a multidisciplinary field utilizing the potential of stem cells and the regenerative capability of the body to restore, maintain, or enhance tissue and organ functions. Stem cells are unspecialized cells that can self-renew but also differentiate into several somatic cells when subjected the appropriate environmental cues. The ability to reliably direct stem cell fate would provide tremendous potential for basic research and clinical therapies. Proper tissue function and regeneration rely on the spatial and temporal control of biophysical and biochemical cues, including soluble molecules, cell-cell contacts, cell-extracellular matrix contacts, and physical forces. The mechanisms involved remain poorly understood. This review focuses on the stem cell-extracellular matrix interactions by summarizing the observations of the effects of material variables (such as overall architecture, surface topography, charge, ζ-potential, surface energy, and elastic modulus) on the stem cell fate. It also deals with the mechanisms underlying the effects of these extrinsic, material variables. Insight in the environmental interactions of the stem cells is crucial for the development of new material-based approaches for cell culture experiments and future experimental and clinical regenerative medicine applications. [ABSTRACT FROM AUTHOR]

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is increasingly being used in analysis of biological samples. For example, it has been applied to distinguish healthy and osteoarthritic human cartilage. This chapter discusses ToF-SIMS principle and instrumentation including the three modes of analysis in ToF-SIMS. ToF-SIMS sets certain requirements for the samples to be analyzed; for example, the samples have to be vacuum compatible. Accordingly, sample processing steps for different biological samples, i.e., proteins, cells, frozen and paraffin-embedded tissues and extracellular matrix for the ToF-SIMS are presented. Multivariate analysis of the ToF-SIMS data and the necessary data preprocessing steps (peak selection, data normalization, mean-centering, and scaling and transformation) are discussed in this chapter.

For therapeutic purposes, an accurate measurement of dopamine level in situ would be highly desirable. A novel strategy for the selective determination of dopamine concentration based on the diamond-like carbon (DLC) electrode is presented in this abstract. The developed DLC electrode is able to detect 10 µM dopamine and has improved sensitivity compared to platinum. Compared to carbon fiber electrodes, the DLC electrode is more stable because the background current is much lower.

Micro-textured biomaterials might enhance cytocompatibility of silicon-based micro-electro-mechanical system (bio-MEMS) dummies. Photolithography-physical vapour deposition was used to produce diamond-like carbon (DLC) or Ti squares and circles on silicon, and also their inverse replicas; then DLC and Ti were compared for their guiding potential, using a SaOS-2 cell model. Scanning electron microscopy at 48 hours indicated cells were well-spread on large-sized patterns (several cells on one pattern) and assumed the geometrical architecture of underlying features. Medium-sized patterns (slightly smaller than solitary indicator cells) were inhabited by singular cells, which stretched from one island to another, assuming longitudinal or branching morphologies. On small-sized patterns (much smaller than individual cells;rpar; cells covered large micro-textured areas, but cellular filopodia bypassed the bare silicon. Immunofluorescence and confocal laser scanning microscopy indicated that the actin cytoskeleton and vinculin-containing adhesion junctions were present on the patterned areas, but not on the bare silicon. Cell density/coverage disclosed a 3.4-3.7-fold preference for the biomaterial patterns over silicon substrate (p 0.001). Differences in the cellular response between materials were lost at 120 hours when cells were confluent. The working hypothesis was proven; enhancement by micro-patterning depends on the pattern size, shape and material and can be used to improve biocompatibility during the initial integration phase of the device.