Katarzyna Pogoda

Cancer cell recognition--mechanical phenotype.

The major characteristics of cancer metastasis is the ability of the primary tumor cells to migrate by way of the blood or lymph vessels and to form tumors at multiple, distant sites. There are evidences that cancer progression is characterized by disruption and/or reorganization of cytoskeleton (i.e. cellular scaffold). This is accompanied by various molecular alterations influencing the overall mechanical resistance of cells. Current approach in diagnosis focuses mainly on microbiological, immunological, and pathological aspects rather than on the biomechanics of diseases. The determination of mechanical properties of an individual living cell has became possible with the development of local measurement techniques, such as atomic force microscopy, magnetic or optical tweezers. The advantage of them lies in the capability to measure living cells at a single cell level and in liquid conditions, close to natural environment. Here, we present the studies on mechanical properties of single cells originating from various cancers. The results show that, independently of the cancer type (bladder, melanoma, prostate, breast and colon), single cells are characterized by the lower Youngs modulus, denoting higher deformability of cancerous cells. However, the obtained Youngs modulus values were dependent on various factors, like the properties of substrates used for cell growth, force loading rate, or indentation depth. Their influence on elastic properties of cells was considered. Based on these findings, the identification of cancerous cells based on their elastic properties was performed. These results proved the AFM capability in recognition of a single, mechanically altered cell, also in cases when morphological changes are not visible. The quantitative analysis of cell deformability carried out using normal (reference) and cancerous cells and, more precisely, their characterization (qualitative and quantitative) can have a significant impact on the development of methodological approaches toward precise identification of pathological cells and would allow for more effective detection of cancer-related changes.

Cancer cell detection in tissue sections using AFM.

Currently, cancer diagnosis relies mostly on morphological examination of exfoliated, aspirated cells or surgically removed tissue. As long as standard diagnosis is concerned, this classical approach seems to be satisfactory. In the recent years, cancer progression has been shown to be accompanied by alterations in mechanical properties of cells. This offers the detection of otherwise unnoticed cancer cell disregarded by histological analysis due to insignificant manifestations. One of techniques, sensitive to changes in mechanical properties, is the atomic force microscopy, which detects cancer cells through their elastic properties. Such measurements were applied to tissue sections collected from patients suffering from various cancers. Despite of heterogeneity and complexity of cancer cell sections, the use of the Youngs modulus as an indicator of cell elasticity allow for detection of cancer cells in tissue slices.

Mid-infrared spectroscopy and microscopy of subcellular structures in eukaryotic cells with atomic force microscopy – infrared spectroscopy

Atomic force microscopy – infrared (AFM-IR) spectroscopy allows spectroscopic studies in the mid-infrared (mid-IR) spectral region with a spatial resolution better than is allowed by the diffraction limit. We show that the high spatial resolution can be used to perform spectroscopic and imaging studies at the subcellular level in fixed eukaryotic cells. We collect AFM-IR images of subcellular structures that include lipid droplets, vesicles and cytoskeletal filaments, by relying on the intrinsic contrast from IR light absorption. We also obtain AFM-IR absorption spectra of individual subcellular structures. Most spectra show features that are recognizable in the IR absorption spectra of cells and tissue obtained with FTIR technology, including absorption bands characteristic of phospholipids and polypeptides. The quality of the spectra and of the images opens the way to structure and composition studies at the subcellular level using mid-IR absorption spectroscopy.

Stiffening of bacteria cells as a first manifestation of bactericidal attack

Development of new antibacterial agents requires generation of new methods that will allow characterization of mechanisms of novel antibacterial actions as well as observation of pathogen susceptibility to antibiotics. A family of synthetic cationic lipids, such as ceragenins, which mimic the action of membrane-targeting natural cationic antibacterial peptides, shows promise as a new family of bactericidal agents. Here we report that treatment of Bacillus subtillis cells with human cathelicidin LL-37 and ceragenins CSA-13 and CSA-131 (1-100μg/mL) leads to physicochemical changes of bacterial cells surface. Especially nanomechanical alternations as quantified by bacterial stiffness and adhesion measurements shows that bacteria respond actively to treatment by tuning their surface properties and this response is tightly controlled by the concentration of bactericidal agents. Following the real-time changes of bacteria stiffness, we observed, that Bacillus subtilis cell envelope can initially stiffen, but in the long-time treatment it softens compare to untreated cells. We postulate, that nanomechanical responses may be considered as a new way to assess some aspects of antimicrobial activity that evolve in time, especially for molecules targeting bacterial membranes.

Assessment of aliphatic poly(ester-carbonate-urea-urethane)s potential as materials for biomedical application

Selected mechanical and biological properties of biodegradable elastomeric poly(ester-carbonate-urea-urethane)s (PECUUs) point towards their potential to be applied as scaffolds in tissue engineering. Here we explore their medical applicability taking into account their hemocompatibility and cytotoxicity. The influence of the ester monomer (derivatives of adipic and succinic acids), as well as diisocyanate type (IPDI and HDI) on the investigated PECUUs properties is presented. The presence of aliphatic diisocyanates, cyclic IPDI or linear HDI, governs the adhesion of Candida cells to these polymers offering the possibility to control the biofilm formation on their surface. In comparison to the linear form, cyclic diisocyanates with pentamethylene succinate or adipate fragments had two to three times lower biofilm mass formation on their surface. Reduced hemoglobin release from red blood cells observed during incubation of tested polymers with human erythrocytes suspension indicates their potential biocompatibility with human tissues. PECUUs were also able to support the growth of human keratinocytes HaCaT on their surface when coated with collagen. In effect, IPDI derivatives might possess a high potential for use in biomedical applications.

Loss of vimentin intermediate filaments decreases peri-nuclear stiffness and enhances cell motility through confined spaces.

The migration of cells through tight constricting spaces or along fibrous tracks in tissues is important for biological processes, such as embryogenesis, wound healing, and cancer metastasis, and depends on the mechanical properties of the cytoskeleton. Migratory cells often express and upregulate the intermediate filament protein vimentin. The viscoelasticity of vimentin networks in shear deformation has been documented, but its role in motility is largely unexplored. Here, we studied the effects of vimentin on cell motility and stiffness using mouse embryo fibroblasts derived from wild-type and vimentin-null mice. We find that loss of vimentin increases motility through small pores and along thin capillaries. Atomic force microscopy measurements reveal that the presence of vimentin enhances the perinuclear stiffness of the cell, to an extent that depends on surface ligand presentation and therefore signaling from extracellular matrix receptors. Together, our results indicate that vimentin hinders three-dimensional motility by providing mechanical resistance against large strains and may thereby protect the structural integrity of cells.

Modification of Polymer Substrates with Extreme Ultraviolet - Potential Application in Cancer Cell Identification

Conformal Invariance and Conserved Quantities for Lagrange Equation of Thin Elastic Rod Peng Wanga,∗, Hui-Rong Feng and Zhi-Mei Lou School of Civil Engineering and Architecture, University of Jinan, Jinan, Shangdong, 250022, P.R. China College of Transportation and Civil Engineering, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, P.R. China Department of Physics, Shaoxing University, Shaoxing, Zhejiang, 312000, P.R. China