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Abstract Rapid and strategic cell placement is necessary for high throughput tissue fabrication. Current adhesive cell patterning systems rely on fluidic shear flow to remove cells outside of the patterned regions, but limitations in washing complexity and uniformity prevent adhesive patterns from being widely applied. Centrifugation is commonly used to study the adhesive strength of cells to various substrates; however, the approach has not been applied to selective cell adhesion systems to create highly organized cell patterns. This study shows centrifugation as a promising method to wash cellular patterns after selective binding of cells to the surface has taken place. After patterning H9C2 cells using biotin-streptavidin as a model adhesive patterning system and washing with centrifugation, there is a significant number of cells removed outside of the patterned areas of the substrate compared to the initial seeding, while there is not a significant number removed from the desired patterned areas. This method is effective in patterning multiple size and linear structures from line widths of 50–200 μm without compromising immediate cell viability below 80%. We also test this procedure on a variety of tube-forming cell lines (MPCs, HUVECs) on various tissue-like surface materials (collagen 1 and Matrigel) with no significant differences in their respective tube formation metrics when the cells were seeded directly on their unconjugated surface versus patterned and washed through centrifugation. This result demonstrates that our patterning and centrifugation system can be adapted to a variety of cell types and substrates to create patterns tailored to many biological applications. 

During epithelial-to-mesenchymal transition (EMT), cancer cells lose their cell–cell adhesion junctions as they become more metastatic, altering cell motility and focal adhesion disassembly associated with increased detachment from the primary tumor and a migratory response into nearby tissue and vasculature. Current in vitro strategies characterizing a cell's metastatic potential heavily favor quantifying the presence of cell adhesion biomarkers through biochemical analysis; however, mechanical cues such as adhesion and motility directly relate to cell metastatic potential without needing to first identify a cell specific biomarker for a particular type of cancer. This paper presents a comprehensive comparison of two functional metrics of cancer aggression, wound closure migration velocity and cell detachment from a culture surface, for three pairs of epithelial cancer cell lines (breast, endometrium, tongue tissue origins). It was found that one functional metric alone was not sufficient to categorize the cancer cell lines; instead, both metrics were necessary to identify functional trends and accurately place cells on the spectrum of metastasis. On average, cell lines with low metastatic potential (MCF-7, Ishikawa, and Cal-27) were more aggressive through wound closure migration compared to loss of cell adhesion. On the other hand, cell lines with high metastatic potential (MDA-MB-231, KLE, and SCC-25) were on average more aggressive through loss of cell adhesion compared to wound closure migration. This trend was true independent of the tissue type where the cells originated, indicating that there is a relationship between metastatic potential and the predominate type of cancer aggression. Our work presents one of the first combined studies relating cell metastatic potential to functional migration and adhesion metrics across cancer cell lines from selected tissue origins, without needing to identify tissue-specific biomarkers to achieve success. Using functional metrics provides powerful clinical relevancy for future predictive tools of cancer metastasis. 

Adhesion of bacteria to oral implant surfaces can lead to oral infections, and the prevention of strong biofilm adherence to implant surfaces can assist in the prevention of these infections like peri-implantitis. In prior studies, single species biofilm adhesion has been quantitatively measured via the laser spallation technique. However, colonizing oral biofilms rarely consists of a single bacteria species. Multiple early colonizer species, including several strains of Streptococci, dominate initial oral biofilm formation. This study aims to characterize the adhesion of a multi-species oral biofilm consisting of S. oralis, S. sanguinis, and S. gordonii on titanium, a common implant material, using the laser spallation technique. Previous work has established these specific Streptococci strains as a multi-species periodontal biofilm model. This study is the first to provide a quantitative adhesion measurement of this multi-species model onto a dental implant surface. First, adhesion strength of the multi-species model is compared to adhesion strength of the single-species streptococci constituents. Fluorescent staining and imaging by fluorescent microscopy are used to identify individual bacteria species within the biofilm. The multi-species biofilm presented in this study provides a more representative model of in vivo early biofilms and provides a more accurate metric for understanding biocompatibility on implant surfaces. 

Increasing antibiotic resistance in bacteria is a critical issue that often leads to infections or other morbidities. Mechanical properties of the bacterial cell wall, such as thickness or elastic modulus, may contribute to the ability of a bacterial cell to resist antibiotics. Techniques like atomic force microscopy (AFM) are used to quantify bacterial cell mechanical properties and image cell structures at nanoscale resolutions. An additional benefit of AFM is the ability to probe samples submerged in liquids, meaning that live bacteria can be imaged or evaluated in environments that more accurately simulate in vivo conditions as compared to other methods like electron microscopy. However, because AFM measurements are highly sensitive to small perturbations in the deflection of the tip of a sensor probe brought into contact with the specimen, immobilization of bacteria prior to measurement is essential for accurate measurements. Traditional chemical fixatives crosslink the molecules within the bacterial cell wall, which prevent the bacteria from locomotion. While effective for imaging, chemical crosslinkers are known to affect the measured stiffness of eukaryotic cells and also may affect the measured stiffness of the bacterial cell wall. Alternative immobilization methods include Cell-Tak™, an adhesive derived from marine mussels that does not interact with the bacterial wall and filters with known pore sizes which entrap bacteria. Previous studies have examined the effect of these immobilization methods on successful imaging of bacteria but have not addressed differences in measured modulus. This study compares the effects of immobilization methods including chemical fixatives, mechanical entrapment in filters, and Cell-Tak™ on the stiffness of the bacterial cell wall as measured by force spectroscopy. 

Bacteria can proliferate orthopedic implants, resulting in infection rates as high as 5%. A consistent problem across implantology is the development of surfaces which successfully promote the adhesion and propagation of healthy fibroblast and osteoblast cells while deterring formation of bacterial biofilms. Selecting surface configurations which favor cell adhesion will lead to decreased infection rates. Progress in identifying appropriate surface configurations is hindered by the lack of quantitative adhesion techniques capable of comparing adhesion of cells and biofilms directly. Recent advancements in adhesion techniques have allowed for quantitatively measured adhesion strengths of both bacterial biofilms and cell monolayers using the laser spallation technique. The quantified stress-based adhesion values allow surface and environmental factors that modulate both bacterial and cell adhesion to implant surfaces to be evaluated. During implantation, blood propagates wound sites completely coating implant surfaces. Quantitatively determining the impact of preconditioning layers that accumulate on the implant surface on cell adhesion is vital to predict implant behavior. Previous work has demonstrated that these preconditioning layers either negatively or neutrally impact bacterial adhesion to titanium implant surfaces. This study focuses on the impact that blood plasma and fibronectin coatings have on the adhesion of osteoblastic (MG 63) cells and fibroblasts to the same titanium surfaces. Adhesion results indicate that preconditioning layers and increased surface roughness positively impact cell adhesion. Incorporating the increased adhesion values for cell adhesion into the Adhesion Index demonstrates that increased surface roughness, coupled with natural wound healing preconditioning of surfaces, yields positive biocompatibility. 

Bacterial biofilms associated with implants remain a significant source of infections in dental, implant, and other healthcare industries due to challenges in biofilm removal. Biofilms consist of bacterial cells surrounded by a matrix of extracellular polymeric substance (EPS), which protects the colony from many countermeasures, including antibiotic treatments. Biofilm EPS composition is also affected by environmental factors. In the oral cavity, the presence of sucrose affects the growth of Streptococcus mutans that produce acids, eroding enamel and forming dental caries. Biofilm formation on dental implants commonly leads to severe infections and failure of the implant. This work determines the effect of sucrose concentration on biofilm EPS formation and adhesion of Streptococcus mutans, a common oral colonizer. Bacterial biofilms are grown with varying concentrations of sucrose on titanium substrates simulating dental implant material. Strategies for measuring adhesion for films such as peel tests are inadequate for biofilms, which have low cohesive strength and will fall apart when tensile loading is applied directly. The laser spallation technique is used to apply a stress wave loading to the biofilm, causing the biofilm to delaminate at a critical tensile stress threshold. Biofilm formation and EPS structures are visualized at high magnification with scanning electron microscopy (SEM). Sucrose enhanced the EPS production of S. mutans biofilms and increased the adhesion strength to titanium, the most prevalent dental implant material. However, there exists a wide range of sucrose concentrations that are conducive for robust formation and adhesion of S. mutans biofilms on implant surfaces.