Flat Control Research Articles

Reducing Cancer Cell Adhesion using Microtextured Surfaces.

For the past century, trypsin has been the primary method of cell dissociation, largely without any major changes to the process. Enzymatic cell detachment strategies for large-scale cell culturing processes are popular but can be labor-intensive, potentially lead to the accumulation of genetic mutations, and produce large quantities of liquid waste. Therefore, engineering surfaces to lower cell adhesion strength could enable the next generation of cell culture surfaces for delicate primary cells and automated, high-throughput workflows. In this study, a process for creating microtextured polystyrene (PS) surfaces to measure the impact of microposts on the adhesion strength of cells is developed. Cell viability and proliferation assays show comparable results in twocancer cell lines between micropost surfaces and standard cell culture vessels. However, cell image analysis on microposts reveals that cell area decreases by half, and leads to an average twofold increase in cell length per area. Using a microfluidic-based method up to a seven times greater percentage of cells are removed from micropost surfaces than the flat control surfaces. These results show that micropost surfaces enable decreased cell adhesion strength while maintaining similar cell viabilities and proliferation as compared to flat PS surfaces.

Study on Extending Range of Artillery Rocket Using Control Surfaces

Extension of range of artillery rocket using gliding technique is very effective way. Extension in the range is achieved by control surfaces without adding any extra charge. Controls are applied after completion of launch phase and trajectory is controlled by canards or flat tail control fins. Contribution in this work is to find the optimum time to initiate the controls during the flight so as to get maximum range. Analysis has been carried out using six degree of freedom equations of motion.

Flatness-based control in successive loops for electropneumatic actuators and robots

The control problem for the nonlinear dynamics of robotic and mechatronic systems with electropneumatic actuation is solved with the use of a flatness-based control approach which is implemented in successive loops. The state-space model of these systems is separated into a series of subsystems, which are connected between them in cascading loops. Each one of these subsystems can be viewed independently as a differentially flat system and control about it can be performed with inversion of its dynamics as in the case of input–output linearized flat systems. In this chain of i=1,2,…,N subsystems, the state variables of the subsequent (i+1th) subsystem become virtual control inputs for the preceding (ith) subsystem, and so on. In turn, exogenous control inputs are applied to the last subsystem and are computed by tracing backwards the virtual control inputs of the preceding N−1 subsystems. The whole control method is implemented in successive loops and its global stability properties are also proven through Lyapunov stability analysis. The validity of the control method is confirmed in two case studies: (a) control of an electropneumatic actuator, (ii) control of a multi-DOF robotic manipulator with electropneumatic actuators.

Synergetic effect of 3D porous microsphere structure and activation of adenosine A2B receptor signal on promoting osteogenic differentiation of BMSCs

Biodegradable microspheres offer great potential as functional building blocks for bottom-up bone tissue engineering. However, it remains challenging to understand and regulate cell behaviors in fabrication of injectable bone microtissues using microspheres. The study aims to develop an adenosine functionalized poly (lactide-co-glycolide) (PLGA) microsphere to enhance cell loading efficiency and inductive osteogenesis potential, and subsequently to investigate adenosine signaling-mediated osteogenic differentiation in cells grown on three-dimensional (3D) microspheres and flat control. Adenosine was loaded on PLGA porous microspheres via polydopamine coating, and the cell adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) were improved on these microspheres. It was found that adenosine A2B receptor (A2BR) was further activated by adenosine treatment, which consequently enhanced osteogenic differentiation of BMSCs. This effect was more obvious on 3D microspheres compared to 2D flats. However, the promotion of osteogenesis on the 3D microspheres was not eliminated by blocking the A2BR with antagonist. Finally, adenosine functionalized microspheres could fabricate injectable microtissues in vitro, and improve cell delivery and osteogenic differentiation after injection in vivo. Therefore, it is considered that adenosine loaded PLGA porous microspheres will be of good value in minimally invasive injection surgery and bone tissue repair.

Submicron-Grooved Films Modulate the Directional Alignment and Biological Function of Schwann Cells.

Topographical cues on material surfaces are crucial for guiding the behavior of nerve cells and facilitating the repair of peripheral nerve defects. Previously, micron-grooved surfaces have shown great potential in controlling nerve cell alignment for studying the behavior and functions of those cells and peripheral nerve regeneration. However, the effects of smaller-sized topographical cues, such as those in the submicron- and nano-scales, on Schwann cell behavior remain poorly understood. In this study, four different submicron-grooved polystyrene films (800/400, 800/100, 400/400, and 400/100) were fabricated to study the behavior, gene expression, and membrane potential of Schwann cells. The results showed that all submicron-grooved films could guide the cell alignment and cytoskeleton in a groove depth-dependent manner. Cell proliferation and cell cycle assays revealed that there was no significant difference between the submicron groove samples and the flat control. However, the submicron grooves can direct the migration of cells and upregulate the expression of critical genes in axon regeneration and myelination (e.g., MBP and Smad6). Finally, the membrane potential of the Schwann cells was significantly altered on the grooved sample. In conclusion, this study sheds light on the role of submicron-grooved patterns in regulating the behavior and function of Schwann cells, which provides unique insights for the development of implants for peripheral nerve regeneration.

Evaluation of focal adhesion mediated subcellular curvature sensing in response to engineered extracellular matrix.

Fibril curvature is bioinstructive to attached cells. Similar to natural healthy tissues, an engineered extracellular matrix can be designed to stimulate cells to adopt desired phenotypes. To take full advantage of the curvature control in biomaterial fabrication methodologies, an understanding of the response to fibril subcellular curvature is required. In this work, we examined morphology, signaling, and function of human cells attached to electrospun nanofibers. We controlled curvature across an order of magnitude using nondegradable poly(methyl methacrylate) (PMMA) attached to a stiff substrate with flat PMMA as a control. Focal adhesion length and the distance of maximum intensity from the geographic center of the vinculin positive focal adhesion both peaked at a fiber curvature of 2.5 μm-1 (both ∼2× the flat surface control). Vinculin experienced slightly less tension when attached to nanofiber substrates. Vinculin expression was also more affected by a subcellular curvature than structural proteins α-tubulin or α-actinin. Among the phosphorylation sites we examined (FAK397, 576/577, 925, and Src416), FAK925 exhibited the most dependance on the nanofiber curvature. A RhoA/ROCK dependance of migration velocity across curvatures combined with an observation of cell membrane wrapping around nanofibers suggested a hybrid of migration modes for cells attached to fibers as has been observed in 3D matrices. Careful selection of nanofiber curvature for regenerative engineering scaffolds and substrates used to study cell biology is required to maximize the potential of these techniques for scientific exploration and ultimately improvement of human health.

A 17.7–19.2-GHz Receiver Front End With an Adaptive Analog Temperature- Compensation Scheme

This article presents an eight-element 17.7–19.2-GHz receiver front end with 1–2 concurrent beams in a 65-nm CMOS technology. Each output beam utilizes a temperature-compensation variable-gain amplifier (TC-VGA) to minimize the temperature-induced gain variation. The concept of the proposed TC-VGA and the realization of corresponding adaptive analog control are introduced in detail. The front-end architecture and circuit-level design to enable a flat wideband gain response, precise phase and amplitude control, low power consumption, and adaptive analog temperature compensation are presented. Wafer probing is conducted to measure the performance of the receiver front end. The measured gain-temperature coefficient is ±0.005 dB/°C from −15°C to 85 °C at 17.7–19.2 GHz, while the counterpart without temperature compensation is −0.1 dB/°C. The chip demonstrates a 28-dB power gain, a 26% 3-dB fractional bandwidth, a 3.2–4.1-dB noise figure (NF), and a −27.4-dBm input 1-dB gain compression point (IP <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$_{\mathrm {1\,dB}}$ </tex-math></inline-formula> ) for each element. In addition, each channel provides 7-bit phase-shifting resolution and 6-bit attenuation for a 15.75-dB gain range with a <1.5° root-mean-square (rms) phase error, and a < 0.22-dB rms amplitude error. The chip occupies <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$4.65\times2.77$ </tex-math></inline-formula> mm2 area with pads, equivalent to 1.61 mm2 per element (for two beams), and consumes 37.2 mW per element per beam. To the best of our knowledge, the receiver front end demonstrates the minimum gain variation with temperature among silicon RF-beamforming front ends.

Optimal placement of sensor and actuator for controlling the piecewise linear Chua circuit via a discretized controller

Controlling the dynamics of chaotic systems is a task which is often addressed in an empirical way, particularly for placing sensors and actuators. Here, we show that selecting the measured variable and placing the actuator can be guided by considering the observability and controllability symbolic coefficients and applying the notion of flatness. This approach is here demonstrated on the piecewise linear Chua circuit, whose specific features are leveraged in constructing a discretized controller with a switch mechanism and optimally placed sensor and actuator. The feedback linearization is compared to a homogeneous and a passivity-based control laws, the flat control laws being more efficient than the others. It is thus shown that the proposed flat control law by feedback linearization is very efficient. The continuous time and discretized Chua circuit, governed by differential and difference equations, respectively, are treated. Most likely, these results could be extendable to a large group of natural and experimental systems.

Binary Colloidal Crystals Promote Cardiac Differentiation of Human Pluripotent Stem Cells via Nuclear Accumulation of SETDB1

Biophysical cues can facilitate the cardiac differentiation of human pluripotent stem cells (hPSCs), yet the mechanism is far from established. One of the binary colloidal crystals, composed of 5 μm Si and 400 nm poly(methyl methacrylate) particles named 5PM, has been applied as a substrate for hPSCs cultivation and cardiac differentiation. In this study, cell nucleus, cytoskeleton, and epigenetic states of human induced pluripotent stem cells on the 5PM were analyzed using atomic force microscopy, molecular biology assays, and the assay for transposase-accessible chromatin sequencing (ATAC-seq). Cells were more spherical with stiffer cell nuclei on the 5PM compared to the flat control. ATAC-seq revealed that chromatin accessibility decreased on the 5PM, caused by the increased entry of histone lysine methyltransferase SETDB1 into the cell nuclei and the amplified level of histone H3K9me3 modification. Reducing cytoskeleton tension using a ROCK inhibitor attenuated the nuclear accumulation of SETDB1 on the 5PM, indicating that the effect is cytoskeleton-dependent. In addition, the knockdown of SETDB1 reversed the promotive effects of the 5PM on cardiac differentiation, demonstrating that biophysical cue-induced cytoskeletal tension, cell nucleus deformation, and then SETDB1 accumulation are critical outside-in signal transformations in cardiac differentiation. Human embryonic stem cells showed similar results, indicating that the biophysical impact of the 5PM surfaces on cardiac differentiation could be universal. These findings contribute to our understanding of material-assistant hPSC differentiation, which benefits materiobiology and stem cell bioengineering.

Robust flat filtering control of a two degrees of freedom helicopter subject to tail rotor disturbances

Robust flat filtering control of a two degrees of freedom helicopter subject to tail rotor disturbances

A Nanobar-Supported Lipid Bilayer System for the Study of Membrane Curvature Sensing Proteins &lt;em&gt;in vitro&lt;/em&gt;

Membrane curvature plays important rolesin various essential processes of cells, such as cell migration, cell division, and vesicle trafficking. It is not only passively generated by cellular activities, but also actively regulates protein interactions and is involved in many intracellular signaling. Thus, it is of great value to examine the role of membrane curvature in regulating the distribution and dynamics of proteins and lipids. Recently, many techniques have been developed to study the relationship between the curved membrane and protein in vitro. Compared to traditional techniques, the newly developed nanobar-supported lipid bilayer (SLB) offers both high-throughput and better accuracy in membrane curvature generation by forming a continuous lipid bilayer on patterned arrays of nanobars with a pre-defined membrane curvature and local flat control. Both the lipid fluidity and protein sensitivity to curved membranes can be quantitatively characterized using fluorescence microscopy imaging. Here, a detailed procedure on how to form a SLB on fabricated glass surfaces containing nanobar arraysand the characterization of curvature-sensitive proteins on such SLB are introduced. In addition, protocols for nanochip reusing and image processing are covered. Beyond the nanobar-SLB, this protocol is readily applicable to all types of nanostructured glass chips for curvature sensing studies.

Effects of arch support doses on the center of pressure and pressure distribution of running using statistical parametric mapping.

Insoles with an arch support have been used to address biomechanical risk factors of running. However, the relationship between the dose of support and running biomechanics remains unclear. The purpose of this study was to determine the effects of changing arch support doses on the center of pressure (COP) and pressure mapping using statistical parametric mapping (SPM). Nine arch support variations (3 heights * 3 widths) and a flat insole control were tested on fifteen healthy recreational runners using a 1-m Footscan pressure plate. The medial-lateral COP (COPML) coordinates and the total COP velocity (COPVtotal) were calculated throughout the entirety of stance. One-dimensional and two-dimensional SPM were performed to assess differences between the arch support and control conditions for time series of COP variables and pressure mapping at a pixel level, respectively. Two-way ANOVAs were performed to test the main effect of the arch support height and width, and their interaction on the peak values of the COPVtotal. The results showed that the COPVtotal during the forefoot contact and forefoot push off phases was increased by arch supports, while the COP medial-lateral coordinates remained unchanged. There was a dose-response effect of the arch support height on peak values of the COPVtotal, with a higher support increasing the first and third valleys but decreasing the third peak of the COPVtotal. Meanwhile, a higher arch support height shifted the peak pressure from the medial forefoot and rearfoot to the medial arch. It is concluded that changing arch support doses, primarily the height, systematically altered the COP velocities and peak plantar pressure at a pixel level during running. When assessing subtle modifications in the arch support, the COP velocity was a more sensitive variable than COP coordinates. SPM provides a high-resolution view of pressure comparisons, and is recommended for future insole/footwear investigations to better understand the underlying mechanisms and improve insole design.

A comparison of vegetable leaves and replicated biomimetic surfaces on the binding of Escherichia coli and Listeria monocytogenes

Biofouling in the food industry is a huge issue, and one possible way to reduce surface fouling is to understand how naturally cleaning surfaces based on biomimetic designs influence bacterial binding. Four self-cleaning leaves (Tenderheart cabbage, Cauliflower, White cabbage and Leek) were analysed for their surface properties and artificial replicates were produced. The leaves and surfaces were subjected to attachment, adhesion and retention assays using Escherichia coli and Listeria monocytogenes. For the attachment assays, the lowest cell numbers occurred on the least hydrophobic and smooth surfaces but were higher than the flat control surface, regardless of the strain. Following the adhesion assays, using L. monocytogenes, the Tenderheart and Cauliflower biomimetic replicated leaves resulted in significantly lowered cell adhesion. Following the retention assays, White cabbage demonstrated lower cell retention for both types of bacteria on the biomimetic replicated surface compared to the flat control surface. The biomimetic surfaces were also more efficient at avoiding bacterial retention than natural leaves, with reductions of about 1 and 2 Log in L. monocytogenes and E. coli retention, respectively, on most of the produced surfaces. Although the surfaces were promising in reducing bacterial binding, the results suggested that different experimental assays exerted different influences on the conclusions. This work demonstrated that consideration needs to be given to the environmental factors where the surface is to be used and that bacterial species influence the propensity of biofouling on a surface.

Laser-processed antiadhesive bionic combs for handling nanofibers inspired by nanostructures on the legs of cribellate spiders.

Nanofibers are drawing the attention of engineers and scientists because their large surface-to-volume ratio is favorable for applications in medicine, filter technology, textile industry, lithium-air batteries, and optical sensors. However, when transferring nanofibers to a technical product in the form of a random network of fibers, referred to as nonwoven fabric, the stickiness of the freshly produced and thus fragile nanofiber nonwoven remains a problem. This is mainly because nanofibers strongly adhere to any surface because of van der Waals forces. In nature, there are animals that are actually able to efficiently produce, process, and handle nanofibers, namely cribellate spiders. For that, the spiders use the calamistrum, a comb-like structure of modified setae on the metatarsus of the hindmost (fourth) legs, to which the 10-30 nm thick silk nanofibers do not stick due to a special fingerprint-like surface nanostructure. In this work, we present a theoretical model of the interaction of linear nanofibers with a sinusoidally corrugated surface. This model allows for a prediction of the adhesive interaction and, thus, the design of a suitable surface structure to prevent sticking of an artificially nonwoven of nanofibers. According to the theoretical prediction, a technical analogon of the nanoripples was produced by ultrashort pulse laser processing on different technically relevant metal surfaces in the form of so-called laser-induced periodic surface structures (LIPSS). Subsequently, by means of a newly established peel-off test, the adhesion of an electrospun polyamide fiber-based nonwoven was quantified on such LIPSS-covered aluminium alloy, steel, and titanium alloy samples, as well as on polished (flat) control samples as reference and, additionally, on samples with randomly rough surfaces. The latter revealed that the adhesion of electrospun nanofiber nonwoven is significantly lowered on the nanostructured surfaces compared with the polished surfaces.

Rocker-profile design shoes improve pendular energy recovery in walking with no effects on total mechanical work

Rocker-profile design shoes are commonly used in clinical settings. Such footwear reduces in-shoe pressure over the forefoot area during the gait, and depending on the rocker-profile type (i.e., toe-only, heel negative, or double rocker), affects lower limb kinematics, kinetics, and muscle electromyographic activity. However, whether wearing rocker-profile shoes influence the dynamics of the body centre of mass (BCoM) is unknown. We used a mathematical procedure combining Lissajous contours and Fourier analysis to describe the 3D trajectory of the BCoM in walking with rocker-profile (RollingSole) and flat (Control) shoes at 0.97, 1.25, and 1.53 m s−1 in 30 participants. Harmonics amplitude and phase were compared using linear and circular statistics, respectively. External (Wext), kinematic internal (Wint,k) and total (Wtot) mechanical works, and the mechanical energy fraction recovered from a pendular exchange of potential and kinetic energy were also calculated. RollingSole shoes yielded greater Wext (1–9 %; P < 0.05) and fractional pendular energy exchange (1–8 %; P < 0.01), with lower Wint,k (2–5 %; P < 0.05) and unchanged Wtot (P ≥ 0.30). RollingSole shoes led also to a greater mean height of the BCoM (1–3 %; P < 0.01), and amplitude of the anteroposterior and vertical symmetric, and mediolateral 2nd-to-5th harmonics (1–30 %; P < 0.01). No differences between shoes were found for the harmonics phase (P ≥ 0.14). Our results indicate that RollingSole shoes enhanced an inverted pendulum-like behaviour of the BCoM during walking with no alterations in total mechanical work. This may result from the combination of rocker-profile design and greater BCoM height (through thicker soles) with such shoes, increasing recovery of mechanical energy in step-to-step transitions and mid-stance.

Experimental study on flat plate skin friction control by porous media based on global fluorescent oil film measurement technology

Skin friction is a primary source of total aircraft drag. It is important, therefore, in science and engineering, to achieve drag reduction control in a boundary layer. In this paper, under the experimental conditions of Reθ = 5909 (x/c = 0.55) and with a zero-pressure gradient, the drag reduction control of a plate boundary layer in porous media is studied. The global skin friction of the plate is measured using fluorescent oil film test technology. The results show that, in contrast with the downstream frictional resistance coefficient of a flat plate that possesses a smooth surface, the coefficient for porous media reduced significantly. Also, the lower the pores per inch (PPI) of the porous media, the greater the drag reduction effect. Among the three porous media with different PPI, porous media with ten PPI has the best drag reduction effect. With increasing distance from the porous media, the drag reduction effect decreases gradually. Porous media significantly increase the slope of the logarithmic region of the velocity profile of the downstream turbulent boundary layer, the dimensionless wall velocity u+ moves upward, and the velocity pulsation in the logarithmic region increases so as to reduce skin friction.

Makrofauna Tanah Sebagai Bioindikator pada Areal Perkebunan yang diberi Limbah Cair Pabrik Kelapa Sawit Sebagai Pupuk

Research on Soil Macrofauna as Bioindicator in Plantation Areas Given Palm Oil Mill Liquid Waste as Fertilizer was conducted in 2021. Samples were taken in Long Bed, Flat Bed, and Control areas using Purposive Sampling and Squares and Hand Sorting methods. Each location is determined by 9 sampling points. Soil macrofauna samples were identified in the USU FMIPA Ecology laboratory. From the results of the study, there were 3 soil macrofauna phyla, namely the Annelida phylum, which consisted of 1 class, 1 order, 2 families, and 2 species. Phylum Arthropoda consists of 5 classes, 10 orders, 16 families, and 27 species. Phylum Mollusca consists of 1 class, 1 order, 1 family, and 1 species. The highest population density (K) was obtained from the species Macrotermes gilvus, which was 81.10 individuals/m2 in the Control area, followed by Ponthoscolex corethrurus at 78.81 individuals/m2 in the Flat Bed area. The highest relative density (KR) was found in the species Macrotermes gilvus, which was 49.1% in the control area, followed by Ponthoscolex corethrurus at 46.84% in the Long Bed area. The highest frequency of presence (FK) was found in the Ponthoscolex corethrurus species, which was 100% in the Control and Flat Bed areas, and 88.88% in the Long Bed areas. Species that can live and reproduce well as bioindicators are Ponthoscolex corethrurus in the Long Bed area, followed by the Flat Bed and Control areas. The highest diversity index value was found in the Flat Bed area, which was 3,831 with the criteria for diversity being high, the distribution of the number of individuals for each species was high, the community stability was good and the soil community had not been polluted, while the Equitabilitas value was 1,258 with the criteria for high uniformity, stable community. The application of palm oil mill effluent as fertilizer to the plantation area can increase the life of the soil macrofauna, and the physical-chemical properties of the soil.

Submicron Topographically Patterned 3D Substrates Enhance Directional Axon Outgrowth of Dorsal Root Ganglia Cultured Ex Vivo.

A peripheral nerve injury results in disruption of the fiber that usually protects axons from the surrounding environment. Severed axons from the proximal nerve stump are capable of regenerating, but axons are exposed to a completely new environment. Regeneration recruits cells that produce and deposit key molecules, including growth factor proteins and fibrils in the extracellular matrix (ECM), thus changing the chemical and geometrical environment. The regenerating axons thus surf on a newly remodeled micro-landscape. Strategies to enhance and control axonal regeneration and growth after injury often involve mimicking the extrinsic cues that are found in the natural nerve environment. Indeed, nano- and micropatterned substrates have been generated as tools to guide axons along a defined path. The mechanical cues of the substrate are used as guides to orient growth or change the direction of growth in response to impediments or cell surface topography. However, exactly how axons respond to biophysical information and the dynamics of axonal movement are still poorly understood. Here we use anisotropic, groove-patterned substrate topography to direct and enhance sensory axonal growth of whole mouse dorsal root ganglia (DRG) transplanted ex vivo. Our results show significantly enhanced and directed growth of the DRG sensory fibers on the hemi-3D topographic substrates compared to a 0 nm pitch, flat control surface. By assessing the dynamics of axonal movement in time-lapse microscopy, we found that the enhancement was not due to increases in the speed of axonal growth, but to the efficiency of growth direction, ensuring axons minimize movement in undesired directions. Finally, the directionality of growth was reproduced on topographic patterns fabricated as fully 3D substrates, potentially opening new translational avenues of development incorporating these specific topographic feature sizes in implantable conduits in vivo.

Complexity-biodiversity relationships on marine urban structures: reintroducing habitat heterogeneity through eco-engineering.

Urbanization is leading to biodiversity loss through habitat homogenization. The smooth, featureless surfaces of many marine urban structures support ecological communities, often of lower biodiversity, distinct from the complex natural habitats they replace. Eco-engineering (design for ecological co-benefits) seeks to enhance biodiversity and ecological functions on urban structures. We assessed the benefits to biodiversity of retrofitting four types of complex habitat panels to an intertidal seawall at patch (versus flat control panels) and site (versus unmodified control seawalls and reference rocky shores) scales. Two years after installation, patch-scale effects of complex panels on biodiversity ranged from neutral to positive, depending on the protective features they provided, though all but one design (honeycomb) supported unique species. Water-retaining features (rockpools) and crevices, which provided moisture retention and cooling, increased biodiversity and supported algae and invertebrates otherwise absent. At the site scale, biodiversity benefits ranged from neutral at the high- and mid-intertidal to positive at the low-intertidal elevation. The results highlight the importance of matching eco-engineering interventions to the niche of target species, and environmental conditions. While species richness was greatest on rockpool and crevice panels, the unique species supported by other panel designs highlights that to maximize biodiversity, habitat heterogeneity is essential.This article is part of the theme issue ‘Ecological complexity and the biosphere: the next 30 years’.

Topography-Mediated Enhancement of Nonviral Gene Delivery in Stem Cells.

Gene delivery holds great promise for bioengineering, biomedical applications, biosensors, diagnoses, and gene therapy. In particular, the influence of topography on gene delivery is considered to be an attractive approach due to low toxicity and localized delivery properties. Even though many gene vectors and transfection systems have been developed to enhance transfection potential and combining it with other forms of stimulations could even further enhance it. Topography is an interesting surface property that has been shown to stimulate differentiation, migration, cell morphology, and cell mechanics. Therefore, it is envisioned that topography might also be able to stimulate transfection. In this study, we tested the hypothesis “topography is able to regulate transfection efficiency”, for which we used nano- and microwave-like topographical substrates with wavelengths ranging from 500 nm to 25 µm and assessed the transfectability of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) and myoblasts. For transfection, Lipofectamine 2000 and a gene encoding plasmid for red-fluorescent protein (m-Cherry) were used and topography-induced cell morphology and transfection efficiency was analyzed. As a result, topography directs cell spreading, elongation, and proliferation as well as the transfection efficiency, which were investigated but were found not to be correlated and dependent on the cell type. A 55% percent improvement of transfection efficiency was identified for hBM-MSCs grown on 2 µm wrinkles (24.3%) as compared to hBM-MSCs cultured on flat controls (15.7%). For myoblast cells, the highest gene-expression efficiency (46.1%) was observed on the 10 µm topography, which enhanced the transfection efficiency by 64% as compared to the flat control (28.1%). From a qualitative assessment, it was observed that the uptake capacity of cationic complexes of TAMRA-labeled oligodeoxynucleotides (ODNs) was not topography-dependent but that the intracellular release was faster, as indicated by the positively stained nuclei on 2 μm for hBM-MSCs and 10 μm for myoblasts. The presented results indicate that topography enhances the gene-delivery capacity and that the responses are dependent on cell type. This study demonstrates the important role of topography on cell stimulation for gene delivery as well as understanding the uptake capacity of lipoplexes and may be useful for developing advanced nonviral gene delivery strategies.