Dynamic duos: the building blocks of dimensional mechanics.

Mechanics studies the relationships between space, time, and matter. These relationships can be expressed in terms of the dimensions of length , time , and mass . Each dimension broadens the scope of mechanics. Historically, mechanics emerged from geometry, which considers quantities like lengths or areas, with dimensions of the form . With the Renaissance, quantities combining space and time were considered, like speed, acceleration and later diffusivity, all of the form . Eventually, mechanics reached its full potential by including "mass-carrying" quantities such as mass, force, momentum, energy, action, power, viscosity, etc. These standard mechanical quantities have dimensions of the form where x and y are integers. In this contribution, we show that, thanks to this dimensional structure, these mass-carrying quantities can be readily arranged into a table such that x and y increase along the row and column, respectively. Ratios of quantities in the same rows provide characteristic lengths, while those in the same columns yield characteristic times, encompassing a great variety of physical phenomena from atomic to astronomical scales. Most generally, we show that selecting duos of mechanical quantities that are neither on the same row nor column of the table yields dynamics, where one mechanical quantity is understood as impelling motion, while the other impedes it. The force and the mass are the prototypes of impelling and impeding factors, but many other duos are possible. We present examples from the physical and biological realms, including planetary motion, sedimentation, explosions, fluid flows, turbulence, diffusion, cell mechanics, capillary and gravity waves, and spreading, pinching, and coalescence of drops and bubbles. This review provides a novel synthesis revealing the power of scaling or dimensional analysis, to understand processes governed by the interplay of two mechanical quantities. This elementary decomposition of space, time and motion into pairs of mechanical factors is the foundation of "dimensional mechanics", a method that this review wishes to promote and advance. Pairs are the fundamental building blocks, but they are only a starting point. Beyond this simple world of mechanical duos, we envision a richer universe that beckons with an interplay of three, four, or more quantities, yielding multiple characteristic lengths, times, and kinematics. This review is complemented by online video lectures, which initiate a discussion on the elaborate interplay of two or more mechanical quantities.

Characterization of Benchtop-Fabricated Arrays of Nanowrinkled Surface Electrodes as a Nitric Oxide Electrochemical Sensor.

In this work, we present an accessible benchtop fabrication technique to obtain a planar array of gold nanowrinkled surface electrodes (ANSE) for the construction of electrochemical cells, specifically to monitor soluble biomarkers of interest in cell culture environments. We present a complete characterization of the array and its response as an electrochemical cell. To validate our sensor, we evaluated the device sensitivity to detect nitric oxide (NO), an important molecule produced by endothelial cells as a response to environmental signals such as mechanics and growth factors. While testing measurements of nitric oxide in aqueous solutions with isotonic salt concentrations, we evidenced the influence of the environmental conditions for such electrochemical measurements, showing that the aqueous medium, usually not accounted for, significantly impacts the outcome. Finally, we present the application of the electrochemical sensor for the detection of nitric oxide released from stimulated endothelial cells as a proof of concept.

Mechanical regulation of the early stages of angiogenesis.

Favouring or thwarting the development of a vascular network is essential in fields as diverse as oncology, cardiovascular disease or tissue engineering. As a result, understanding and controlling angiogenesis has become a major scientific challenge. Mechanical factors play a fundamental role in angiogenesis and can potentially be exploited for optimizing the architecture of the resulting vascular network. Largely focusing on in vitro systems but also supported by some in vivo evidence, the aim of this Highlight Review is dual. First, we describe the current knowledge with particular focus on the effects of fluid and solid mechanical stimuli on the early stages of the angiogenic process, most notably the destabilization of existing vessels and the initiation and elongation of new vessels. Second, we explore inherent difficulties in the field and propose future perspectives on the use of in vitro and physics-based modelling to overcome these difficulties.

Importance of the Microenvironment and Mechanosensing in Adipose Tissue Biology.

The expansion of adipose tissue is an adaptive mechanism that increases nutrient buffering capacity in response to an overall positive energy balance. Over the course of expansion, the adipose microenvironment undergoes continual remodeling to maintain its structural and functional integrity. However, in the long run, adipose tissue remodeling, typically characterized by adipocyte hypertrophy, immune cells infiltration, fibrosis and changes in vascular architecture, generates mechanical stress on adipose cells. This mechanical stimulus is then transduced into a biochemical signal that alters adipose function through mechanotransduction. In this review, we describe the physical changes occurring during adipose tissue remodeling, and how they regulate adipose cell physiology and promote obesity-associated dysfunction in adipose tissue.

Spreading, pinching, and coalescence: the Ohnesorge units.

Understanding the kinematics and dynamics of spreading, pinching, and coalescence of drops is critically important for a diverse range of applications involving spraying, printing, coating, dispensing, emulsification, and atomization. Hence experimental studies visualize and characterize the increase in size over time for drops spreading over substrates, or liquid bridges between coalescing drops, or the decrease in the radius of pinching necks during drop formation. Even for Newtonian fluids, the interplay of inertial, viscous, and capillary stresses can lead to a number of scaling laws, with three limiting self-similar cases: visco-inertial (VI), visco-capillary (VC) and inertio-capillary (IC). Though experiments are presented as examples of the methods of dimensional analysis, the lack of precise values or estimates for pre-factors, transitions, and scaling exponents presents difficulties for quantitative analysis and material characterization. In this tutorial review, we reanalyze and summarize an elaborate set of landmark published experimental studies on a wide range of Newtonian fluids. We show that moving beyond VI, VC, and IC units in favor of intrinsic timescale and lengthscale determined by all three material properties (viscosity, surface tension and density), creates a complementary system that we call the Ohnesorge units. We find that in spite of large differences in topological features, timescales, and material properties, the analysis of spreading, pinching and coalescing drops in the Ohnesorge units results in a remarkable collapse of the experimental datasets, highlighting the shared and universal features displayed in such flows.

Hierarchical Modeling of the Liver Vascular System.

The liver plays a key role in the metabolic homeostasis of the whole organism. To carry out its functions, it is endowed with a peculiar circulatory system, made of three main dendritic flow structures and lobules. Understanding the vascular anatomy of the liver is clinically relevant since various liver pathologies are related to vascular disorders. Here, we develop a novel liver circulation model with a deterministic architecture based on the constructal law of design over the entire scale range (from macrocirculation to microcirculation). In this framework, the liver vascular structure is a combination of superimposed tree-shaped networks and porous system, where the main geometrical features of the dendritic fluid networks and the permeability of the porous medium, are defined from the constructal viewpoint. With this model, we are able to emulate physiological scenarios and to predict changes in blood pressure and flow rates throughout the hepatic vasculature due to resection or thrombosis in certain portions of the organ, simulated as deliberate blockages in the blood supply to these sections. This work sheds light on the critical impact of the vascular network on mechanics-related processes occurring in hepatic diseases, healing and regeneration that involve blood flow redistribution and are at the core of liver resilience.

Three-Dimensional Porous Scaffolds Derived from Bovine Cancellous Bone Matrix Promote Osteoinduction, Osteoconduction, and Osteogenesis.

The use of three-dimensional porous scaffolds derived from decellularized extracellular matrix (ECM) is increasing for functional repair and regeneration of injured bone tissue. Because these scaffolds retain their native structures and bioactive molecules, in addition to showing low immunogenicity and good biodegradability, they can promote tissue repair and regeneration. Nonetheless, imitating these features in synthetic materials represents a challenging task. Furthermore, due to the complexity of bone tissue, different processes are necessary to maintain these characteristics. We present a novel approach using decellularized ECM material derived from bovine cancellous bone by demineralization, decellularization, and hydrolysis of collagen to obtain a three-dimensional porous scaffold. This study demonstrates that the three-dimensional porous scaffold obtained from bovine bone retained its osteoconductive and osteoinductive properties and presented osteogenic potential when seeded with human Wharton's jelly mesenchymal stromal cells (hWJ-MSCs). Based on its characteristics, the scaffold described in this work potentially represents a therapeutic strategy for bone repair.

Development of a Diagnostic Biosensor Method of Hypersensitivity Pneumonitis towards a Point-of-Care Biosensor.

In spite of a current increasing trend in the development of miniaturized, standalone point-of-care (PoC) biosensing platforms in the literature, the actual implementation of such systems in the field is far from being a reality although deeply needed. In the particular case of the population screenings for local or regional diseases related to specific pathogens, the diagnosis of the presence of specific antibodies could drastically modify therapies and even the organization of public policies. The aim of this work was to develop a fast, cost-effective detection method based on the manipulation of functionalized magnetic beads for an efficient diagnosis of hypersensitivity pneumonitis (HP), looking for the presence of anti-pigeon antigen antibodies (APAA) in a patient's serum. We presented a Diagnostic Biosensor Method (DBM) in detail, with validation by comparison with a traditional high-throughput platform (ELISA assay). We also demonstrated that it was compatible with a microfluidic chip that could be eventually incorporated into a PoC for easy and broad deployment using portable optical detectors. After standardization of the different reaction steps, we constructed and validated a plastic chip that could easily be scaled to high-volume manufacturing in the future. The solution proved comparable to conventional ELISA assays traditionally performed by the clinicians in their laboratory and should be compatible with other antibody detection directly from patient samples.

Determination by Relaxation Tests of the Mechanical Properties of Soft Polyacrylamide Gels Made for Mechanobiology Studies.

Following the general aim of recapitulating the native mechanical properties of tissues and organs in vitro, the field of materials science and engineering has benefited from recent progress in developing compliant substrates with physical and chemical properties similar to those of biological materials. In particular, in the field of mechanobiology, soft hydrogels can now reproduce the precise range of stiffnesses of healthy and pathological tissues to study the mechanisms behind cell responses to mechanics. However, it was shown that biological tissues are not only elastic but also relax at different timescales. Cells can, indeed, perceive this dissipation and actually need it because it is a critical signal integrated with other signals to define adhesion, spreading and even more complicated functions. The mechanical characterization of hydrogels used in mechanobiology is, however, commonly limited to the elastic stiffness (Young's modulus) and this value is known to depend greatly on the measurement conditions that are rarely reported in great detail. Here, we report that a simple relaxation test performed under well-defined conditions can provide all the necessary information for characterizing soft materials mechanically, by fitting the dissipation behavior with a generalized Maxwell model (GMM). The simple method was validated using soft polyacrylamide hydrogels and proved to be very useful to readily unveil precise mechanical properties of gels that cells can sense and offer a set of characteristic values that can be compared with what is typically reported from microindentation tests.

Fabrication of Adhesive Substrate for Incorporating Hydrogels to Investigate the Influence of Stiffness on Cancer Cell Behavior.

Stiffness control of cell culture platforms provides researchers in cell biology with the ability to study different experimental models in conditions of mimicking physiological or pathological microenvironments. Nevertheless, the signal transduction pathways and drug sensibility of cancer cells have been poorly characterized widely using biomimetic platforms because the limited experience of cancer cell biology groups about handling substrates with specific mechanical properties. The protein cross-linking and stiffening control are crucial checkpoints that could strongly affect cell adhesion and spreading, misrepresenting the data acquired, and also generating inaccurate cellular models. Here, we introduce a simple method to adhere to polyacrylamide (PAA) hydrogels on glass coverslips without any special treatment for mechanics studies in cancer cell biology. By using a commercial photosensitive glue, Loctite 3525, it is possible to polymerize PAA hydrogels directly on glass surfaces. Furthermore, we describe a cross-linking reaction method to attach proteins to PAA as an alternative method to Sulfo-SANPAH cross-linking, which is sometimes difficult to implement and reproduce. In this chapter, we describe a reliable procedure to fabricate ECM protein-cross-linked PAA hydrogels for mechanotransduction studies on cancer cells.

The liver, a functionalized vascular structure.

The liver is not only the largest organ in the body but also the one playing one of the most important role in the human metabolism as it is in charge of transforming toxic substances in the body. Understanding the way its blood vasculature works is key. In this work we show that the challenge of predicting the hepatic multi-scale vascular network can be met thanks to the constructal law of design evolution. The work unveils the structure of the liver blood flow architecture as a combination of superimposed tree-shaped networks and porous system. We demonstrate that the dendritic nature of the hepatic artery, portal vein and hepatic vein can be predicted, together with their geometrical features (diameter ratio, duct length ratio) as the entire blood flow architectures follow the principle of equipartition of imperfections. At the smallest scale, the shape of the liver elemental systems-the lobules-is discovered, while their permeability is also predicted. The theory is compared with good agreement to anatomical data from the literature.

Fibrillar Collagen Type I Participates in the Survival and Aggregation of Primary Hepatocytes Cultured on Soft Hydrogels.

Liver is an essential organ that carries out multiple functions such as glycogen storage, the synthesis of plasma proteins, and the detoxification of xenobiotics. Hepatocytes are the parenchyma that sustain almost all the functions supported by this organ. Hepatocytes and non-parenchymal cells respond to the mechanical alterations that occur in the extracellular matrix (ECM) caused by organogenesis and regenerating processes. Rearrangements of the ECM modify the composition and mechanical properties that result in specific dedifferentiation programs inside the hepatic cells. Quiescent hepatocytes are embedded in the soft ECM, which contains an important concentration of fibrillar collagens in combination with a basement membrane-associated matrix (BM). This work aims to evaluate the role of fibrillar collagens and BM on actin cytoskeleton organization and the function of rat primary hepatocytes cultured on soft elastic polyacrylamide hydrogels (PAA HGs). We used rat tail collagen type I and Matrigel® as references of fibrillar collagens and BM respectively and mixed different percentages of collagen type I in combination with BM. We also used peptides obtained from decellularized liver matrices (dECM). Remarkably, hepatocytes showed a poor adhesion in the absence of collagen on soft PAA HGs. We demonstrated that collagen type I inhibited apoptosis and activated extracellular signal-regulated kinases 1/2 (ERK1/2) in primary hepatocytes cultured on soft hydrogels. Epidermal growth factor (EGF) was not able to rescue cell viability in conjugated BM but affected cell aggregation in soft PAA HGs conjugated with combinations of different proportions of collagen and BM. Interestingly, actin cytoskeleton was localized and preserved close to plasma membrane (cortical actin) and proximal to intercellular ducts (canaliculi-like structures) in soft conditions; however, albumin protein expression was not preserved, even though primary hepatocytes did not remodel their actin cytoskeleton significantly in soft conditions. This investigation highlights the important role of fibrillar collagens on soft hydrogels for the maintenance of survival and aggregation of the hepatocytes. Data suggest evaluating the conditions that allow the establishment of optimal biomimetic environments for physiology and cell biology studies, where the phenotype of primary cells may be preserved for longer periods of time.

Hepatic C9 cells switch their behaviour in short or long exposure to soft substrates.

BACKGROUND INFORMATION: There have been several studies to understand the influence of stiffness of the culture substrates for different types of adherent cells. It is generally accepted that cell proliferation, spreading and focal adhesions increase with higher matrix stiffness. However, what remains unclear is whether this kind of cell behaviour may be reverted by culturing on soft substrates those cell lines that were originally selected or primed on stiff surfaces. RESULTS: Here, we studied the influence of substrate softness on proliferation, adhesion and morphology of the highly proliferative hepatic C9 cell line. We cultured C9 cells on soft and stiff polydimethylsiloxane (PDMS) substrates prepared with two different elastic moduli in the range of 10 and 200 kPa, respectively. Lower cell proliferation was observed on substrates with lower stiffness without affecting cell viability. The proliferation rate of C9 cell line along with extracellular growth-regulated kinase (ERK) phosphorylation was decreased on soft PDMS. Despite this cell line has been described as a hepatic epithelial cell, our characterisation demonstrated that the origin of C9 cells is uncertain, although clearly epithelial, with the expression of markers of several hepatic cells. Surprisingly, consecutive passages of C9 cells on soft PDMS did not alter this mesenchymal phenotype, vimentin expression did not decrease when culturing cells in soft substrates, even though the ERK phosphorylation levels eventually were increased after several passages on soft PDMS, triggering again an increase of cell proliferation. CONCLUSIONS AND SIGNIFICANCE: This study shows that the exposure of C9 cells to soft substrates promoted a decrease of cell proliferation rate, as reported for other types of cells on PDMS, whereas a much longer term exposure caused cells to adapt to softness after trained for several passages, reactivating proliferation. During this phenomenon, the morphology and phenotype of trained cells was modified accompanying the increase of cell proliferation rate contrary to the effect observed in short periods of cell culture. In contrast to previous reports, cell death was not observed during these experiments, discarding a cell selection mechanism and suggesting soft cell adaptation may be limited in time in C9 cells.

Building a microfluidic cell culture platform with stiffness control using Loctite 3525 glue.

The study of mechanotransduction signals and cell response to mechanical properties requires designing culture substrates that possess some, or ideally all, of the following characteristics: (1) biological compatibility and adhesive properties, (2) stiffness control or tunability in a dynamic mode, (3) patternability on the microscale and (4) integrability in microfluidic chips. The most common materials used to address cell mechanotransduction are hydrogels, due to their softness. However, they may be impractical when complex scaffolds are sought and they lack viscous dissipative properties that are very important in mechanobiology. In this work, we show that an off-the-shelf, biocompatible photosensitive glue, Loctite 3525, may be used readily in mechanobiology assays without any special treatment prior to fabrication of cell culture platforms. Despite a high (MPa) stiffness easily tunable by UV exposure time at a fixed dose, 3T3 fibroblasts showed a response to the mechanics of the material similar to that obtained on much softer (kPa) hydrogels. Loctite's viscous dissipation properties indeed seemed to be responsible for such cell mechanical response, as suggested by recent works where more complex two-phase hydrogels were employed. More interestingly, it was possible to stiffen soft Loctite substrates by post-exposing them during cell culture, to observe changes in cell spreading caused by a dynamic stiffness modification. Thanks to Loctite 3525's patternability, micropillars were also fabricated to demonstrate the compatibility with traction force microscopy studies. Finally, the glue was used as an excellent adhesion layer for hydrogels on glass or PDMS, without the need for additional treatment, enabling the easy fabrication of microfluidic chips integrating hydrogels.

Micro-Macro: Selective Integration of Microfeatures Inside Low-Cost Macromolds for PDMS Microfluidics Fabrication.

Microfluidics has become a very promising technology in recent years, due to its great potential to revolutionize life-science solutions. Generic microfabrication processes have been progressively made available to academic laboratories thanks to cost-effective soft-lithography techniques and enabled important progress in applications like lab-on-chip platforms using rapid- prototyping. However, micron-sized features are required in most designs, especially in biomimetic cell culture platforms, imposing elevated costs of production associated with lithography and limiting the use of such devices. In most cases, however, only a small portion of the structures require high-resolution and cost may be decreased. In this work, we present a replica-molding method separating the fabrication steps of low (macro) and high (micro) resolutions and then merging the two scales in a single chip. The method consists of fabricating the largest possible area in inexpensive macromolds using simple techniques such as plastics micromilling, laser microfabrication, or even by shrinking printed polystyrene sheets. The microfeatures were made on a separated mold or onto existing macromolds using photolithography or 2-photon lithography. By limiting the expensive area to the essential, the time and cost of fabrication can be reduced. Polydimethylsiloxane (PDMS) microfluidic chips were successfully fabricated from the constructed molds and tested to validate our micro-macro method.

Cell Culture Platforms with Controllable Stiffness for Chick Embryonic Cardiomyocytes.

For several years, cell culture techniques have been physiologically relevant to understand living organisms both structurally and functionally, aiming at preserving as carefully as possible the in vivo integrity and function of the cells. However, when studying cardiac cells, glass or plastic Petri dishes and culture-coated plates lack important cues that do not allow to maintain the desired phenotype, especially for primary cell culture. In this work, we show that microscaffolds made with polydimethylsiloxane (PDMS) enable modulating the stiffness of the surface of the culture substrate and this originates different patterns of adhesion, self-organization, and synchronized or propagated activity in the culture of chick embryonic cardiomyocytes. Thanks to the calcium imaging technique, we found that the substrate stiffness affected cardiomyocyte adhesion, as well as the calcium signal propagation in the formed tissue. The patterns of activity shown by the calcium fluorescence variations are reliable clues of the functional organization achieved by the cell layers. We found that PDMS substrates with a stiffness of 25 kPa did not allow the formation of cell layers and therefore the optimal propagation of the intracellular calcium signals, while softer PDMS substrates with Young's modulus within the physiological in vivo reported range did permit synchronized and coordinated contractility and intracellular calcium activity. This type of methodology allows us to study phenomena such as arrhythmias. For example, the occurrence of synchronized activity or rotors that can initiate or maintain cardiac arrhythmias can be reproduced on different substrates for study, so that replacement tissues or patches can be better designed.

Influence of the PLGA/gelatin ratio on the physical, chemical and biological properties of electrospun scaffolds for wound dressings.

Chronic wounds are a global health problem, and their treatments are difficult and long lasting. The development of medical devices through tissue engineering has been conducted to heal this type of wound. In this study, it was demonstrated that the combination of natural and synthetic polymers, such as poly (D-L lactide-co-glycolide) (PLGA) and gelatin (Ge), were useful for constructing scaffolds for wound healing. The aim of this study was to evaluate the influence of different PLGA/gelatin ratios (9:1, 7:3 and 5:5 (v/v)) on the physical, chemical and biological properties of electrospun scaffolds for wound dressings. These PLGA/Ge scaffolds had randomly oriented fibers with smooth surfaces and exhibited distances between fibers of less than 10 µm. The 7:3 and 5:5 PLGA/Ge scaffolds showed higher swelling, hydrophilicity and degradation rates than pure PLGA and 9:1 (v/v) PLGA/Ge scaffolds. Young's moduli of the scaffolds were 72 ± 10, 48 ± 6, 58 ± 6 and 6 ± 1 MPa for the pure PLGA scaffold and the 9:1, 7:3 and 5:5 (v/v) PLGA/Ge scaffolds, respectively. Mesenchymal stem cells (MSCs) seeded on all the PLGA/Ge scaffolds were viable, and the cells were attached to the fibers at the different analyzed timepoints. The most significant proliferation rate was observed for cells on the 7:3 PLGA/Ge scaffolds. Biocompatibility analysis showed that all the scaffolds produced inflammation at the first week postimplantation; however, the 7:3 and 5:5 (v/v) PLGA/Ge scaffolds were degraded completely, and there was no inflammatory reaction observed at the fourth week after implantation. In contrast, the 9:1 PLGA/Ge scaffolds persisted in the tissue for more than four weeks; however, at the eighth week, no traces of the scaffolds were found. In conclusion, the scaffolds with the 7:3 PLGA/Ge ratio showed suitable physical, chemical and biological properties for applications in chronic wound treatments.

Method for the Direct Fabrication of Polyacrylamide Hydrogels with Controlled Stiffness in Polystyrene Multiwell Plates for Mechanobiology Assays.

Polyacrylamide (PAA) hydrogels are now widely used in mechanobiology because the well-defined available protocols allow a robust and reproducible control of substrate stiffness within a physiological range. However, several assays require hydrogels inside traditional plastic substrates and the current methods remain relatively tedious. Here, we present a simple and direct fabrication technique that successfully attaches PAA hydrogels inside polystyrene multiwell plates and Petri dishes of different sizes. It permits a control of the Young's modulus of the gels, within the desired range for mechanobiology. Some critical steps, that had to be overcome to guarantee protein conjugation and cell attachment, are detailed, as they differ from the standardized preparation on glass substrates. To validate our process, we demonstrated that HepG2 and 3T3L1 cell lines as well as primary hepatocytes seeded on PAA gels of different stiffnesses in plastics showed a mechanical response identical to the cells cultured on traditional gels.

Progress on the Use of Commercial Digital Optical Disc Units for Low-Power Laser Micromachining in Biomedical Applications.

The development of organ-on-chip and biological scaffolds is currently requiring simpler methods for microstructure biocompatible materials in three dimensions, to fabricate structural and functional elements in biomaterials, or modify the physicochemical properties of desired substrates. Aiming at addressing this need, a low-power CD-DVD-Blu-ray laser pickup head was mounted on a programmable three-axis micro-displacement system in order to modify the surface of polymeric materials in a local fashion. Thanks to a specially-designed method using a strongly absorbing additive coating the materials of interest, it has been possible to establish and precisely control processes useful in microtechnology for biomedical applications. The system was upgraded with Blu-ray laser for additive manufacturing and ablation on a single platform. In this work, we present the application of these fabrication techniques to the development of biomimetic cellular culture platforms thanks to the simple integration of several features typically achieved with traditional, less cost-effective microtechnology methods in one step or through replica-molding. Our straightforward approach indeed enables great control of local laser microablation or polymerization for true on-demand biomimetic micropatterned designs in transparent polymers and hydrogels and is allowing integration of microfluidics, microelectronics, surface microstructuring, and transfer of superficial protein micropatterns on a variety of biocompatible materials.

Fabrication of low-cost micropatterned polydimethyl-siloxane scaffolds to organise cells in a variety of two-dimensioanl biomimetic arrangements for lab-on-chip culture platforms.

We present the rapid-prototyping of type I collagen micropatterns on poly-dimethylsiloxane substrates for the biomimetic confinement of cells using the combination of a surface oxidation treatment and 3-aminopropyl triethoxysilane silanisation followed by glutaraldehyde crosslinking. The aim of surface treatment is to stabilise microcontact printing transfer of this natural extracellular matrix protein that usually wears out easily from poly-dimethylsiloxane, which is not suitable for biomimetic cell culture platforms and lab-on-chip applications. A low-cost CD-DVD laser was used to etch biomimetic micropatterns into acrylic sheets that were in turn replicated to poly-dimethylsiloxane slabs with the desired features. These stamps were finally inked with type I collagen for microcontact printing transfer on the culture substrates in a simple manner. Human hepatoma cells (HepG2) and rat primary hepatocytes, which do not adhere to bare poly-dimethylsiloxane, were successfully seeded and showed optimal adhesion and survival on simple protein micropatterns with a hepatic cord geometry in order to validate our technique. HepG2 cells also proliferated on the stamps. Soft and stiff poly-dimethylsiloxane layers were also tested to demonstrate that our cost-effective process is compatible with biomimetic organ-on-chip technology integrating tunable stiffness with a potential application to drug testing probes development where such cells are commonly used.