ABSTRACT
Cell-matrix interactions govern cell behavior and tissue function by facilitating transduction of biomechanical cues. Engineered tissues often incorporate these interactions by employing cell-adhesive materials. However, using constitutively active cell-adhesive materials impedes control over cell fate and elicits inflammatory responses upon implantation. Here, an alternative cell-material interaction strategy that provides mechanotransducive properties via discrete inducible on-cell crosslinking (DOCKING) of materials, including those that are inherently non-cell-adhesive, is introduced. Specifically, tyramine-functionalized materials are tethered to tyrosines that are naturally present in extracellular protein domains via enzyme-mediated oxidative crosslinking. Temporal control over the stiffness of on-cell tethered 3D microniches reveals that DOCKING uniquely enables lineage programming of stem cells by targeting adhesome-related mechanotransduction pathways acting independently of cell volume changes and spreading. In short, DOCKING represents a bioinspired and cytocompatible cell-tethering strategy that offers new routes to study and engineer cell-material interactions, thereby advancing applications ranging from drug delivery, to cell-based therapy, and cultured meat.
Subject(s)
Biocompatible Materials/chemistry , Mechanotransduction, Cellular , Animals , Biocompatible Materials/metabolism , Biocompatible Materials/pharmacology , Cell Adhesion/drug effects , Cell Differentiation/drug effects , Cell Lineage , Dextrans/chemistry , Horseradish Peroxidase/metabolism , Humans , Hydrogels/chemistry , Integrins/metabolism , Mechanotransduction, Cellular/drug effects , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Mice , Mice, Inbred C57BL , Oligopeptides/chemistry , Oxidation-Reduction , Tyramine/chemistryABSTRACT
Cell-laden micrometer-sized hydrogels (microgels) hold great promise for improving high throughput ex-vivo drug screening and engineering biomimetic tissues. Microfluidics is a powerful tool to produce microgels. However, only a limited amount of biomaterials have been reported to be compatible with on-chip microgel formation. Moreover, these biomaterials are often associated with mechanical instability, cytotoxicity, and cellular senescence. To resolve this challenge, dextran-tyramine has been explored as a novel biomaterial for on-chip microgel formation. In particular, dextran-tyramine is compared with two commonly used biomaterials, namely, polyethylene-glycol diacrylate (PEGDA) and alginate, which crosslink through enzymatic reaction, UV polymerization, and ionic interaction, respectively. Human mesenchymal stem cells (hMSCs) encapsulated in dextran-tyramine microgels demonstrate significantly higher (95%) survival as compared to alginate (81%) and PEGDA (69%). Long-term cell cultures demonstrate that hMSCs in PEGDA microgels become senescent after 7 d. Alginate microgels dissolve within 7 d due to Ca2+ loss. In contrast, dextran-tyramine based microgels remain stable, sustain hMSCs metabolic activity, and permit for single-cell level analysis for at least 28 d of culture. In conclusion, enzymatically crosslinking dextran-tyramine conjugates represent a novel biomaterial class for the on-chip production of cell-laden microgels, which possesses unique advantages as compared to the commonly used UV and ionic crosslinking biomaterials.
Subject(s)
Alginates/chemistry , Cross-Linking Reagents/chemistry , Lab-On-A-Chip Devices , Mesenchymal Stem Cells/metabolism , Microfluidic Analytical Techniques/methods , Polyethylene Glycols/chemistry , Cell Culture Techniques/methods , Cells, Cultured , Cells, Immobilized/cytology , Cells, Immobilized/metabolism , Gels , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Humans , Mesenchymal Stem Cells/cytology , Time FactorsABSTRACT
Mechanical properties of hydrogel particles are of importance for their interactions with cells or tissue, apart from their relevance to other applications. While so far the majority of works aiming at tuning particle mechanics relied on chemical cross-linking, we report a novel approach using inwards interweaving self-assembly of poly(allylamine) (PA) and poly(styrenesulfonic acid) (PSSA) on agarose gel beads. Using this technique, shell thicknesses up to tens of micrometers can be achieved from single-polymer incubations and accurately controlled by varying the polymer concentration or incubation period. We quantified the changes in mechanical properties of hydrogel core-shell particles. The effective elastic modulus of core-shell particles was determined from force spectroscopy measurements using the colloidal probe-AFM (CP-AFM) technique. By varying the shell thickness between 10 and 24 µm, the elastic modulus of particles can be tuned in the range of 10-190 kPa and further increased by increasing the layer number. Through fluorescence quantitative measurements, the polymeric shell density was found to increase together with shell thickness and layer number, hence establishing a positive correlation between elastic modulus and shell density of core-shell particles. This is a valuable method for constructing multidensity or single-density shells of tunable thickness and is particularly important in mechanobiology as studies have reported enhanced cellular uptake of particles in the low-kilopascal range (<140 kPa). We anticipate that our results will provide the first steps toward the rational design of core-shell particles for the separation of biomolecules or systemic study of stiffness-dependent cellular uptake.
Subject(s)
Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Mechanical Phenomena , Polymers/chemistry , Hydrogel, Polyethylene Glycol Dimethacrylate/chemical synthesis , Particle Size , Polyamines/chemistry , Polymers/chemical synthesis , Polystyrenes/chemistry , Surface PropertiesABSTRACT
A fabrication method of a multifunctional hybrid material is achieved by using the insoluble organic nacre matrix of the Haliotis laevigata shell infiltrated with gelatin as a confined reaction environment. Inside this organic scaffold magnetite nanoparticles (MNPs) are synthesized. The amount of MNPs can be controlled through the synthesis protocol therefore mineral loadings starting from 15 wt % up to 65 wt % can be realized. The demineralized organic nacre matrix is characterized by small-angle and very-small-angle neutron scattering (SANS and VSANS) showing an unchanged organic matrix structure after demineralization compared to the original mineralized nacre reference. Light microscopy and confocal laser scanning microscopy studies of stained samples show the presence of insoluble proteins at the chitin surface but not between the chitin layers. Successful and homogeneous gelatin infiltration in between the chitin layers can be shown. The hybrid material is characterized by TEM and shows a layered structure filled with MNPs with a size of around 10 nm. Magnetic analysis of the material demonstrates superparamagnetic behavior as characteristic for the particle size. Simulation studies show the potential of collagen and chitin to act as nucleators, where there is a slight preference of chitin over collagen as a nucleator for magnetite. Colloidal-probe AFM measurements demonstrate that introduction of a ferrogel into the chitin matrix leads to a certain increase in the stiffness of the composite material.
ABSTRACT
The physico-chemical properties of colloidal particles determine their uptake into cells. For a series of microparticles only one parameter, the mechanical stiffness, was varied, whereas other parameters such as size, shape, and charge were kept constant. The uptake was monitored in situ by analyzing individual particle trajectories including the progress of endocytosis, derived from local pH measurements around each particle. Evidence is presented that soft particles with low stiffness are transported faster to lysosomes than stiffer ones.
Subject(s)
Colloids/metabolism , Lysosomes/metabolism , Capsules/chemistry , Capsules/metabolism , Cell Line , Colloids/chemistry , Elastic Modulus , Electrolytes/chemistry , Endocytosis , HeLa Cells , Humans , Hydrogen-Ion Concentration , MCF-7 Cells , Microscopy, Atomic Force , Nanoparticles/chemistryABSTRACT
Microcapsules are reviewed with special emphasis on the relevance of controlled mechanical properties for functional aspects. At first, assembly strategies are presented that allow control over the decisive geometrical parameters, diameter and wall thickness, which both influence the capsule's mechanical performance. As one of the most powerful approaches the layer-by-layer technique is identified. Subsequently, ensemble and, in particular, single-capsule deformation techniques are discussed. The latter generally provide more in-depth information and cover the complete range of applicable forces from smaller than pN to N. In a theory chapter, we illustrate the physics of capsule deformation. The main focus is on thin shell theory, which provides a useful approximation for many deformation scenarios. Finally, we give an overview of applications and future perspectives where the specific design of mechanical properties turns microcapsules into (multi-)functional devices, enriching especially life sciences and material sciences.
Subject(s)
Capsules/chemistry , Drug Compounding , Drug Delivery Systems , Models, Chemical , Chemical Phenomena , Drug Compounding/trends , Mechanical Phenomena , Nanocapsules/chemistry , Nanotechnology/trendsABSTRACT
Droplet microfluidics is combined with bio-orthogonal thiol-ene click chemistry to fabricate micrometer-sized, monodisperse fibrinogen-containing hyaluronic acid hydrogel microbeads in a mild, radical-free procedure in the presence of human mesenchymal stem cells (hMSCs). The gel beads serve as microniches for the 3D culture of single hMSCs, containing hyaluronic acid and additional fibrinogen for cell surface binding, and they are porous and stable in tissue culture medium for up to 4 weeks with mechanical properties right in the range of soft solid tissues (0.9-9.2 kPa). The encapsulation procedure results in 70% viable hMSCs in the microbeads after 24 hours of culture and a very high degree of viability of the cells after long term culture of 2 weeks. hMSCs embedded in the microniches display an overall rounded morphology, consistent with those previously observed in 3D culture. Upon induction, the multipotency and differentiation potential of the hMSCs are characterized by staining of corresponding biomarkers, demonstrating a clear heterogeneity in the cell population. These hydrogel microbeads represent a versatile microstructured material platform with great potential for studying the differences of material cues and soluble factors in stem cell differentiation in a 3D tissue-like environment at the single cell level.
ABSTRACT
While soft hydrogel nano- and microstructures hold great potential for therapeutic treatments and in vivo applications, their nanomechanical characterization remains a challenge. In this paper, soft, single-component, supported hydrogel films were fabricated using pendant-thiol-modified poly(methacrylic acid) (PMASH). The influence of hydrogel architecture on deformation properties was studied by fabricating films on particle supports and producing free-standing capsules. The influence of the degree of thiol-based cross-linking on the mechanical properties of the soft hydrogel systems (core-shell and capsules) was studied using a colloidal-probe (CP) AFM technique. It was found that film mechanical properties, stability, and capsule swelling could be finely tuned by controlling the extent of poly(methacrylic acid) thiol modification. Furthermore, switching the pH from 7.4 to 4.0 led to film densification due to increased hydrogen bonding. Hydrogel capsule systems were found to have stiffness values ranging from 0.9 to 16.9 mN m(-1) over a thiol modification range of 5 to 20 mol %. These values are significantly greater than those for previously reported PMASH planar films of 0.7-5.7 mN m(-1) over the same thiol modification range (Best et al., Soft Matter 2013, 9, 4580-4584). Films on particle substrates had comparable mechanical properties to planar films, demonstrating that while substrate geometry has a negligible effect, membrane and tension effects may play an important role in capsule force resistance. Further, when transitioning from solid-supported films to free-standing capsules, simple predictions of shell stiffness based on modulus changes found for supported films are not valid. Rather, additional effects like diameter increases (geometrical changes) as well as tension buildup need to be taken into account. These results are important for research related to the characterization of soft hydrogel materials and control over their mechanical properties.
Subject(s)
Hydrogel, Polyethylene Glycol Dimethacrylate/chemistry , Capsules/chemistry , Hydrogen-Ion Concentration , Molecular Structure , Particle Size , Polymethacrylic Acids/chemistry , Sulfhydryl Compounds/chemistry , Surface PropertiesABSTRACT
A droplet-based microfluidics technique is used to produce monodisperse, 80 µm collagen-gelatin beads with tunable mechanical properties in the range of 1-10 kPa after photo-crosslinking. The gel beads are porous, mechanically robust and stable in buffer, but can be degraded enzymatically. Encapsulated fibroblast cells maintain 70% viability after one-week encapsulation and preliminary results show that the degree of spreading of cells in gels is correlated with the stiffness of the material.
ABSTRACT
Spider silk fibers are well known for their mechanical properties, and they are therefore in the focus of materials scientists. Additionally, silks display biocompatibility making them interesting materials for applications in medicine or cosmetics. Due to the low abundance of natural spider silk proteins because of the spider's cannibalism, the recombinant spider silk protein eADF4 has been established for material science applications. Once processed into micron-sized particles by controlled salting-out, these particles can be used as drug delivery vehicles. For any application of the silk particles it is important to know their mechanical characteristics for processing and storage reasons. Here, we examine the swelling behavior and mechanics of these particles. Upon hydration, a drastic drop in elastic modulus occurs by orders of magnitude, from 0.8 GPa in the dry state to 2.99 MPa in the wet state. Importantly, the elastic modulus of recombinant silk particles can be tuned by varying the molecular weight of the used proteins, as well as chemical crosslinking thereof.
ABSTRACT
This work aims at establishing a link between process conditions and resulting micromechanical properties for aminoplast core/shell microcapsules. The investigated capsules were produced by the in situ polymerization of melamine formaldehyde resins, which represents a widely used and industrially relevant approach in the field of microencapsulation. Within our study, we present a quantitative morphological analysis of the capsules' size and shell thickness. The diameter of the investigated capsules ranged from 10 to 50 µm and the shell thickness was found in a range between 50 and 200 nm. As key parameter for the control of the shell thickness, we identified the amount of amino resin per total surface area of the dispersed phase. Mechanical properties were investigated using small deformations on the order of the shell thickness by atomic force microscopy with a colloidal probe setup. The obtained capsule stiffness increased with an increasing shell thickness from 2 to 30 N/m and thus showed the same trend on the process parameters as the shell thickness. A simple analytical model was adopted to explain the relation between capsules' geometry and mechanics and to estimate the elastic modulus of the shell about 1.7 GPa. Thus, this work provides strategies for a rational design of microcapsule mechanics.