Analytical model of friction at low shear rates for soft materials in 3D printing.

BACKGROUND: The biological properties of silicone elastomers such as polydimethylsiloxane (PDMS) have widespread use in biomedicine for soft tissue implants, contact lenses, soft robots, and many other small medical devices, due to its exceptional biocompatibility. Additive manufacturing of soft materials still has significant challenges even with major advancements that have occurred in development of these technologies for customized medical devices and tissue engineering. OBJECTIVE: The aim of this study was to develop a mathematical model of tangential stress in relation to shear stress, shear rate, 3D printing pressure and velocity, for non-Newtonian gels and fluids that are used as materials for 3D printing. METHOD: This study used FENE (finitely extensible nonlinear elastic model) model, for non-Newtonian gels and fluids to define the dependences between tangential stress, velocity, and pressure, considering viscosity, shear stress and shear rates as governing factors in soft materials friction and adhesion. Experimental samples were fabricated as showcases, by SLA and FDM 3D printing technologies: elastic polymer samples with properties resembling elastic properties of PDMS and thermoplastic polyurethane (TPU) samples. Experimental 3D printing parameters were used in the developed analytical solution to analyse the relationships between governing influential factors (tangential stress, printing pressure, printing speed, shear rate and friction coefficient). Maple software was used for numerical modelling. RESULTS: Analytical model applied on a printed elastic polymer, at low shear rates, exhibited numerical values of tangential stress of 0.208-0.216 N m - 2 at printing velocities of 0.9 to 1.2 mm s - 1, while the coefficient of friction was as low as 0.09-0.16. These values were in accordance with experimental data in literature. Printing pressure did not significantly influence tangential stress, whereas it was slightly influenced by shear rate changes. Friction coefficient linearly increased with tangential stress. CONCLUSION: Simple analytical model of friction for elastic polymer in SLA 3D printing showed good correspondence with experimental literature data for low shear rates, thus indicating possibility to use it for prediction of printing parameters towards desired dimensional accuracy of printed objects. Further development of this analytical model should enable other shear rate regimes, as well as additional soft materials and printing parameters.

Hydrophobic-Interaction-Induced Stiffening of α-Synuclein Fibril Networks.

In water, networks of semiflexible fibrils of the protein α-synuclein stiffen significantly with increasing temperature. We make plausible that this reversible stiffening is a result of hydrophobic contacts between the fibrils that become more prominent with increasing temperature. The good agreement of our experimentally observed temperature dependence of the storage modulus of the network with a scaling theory linking network elasticity with reversible cross-linking enables us to quantify the endothermic binding enthalpy and estimate the effective size of hydrophobic patches on the fibril surface. Our findings may not only shed light on the role of amyloid deposits in disease conditions, but can also inspire new approaches for the design of thermoresponsive materials.

Oligomers of Parkinson's Disease-Related α-Synuclein Mutants Have Similar Structures but Distinctive Membrane Permeabilization Properties.

Single-amino acid mutations in the human α-synuclein (αS) protein are related to early onset Parkinson's disease (PD). In addition to the well-known A30P, A53T, and E46K mutants, recently a number of new familial disease-related αS mutations have been discovered. How these mutations affect the putative physiological function of αS and the disease pathology is still unknown. Here we focus on the H50Q and G51D familial mutants and show that like wild-type αS, H50Q and G51D monomers bind to negatively charged membranes, form soluble partially folded oligomers with an aggregation number of ~30 monomers under specific conditions, and can aggregate into amyloid fibrils. We systematically studied the ability of these isolated oligomers to permeabilize membranes composed of anionic phospholipids (DOPG) and membranes mimicking the mitochondrial phospholipid composition (CL:POPE:POPC) using a calcein release assay. Small-angle X-ray scattering studies of isolated oligomers show that oligomers formed from wild-type αS and the A30P, E46K, H50Q, G51D, and A53T disease-related mutants are composed of a similar number of monomers. However, although the binding affinity of the monomeric protein and the aggregation number of the oligomers formed under our specific protocol are comparable for wild-type αS and H50Q and G51D αS, G51D oligomers cannot disrupt negatively charged and physiologically relevant model membranes. Replacement of the membrane-immersed glycine with a negatively charged aspartic acid at position 51 apparently abrogates membrane destabilization, whereas a mutation in the proximal but solvent-exposed part of the membrane-bound α-helix such as that found in the H50Q mutant has little effect on the bilayer disrupting properties of oligomers.

Alpha-synuclein amyloid oligomers act as multivalent nanoparticles to cause hemifusion in negatively charged vesicles.

Multivalent membrane binding sites on the α-synuclein oligomer result in clustering of vesicles and hemifusion of negatively charged model membranes. These multivalent, biological nanoparticles are reminiscent of inorganic nanoparticles in their interactions with membranes. Alpha-synuclein oligomers induce lipid exchange efficiently, with fewer than 10 oligomers/vesicle required to complete hemifusion. No full fusion or vesicle content mixing is observed.

Membrane interactions and fibrillization of α-synuclein play an essential role in membrane disruption.

We studied α-synuclein (αS) aggregation in giant vesicles, and observed dramatic membrane disintegration, as well as lipid incorporation into micrometer-sized suprafibrillar aggregates. In the presence of dye-filled vesicles, dye leakage and fibrillization happen concurrently. However, growing fibrils do not impair the integrity of phospholipid vesicles that have a low affinity for αS. Seeding αS aggregation accelerates dye leakage, indicating that oligomeric species are not required to explain the observed effect. The evolving picture suggests that fibrils that appear in solution bind membranes and recruit membrane-bound monomers, resulting in lipid extraction, membrane destabilization and the formation of lipid-containing suprafibrillar aggregates.

α-Synuclein oligomers distinctively permeabilize complex model membranes.

α-Synuclein oligomers are increasingly considered to be responsible for the death of dopaminergic neurons in Parkinson's disease. The toxicity mechanism of α-synuclein oligomers likely involves membrane permeabilization. Even though it is well established that α-synuclein oligomers bind and permeabilize vesicles composed of negatively-charged lipids, little attention has been given to the interaction of oligomers with bilayers of physiologically relevant lipid compositions. We investigated the interaction of α-synuclein with bilayers composed of lipid mixtures that mimic the composition of plasma and mitochondrial membranes. In the present study, we show that monomeric and oligomeric α-synuclein bind to these membranes. The resulting membrane leakage differs from that observed for simple artificial model bilayers. Although the addition of oligomers to negatively-charged lipid vesicles displays fast content release in a bulk permeabilization assay, adding oligomers to vesicles with compositions mimicking mitochondrial membranes shows a much slower loss of content. Oligomers are unable to induce leakage in the artificial plasma membranes, even after long-term incubation. CD experiments indicate that binding to lipid bilayers initially induces conformational changes in both oligomeric and monomeric α-synuclein, which show little change upon long-term incubation of oligomers with membranes. The results of the present study demonstrate that the mitochondrial model membranes are more vulnerable to permeabilization by oligomers than model plasma membranes reconstituted from brain-derived lipids; this preference may imply that increasingly complex membrane components, such as those in the plasma membrane mimic used in the present study, are less vulnerable to damage by oligomers.