Gustavo R. Plaza

Stretching of supercontracted fibers: a link between spinning and the variability of spider silk.

SUMMARY The spinning of spider silk requires a combination of aqueous environment and stretching, and the aim of this work was to explore the role of stretching silk fibers in an aqueous environment and its effect on the tensile properties of spider silk. In particular, the sensitivity of the spider silk tensile behaviour to wet-stretching could be relevant in the search for a relationship between processing and the variability of the tensile properties. Based on this idea and working with MAS silk from Argiope trifasciata orb-web building spiders, we developed a novel procedure that permits modification of the tensile properties of spider silk: silk fibers were allowed to supercontract and subsequently stretched in water. The ratio between the length after stretching and the initial supercontracted length was used to control the process. Tensile tests performed in air, after drying, demonstrated that this simple procedure allows to predictable reproduction of the stress-strain curves of either naturally spun or forcibly silked fibers. These results suggest that the supercontracted state has a critical biological function during the spinning process of spider silk.

Self-tightening of spider silk fibers induced by moisture

Abstract Spider dragline silk has a unique combination of desirable mechanical properties—low density, high tensile strength and large elongation until breaking—that makes it attractive from an engineering perspective [Nature 410 (2001) 541]. Nevertheless, this outstanding performance is threatened by the way mechanical properties are affected by a wet environment, particularly if the stress of these fibers can relax when exposed to moisture. Tests on spider dragline silk ( Argiope trifasciata ) performed by the authors have shown that when the fiber is clamped and exposed to a wet enough environment non-vanishing supercontraction forces develop. When the moisture is removed the residual stresses increase, and this effect has proven long lasting, as the fiber remains stressed for hours. In addition, the tensile properties of the fiber remain unaffected by the residual stresses build up after removing the moisture or after a wetting and drying cycle. These tests give support to the thesis that supercontraction helps to keep the spider webs tight and opens new applications for synthetic analogs.

The effect of spinning forces on spider silk properties.

SUMMARY A new forced silking procedure has been developed that allows measurement of the low forces involved in the silking process and, subsequently, retrieval and tensile testing of the samples spun at the measured silking forces. A strong correlation between silking force and tensile behaviour of spider silk has been established. Fibres spun at high silking force – compared with the conventional yield stress – are stiff and show stress–strain curves previously found in forcibly silked fibres. By contrast, fibres spun at low and very low silking forces are more compliant, and their tensile behaviour corresponds to that of fibres naturally spun by the spider or to fibres subjected to maximum supercontraction, respectively. It has also been found that samples retrieved from processes with significant variations in the silking force are largely variable in terms of force–displacement curves, although reproducibility improves if force is re-scaled into stress. Fibres retrieved from processes with constant silking force show similar tensile properties both in terms of force–displacement and stress–strain curves.

The hidden link between supercontraction and mechanical behavior of spider silks.

The remarkable properties of spider silks have stimulated an increasing interest in understanding the roles of their composition and processing, as well as in the mass-production of these fibers. Previously, the variability in the mechanical properties of natural silk fibers was a major drawback in the elucidation of their behavior, but the authors have found that supercontraction of these fibers allows one to characterize and reproduce the whole range of tensile properties in a consistent way. The purpose of this review is to summarize these findings. After a review of the pertinent mechanical properties, the role of supercontraction in recovering and tailoring the tensile properties is explained, together with an alignment parameter to characterize silk fibers. The concept of the existence of a mechanical ground state is also mentioned. These behaviors can be modeled, and two such models-at the molecular and macroscopic levels-are briefly outlined. Finally, the assessment of the existence of supercontraction in bio-inspired fibers is considered, as this property may have significant consequences in the design and production of artificial fibers.

Mechanical Behavior of Silk During the Evolution of Orb-Web Spinning Spiders

The development of an accurate and reproducible approach to measuring the tensile behavior of spider silk has allowed characterizing and comparing the range of mechanical properties exhibited by different spider species with unprecedented detail. The comparison of silks spun by spiders belonging to different phylogenetic groups has revealed that evolution locked in many of the important properties of spider silks very early in the history of orb-web weaving spiders, despite the fact that the silk gland system is relatively isolated in physiological terms from the rest of the organism and should thus mutate quickly. The variations observed between species may be grouped in at least two patterns that are shown not to be related to phylogeny. Beyond the relevance of these results for the evolutionary biology of spiders and silks, the conservation of the basic traits observed in the mechanical behavior of spider silks is likely to set a limit to the range of properties that can be expected from artificial fibers bioinspired in natural silks.

Relationship between microstructure and mechanical properties in spider silk fibers: identification of two regimes in the microstructural changes

The relationship between microstructure and mechanical properties has been investigated in Argiope trifasciata dragline silk fibers (major ampullate silk, MAS) by X-ray diffraction, Raman spectroscopy and tensile testing. We have analyzed three fractions of the material, i.e. amorphous, highly oriented nanocrystals and weakly oriented material, for different values of the macroscopic alignment parameter α, calculated as the relative difference between the length of the fiber and its length when supercontracted. Two distinct regimes have been identified: for low values of the alignment parameter α, microstructural changes are dominated by the reorientation of the nanocrystals; however, at high values (α > 0.5) of the alignment parameter, an increase in the fraction of the crystalline phase is revealed. The two regimes are also reflected in the mechanical behaviour, which can be explained by microstructural changes. This finding of the two distinct regimes in the microstructural evolution, which separates the reorientation and the increase in the crystalline phase, will be valuable to develop and validate molecular models of natural and artificial silk fibers, as well as to deepen our present knowledge of the origin of the outstanding properties of MAS fibers. In addition, we have analyzed the characteristics of the crystal lattice, and discussed the relationship between the percentage of short side-chain residues and the unit cell dimensions in different silks.

Sequential origin in the high performance properties of orb spider dragline silk

Major ampullate (MA) dragline silk supports spider orb webs, combining strength and extensibility in the toughest biomaterial. MA silk evolved ~376 MYA and identifying how evolutionary changes in proteins influenced silk mechanics is crucial for biomimetics, but is hindered by high spinning plasticity. We use supercontraction to remove that variation and characterize MA silk across the spider phylogeny. We show that mechanical performance is conserved within, but divergent among, major lineages, evolving in correlation with discrete changes in proteins. Early MA silk tensile strength improved rapidly with the origin of GGX amino acid motifs and increased repetitiveness. Tensile strength then maximized in basal entelegyne spiders, ~230 MYA. Toughness subsequently improved through increased extensibility within orb spiders, coupled with the origin of a novel protein (MaSp2). Key changes in MA silk proteins therefore correlate with the sequential evolution high performance orb spider silk and could aid design of biomimetic fibers.

Minor ampullate silks from Nephila and Argiope spiders: tensile properties and microstructural characterization.

The mechanical behavior and microstructure of minor ampullate gland silk (miS) of two orb-web spinning species, Argiope trifasciata and Nephila inaurata, were extensively characterized, enabling detailed comparison with other silks. The similarities and differences exhibited by miS when compared with the intensively studied major ampullate gland silk (MAS) and silkworm (Bombyx mori) silk offer a genuine opportunity for testing some of the hypotheses proposed to correlate microstructure and tensile properties in silk. In this work, we show that miSs of different species show similar properties, even when fibers spun by spiders that diverged over 100 million years are compared. The tensile properties of miS are comparable to those of MAS when tested in air, significantly in terms of work to fracture, but differ considerably when tested in water. In particular, miS does not show a supercontraction effect and an associated ground state. In this regard, the behavior of miS in water is similar to that of B. mori silk, and it is shown that the initial elastic modulus of both fibers can be explained using a common model. Intriguingly, the microstructural parameters measured in miS are comparable to those of MAS and considerably different from those found in B. mori. This fact suggests that some critical microstructural information is still missing in our description of silks, and our results suggest that the hydrophilicity of the lateral groups or the large scale organization of the sequences might be routes worth exploring.

Biomimetic spinning of artificial spider silk from a chimeric minispidroin

Herein we present a chimeric recombinant spider silk protein (spidroin) whose aqueous solubility equals that of native spider silk dope and a spinning device that is based solely on aqueous buffers, shear forces and lowered pH. The process recapitulates the complex molecular mechanisms that dictate native spider silk spinning and is highly efficient; spidroin from one liter of bacterial shake-flask culture is enough to spin a kilometer of the hitherto toughest as-spun artificial spider silk fiber.

Supercontraction of dragline silk spun by lynx spiders (Oxyopidae).

Supercontraction is commonly considered as a functional adaptation of major ampullate gland (MA) silk to its role as the main structural material in orb-webs. However, the observation of supercontraction in the dragline silk of a lynx spider species, as it is shown in this work, offers a strong support to the hypothesis that the appearance of supercontraction preceded the advent of capture webs. Moreover, the absence of proline in the sequence of dragline silk spidroin in Oxyopidae and related spiders indicates that the presence of this amino acid may not be required for the existence of supercontraction. In this regard, the presence of particular subrepeats--in orb-web and non-orb-web building spiders--adds new clues for the understanding of supercontraction and associated effects.