Spidroin-Based Biomaterials in Tissue Engineering: General Approaches and Potential Stem Cell Therapies

Spider silks are increasingly gaining interest for potential use as biomaterials in tissue engineering and biomedical applications. Owing to their facile and versatile processability in native and regenerated forms, they can be easily tuned via chemical synthesis or recombinant technologies to address specific issues required for applications. In the past few decades, native spider silk and recombinant silk materials have been explored for a wide range of applications due to their superior strength, toughness, and elasticity as well as biocompatibility, biodegradation, and nonimmunogenicity. Herein, we present an overview of the recent advances in spider silk protein that fabricate biomaterials for tissue engineering and regenerative medicine. Beginning with a brief description of biological and mechanical properties of spidroin-based materials and the cellular regulatory mechanism, this review summarizes various types of spidroin-based biomaterials from genetically engineered spider silks and their prospects for specific biomedical applications (e.g., lung tissue engineering, vascularization, bone and cartilage regeneration, and peripheral nerve repair), and finally, we prospected the development direction and manufacturing technology of building more refined and customized spidroin-based protein scaffolds.

Critical role of minor eggcase silk component in promoting spidroin chain alignment and strong fiber formation

Natural spider silk with extraordinary mechanical properties is typically spun from more than one type of spidroin. Although the main components of various spider silks have been widely studied, little is known about the molecular role of the minor silk components in spidroin self-assembly and fiber formation. Here, we show that the minor component of spider eggcase silk, TuSp2, not only accelerates self-assembly but remarkably promotes molecular chain alignment of spidroins upon physical shearing. NMR structure of the repetitive domain of TuSp2 reveals that its dimeric structure with unique charged surface serves as a platform to recruit different domains of the main eggcase component TuSp1. Artificial fiber spun from the complex between TuSp1 and TuSp2 minispidroins exhibits considerably higher strength and Young’s modulus than its native counterpart. These results create a framework for rationally designing silk biomaterials based on distinct roles of silk components.

Comparing Modern and Classical Perspectives on Spider Silks and Webs

Spiders have always fascinated humankind as whilst they are often reviled, their product, the web and its silk, are commonly viewed in awe. As such, silks’ material properties and the fear and fascination surrounding the animals that spin it are seen to play an important role in the development of many cultures and societies. More recently this is even more so with the formalization of this inspiration in scientific and technical communities through biomimetics. The aim of this work is to reflect on the beginnings of our relationship with silk and discuss concepts associated with spider silks and webs in ancient Greek and Roman times whilst comparing this with our current understanding of the field. In this way, ancient texts, namely Greek and Latin ones, are found to intersect with modern advanced disciplines, ranging from architecture to medicine to physics. This allows us not only to understand how natural observation has evolved from antiquity to today, but also how such a highly interdisciplinary research network has been spun by some shared conceptual threads.

Identification and quantification of fatty acids in hunting web of adult Steatoda grossa (Theridiidae) female spiders

This is the first study on composition of fatty acids in hunting web of Steatoda grossa (Theridiidae) spiders and one of only four similar studies ever made. Its main contribution is a discovery that fatty acids not only cover an outside of the web fibers, but they are even more abundantly represented in the fibers’ inner structure. Although little attention has been so far attributed to the contents of fatty acids in spider silks, one has to remember that their biocompatibility combined with an extraordinary tensile strength make them a worth investigating template for material bioengineering studies.

Golden orb-weaving spider (Trichonephila clavipes) silk genes with sex-biased expression and atypical architectures

Spider silks are renowned for their high-performance mechanical properties. Contributing to these properties are proteins encoded by the spidroin (spider fibroin) gene family. Spidroins have been discovered mostly through cDNA studies of females based on the presence of conserved terminal regions and a repetitive central region. Recently, genome sequencing of the golden orb-web weaver, Trichonephila clavipes, provided a complete picture of spidroin diversity. Here, we refine the annotation of T. clavipes spidroin genes including the reclassification of some as non-spidroins. We rename these non-spidroins as spidroin-like (SpL) genes because they have repetitive sequences and amino acid compositions like spidroins, but entirely lack the archetypal terminal domains of spidroins. Insight into the function of these spidroin and SpL genes was then examined through tissue- and sex-specific gene expression studies. Using qPCR, we show that some silk genes are upregulated in male silk glands compared to females, despite males producing less silk in general. We also find that an enigmatic spidroin that lacks a spidroin C-terminal domain is highly expressed in silk glands, suggesting that spidroins could assemble into fibers without a canonical terminal region. Further, we show that two SpL genes are expressed in silk glands, with one gene highly evolutionarily conserved across species, providing evidence that particular SpL genes are important to silk production. Together, these findings challenge long-standing paradigms regarding the evolutionary and functional significance of the proteins and conserved motifs essential for producing spider silks.

Spider Silks: An Overview of Their Component Proteins for Hydrophobicity and Biomedical Applications

: Spider silks have received extensive attention from scientists and industries around the world because of their remarkable mechanical properties, which include high tensile strength and extensibility. It is a leading-edge biomaterial resource, with a wide range of potential applications. Spider silks are composed of silk proteins, which are usually very large molecules, yet many silk proteins still remain largely underexplored. While there are numerous reviews on spider silks from diverse perspectives, here we provide a most up-to-date overview of the spider silk component protein family in terms of its molecular structure, evolution, hydrophobicity, and biomedical applications. Given the confusion regarding spidroin naming, we emphasize the need for coherent and consistent nomenclature for spidroins and provide recommendations for preexisting spidroin names that are inconsistent with nomenclature. We then review recent advances in the components, identification, and structures of spidroin genes. We next discuss the hydrophobicity of spidroins, with particular attention on the unique aquatic spider silks. Aquatic spider silks are less known but may inspire innovation in biomaterials. Furthermore, we provide new insights into antimicrobial peptides from spider silk glands. Finally, we present possibilities for future uses of spider silks.

Tensegrity Modelling and the High Toughness of Spider Dragline Silk

This work establishes a tensegrity model of spider dragline silk. Tensegrity systems are ubiquitous in nature, being able to capture the mechanics of biological shapes through simple and effective modes of deformation via extension and contraction. Guided by quantitative microstructural characterization via air plasma etching and low voltage scanning electron microscopy, we report that this model is able to capture experimentally observed phenomena such as the Poisson effect, tensile stress-strain response, and fibre toughness. This is achieved by accounting for spider silks’ hierarchical organization into microfibrils with radially variable properties. Each fibril is described as a chain of polypeptide tensegrity units formed by crystalline granules operating under compression, which are connected to each other by amorphous links acting under tension. Our results demonstrate, for the first time, that a radial variability in the ductility of tensegrity chains is responsible for high fibre toughness, a defining and desirable feature of spider silk. Based on this model, a discussion about the use of graded tensegrity structures for the optimal design of next-generation biomimetic fibres is presented.

Synthetic Spider Silk Provides New Insights into the Mechanisms of Flagelliform Silk Fiber Assemble and Elastomeric Behavior

<p>In order to better understand the relationship between the elastomeric behavior of Flagelliform (Flag) spider silks and its molecular structure, it was designed and produced the <i>Nephilengys cruentata</i> Flageliform (Flag) spidroin analogue rNcFlag2222. The recombinant proteins are composed by the elastic repetitive glycine-rich motifs (GPGGX/GGX) and the spacer region, rich in hydrophilic charged amino acids, present at the native silk spidroin. Using different approaches for nanomolecular protein analysis, the structural data of rNcFlag2222 recombinant proteins were compared in its fibrillar and in its fully solvated states. Based on the results and previous published data, it was possible to propose a model for the molecular dynamics of Flag spidroins’ repetitive core, during gland storage and fiber formation, and their contribution to its exceptional mechanoelastic properties. This model assumes that the Flag silk proteins acquire elastomeric behavior through a mechanism similar to collagen proteins, with the repetitive glycine-rich and the spacer regions, together with water, playing important roles in fiber assemble and elastomeric behavior.</p>

Synthetic Spider Silk Provides New Insights into the Mechanisms of Flagelliform Silk Fiber Assemble and Elastomeric Behavior

<p>In order to better understand the relationship between the elastomeric behavior of Flagelliform (Flag) spider silks and its molecular structure, it was designed and produced the <i>Nephilengys cruentata</i> Flageliform (Flag) spidroin analogue rNcFlag2222. The recombinant proteins are composed by the elastic repetitive glycine-rich motifs (GPGGX/GGX) and the spacer region, rich in hydrophilic charged amino acids, present at the native silk spidroin. Using different approaches for nanomolecular protein analysis, the structural data of rNcFlag2222 recombinant proteins were compared in its fibrillar and in its fully solvated states. Based on the results and previous published data, it was possible to propose a model for the molecular dynamics of Flag spidroins’ repetitive core, during gland storage and fiber formation, and their contribution to its exceptional mechanoelastic properties. This model assumes that the Flag silk proteins acquire elastomeric behavior through a mechanism similar to collagen proteins, with the repetitive glycine-rich and the spacer regions, together with water, playing important roles in fiber assemble and elastomeric behavior.</p>