Engineered Spider Silk Proteins for Biomimetic Spinning of Fibers with Toughness Equal to Dragline Silks.

Spider silk is the toughest fiber found in nature, and bulk production of artificial spider silk that matches its mechanical properties remains elusive. Development of miniature spider silk proteins (mini-spidroins) has made large-scale fiber production economically feasible, but the fibers' mechanical properties are inferior to native silk. The spider silk fiber's tensile strength is conferred by poly-alanine stretches that are zipped together by tight side chain packing in ß-sheet crystals. Spidroins are secreted so they must be void of long stretches of hydrophobic residues, since such segments get inserted into the endoplasmic reticulum membrane. At the same time, hydrophobic residues have high ß-strand propensity and can mediate tight inter-ß-sheet interactions, features that are attractive for generation of strong artificial silks. Protein production in prokaryotes can circumvent biological laws that spiders, being eukaryotic organisms, must obey, and the authors thus design mini-spidroins that are predicted to more avidly form stronger ß-sheets than the wildtype protein. Biomimetic spinning of the engineered mini-spidroins indeed results in fibers with increased tensile strength and two fiber types display toughness equal to native dragline silks. Bioreactor expression and purification result in a protein yield of ≈9 g L-1 which is in line with requirements for economically feasible bulk scale production.

Image-guided thermal ablation in autonomously functioning thyroid nodules. A retrospective multicenter three-year follow-up study from the Italian Minimally Invasive Treatment of the Thyroid (MITT) Group.

OBJECTIVES: To report the results of a multicenter retrospective evaluation of the clinical outcomes of thermal ablation (TA) in a large series of autonomously functioning thyroid nodules (AFTN) with a follow-up protracted up to 3 years. METHODS: Patients treated with single TA for an AFTN in Italy were included. Changes in nodule volume, TSH values, and ongoing anti-thyroid therapy were assessed at the 2-, 6-, 12-, 24-, and 36-month follow-up controls. Complications and need of any additional therapy after TA were also registered. RESULTS: A total of 361 patients (244 females, 117 males, median age 58 years, IQR 46-70 years) were included. Nodule volume was significantly reduced at all time points (p < 0.001) (median volume reduction 58% at 6-month and 60% at 12-month). Serum TSH values increased significantly at all time points. After TA, anti-thyroid therapy was withdrawn in 32.5% of patients at 2 months, in 38.9% at 6 months, and in 41.3% at 12 months. A significant difference in the rate of patients who withdrawn medical therapy at 12 months was registered between small (< 10 mL) (74%), medium (49%), or large (> 30 mL) nodules (19%). A single major complication occurred (0.25%). Additional treatments were needed in 34/361 (9.4%) of cases including 4 (1.1%) surgical treatment. CONCLUSIONS: Image-guided thermal ablation offers a further safe and effective therapeutic option in patients with AFTN. Clinical outcomes are significantly more favorable in small than in large size AFTN. KEY POINTS: • Thermal ablations (TA) can be safely and effectively used in patients with autonomously functioning thyroid nodules (AFTN). • TA results in a clinically significant nodule volume reduction that is paralleled by TSH level normalization and anti-thyroid drug therapy discontinuation (after TA anti-thyroid therapy was withdrawn in 41.3% at 12 months). • Clinical outcomes after TA are more favorable in small nodules, and when a large amount of thyroid nodule tissue is ablated.

Mechanical Properties and Weibull Scaling Laws of Unknown Spider Silks.

Spider silks present extraordinary mechanical properties, which have attracted the attention of material scientists in recent decades. In particular, the strength and the toughness of these protein-based materials outperform the ones of many man-made fibers. Unfortunately, despite the huge interest, there is an absence of statistical investigation on the mechanical properties of spider silks and their related size effects due to the length of the fibers. Moreover, several spider silks have never been mechanically tested. Accordingly, in this work, we measured the mechanical properties and computed the Weibull parameters for different spider silks, some of them unknown in the literature. We also measured the mechanical properties at different strain rates for the dragline of the species Cupiennius salei. For the same species, we measured the strength and Weibull parameters at different fiber lengths. In this way, we obtained the spider silk scaling laws directly and according to Weibull's prediction. Both length and strain rates affect the mechanical properties of spider silk, as rationalized by Weibull's statistics.

Properties of Biomimetic Artificial Spider Silk Fibers Tuned by PostSpin Bath Incubation.

Efficient production of artificial spider silk fibers with properties that match its natural counterpart has still not been achieved. Recently, a biomimetic process for spinning recombinant spider silk proteins (spidroins) was presented, in which important molecular mechanisms involved in native spider silk spinning were recapitulated. However, drawbacks of these fibers included inferior mechanical properties and problems with low resistance to aqueous environments. In this work, we show that ≥5 h incubation of the fibers, in a collection bath of 500 mM NaAc and 200 mM NaCl, at pH 5 results in fibers that do not dissolve in water or phosphate buffered saline, which implies that the fibers can be used for applications that involve wet/humid conditions. Furthermore, incubation in the collection bath improved the strain at break and was associated with increased ß-sheet content, but did not affect the fiber morphology. In summary, we present a simple way to improve artificial spider silk fiber strain at break and resistance to aqueous solvents.

Quality of life and outcomes in patients with a large toxic adenoma undergoing laser ablation plus radioiodine vs lobectomy.

PURPOSE: Patients with toxic adenomas (TAs) that are too large to undergo radioactive iodine (RAI) treatment aimed at resolving hyperthyroidism and/or relieving mechanical pressure symptoms are referred to surgery. This prospective study aimed to assess the outcomes of combining laser ablation (LA) plus RAI vs lobectomy to treat large TAs in terms of clinical efficacy and the health-related quality of life (HRQoL). PATIENTS AND METHODS: Patients with TAs of volumes greater than 20 mL and a calculated therapeutic activity exceeding 600 Mbq were randomly assigned to undergo LA + RAI (Group A) or lobectomy (Group B). The HRQoL was assessed using 12-item Short Form Health Survey questionnaire before and 6 months after treatment. RESULTS: Twenty-seven patients entered the study. After completing treatment, patients in Group A showed a TA reduction by a mean of 68% compared to baseline. Two of 14 patients (14.3%) in Group A and 2 of 13 (15.4%) in Group B became subclinically hypothyroid, whereas the remaining patients were euthyroid. HRQoL significantly improved in both groups after treatment. CONCLUSIONS: For patients with large TAs, a combination of LA and RAI is a feasible alternative to surgery. Similar to surgery, LA + RAI resolves the mechanical discomfort induced by nodule pressure and effectively treats the hyperthyroidism. This procedure also avoids the potential complications associated with surgery while guaranteeing a similar HRQoL benefit.

Symposium review: The importance of the ruminal epithelial barrier for a healthy and productive cow.

The stratified squamous ruminal epithelium is the main site for absorption of key nutrients (e.g., short-chain fatty acids; SCFA) and electrolytes (e.g., sodium and magnesium). The absorptive function has to be highly selective to prevent simultaneous entry of microbes and toxins from the rumen into the blood. As such, epithelial absorption is primarily transcellular, whereas the paracellular pathway appears rather tightly sealed. A network of tight junction (claudin-1, claudin-4, and occludin) and tight junction-associated proteins (e.g., zonula occludens) accomplishes the latter. When microbial fermentation activity is high such as with highly fermentable diets, rumen epithelial functions are often challenged by acidity, high osmolarity, toxins (e.g., endotoxin and histamine), and immune mediators (inflammatory mediators and cytokines) released during local and systemic inflammation. Epithelial damage by low pH in combination with high luminal SCFA concentrations is not immediately reversible and may initially aggravate upon return to physiological pH. In contrast, barrier opening upon hyperosmolarity is acutely transient. The initial insults set by luminal acidity and SCFA and the increasing concentrations of microbial-associated molecular patterns such as lipopolysaccharides are key factors that trigger inflammation not only in the rumen but also in the hindgut (cecum and colon), which reach out to the liver and other organs, causing systemic inflammation. Low feed intake during parturition, transportation, heat stress, or disease is the second most relevant challenge for the ruminal epithelial barrier. The barrier opening is usually only transient and quickly restored upon refeeding. Due to a rapid, dose-dependent, and prolonged decrease in absorption capacity for SCFA, however, any feed restriction increases the odds for postrestriction subacute ruminal acidosis. Inflammation due to acidosis can be alleviated by supplemental thiamine, yeasts, and plant bioactive (phytogenic) compounds. Butyrate is used in weaning calves to support ruminal barrier development; however, excess butyrate may promote hyperkeratosis, parakeratosis, and epithelial injury in the fully developed rumen of adult cows. Further research is needed to enhance the understanding of the various factors that counteract barrier impairment and help barrier restoration during acidogenic feeding, especially when concurring with unavoidable periods of feed restriction.

Effects of glucagon-like peptides 1 and 2 and epidermal growth factor on the epithelial barrier of the rumen of adult sheep.

Epidermal growth factor (EGF) and glucagon-like peptides (GLP) modulate the tight junctions (TJ) of the intestinal epithelial barrier (EB) of monogastric animals. This work tried to elucidate whether GLP-1, GLP-2 and EGF can affect the EB of the rumen. Ovine ruminal epithelia were incubated in Ussing chambers for 7 hr with 25 or 250 nM of either GLP-1 or GLP-2 on the serosal side, with 2.5 nM of EGF on the serosal side or with 0.25 or 2.5 nM EGF on the mucosal side. No treatment affected tissue conductance. Short-circuit current (Isc ) was affected by time and treatment and their interactions. Only 250 nM of either GLP-1 or GLP-2 decreased Isc in certain periods compared with 25 nM GLP-1 or 0.25 nM mucosally applied EGF; however, not when compared to control epithelia. Fluorescein flux rates (Jfluor ) of ruminal epithelia were affected by treatment, time and time × treatment interaction. The time × treatment interaction was based on an increase in Jfluor between the first and last hour in epithelia incubated with 25 nM GLP-1 or GLP-2 and in epithelia incubated with EGF. After 7 hr incubation, claudin-7 mRNA expression was downregulated in all treatments. Claudin-1 mRNA was upregulated after incubation with 2.5 nM EGF on the serosal side, claudin-4 mRNA was downregulated by 2.5 nM EGF on the mucosal side, and occludin mRNA was increased after incubation with 250 nM GLP-2. The protein abundance of all tested TJ proteins was not influenced by treatment. We conclude that GLP-1, GLP-2, and EGF have no obvious acute effects on the EB of ruminal epithelia under simulated physiological conditions ex vivo. However, by decreasing the mRNA expression of claudin-7 and partly affecting other TJ proteins, they may modulate EB in the longer term or under certain conditions.

Key role of short-chain fatty acids in epithelial barrier failure during ruminal acidosis.

Subacute ruminal acidosis is induced by high concentrations of short-chain fatty acids (SCFA, mainly acetate, propionate, and butyrate) that release protons to decrease the pH of the ruminal digesta. This low pH, in turn, is thought to damage epithelial barrier function. The present study applied a model of simulated ruminal acidosis ex vivo to investigate if SCFA directly contribute to epithelial barrier failure beyond their role as proton donors. Epithelial tissues from the rumen of slaughtered sheep were mounted in Ussing chambers and incubated under 3 different conditions. Two groups were incubated in the absence of SCFA at mucosal pH 6.1 (control) and pH 5.1, respectively, for 7 h. A third group was first incubated in a mucosal solution containing 100 mM SCFA at pH 5.1 for 2 h and, thereafter, in a mucosal solution without SCFA at pH 6.1 for the remaining 5 h. Transepithelial conductance (Gt), short-circuit current (Isc), and fluorescein fluxes were determined. After 7 h of incubation, the expression levels of claudin-1, claudin-4, claudin-7, and occludin were measured by quantitative reverse-transcription PCR and Western blot. Furthermore, the local distribution of these tight junction (TJ) proteins was examined by confocal laser scanning microscopy. A 7-h incubation at pH 5.1 in the absence of SCFA did not influence either Gt or fluorescein flux rates of ruminal tissues ex vivo compared with the control. In contrast, incubation at pH 5.1 with SCFA for only 2 h induced increases in Gt and fluorescein flux rates that continued even after tissues were returned back to pH 6.1. Expression analysis showed that pH 5.1 without SCFA for 7 h induced no changes in mRNA expression of claudin-1, claudin-4, claudin-7, and occludin and a selective decrease in protein expression of only claudin-4 compared with the control. However, a 2-h incubation at pH 5.1 in the presence of SCFA decreased the mRNA-expression of claudin-7, as well as the protein expression of claudin-4, claudin-7, and occludin. The decreased expression of these TJ proteins in the group incubated with SCFA was also evident in immunohistochemistry. Immunohistochemistry additionally evidenced a considerable retraction of all tested TJ proteins out of the TJ in that group. We conclude that a low mucosal pH of 5.1 is tolerated well by ruminal epithelia for several hours. However, a low pH in combination with SCFA induces damage to the TJ and disturbs barrier function, which is not immediately reversible upon the removal of the acidotic insult.

Laser ablation is more effective for spongiform than solid thyroid nodules. A 4-year retrospective follow-up study.

PURPOSE: Laser ablation (LA) is used as therapeutic modality for reducing the volume of large benign thyroid nodules. The aim of this retrospective study was to assess the efficacy of LA therapy in patients with benign non-functioning thyroid nodules in a 4-year follow-up and evaluate whether different compactness of nodules may influence the final shrinkage. PATIENTS AND METHODS: Fifty-six euthyroid patients (42 females; mean age 54.7 ± 11.7 years) with benign cold thyroid solitary nodules or a dominant nodule within a multinodular goitre underwent LA between July 2009 and March 2012. Nodule volume, thyroid function test and ultrasound were monitored at baseline, and at 3, 6 and 12 months after the procedure, then annually. RESULTS: With a mean baseline volume of 15.7 ± 11.7 mL, nodule volume decreased by 55.5% (6.5 ± 5.7 mL) 4 years after LA (p < .01). Nodules had a significant decrease at 3 and 6 months, thereafter they remained stable, with an insignificant tendency to re-growth at 48 months. Thyroid functions and antibodies were unchanged throughout the follow-up. When dividing nodules into solid and spongiform, the former decreased at 6 months, remained stable up to 24 months, but showed a non-significant tendency to increase thereafter. Spongiform nodules progressively decreased up to 48 months. The difference in nodules' reduction between solid and spongiform nodules was significant from 3 months (p = .04) and became even more significant up to 48 months (p = .001). CONCLUSIONS: The LA technique succeeded in reducing thyroid nodules by about 50% at 4 years, but was more effective for spongiform than solid nodules.

Rates of diagnostic and therapeutic procedures in a cohort of patients undergoing first endocrine consultation for thyroid nodular disease.

PURPOSE: This study aimed to evaluate the rates of necessary diagnostic and therapeutic procedures in a cohort of patients undergoing their first endocrine consultation for thyroid nodular disease. METHODS: This was an observational study conducted between January 1 and June 30, 2023, on patients undergoing their first endocrine consultation for thyroid nodular disease. Data were collected, including age, thyroid-stimulating hormone (TSH) concentration, reasons for performing thyroid ultrasound (US), and thyroid US reports. The US was performed at the time of the endocrine consultation according to the American College of Radiology Thyroid Imaging, Reporting and Data System (ACR-TIRADS) risk stratification system. Patients underwent further investigations/treatment or simple US follow-up as necessary. RESULTS: A total of 373 patients with thyroid nodules were evaluated. Reasons for undergoing thyroid US were unrelated to suspected thyroid disease in 126 (33.8%), incidentalomas in 91 (24.4%), dysfunction or positivity for thyroid antibodies in 67 (18%), symptoms or visible nodules in 61 (16.3%), and family history of thyroid disease in 28 (7.5%). A total of 193 diagnostic or therapeutic procedures were performed in 133 (35.7%) patients [fine-needle aspiration (FNA): 121 (62.7%), surgery: 28 (14.5%), percutaneous ethanol injection: 20 (10.4%), scintigraphy: 10 (5.2%); thermal ablation: 7 (3.6%), and radioactive treatment: 7 (3.6%)]. CONCLUSIONS: In the present study only one-third of the patients undergoing endocrine consultation with first detected thyroid nodules required any diagnostic or therapeutic procedures.

Laser ablation for spongiform thyroid nodules: identifying baseline nodule volume and energy to be delivered for successful treatment.

BACKGROUND: Laser ablation (LA) is a minimally invasive treatment. It has been widely used since the early 2000s to induce volume reduction of symptomatic benign thyroid nodules. Up to 40% of laser-treated nodules have been reported to achieve a volume reduction of <50% (technique inefficacy) at 12 months and tend to regrow over time. OBJECTIVE: This study aimed to assess the optimal baseline volume and energy to be delivered to minimize technique inefficacy. METHODS: This was a retrospective study. Data were collected, including baseline volume, energy delivered, and 12-month volume reduction ratio (VRR) of spongiform nodules (EU-TIRADS 2) treated with LA between 2010 and 2020. Based on these data, the optimal baseline volume and energy to be delivered were calculated to maximize the rate of nodules with technique efficacy (VRR ≥ 50% at 12-month follow-up). RESULTS: A total of 205 patients with spongiform nodules were included in this study. The energy delivered was positively associated with VRR. However, no association was observed between baseline volume and VRR. Delivering energy ≥500 J/mL to nodules with a mean baseline volume of 11.4 ± 4 mL resulted in technique efficacy in 83% of cases. CONCLUSION: Treating spongiform nodules with a baseline volume of ≤15 mL and delivering energy ≥500 J/mL are key factors for achieving a relevant rate of technique efficacy.

Patients undergoing endocrine consultation and first diagnosis of nodular disease: Indications of thyroid ultrasound and completeness of ultrasound reports.

PURPOSE: To evaluate reasons for performing ultrasonography (US) and completeness of US reports in patients undergoing endocrine consultation with the first diagnosis of nodular disease. METHODS: Since January 1 to June 30, 2021, we prospectively collected patient data (age and thyroid-stimulating hormone concentrations), reasons for performing thyroid US, and completeness of reports regarding the description of the thyroid gland and nodules. In the case of multiple nodules, we considered the nodule suspected of malignancy and the largest one. To evaluate the accuracy of thyroid nodule description, we referred to the five characteristics suggested by the ACR TI-RADS system. RESULTS: A total of 341 patients with thyroid nodules received endocrine consultation (female, 78%). The most frequent reasons for performing thyroid US were unrelated to a suspected thyroid disease (31.7%), followed by incidentaloma (23.5%), dysfunction or positivity for thyroid antibodies (19.1%), symptomatic or visible nodules (17.6%), and family history of any thyroid disease (8.2%). Gland texture was not reported in 41.9%. The depth of the lobes was the dimension reported most frequently (42.2%), but any diameter was not reported in 57.8% of the cases. As regards the description of the most relevant nodule, length was reported more frequently (75.9%). Margins and echogenicity were more frequently described (54.5% and 44.3%, respectively) than other characteristics (composition: 27%; shape: 8.8%; echogenic foci: 6.7%). No reports had indicated the malignancy risk stratification. CONCLUSIONS: The results of the study demonstrate that in patients undergoing endocrine consultation with first detected thyroid nodules, US was mostly performed in asymptomatic cases, US reports were incomplete, and no risk stratification system was reported.

Influence of experimental methods on the mechanical properties of silk fibers: A systematic literature review and future road map.

Spider silk fibers are of scientific and industrial interest because of their extraordinary mechanical properties. These properties are normally determined by tensile tests, but the values obtained are dependent on the morphology of the fibers, the test conditions, and the methods by which stress and strain are calculated. Because of this, results from many studies are not directly comparable, which has led to widespread misconceptions in the field. Here, we critically review most of the reports from the past 50 years on spider silk mechanical performance and use artificial spider silk and native silks as models to highlight the effect that different experimental setups have on the fibers' mechanical properties. The results clearly illustrate the importance of carefully evaluating the tensile test methods when comparing the results from different studies. Finally, we suggest a protocol for how to perform tensile tests on silk and biobased fibers.

The mechanical characterization of the legs, fangs, and prosoma in the spider Harpactira curvipes (Pocock 1897).

The exoskeleton of spiders is the primary structure that interacts with the external mechanical stimuli, thus playing a crucial role in spider life. In particular, fangs, legs, and prosoma are the main rigid structures of the exoskeleton and their properties must be measured to better understand their mechanical behaviours. Here we investigate, by means of nanoindentation, the mechanical properties of the external sclerotized cuticles of such parts in the spider Harpactira curvipes. Interestingly, the results show that the leg's cuticle is stiffer than the prosoma and has a stiffness similar to the one of the tip fangs. This could be explained by the legs' function in perceiving vibrations that could be facilitated by higher stiffness. From a broader perspective, this characterization could help to understand how the same basic material (the cuticle, i.e. mainly composed of chitin) can be tuned to achieve different mechanical functions, which improves the animal's adaptation to specific evolutive requirements. We, thus, hope that this work stimulates further comparative analysis. Moreover, these results may also be potentially important to inspire the design of graded materials with superior mechanical properties.

Artificial and natural silk materials have high mechanical property variability regardless of sample size.

Silk fibres attract great interest in materials science for their biological and mechanical properties. Hitherto, the mechanical properties of the silk fibres have been explored mainly by tensile tests, which provide information on their strength, Young's modulus, strain at break and toughness modulus. Several hypotheses have been based on these data, but the intrinsic and often overlooked variability of natural and artificial silk fibres makes it challenging to identify trends and correlations. In this work, we determined the mechanical properties of Bombyx mori cocoon and degummed silk, native spider silk, and artificial spider silk, and compared them with classical commercial carbon fibres using large sample sizes (from 10 to 100 fibres, in total 200 specimens per fibre type). The results confirm a substantial variability of the mechanical properties of silk fibres compared to commercial carbon fibres, as the relative standard deviation for strength and strain at break is 10-50%. Moreover, the variability does not decrease significantly when the number of tested fibres is increased, which was surprising considering the low variability frequently reported for silk fibres in the literature. Based on this, we prove that tensile testing of 10 fibres per type is representative of a silk fibre population. Finally, we show that the ideal shape of the stress-strain curve for spider silk, characterized by a pronounced exponential stiffening regime, occurs in only 25% of all tested spider silk fibres.

Prey localization in spider orb webs using modal vibration analysis.

Spider webs are finely tuned multifunctional structures, widely studied for their prey capture functionalities such as impact strength and stickiness. However, they are also sophisticated sensing tools that enable the spider to precisely determine the location of impact and capture the prey before it escapes. In this paper, we suggest a new mechanism for this detection process, based on potential modal analysis capabilities of the spider, using its legs as distinct distributed point sensors. To do this, we consider a numerical model of the web structure, including asymmetry in the design, prestress, and geometrical nonlinearity effects. We show how vibration signals deriving from impacts can be decomposed into web eigenmode components, through which the spider can efficiently trace the source location. Based on this numerical analysis, we discuss the role of the web structure, asymmetry, and prestress in the imaging mechanism, confirming the role of the latter in tuning the web response to achieve an efficient prey detection instrument. The results can be relevant for efficient distributed impact sensing applications.

An Image-Analysis-Based Method for the Prediction of Recombinant Protein Fiber Tensile Strength.

Silk fibers derived from the cocoon of silk moths and the wide range of silks produced by spiders exhibit an array of features, such as extraordinary tensile strength, elasticity, and adhesive properties. The functional features and mechanical properties can be derived from the structural composition and organization of the silk fibers. Artificial recombinant protein fibers based on engineered spider silk proteins have been successfully made previously and represent a promising way towards the large-scale production of fibers with predesigned features. However, for the production and use of protein fibers, there is a need for reliable objective quality control procedures that could be automated and that do not destroy the fibers in the process. Furthermore, there is still a lack of understanding the specifics of how the structural composition and organization relate to the ultimate function of silk-like fibers. In this study, we develop a new method for the categorization of protein fibers that enabled a highly accurate prediction of fiber tensile strength. Based on the use of a common light microscope equipped with polarizers together with image analysis for the precise determination of fiber morphology and optical properties, this represents an easy-to-use, objective non-destructive quality control process for protein fiber manufacturing and provides further insights into the link between the supramolecular organization and mechanical functionality of protein fibers.

Silk fibroin films with embedded magnetic nanoparticles: evaluation of the magneto-mechanical stimulation effect on osteogenic differentiation of stem cells.

We report about a biomaterial in the form of film ∼10 µm thick, consisting of a silk fibroin matrix with embedded iron oxide superparamagnetic nanoparticles, for prospective applications as bioactive coating in regenerative medicine. Films with different load of magnetic nanoparticles are produced (nanoparticles/silk fibroin nominal ratio = 5, 0.5 and 0 wt%) and the structural, mechanical and magnetic properties are studied. The nanoparticles form aggregates in the silk fibroin matrix and the film stiffness, as tested by nanoindentation, is spatially inhomogeneous, but the protein structure is not altered. In vitro biological tests are carried out on human bone marrow-derived mesenchymal stem cells cultured on the films up to 21 days, with and without an applied static uniform magnetic field. The sample with the highest nanoparticles/silk fibroin ratio shows the best performance in terms of cell proliferation and adhesion. Moreover, it promotes a faster and better osteogenic differentiation, particularly under magnetic field, as indicated by the gene expression level of typical osteogenic markers. These findings are explained in light of the results of the physical characterization, combined with numerical calculations. It is established that the applied magnetic field triggers a virtuous magneto-mechanical mechanism in which dipolar magnetic forces between the nanoparticle aggregates give rise to a spatial distribution of mechanical stresses in the silk fibroin matrix. The film with the largest nanoparticle load, under cell culture conditions (i.e. in aqueous environment), undergoes matrix deformations large enough to be sensed by the seeded cells as mechanical stimuli favoring the osteogenic differentiation.

1000 spider silkomes: Linking sequences to silk physical properties.

Spider silks are among the toughest known materials and thus provide models for renewable, biodegradable, and sustainable biopolymers. However, the entirety of their diversity still remains elusive, and silks that exceed the performance limits of industrial fibers are constantly being found. We obtained transcriptome assemblies from 1098 species of spiders to comprehensively catalog silk gene sequences and measured the mechanical, thermal, structural, and hydration properties of the dragline silks of 446 species. The combination of these silk protein genotype-phenotype data revealed essential contributions of multicomponent structures with major ampullate spidroin 1 to 3 paralogs in high-performance dragline silks and numerous amino acid motifs contributing to each of the measured properties. We hope that our global sampling, comprehensive testing, integrated analysis, and open data will provide a solid starting point for future biomaterial designs.

How spiders hunt heavy prey: the tangle web as a pulley and spider's lifting mechanics observed and quantified in the laboratory.

The spiders of Theridiidae's family display a peculiar behaviour when they hunt extremely large prey. They lift the quarry, making it unable to escape, by attaching pre-tensioned silk threads to it. In this work, we analysed for the first time in the laboratory the lifting hunting mechanism and, in order to quantify the phenomenon, we applied the lifting mechanics theory. The comparison between the experiments and the theory suggests that, during the process, spiders do not stretch the silk too much by keeping it in the linear elastic regime. We thus report here further evidence for the strong role of silk in spiders' evolution, especially how spiders can stretch and use it as an external tool to overcome their muscles' limits and capture prey with large mass, e.g. 50 times the spider's mass.