Silk, “pattern” of natural fibrous polymer : vibrationnal and nano/micromechanical analysis from fibre to composite

Les protéines fibreuses (kératine, élastine, collagène, fibroïne…) représentent 1/3 des protéinesconstitutives des mammifères et des oiseaux. Ce sont des protéines qui ont une fonction de protection et/oumécanique. La soie apparait comme le système le plus « simple » car elle est principalement constituée demotifs de répétition à base d’alanine et de glycine, deux petits acides aminés. Certaines soies présentent despropriétés mécaniques comparables ou supérieures à celles des fibres synthétiques et seraient susceptiblesd’être de nouveau largement utilisées dans des applications techniques (par exemple biomédicales) si lavariabilité de leurs propriétés était maîtrisée. Ce travail porte sur la structure des soies grèges ou décreuséesde Bombyx mori (ver à soie domestique), de Nephila madagascariensis (araignée sauvage, fibre sansenveloppe de séricine), de Bombyx mori génétiquement modifié (incluant un gène de Nephila) et sur unesoie recombinante 4RepCT (Escherichia coli). La soie est analysée par spectrométrie Raman (et IRTF) ettraction uni-axiale, ainsi que par le couplage de ces méthodes. L’analyse de la région des bas nombresd’onde en spectroscopie Raman a permis de caractériser des régions ordonnées de 2 à 3 μm de long etdistantes d’environ 60 μm. Il s’agit de la première mise en évidence d’une hétérogénéité de structure de lasoie. Le couplage avec la traction uni-axiale montre une sollicitation de ces régions ordonnées sousdéformation, suggérant une organisation de la soie selon le modèle de Prevorsek, c’est à dire qu’une mêmechaîne macromoléculaire appartient à la fois à des régions amorphes et à des régions ordonnées. L’étudestatistique des propriétés mécaniques de la soie de ver et d’araignée montre une grande distribution, maisune bonne stabilité dans le temps (dizaines d’années). La modification génétique ne procure pasd’amélioration des propriétés mécaniques de la fibre, seulement une légère diminution de la variabilité.Diverses stratégies sont mises en oeuvre pour tenter d’échapper à cette variabilité : production bactérienne,solubilisation de la soie et régénération sous forme de films. Le rôle de l’eau lors de la biosynthèse de lasoie, ainsi que l’effet de divers paramètres (filtration, pH, séchage…) lors de la préparation des films ont étéétudiés. Nous avons pu confirmer que la présence d’agrégats de protéines favorise l’organisation dans lesfilms et 2 types de films ont donc été préparés. Les films les plus amorphes présentent les propriétésmécaniques les plus intéressantes, même si elles n’atteignent de quelques % de celles des fibres. Lafabrication de composites à matrice de soie régénérée renforcée par des fibres de soie permet d’augmenterla résistance et la déformation à rupture. Ces premiers résultats sont encourageants pour le développementde matériaux composites fibres de soie/matrice de soie régénérée.Fibrous proteins (keratin, elastin, collagen, fibroin ...) make up to one third of the proteins ofmammals and birds. They are structural proteins with a protective and/or mechanical function. Silk appearsto be the ‘simplest’ model because it mainly consists of two small amino acids residues (alanine andglycine). Some silks have comparable or superior mechanical properties compared to those of syntheticfibres and could be used in technical applications (e.g. biomedical) if the variability of their properties canbe controlled. This work focuses on the structure of silks from: Bombyx mori (domestic silkworm)degummed or not, Nephila madagascariensis (wild spider, no sericin coating), GM Bombyx mori (includinga gene of Nephila) a recombinant spider silk 4RepCT (Escherichia Coli). Silk is analyzed by Ramanspectroscopy (and FTIR), uni-axial tensile testing, and also by the coupling of these methods. The analysisof the low wavenumbers region in Raman spectroscopy allowed the characterization of ordered regions of 2to 3 microns separated by about 60 microns. This is the first evidence of the heterogeneous structure ofsilk. Coupling with the uni-axial tensile test shows that these ordered regions are stressed under macroscopicdeformation, suggesting silk organization according to Prevorsek’s model, i.e. that the samemacromolecular chain belongs to both amorphous and ordered regions. The statistical study of themechanical properties of silkworm and spider silks shows great dispersion, but a good stability over time(decades). Genetic modification does not improve the fibres mechanical properties but a slight decrease intheir variability. Various strategies have been investigated to control the variability: bacterial production,solubilization of silk and films regeneration. The role of water in silk biosynthesis, as well as the effect ofvarious parameters (filtration, pH, drying ...) during the preparation of the films were studied. It wasconfirmed that the presence of protein aggregates promotes the organization in film and two types of filmswere prepared. The most amorphous ones have the most interesting mechanical properties, though only afew percent of those from the starting fibres. The fabrication of regenerated silk matrix compositesreinforced by silk fibres increases the strength and strain to failure. These initial results are encouraging forthe development of silk fibres/regenerated silk matrix composite materials

La soie, "modèle" de polymères naturels fibreux : analyse vibrationnelle et nano/micromécanique, de la fibre au composite

Fibrous proteins (keratin, elastin, collagen, fibroin ...) make up to one third of the proteins ofmammals and birds. They are structural proteins with a protective and/or mechanical function. Silk appearsto be the ‘simplest’ model because it mainly consists of two small amino acids residues (alanine andglycine). Some silks have comparable or superior mechanical properties compared to those of syntheticfibres and could be used in technical applications (e.g. biomedical) if the variability of their properties canbe controlled. This work focuses on the structure of silks from: Bombyx mori (domestic silkworm)degummed or not, Nephila madagascariensis (wild spider, no sericin coating), GM Bombyx mori (includinga gene of Nephila) a recombinant spider silk 4RepCT (Escherichia Coli). Silk is analyzed by Ramanspectroscopy (and FTIR), uni-axial tensile testing, and also by the coupling of these methods. The analysisof the low wavenumbers region in Raman spectroscopy allowed the characterization of ordered regions of 2to 3 microns separated by about 60 microns. This is the first evidence of the heterogeneous structure ofsilk. Coupling with the uni-axial tensile test shows that these ordered regions are stressed under macroscopicdeformation, suggesting silk organization according to Prevorsek’s model, i.e. that the samemacromolecular chain belongs to both amorphous and ordered regions. The statistical study of themechanical properties of silkworm and spider silks shows great dispersion, but a good stability over time(decades). Genetic modification does not improve the fibres mechanical properties but a slight decrease intheir variability. Various strategies have been investigated to control the variability: bacterial production,solubilization of silk and films regeneration. The role of water in silk biosynthesis, as well as the effect ofvarious parameters (filtration, pH, drying ...) during the preparation of the films were studied. It wasconfirmed that the presence of protein aggregates promotes the organization in film and two types of filmswere prepared. The most amorphous ones have the most interesting mechanical properties, though only afew percent of those from the starting fibres. The fabrication of regenerated silk matrix compositesreinforced by silk fibres increases the strength and strain to failure. These initial results are encouraging forthe development of silk fibres/regenerated silk matrix composite materials.Les protéines fibreuses (kératine, élastine, collagène, fibroïne…) représentent 1/3 des protéinesconstitutives des mammifères et des oiseaux. Ce sont des protéines qui ont une fonction de protection et/oumécanique. La soie apparait comme le système le plus « simple » car elle est principalement constituée demotifs de répétition à base d’alanine et de glycine, deux petits acides aminés. Certaines soies présentent despropriétés mécaniques comparables ou supérieures à celles des fibres synthétiques et seraient susceptiblesd’être de nouveau largement utilisées dans des applications techniques (par exemple biomédicales) si lavariabilité de leurs propriétés était maîtrisée. Ce travail porte sur la structure des soies grèges ou décreuséesde Bombyx mori (ver à soie domestique), de Nephila madagascariensis (araignée sauvage, fibre sansenveloppe de séricine), de Bombyx mori génétiquement modifié (incluant un gène de Nephila) et sur unesoie recombinante 4RepCT (Escherichia coli). La soie est analysée par spectrométrie Raman (et IRTF) ettraction uni-axiale, ainsi que par le couplage de ces méthodes. L’analyse de la région des bas nombresd’onde en spectroscopie Raman a permis de caractériser des régions ordonnées de 2 à 3 μm de long etdistantes d’environ 60 μm. Il s’agit de la première mise en évidence d’une hétérogénéité de structure de lasoie. Le couplage avec la traction uni-axiale montre une sollicitation de ces régions ordonnées sousdéformation, suggérant une organisation de la soie selon le modèle de Prevorsek, c’est à dire qu’une mêmechaîne macromoléculaire appartient à la fois à des régions amorphes et à des régions ordonnées. L’étudestatistique des propriétés mécaniques de la soie de ver et d’araignée montre une grande distribution, maisune bonne stabilité dans le temps (dizaines d’années). La modification génétique ne procure pasd’amélioration des propriétés mécaniques de la fibre, seulement une légère diminution de la variabilité.Diverses stratégies sont mises en oeuvre pour tenter d’échapper à cette variabilité : production bactérienne,solubilisation de la soie et régénération sous forme de films. Le rôle de l’eau lors de la biosynthèse de lasoie, ainsi que l’effet de divers paramètres (filtration, pH, séchage…) lors de la préparation des films ont étéétudiés. Nous avons pu confirmer que la présence d’agrégats de protéines favorise l’organisation dans lesfilms et 2 types de films ont donc été préparés. Les films les plus amorphes présentent les propriétésmécaniques les plus intéressantes, même si elles n’atteignent de quelques % de celles des fibres. Lafabrication de composites à matrice de soie régénérée renforcée par des fibres de soie permet d’augmenterla résistance et la déformation à rupture. Ces premiers résultats sont encourageants pour le développementde matériaux composites fibres de soie/matrice de soie régénérée

The effect of heat on keratin and implications for the archaeological record

A horncore feature was encountered during excavations at Border Cave, in Member 2 BSL, dated 60–49 ka. The basal half of the horncore lay towards the centre of a combustion feature and was calcined. The tip half lay on a mat of burnt grass bedding towards the edge of the fireplace. It was covered with a black shiny residue, which was also present on nearby stone tools. The feature was jacketed and excavated later under a microscope in the laboratory. Raman spectroscopy identified the residue only as amorphous carbon, indicating the presence of a heated organic substance. Knowing that variation exists in modern and archaeological glue recipes, we wanted to know if the black residue was melted keratin, and if it was, whether it had been used as a substitute for beeswax or latex for hafting. We set out to test the hypothesis by conducting a heating experiment with a modern sheathed horncore. Experimental results showed that keratin does not transform into viable glue when heated. Instead, it reduces immediately to a glassy brittle residue. Our finding strongly supports a scenario of incidental deposition of residue on the archaeological stone tools. Previous combustion experiments have shown that the sediment below a fire can reach 300+ °C through vertical heat transfer. This implies that horn sheaths in archaeological deposits, separated from overlying combustion features by thousands of years, can transform into a black residue that may deposit on nearby stone and bone tools.Fil: Backwell, Lucinda Ruth. University of the Witwatersrand; Sudáfrica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto Superior de Estudios Sociales. Universidad Nacional de Tucumán. Instituto Superior de Estudios Sociales; ArgentinaFil: Wojcieszak, Marine. University of the Witwatersrand; Sudáfrica. Royal Institute For Cultural Heritage; BélgicaFil: Wadley, Lyn. University of the Witwatersrand; Sudáfric

Was yellow lead chromate pigment used during Middle Stone Age at Sibudu rock shelter (South Africa)?

A recent study using the combination of optical microscopy and Raman spectroscopy revealed the presence of bright yellow residues of lead (II) chromate associated with grinding striations on a 58,000-year-old grindstone excavated at Sibudu rock shelter in South Africa. Lead (II) chromate (PbCrO4) exists in nature as the rare mineral crocoite, however was available as the synthetic pigment, chrome yellow, from the 19th century. At the time of the residue analysis on the grindstone, it was not possible to determine whether the yellow residues were due to modern contamination or a result of ancient use. Careful analyses of the excavation protocol followed at Sibudu rock shelter and all implements that might possibly have been in contact with the grindstone after excavation identified yellow spray paint as a possible origin of the contamination. Here, we performed experimental grinding of the mineral crocoite on a sandstone slab, sprayed yellow paint on rhyolite and compared optical microscopy and Raman spectroscopic analysis results to the archaeological sample. The optical appearance of the archaeological residues, the absence of associated minerals of crocoite, and the presence of organic matter mixed with lead chromate demonstrate that the residues on the archaeological grindstone stem from modern contamination. This paper draws attention to contamination issues originating from excavation practices

Micro-Mechanics of Fresh and 30-year-old Nephila Inaurata Madagascariensis Dragline Silk

International audienc

Evidence for large land snail cooking and consumption at Border Cave c. 170–70 ka ago. Implications for the evolution of human diet and social behaviour

Fragments of land snail (Achatinidae) shell were found at Border Cave in varying proportions in all archaeological members, with the exception of the oldest members 5 WA and 6 BS (>227,000 years ago). They were recovered in relatively high frequencies in Members 4 WA, 4 BS, 1 RGBS and 3 WA. The shell fragments present a range of colours from lustrous beige to brown and matt grey. The colour variability can occur when shell is heated. This possibility was explored here through experimental heating of giant land snail shell fragments (Achatinidae, Metachatina kraussi - brown lipped agate snail) in a muffle furnace from 200 to 550 °C for different lengths of time. Colour change, weight loss, and shattering of the heated samples were recorded. Transformation of aragonite into calcite and the occurrence of organic material was investigated by means of Infrared and Raman spectroscopy. Scanning electron microscopy was also used on selected specimens to help identify heat-induced transformation as opposed to taphonomic alteration. The identification on archaeological fragments of features produced by experimentally heating shells at high temperatures or for long periods has led us, after discarding alternative hypotheses, to conclude that large African land snails were systematically brought to the site by humans, roasted and consumed, starting from 170,000 years ago and, more intensively between 160,000 and 70,000 years ago. Border Cave is at present the earliest known site at which this subsistence strategy is recorded. Previous research has shown that charred whole rhizomes and fragments of edible Hypoxis angustifolia were also brought to Border Cave to be roasted and shared at the site. Thus, evidence from both the rhizomes and snails in Border Cave supports an interpretation of members of the group provisioning others at a home base, which gives us a glimpse into the complex social life of early Homo sapiens.Fil: Wojcieszak, Marine. University of the Witwatersrand; SudáfricaFil: Backwell, Lucinda Ruth. Grupo de Investigacion En Arqueologia Andina (arqand) ; Facultad de Cs.naturales E Instituto Miguel Lillo ; Universidad Nacional de Tucuman; . Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; ArgentinaFil: d’Errico, Francesco. Universite de Bordeaux; FranciaFil: Wadley, Lyn. University of the Witwatersrand; Sudáfric

Fire and grass-bedding construction 200 thousand years ago at Border Cave, South Africa

Early plant use is seldom described in the archaeological record because of poor preservation. We report the discovery of grass bedding used to create comfortable areas for sleeping and working by people who lived in Border Cave at least 200,000 years ago. Sheaves of grass belonging to the broad-leafed Panicoideae subfamily were placed near the back of the cave on ash layers that were often remnants of bedding burned for site maintenance. This strategy is one forerunner of more-complex behavior that is archaeologically discernible from ~100,000 years ago

Fire and grass-bedding construction 200 thousand years ago at Border Cave, South Africa

Early plant use is seldom described in the archaeological record because of poor preservation. We report the discovery of grass bedding used to create comfortable areas for sleeping and working by people who lived in Border Cave at least 200,000 years ago. Sheaves of grass belonging to the broad-leafed Panicoideae subfamily were placed near the back of the cave on ash layers that were often remnants of bedding burned for site maintenance. This strategy is one forerunner of more-complex behavior that is archaeologically discernible from ~100,000 years ago

Beads and bead residues as windows to past behaviours and taphonomy: a case study from Grassridge Rockshelter, Eastern Cape, South Africa

© 2020, Springer-Verlag GmbH Germany, part of Springer Nature. Ostrich eggshell and gastropod shell beads provide important evidence for understanding how past peoples decorated and cultured their bodies and may also be used as proxy evidence for interpreting the nature and extent of past social networks. This study focuses on the ostrich eggshell and gastropod shell bead assemblages from the terminal Pleistocene (~ 13.5 to 11.6 ka) and mid-Holocene (~ 7.3 to 6.7 ka) occupations from Grassridge Rockshelter, South Africa. We present results from a multi-method approach to understanding bead manufacture and use that combines a technological analysis of the bead assemblages with Raman spectroscopy. Raman spectroscopy analyses were conducted on surface residues identified on the beads, ochre pieces, a grooved stone, and sediment samples, and provide further insight into past behaviours and taphonomy, as well as modern contaminants. Results indicate that ostrich eggshell beads were manufactured at Grassridge during both occupations, and that bead size changed through time. Use-wear and residue analyses demonstrate the complex taphonomy associated with bead studies from archaeological contexts, and the need for further taphonomic research. These analyses also suggest that some ostrich eggshell and Nassarius beads were potentially worn against ochred surfaces, such as skin or hide, as evidenced by the amount and location of the ochreous residues identified on the beads

Water dependent structural changes of silk from Bombyx mori gland to fibre as evidenced by Raman and IR spectroscopies

International audienceDSC, attenuated total reflexion infrared (ATR-FTIR), and Raman microspectroscopy were used for the first time in a close to in vivo environment to study ready-to-spin Bombyx mori silkworms. The aim was to understand the change of the fibroin backbone organisation from the gland to the fibre. Raman shifts of the Amide I components reveal a strong change of organisation in the middle part of the hydrated gland, as anticipated previously measured modifications of salts concentrations and pH. Series of bands characteristics of the fully hydrated silk disappear, as observed for spider silk, despite the different aminoacid sequence. Confirmation is obtained from IR spectra taking into account the superimposed water component. The change of the silk water interaction in the central part of the gland, from a hydrophobic to hydrophilic behaviour, is related to the water content decrease along the gland. pH sensitive carboxylate side chains markers confirm the modification. Fibroin organisation was also studied in the dried gland and in the spun fibre. The fibre extrusion by orients the fibroin chains along the fibre axis, with intercalated water molecules, leading to a material with specific mechanical properties, compared to the amorphous dried gland