Search bioRxiv⌕ Search

Biology subjects

Spaeker, O.

Publications and source records attributed to Spaeker, O..

4 recordsLinked to original sources

Material-driven fibronectin assembly enables low-dose BMP-2 to regenerate structurally mature bone in critical-sized load-bearing defects

Bone grafts for critical-sized load-bearing defects are usually judged by how much bone forms, and rarely by whether that bone is structurally and mechanically competent. Here, degradable ceramic monetites coated with poly(ethyl acrylate) (PEA) and functionalised with fibronectin (FN) and rhBMP-2 at 10-25 g/mL, 60-150-fold below the approved clinical concentration, are assessed across length scales. MicroCT showed that only monetites functionalised with rhBMP-2 bridged the defect, with a significant increase in new bone volume in both the monetite vicinity and the whole defect volume. In contrast, platelet-rich plasma (PRP) functionalisation significantly reduced bone formation relative to FN, showing that the identity of the adhesion cue, and not merely its presence, determines the outcome. Monetite degradation and increased interconnected porosity favoured bone ingrowth and produced a continuous interface between monetite and new bone. The multiscale analyses of quantitative polarised light microscopy, SAXS and nanoindentation show that PEA-coated monetites functionalised with FN and 25 g/mL rhBMP-2 promoted structurally and mechanically more mature bone, with a hierarchical organisation and local mechanical properties approaching those of native cortical tissue. In the bridged region this group recapitulated the preferential collagen fibre orientation of native cortical bone and reached an elastic modulus and hardness that were not statistically distinguishable from it, although mineral particles throughout the regenerated tissue remained thicker and less aligned than in cortical bone. Synchrotron microCT in an ovine model extended these findings to a clinically relevant scale, supporting the translational potential of the system. Taken together, these findings position PEA-coated monetites functionalised with FN and low-dose rhBMP-2 as a strategy for the structural and functional regeneration of large bone defects.

bioengineering↗

Gradients of orientation, composition and hydration of proteins for efficient light collection by the cornea of the horseshoe crab

The lateral eyes of the horseshoe crab, Limulus polyphemus, are the largest compound eyes within recent Arthropoda. While this visual system has been extensively described before, the precise mechanism allowing vision has remained controversial. Correlating quantitative refractive index (RI) mapping and detailed structural analysis, we demonstrate how gradients of RI in the cornea result from the hierarchical organization of chitin-protein fibers, heterogeneity in protein composition and bromine doping, as well as spatial variation in water content. Combining the realistic cornea structure and measured RI gradients with full-wave optical modelling and ray-tracing approaches, we show that the light collection mechanism depends on both refraction-based graded index (GRIN) optics and total internal reflection. The optical properties of the cornea are governed by different mechanisms at different hierarchical levels, demonstrating the remarkable versatility of arthropod cuticle. One-sentence summaryStructural hierarchy and protein hydration determine the optical performance of the cornea of L. polyphemus.

biophysics↗

Spores and extracellular matrix components impart molecular order in Bacillus subtilis biofilms.

Biofilms are multicellular microbial communities that encase themselves in an extracellular matrix (ECM) of secreted biopolymers and attach to surfaces and interfaces. Bacterial biofilms are detrimental in hospital and industrial settings, but they can be beneficial in agricultural contexts. An essential property of biofilms that grants them with increased survival relative to planktonic cells is phenotypic heterogeneity; the division of the biofilm population into functionally distinct subgroups of cells. Phenotypic heterogeneity in biofilms can be traced to the cellular level, however, the molecular structures and elemental distribution across whole biofilms as well as possible linkages between them remain unexplored. Mapping X-ray diffraction (XRD) across intact biofilms in time and space, we revealed the dominant structural features in Bacillus subtilis biofilms, stemming from matrix components, spores and water. By simultaneously following the X-ray fluorescence (XRF) signal of biofilms and isolated matrix components, we discovered that the ECM preferentially binds calcium ions over other metal ions, specifically, zinc, manganese and iron. These ions, remaining free to flow below macroscopic wrinkles that act as water channels, eventually accumulate and lead to sporulation. The possible link between ECM properties, regulation of metal ion distribution and sporulation across whole intact biofilms unravels the importance of molecular-level heterogeneity in shaping biofilm physiology and development. Significance StatementBiofilms are multicellular soft microbial communities that are able to colonize synthetic surfaces as well as living organisms. To survive sudden environmental changes and efficiently share their common resources, cells in a biofilm divide into subgroups with distinct functions, leading to phenotypic heterogeneity. Here, by studying intact biofilms by synchrotron X-ray diffraction and fluorescence, we revealed correlations between biofilm macroscopic architectural heterogeneity and the spatio-temporal distribution of extracellular matrix, spores, water and metal ions. Our findings demonstrate that biofilm heterogeneity is not only affected by local genetic expression and cellular differentiation, but also by passive effects resulting from the physicochemical properties of the molecules secreted by the cells, leading to differential distribution of nutrients that propagates through macroscopic length scales.

microbiology↗

Epidermal cell surface structure and chitin-protein co-assembly determine fiber architecture in the Locust cuticle

The geometrical similarity of helicoidal fiber arrangement in many biological fibrous extracellular matrices, such as bone, plant cell wall or arthropod cuticle, to that of cholesteric liquid mesophases has led to the hypothesis that they may form passively through a mesophase precursor rather than by direct cellular control. In search of direct evidence to support or refute this hypothesis, here, we studied the process of cuticle formation in the tibia of the migratory locust, Locusta migratoria, where daily growth layers arise by the deposition of fiber arrangements alternating between unidirectional and helicoidal structures. Using FIB/SEM volume imaging and scanning X-ray scattering, we show that the epidermal cells determine an initial fiber orientation from which the final architecture emerges by the self-organized co-assembly of chitin and proteins. Fiber orientation in the locust cuticle is therefore determined by both active and passive processes.

biophysics↗