Tailoring of Architecture and Intrinsic Structure of Electrospun Nanofibers by Process Parameters for Tissue Engineering Applications

Electrospinning process is commercially used to form nanofibers as scaffolds in tissue engineering. Similarities in morphology of electrospun nanofibers to the natural extracellular matrix, flexibility, and low cost of the process contribute to their use in regeneration of cartilage, ligaments/tendons, muscles, and bones. Required properties are tailored by the use of appropriate polymers: polyesters, their copolymers, blends with natural biopolymers such as gelatin, collagen, chitosan, or composites with nanoparticles

Shared patterns in body size declines among crinoids during the Palaeozoic extinction events

Crinoids were among the most abundant marine benthic animals throughout the Palaeozoic, but their body size evolution has received little attention. Here, we compiled a comprehensive database on crinoid calyx biovolumes throughout the Palaeozoic. A model comparison approach revealed contrasting and complex patterns in body size dynamics between the two major crinoid clades (Camerata and Pentacrinoidea). Interestingly, two major drops in mean body size at around two mass extinction events (during the late Ordovician and the late Devonian respectively) are observed, which is reminiscent of current patterns of shrinking body size of a wide range of organisms as a result of climate change. The context of some trends (marked declines during extinctions) suggests the cardinal role of abiotic factors (dramatic climate change associated with extinctions) on crinoid body size evolution; however, other patterns (two intervals with either relative stability or steady size increase in periods between mass extinctions) are more consistent with biotic drivers

Experimental neoichnology of post-autotomy arm movements of sea lilies and possible evidence of thrashing behaviour in Triassic holocrinids

Echinoderms exhibit remarkable powers of autotomy. For instance, crinoids can shed arm and stalk portions when attacked by predators. In some species, it has been reported that the autotomized arms display vigorous movements, which are thought to divert the attention of predators. This phenomenon, however, has not been well explored. Here we present results of experiments using the shallowest water species of living stalked crinoid (Metacrinus rotundus) collected at 140 m depth. A wide range of movements of detached arms, from sluggish writhing to violent flicks, was observed. Interestingly, autotomized arms produce distinct traces on the sediment surface. They are composed of straight or arched grooves usually arranged in radiating groups and shallow furrows. Similar traces were found associated with detached arms of the oldest (Early Triassic) stem-group isocrinid (Holocrinus). This finding may suggest that the origins of autotomy-related thrashing behaviour in crinoids could be traced back to at least the Early Triassic, underscoring the magnitude of anti-predatory traits that occurred during the Mesozoic Marine Revolution. A new ethological category, autotomichnia, is proposed for the traces produced by thrashing movements of shed appendages

Novel 3D Hybrid Nanofiber Scaffolds for Bone Regeneration

Development of hybrid scaffolds and their formation methods occupies an important place in tissue engineering. In this paper, a novel method of 3D hybrid scaffold formation is presented as well as an explanation of the differences in scaffold properties, which were a consequence of different crosslinking mechanisms. Scaffolds were formed from 3D freeze-dried gelatin and electrospun poly(lactide-co-glicolide) (PLGA) fibers in a ratio of 1:1 w/w. In order to enhance osteoblast proliferation, the fibers were coated with hydroxyapatite nanoparticles (HAp) using sonochemical processing. All scaffolds were crosslinked using an EDC/NHS solution. The scaffolds’ morphology was imaged using scanning electron microscopy (SEM). The chemical composition of the scaffolds was analyzed using several methods. Water absorption and mass loss investigations proved a higher crosslinking degree of the hybrid scaffolds than a pure gelatin scaffold, caused by additional interactions between gelatin, PLGA, and HAp. Additionally, mechanical properties of the 3D hybrid scaffolds were higher than traditional hydrogels. In vitro studies revealed that fibroblasts and osteoblasts proliferated and migrated well on the 3D hybrid scaffolds, and also penetrated their structure during the seven days of the experiment

Impact of seawater Mg2+/Ca2+ on Mg/Ca of asterozoan skeleton – Evidence from culturing and the fossil record

Skeletal Mg/Ca in echinoderms is thought to be related to ambient seawater Mg2+/Ca2+ ratio, which prompts the use of well-preserved fossil echinoderms as paleoseawater Mg2+/Ca2+ proxies. Nonetheless, experimental studies testing the effect of seawater Mg2+/Ca2+ ratio on echinoderm skeleton are limited to one echinoderm class, i.e. sea urchins (echinoids). Here, we investigated the effect of decreased seawater Mg2+/Ca2+ ratio on skeletal composition (Mg/Ca ratio) of a sea star Asterias rubens and a brittle star Ophiocomina nigra. Specimens were tagged with manganese and then cultured under three different Mg2+/Ca2+ molar ratios (~5.2, ~2.5, ~1.5 mol/mol) in order to simulate variations of ambient seawater Mg2+/Ca2+ ratio that existed throughout the Phanerozoic. Decreased Mg2+/Ca2+ in seawater did not affect the respiration rates or inhibit calcification of sea stars, which suggests that they were not significantly stressed by the treatment. It did, however, clearly affect growth and mortality in brittle stars, for which reliable geochemical data could not be obtained. Under decreased seawater Mg2+/Ca2+ sea stars produced a skeleton with decreased Mg/Ca ratio. This is consistent with the hypothesis that the skeletal chemistry in echinoderms is influenced by seawater chemistry and, consequently, that fossil echinoderms may preserve a record of paleoseawater Mg2+/Ca2+. Although our data from well-preserved fossil sea stars from the Middle Jurassic and Miocene are consistent with low Mg2+/Ca2+ ratio of the Jurassic “calcite sea” and high Mg2+/Ca2+ ratio of the Miocene "aragonite sea", the accuracy of paleoseawater Mg2+/Ca2+ reconstructions from fossil echinoderms is limited by the fact that Mg partition coefficients vary significantly between different echinoderm species.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Immobilization of Gelatin on Fibers for Tissue Engineering Applications: A Comparative Study of Three Aliphatic Polyesters

Immobilization of cell adhesive proteins on the scaffold surface has become a widely reported method that can improve the interaction between scaffold and cells. In this study, three nanofibrous scaffolds obtained by electrospinning of poly(caprolactone) (PCL), poly(L-lactide-co-caprolactone) (PLCL) 70:30, or poly(L-lactide) (PLLA) were subjected to chemical immobilization of gelatin based on aminolysis and glutaraldehyde cross-linking, as well as physisorption of gelatin. Two sets of aminolysis conditions were applied to evaluate the impact of amine group content. Based on the results of the colorimetric bicinchoninic acid (BCA) assay, it was shown that the concentration of gelatin on the surface is higher for the chemical modification and increases with the concentration of free NH2 groups. XPS (X-ray photoelectron spectroscopy) analysis confirmed this outcome. On the basis of XPS results, the thickness of the gelatin layer was estimated to be less than 10 nm. Initially, hydrophobic scaffolds are completely wettable after coating with gelatin, and the time of waterdrop absorption was correlated with the surface concentration of gelatin. In the case of all physically and mildly chemically modified samples, the decrease in stress and strain at break was relatively low, contrary to strongly aminolyzed PLCL and PLLA samples. Incubation testing performed on the PCL samples showed that a chemically immobilized gelatin layer is more stable than a physisorbed one; however, even after 90 days, more than 60% of the initial gelatin concentration was still present on the surface of physically modified samples. Mouse fibroblast L929 cell culture on modified samples indicates a positive effect of both physical and chemical modification on cell morphology. In the case of PCL and PLCL, the best morphology, characterized by stretched filopodia, was observed after stronger chemical modification, while for PLLA, there was no significant difference between modified samples. Results of metabolic activity indicate the better effect of chemical immobilization than of physisorption of gelatin

Effects of seawater chemistry (Mg 2+ /Ca 2+ ratio) and diet on the skeletal Mg/Ca ratio in the common sea urchin Paracentrotus lividus

It has been argued that concentration of major metallic ions such as Mg 2+ and Ca 2+ plays a role in determining the composition of the echinoderm skeleton. Consequently, in several studies Mg/Ca ratio from modern and fossil echinoderm ossicles was used as a proxy of secular Mg 2+ /Ca 2+ changes of Phanerozoic seawater. However, although significant progress has been made in understanding biomineralization of echinoderms, it is still largely unknown what are the sources and physiological pathways of major ions that contribute to skeleton formation. Herein we tested the effects of modifications of ambient seawater Mg 2+ /Ca 2+ ratio (which is typically ∼5) and Mg-enrichment of the diet on the Mg/Ca ratio in regenerating spines of sea urchin Paracentrotus lividus under experimental conditions. We found that sea urchins cultured in seawater with Mg 2+ /Ca 2+ ratio decreased to ∼1.9 produced a skeleton with also decreased Mg/Ca ratio. However, the skeleton of specimens fed on a Mg-enriched diet showed significantly higher Mg/Ca ratio. This suggests that the seawater is an important but not the only source of ions that contributes to the Mg/Ca ratio of the skeleton. Consequently, the reliability of geochemical models that link directly seawater chemistry with the Mg/Ca ratio of the skeleton should be reevaluated.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Development of Poly(methyl methacrylate)/nano-hydroxyapatite (PMMA/nHA) Nanofibers for Tissue Engineering Regeneration Using an Electrospinning Technique

The study explores the in vitro biocompatibility and osteoconductivity of poly(methyl methacrylate)/nano-hydroxyapatite (PMMA/nHA) composite nanofibrous scaffolds for bone tissue engineering (BTE). Electrospun scaffolds, exhibiting both low and high fiber orientation, were investigated. The inclusion of hydroxyapatite nanoparticles enhances the osteoconductivity of the scaffolds while maintaining the ease of fabrication through electrospinning. SEM analysis confirms the high-quality morphology of the scaffolds, with successful incorporation of nHA evidenced by SEM-EDS and FTIR methods. DSC analysis indicates that nHA addition increases the PMMA glass transition temperature (Tg) and reduces stress relaxation during electrospinning. Furthermore, higher fiber orientation affects PMMA Tg and stress relaxation differently. Biological studies demonstrate the composite material’s non-toxicity, excellent osteoblast viability, attachment, spreading, and proliferation. Overall, PMMA/nHA composite scaffolds show promise for BTE applications

Advanced Graft Development Approaches for ACL Reconstruction or Regeneration

The Anterior Cruciate Ligament (ACL) is one of the major knee ligaments, one which is greatly exposed to injuries. According to the British National Health Society, ACL tears represent around 40% of all knee injuries. The number of ACL injuries has increased rapidly over the past ten years, especially in people from 26–30 years of age. We present a brief background in currently used ACL treatment strategies with a description of surgical reconstruction techniques. According to the well-established method, the PubMed database was then analyzed to scaffold preparation methods and materials. The number of publications and clinical trials over the last almost 30 years were analyzed to determine trends in ACL graft development. Finally, we described selected ACL scaffold development publications of engineering, medical, and business interest. The systematic PubMed database analysis indicated a high interest in collagen for the purpose of ACL graft development, an increased interest in hybrid grafts, a numerical balance in the development of biodegradable and nonbiodegradable grafts, and a low number of clinical trials. The investigation of selected publications indicated that only a few suggest a real possibility of creating healthy tissue. At the same time, many of them focus on specific details and fundamental science. Grafts exhibit a wide range of mechanical properties, mostly because of polymer types and graft morphology. Moreover, most of the research ends at the in vitro stage, using non-certificated polymers, thus requiring a long time before the medical device can be placed on the market. In addition to scientific concerns, official regulations limit the immediate introduction of artificial grafts onto the market