Engineering of Metabolic Pathways by Artificial Enzyme Channels.

Application of industrial enzymes for production of valuable chemical compounds has greatly benefited from recent developments in Systems and Synthetic Biology. Both, in vivo and in vitro systems have been established, allowing conversion of simple into complex compounds. Metabolic engineering in living cells needs to be balanced which is achieved by controlling gene expression levels, translation, scaffolding, compartmentation, and flux control. In vitro applications are often hampered by limited protein stability/half-life and insufficient rates of substrate conversion. To improve stability and catalytic activity, proteins are post-translationally modified and arranged in artificial metabolic channels. Within the review article, we will first discuss the supramolecular organization of enzymes in living systems and second summarize current and future approaches to design artificial metabolic channels by additive manufacturing for the efficient production of desired products

Structure optimisation and biological evaluation of bone scaffolds prepared by co-sintering of silicate and phosphate glasses

A degradable phosphate glass (ICEL) and a bioactive silicate glass (CEL2) were mixed in different ratios (wt-%: 100%ICEL, 70%ICEL-30%CEL2, 30%ICEL-70%CEL2, 100%CEL2; codes 100-0, 70-30, 30-70, 0-100) and then co-sintered to obtain three-dimensional porous scaffolds by gel casting foaming. Thermal analyses were carried out on the glass mixtures and were used as a starting point for the optimisation of the scaffold sintering treatment. The microcomputed tomography and field emission scanning electron microscope analyses allowed the selection of the optimal sintering temperature to obtain an adequate structure in terms of total and open porosity. The scaffolds showed an increasing solubility with increasing ICEL glass content, and for 30-70 and 0-100, the precipitation of hydroxyapatite in simulated body fluid was observed. In vitro tests indicated that all the scaffolds showed no cytotoxic effect. The co-sintering of silicate and phosphate glasses showed to be a promising strategy to tailor the scaffold osteoconductivity, degradation and bioactivit

Bioactivity in silica/poly(γ-glutamic acid) sol–gel hybrids through calcium chelation

Bioactive glasses and inorganic/organic hybrids have great potential as biomedical implant materials. Sol–gel hybrids with interpenetrating networks of silica and biodegradable polymers can combine the bioactive properties of a glass with the toughness of a polymer. However, traditional calcium sources such as calcium nitrate and calcium chloride are unsuitable for hybrids. In this study calcium was incorporated by chelation to the polymer component. The calcium salt form of poly(γ-glutamic acid) (γCaPGA) was synthesized for use as both a calcium source and as the biodegradable toughening component of the hybrids. Hybrids of 40 wt.% γCaPGA were successfully formed and had fine scale integration of Ca and Si ions, according to secondary ion mass spectrometry imaging, indicating a homogeneous distribution of organic and inorganic components. 29Si magic angle spinning nuclear magnetic resonance data demonstrated that the network connectivity was unaltered with changing polymer molecular weight, as there was no perturbation to the overall Si speciation and silica network formation. Upon immersion in simulated body fluid a hydroxycarbonate apatite surface layer formed on the hybrids within 1 week. The polymer molecular weight (Mw 30–120 kDa) affected the mechanical properties of the resulting hybrids, but all hybrids had large strains to failure, >26%, and compressive strengths, in excess of 300 MPa. The large strain to failure values showed that γCaPGA hybrids exhibited non-brittle behaviour whilst also incorporating calcium. Thus calcium incorporation by chelation to the polymer component is justified as a novel approach in hybrids for biomedical materials

Cytotoxicity, chemical stability, and surface properties of ferroelectric ceramics for biomaterials

© 2017 The American Ceramic Society Surface chemistry and topo-physical properties determine the interactions of biomaterials with their physiological environment. Ferroelectrics hold great promise as the next generation of scaffolds for tissue repair since they feature tunable surface electrical charges, piezoelectricity, and sensing capabilities. We investigate the topography, wettability, chemical stability, and cytotoxicity in salient ferroelectric systems such as (1−x) (Na1/2Bi1/2)TiO3–xBaTiO3, (1−x)Ba(Zr0.2Ti0.8)O3−x(Ba0.7Ca0.3)TiO3, and Pb(Zr,Ti)O3 to test their suitability as biomaterials. The lead-free ferroelectrics promote in vitro cell viability and proliferation to a considerably high extent. 0.94 mol % (Na1/2Bi1/2)TiO3–0.06 mol% BaTiO3 showed the greatest potential leading to a cell viability of (149 ± 30)% and DNA synthesis of (299 ± 85)% in comparison to the reference. Lead leaching from Pb(Zr,Ti)O3 negatively affected the cultured cells. Wettability and chemical stability are key factors that determine the cytotoxicity of ferroelectrics. These variables have to be considered in the design of novel electroactive scaffolds based on ferroelectric ceramics

On the reproducibility of extrusion-based bioprinting: round robin study on standardization in the field

The outcome of three-dimensional (3D) bioprinting heavily depends, amongst others, on the interaction between the developed bioink, the printing process, and the printing equipment. However, if this interplay is ensured, bioprinting promises unmatched possibilities in the health care area. To pave the way for comparing newly developed biomaterials, clinical studies, and medical applications (i.e. printed organs, patient-specific tissues), there is a great need for standardization of manufacturing methods in order to enable technology transfers. Despite the importance of such standardization, there is currently a tremendous lack of empirical data that examines the reproducibility and robustness of production in more than one location at a time. In this work, we present data derived from a round robin test for extrusion-based 3D printing performance comprising 12 different academic laboratories throughout Germany and analyze the respective prints using automated image analysis (IA) in three independent academic groups. The fabrication of objects from polymer solutions was standardized as much as currently possible to allow studying the comparability of results from different laboratories. This study has led to the conclusion that current standardization conditions still leave room for the intervention of operators due to missing automation of the equipment. This affects significantly the reproducibility and comparability of bioprinting experiments in multiple laboratories. Nevertheless, automated IA proved to be a suitable methodology for quality assurance as three independently developed workflows achieved similar results. Moreover, the extracted data describing geometric features showed how the function of printers affects the quality of the printed object. A significant step toward standardization of the process was made as an infrastructure for distribution of material and methods, as well as for data transfer and storage was successfully established

On the reproducibility of extrusion-based bioprinting: round robin study on standardization in the field

The outcome of three-dimensional (3D) bioprinting heavily depends, amongst others, on the interaction between the developed bioink, the printing process, and the printing equipment. However, if this interplay is ensured, bioprinting promises unmatched possibilities in the health care area. To pave the way for comparing newly developed biomaterials, clinical studies, and medical applications (i.e. printed organs, patient-specific tissues), there is a great need for standardization of manufacturing methods in order to enable technology transfers. Despite the importance of such standardization, there is currently a tremendous lack of empirical data that examines the reproducibility and robustness of production in more than one location at a time. In this work, we present data derived from a round robin test for extrusion-based 3D printing performance comprising 12 different academic laboratories throughout Germany and analyze the respective prints using automated image analysis (IA) in three independent academic groups. The fabrication of objects from polymer solutions was standardized as much as currently possible to allow studying the comparability of results from different laboratories. This study has led to the conclusion that current standardization conditions still leave room for the intervention of operators due to missing automation of the equipment. This affects significantly the reproducibility and comparability of bioprinting experiments in multiple laboratories. Nevertheless, automated IA proved to be a suitable methodology for quality assurance as three independently developed workflows achieved similar results. Moreover, the extracted data describing geometric features showed how the function of printers affects the quality of the printed object. A significant step toward standardization of the process was made as an infrastructure for distribution of material and methods, as well as for data transfer and storage was successfully established

Emergence of spatially heterogeneous burst suppression in a neural field model of electrocortical activity

Burst suppression in the electroencephalogram (EEG) is a well-described phenomenon that occurs during deep anesthesia, as well as in a variety of congenital and acquired brain insults. Classically it is thought of as spatially synchronous, quasi-periodic bursts of high amplitude EEG separated by low amplitude activity. However, its characterization as a “global brain state” has been challenged by recent results obtained with intracranial electrocortigraphy. Not only does it appear that burst suppression activity is highly asynchronous across cortex, but also that it may occur in isolated regions of circumscribed spatial extent. Here we outline a realistic neural field model for burst suppression by adding a slow process of synaptic resource depletion and recovery, which is able to reproduce qualitatively the empirically observed features during general anesthesia at the whole cortex level. Simulations reveal heterogeneous bursting over the model cortex and complex spatiotemporal dynamics during simulated anesthetic action, and provide forward predictions of neuroimaging signals for subsequent empirical comparisons and more detailed characterization. Because burst suppression corresponds to a dynamical end-point of brain activity, theoretically accounting for its spatiotemporal emergence will vitally contribute to efforts aimed at clarifying whether a common physiological trajectory is induced by the actions of general anesthetic agents. We have taken a first step in this direction by showing that a neural field model can qualitatively match recent experimental data that indicate spatial differentiation of burst suppression activity across cortex

Avaliação da qualidade de vida, da dor nas costas, da funcionalidade e de alterações da coluna vertebral de estudantes de fisioterapia

O objetivo deste estudo foi avaliar a qualidade de vida, dor nas costas, funcionalidade e as alterações da coluna vertebral de estudantes de fisioterapia. Participaram 42 universitários, sendo avaliada a coluna vertebral por meio do arcômetro, a dor nas costas por meio de um questionário multidimensional de dor, a qualidade de vida por meio do questionário SF-36 e funcionalidade a partir do Roland-Morris. Foi realizada estatística descritiva e teste qui-quadrado (α=0,05). Os resultados demonstraram: prevalência de alterações nas curvaturas da coluna vertebral de 38,1%; (n=16); alta prevalência de dor nas costas (69%; n=29); baixa prevalência de comprometimento da funcionalidade (3,4%; n=1); que não há associação entre as alterações nas curvaturas da coluna vertebral e funcionalidade e dor nas costas; e que os escores dos domínios do SF-36 foram maiores que 45, exceto o domínio "dor", cujo escore aproximado foi de 35. Conclui-se que quanto menor os níveis de intensidade da dor melhor o nível de qualidade de vida