Biological evaluations of novel 2,3,3-Trisphosphonate in osteoclastic and osteoblastic activities

Bisphosphonates (BPs) are the first line treatment for many bone diseases including hypercalcimia associated with bone malignancies. In this paper, we introduce a new analogue of bisphosphonate called the 2,3,3-Trisphosphonate (2,3,3-TriPP) that was synthesised in a two steps reaction. In vitro investigations using a medically known bisphosphonate (Etidronate) and the 2,3,3-TrisPP were performed with an aim to evaluate biological effect of this novel compound in major bone cells. 2,3,3-TrisPP showed to have potential to supress the bone resorption process, as our data found that this novel compound exhibited cytotoxic effect in osteoclastic cells at a low concentration of 0.172 mg/mL (LC50). A molecular docking computational simulation calculated a high level of binding affinity between the human farnesyl pyrophosphate synthase (hFPPS) and 2,3,3-TrisPP. This calculation suggested 2,3,3TrisPP may have undergone the mevalonate pathway to prevent the prenylation step during biosynthesis and subsequently resulted in the deactivation of osteoclastic cells. Finally, high levels of osteoblast mineralisation potentials were recorded upon treatments with 2,3,3-TrisPP (0.01-0.1 mg/ml), which implied 2,3,3-TrsiPP may also facilitate bone regeneration.Peer reviewe

Biofabrication of 3D hard-soft and composite constructs for bone regeneration

Biofabrication encompasses the use of additive manufacturing techniques for fabricating complex constructs from a wide range of biomaterials, cells and bioactive substances as well as their maturation for the formation of tissue. The fabricated constructs should provide mechanical stability, porosity, and accurate positioning of cells. The aim of this work was the creation of hybrid constructs consisting of a combination of a thermoplastic hard polymer with and without addition of bioactive glass particles and a soft hydrogel matrix with immobilised cells. The hard phase should enhance the limited mechanical performance of the soft hydrogel phase. Moreover the addition of bioactive glass will enhance the local bioactivity of the scaffolds, of relevance for bone tissue engineering [1]. The hydrogel composition, based on alginate, was tailored to enable the proliferation, migration and differentiation of cells. The mechanical properties and the degradation kinetic of the constructs were investigated. Alginate-dialdehyde (ADA) gelatine (GEL) hydrogel (= ADA-GEL) containing murine bone marrow derived stroma cells (ST2) and polycaprolactone (PCL), polyethylene glycol (PEG) blends were used. Processing was done by additive manufacturing using a dispense plotter equipped with multiple cartridges. Process parameters like plotting speed, pressure and temperature were optimized for the two material systems. Porosity, degradation behaviour and mechanical stability of the PCL-PEG frame structure scaffolds were tested as well as the response of ST2 cells. The presence of bioactive glass leading to enhance local formation of hydroxyapatite was investigated. The cell behaviour and cell development were characterized by assessing the morphology and by measuring the viability of the immobilized cells in the ADA-GEL over an incubation period of 28 days. Both materials could be processed in a defined manner with optimized process parameters. The PEG phase could be dissolved and porous (bioactive) struts forming a framework structure were created. The viability of immobilized ST2 cells after hydrogel plotting was proven as well as their attachment, migration and proliferation by SEM and fluorescence microscopy images. Thus, two promising material systems for creating hybrid constructs were successfully evaluated. The two phase plotting approach enables the fabrication of hydrogel constructs with improved mechanical properties and bioactivity, which exhibit high potential for applications in bone regeneration. [1] A. J. Leite, et al., Bioplotting of a bioactive alginate dialdehyde-gelatin composite hydrogel containing bioactive glass nanoparticles, Biofabrication, 8, pp. 035005 (2016

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

Fabrication and characterization of alginate-keratin based composite microspheres containing bioactive glass for tissue engineering applications

3D cell encapsulation within hydrogels has attracted more and more attention in tissue engineering applications because hydrogels provide a hydrated environment closely mimicking the in vivo environment for cell and tissue growth1. This present study considers the fabrication of alginate-keratin based composite microspheres containing bioactive glass (BG) of 45S5 composition for cell encapsulation. We propose the use of alginate di-aldehyde (ADA) synthesized via periodate oxidation of alginate to enhance the biodegradability of alginate, and the incorporation of keratin into the alginate based hydrogel to improve cellular interaction of the hydrogel. Keratins extracted from wool contain cell adhesive peptide sequences including RGD (arginine-glycine-aspartic acid), and LDV (leucine-aspartic acid-valine)2. BG particles, well known for promoting calcium phosphate deposition, were incorporated into the microspheres to enhance osseointegration3. The microspheres were prepared via a pressure-driven extrusion technique. Weight loss, protein release measurements, and FTIR spectroscopy of the fabricated microspheres were carried out. The morphology and microstructure of the microspheres were investigated by light microscopy and scanning electron microscopy (SEM), respectively. The results demonstrated that the composition of the hydrogels had a significant effect on their physical properties. Biological properties of ADA-keratin based microspheres were evaluated by encapsulating MG-63 osteosarcoma cells into the microspheres. Cell viability of MG-63 cells in ADA-keratin-1%BG hydrogels was found to be comparable to that of alginate-keratin and ADA-keratin after culturing for 21 days. The results proved that such novel composite hydrogel might be a promising material for biofabrication in bone healing approaches. Please click Additional Files below to see the full abstract

Model with automatic learning for temperature forecast and control in tunel-ventilated broiler houses

Orientadores: José Mário Martínez, Maria Aparecida Diniz EhrhardtTese (doutorado) Universidade Estadual de Campinas, Instituto de Matemática, Estatística e Computação CientíficaResumo: A avicultura de corte possui grande importância no setor econômico e social brasileiro, destacando o país como um dos três maiores produtores mundiais e principal exportador de carne de frango. Um dos desafios desta área é o estresse térmico ambiental, que exerce um impacto negativo na produção e no bem-estar animal. A ventilação é o principal mecanismo controlador das condições climáticas às quais as aves são expostas. Decisões simples como ligar ou desligar exautores são essenciais, e escolhas erradas podem ser as causas de pouco ganho de peso, doenças e, em um extremo, a morte do frango. Softwares de CFD (Dinâmica dos Fluidos Computacional) vêm sendo aplicados em estudos de ambiência dos aviários, porém devido ao seu alto custo computacional não auxiliam em tomadas de decisões imediatas. Este trabalho tem como objetivo apresentar um sistema de computador automático para controlar a temperatura de aviários. O sistema é baseado em um modelo de autocorreção que toma decisões em tempo real sobre o funcionamento dos equipamentos de ventilação e refrigeração. O modelo utiliza equações diferenciais parciais, cujos parâmetros são atualizados através de métodos de otimização sem derivadas, de acordo com dados online recolhidos. Os testes revelam que o sistema é consistente e pode ser utilizado em muitas situações que envolvam controle ambientalAbstract: The poultry production has great importance in the Brazilian economic and social sector, highlighting the country as one of the three world's largest producers and main exporter of chicken meat. One of the challenges in this area is the environmental heat stress, which has a negative impact on production and animal welfare. Ventilation is the primary mechanism controlling the climatic conditions to which the broilers are exposed. Simple decisions such as turn exhaust fans on or off are essential, and wrong choices can lead to poor weight gain, illness and death of the broiler. CFD software (Computational Fluid Dynamics) has been applied in aviaries design, but due to its high computational cost it does not help in real time decisions. This research aims to present an automatic computer system for controlling temperature of broiler houses. The system is based on a self-correcting model that takes real-time decisions on the operation of the ventilation, or cooling equipments. The model uses partial differential equations, whose parameters are updated by derivative-free optimization methods, according to on-line collected data. Experiments reveal that the system is consistent and can be used in many situations that involve environmental controlDoutoradoMatematica AplicadaDoutora em Matemática Aplicada144669/2013-7CAPESCNP

Magnetic Glass Ceramics by Sintering of Borosilicate Glass and Inorganic Waste

Ceramics and glass ceramics based on industrial waste have been widely recognized as competitive products for building applications; however, there is a great potential for such materials with novel functionalities. In this paper, we discuss the development of magnetic sintered glass ceramics based on two iron-rich slags, coming from non-ferrous metallurgy and recycled borosilicate glass. The substantial viscous flow of the glass led to dense products for rapid treatments at relatively low temperatures (900–1000 °C), whereas glass/slag interactions resulted in the formation of magnetite crystals, providing ferrimagnetism. Such behavior could be exploited for applying the obtained glass ceramics as induction heating plates, according to preliminary tests (showing the rapid heating of selected samples, even above 200 °C). The chemical durability and safety of the obtained glass ceramics were assessed by both leaching tests and cytotoxicity tests