10 research outputs found
Targeted protein delivery: carbodiimide crosslinking influences protein release from microparticles incorporated within collagen scaffolds
open access articleTissue engineering response may be tailored via controlled, sustained release of active agents from protein-loaded degradable microparticles incorporated directly within three-dimensional (3D) ice-templated collagen scaffolds. However, the effects of covalent crosslinking during scaffold preparation on the availability and release of protein from the incorporated microparticles have not been explored. Here, we load 3D ice-templated collagen scaffolds with controlled additions of poly-(DL-lactide-co-glycolide) microparticles. We probe the effects of subsequent N-(3-dimethylaminopropyl)- N0-ethylcarbodiimide hydrochloride crosslinking on protein release, using microparticles with different internal protein distributions. Fluorescein isothiocyanate labelled bovine serum albumin is used as a model protein drug. The scaffolds display a homogeneous microparticle distribution,
and a reduction in pore size and percolation diameter with increased microparticle addition, although these values did not fall below those reported as necessary for cell invasion. The protein distribution within the microparticles, near the surface or more deeply located within the microparticles, was important in determining the release profile and effect of crosslinking, as the surface
was affected by the carbodiimide crosslinking reaction applied to the scaffold. Crosslinking of microparticles with a high proportion of protein at the surface caused both a reduction and delay in protein release. Protein located within the bulk of the microparticles, was protected from the crosslinking reaction and no delay in the overall release profile was seen
Targeted protein delivery: carbodiimide crosslinking influences protein release from microparticles incorporated within collagen scaffolds.
Tissue engineering response may be tailored via controlled, sustained release of active agents from protein-loaded degradable microparticles incorporated directly within three-dimensional (3D) ice-templated collagen scaffolds. However, the effects of covalent crosslinking during scaffold preparation on the availability and release of protein from the incorporated microparticles have not been explored. Here, we load 3D ice-templated collagen scaffolds with controlled additions of poly-(DL-lactide-co-glycolide) microparticles. We probe the effects of subsequent N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride crosslinking on protein release, using microparticles with different internal protein distributions. Fluorescein isothiocyanate labelled bovine serum albumin is used as a model protein drug. The scaffolds display a homogeneous microparticle distribution, and a reduction in pore size and percolation diameter with increased microparticle addition, although these values did not fall below those reported as necessary for cell invasion. The protein distribution within the microparticles, near the surface or more deeply located within the microparticles, was important in determining the release profile and effect of crosslinking, as the surface was affected by the carbodiimide crosslinking reaction applied to the scaffold. Crosslinking of microparticles with a high proportion of protein at the surface caused both a reduction and delay in protein release. Protein located within the bulk of the microparticles, was protected from the crosslinking reaction and no delay in the overall release profile was seen.This work was supported by the European Research Council [ERC Advanced Grant 320598 3D-E] and was also funded by a grant from the Medical Research Council, Arthritis Research UK, Reumafonds and the UKRM
New Microfluidic System for Electrochemical Impedance Spectroscopy Assessment of Cell Culture Performance: Design and Development of New Electrode Material
Electrochemical impedance spectroscopy (EIS) is widely accepted as an effective and non-destructive method to assess cell health during cell-culture. However, there is a lack of compact devices compatible with microfluidic integration and microscopy that could provide the real-time and non-invasive monitoring of cell-cultures using EIS. In this paper, we reported the design and characterization of a modular EIS testing system based on a patented technology. This device was fabricated using easily processable methodologies including screen-printing of the impedance electrodes and molding or micromachining of the cell culture chamber with an easy assembly procedure. Accordingly, to obtain processable, biocompatible and sterilizable electrode materials that lower the impact of interfacial impedance on TEER (Transepithelial electrical resistance) measurements, and to enable concomitant microscopy observations, we optimized the formulation of the electrode inks and the design of the EIS electrodes, respectively. First, electrode materials were based on carbon biocompatible inks enriched with IrOx particles to obtain low interfacial impedance electrodes approaching the performances of classical non-biocompatible Ag/AgCl second-species electrodes. Secondly, we proposed three original electrode designs, which were compared to classical disk electrodes that were optically compatible with microscopy. We assessed the impact of the electrode design on the response of the impedance sensor using COMSOL Multiphysics. Finally, the performance of the impedance spectroscopy devices was assessed in vitro using human airway epithelial cell cultures
Development and characterisation of microporous biomimetic scaffolds loaded with magnetic nanoparticles as bone repairing material
Fine-tuning of the scaffolds structural features for bone tissue engineering can be an efficient approach to
regulate the specific response of the osteoblasts. Here, we loaded magnetic nanoparticles aka superparamagnetic
iron oxide nanoparticles (SPIONs) into 3D composite scaffolds based on biological macromolecules (chitosan,
collagen, hyaluronic acid) and calcium phosphates for potential applications in bone regeneration, using a
biomimetic approach. We assessed the effects of organic (chitosan/collagen/hyaluronic acid) and inorganic
(calcium phosphates, SPIONs) phase over the final features of the magnetic scaffolds (MS). Mechanical properties,
magnetic susceptibility and biological fluids retention are strongly dependent on the final composition of
MS and within the recommended range for application in bone regeneration. The MS architecture/pore size can
be made bespoken through changes of the final organic/inorganic ratio. The scaffolds undertake mild degradation
as the presence of inorganic components hinders the enzyme catalytic activity. In vitro studies indicated
that osteoblasts (SaOS-2) on MS9 had similar cell behaviour activity in comparison with the TCP control. In vivo
data showed an evident development of integration and resorption of the MS composites with low inflammation
activity. Current findings suggest that the combination of SPIONs into 3D composite scaffolds can be a promising
toolkit for bone regeneration
Development of a multiparametric (bio)sensing platform for continuous monitoring of stress metabolites
There is a growing need for real-time monitoring of metabolic products that could reflect cell damages over extended periods. In this paper, we report the design and development of an original multiparametric (bio)sensing platform that is tailored for the real-time monitoring of cell metabolites derived from cell cultures. Most attractive features of our developed electrochemical (bio)sensing platform are its easy manufacturing process, that enables seamless scale-up, modular and versatile approach, and low cost. In addition, the developed platform allows a multiparametric analysis instead of single-analyte analysis. Here we provide an overview of the sensors-based analysis of four main factors that can indicate a possible cell deterioration problem during cell-culture: pH, hydrogen peroxide, nitric oxide/nitrite and lactate. Herein, we are proposing a sensors platform based on thick-film coupled to microfluidic technology that can be integrated into any microfluidic system using Luer-lock connectors. This platform allows obtaining an accurate analysis of the secreting stress metabolites during cell/tissues culture
PLA/chitosan/keratin composites for biomedical applications
Novel composites based on PLA, chitosan and keratin was obtained via blend preparation. The goal of this contribution was to evaluate mechanical and in vitro behavior of the composites. The results point out composites with improved Young modulus and decreased tensile strength, significant increase in hardness (compared to PLA) and a good uptake of the surface properties. Biological assessments using human osteosarcoma cell line on these composites indicate a good viability/proliferation outcome. Hence preliminary results regarding mechanical behavior and in vitro osteoblast response suggest that these composites might have prospective application in medical field
Design, characterization and preliminary biological evaluation of new lignin-PLA biocomposites
The study focuses on the obtainment of new poly (lactic acid)-lignin biocomposites. The effect of lignin loading on the morphology and mechanical properties, as well as the water uptake behaviour of the obtained biocomposites, was investigated in order to elucidate the influence of lignin incorporation into a poly (lactic acid) matrix. The addition of 7% lignin improved the Young modulus and led to a decrease in the tensile strength in comparison with the corresponding values of the poly (lactic acid) matrix, while the water sorption capacity slowly decreased. A subsequent increment in lignin loading from 7 to 15wt% resulted in an increase in tensile strength, as well as in a decline in the water sorption capacity. These results show the importance of the lignin content in controlling the properties of such composites. Furthermore, the behaviour of the PLA-lignin biocomposites in SBF was another concern for evaluation of mechanical performance and biological activity. The mechanical performance declined after immersion in simulated body fluid, but the properties of the biomaterials remained sufficiently high for the perspective of their use in medical applications. In-vitro biocompatibility studies evidenced that the addition of lignin to a poly (lactic acid) matrix can allow tailoring the final properties of the composites without inducing any significant change in cell metabolic activity (compared to poly (lactic acid) itself)
Biomimetic Composites Based on Calcium Phosphates and Chitosan - Hyaluronic Acid with Potential Application in Bone Tissue Engineering
Poly(Acrylic Acid)–Poly(Ethylene Glycol) Nanoparticles Designed for Ophthalmic Drug Delivery
Poly(acrylic acid) (PAA) and poly(ethylene glycol) (PEG), four-arm, amine-terminated particles with nanometer size and spherical shape were obtained by the polymers cross-linking, via activation with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride, in a w/o emulsion. The morphology and surface charge of the final particles are strongly dependent on the molar ratio of PAA–PEG and the PAA concentration. The physicochemical characteristics correlated with the drug-loading capacity, in vitro and ex vivo release kinetics of pilocarpine hydrochloride and biocompatibility results indicate that these nanoparticles exhibit the prerequisite behavior for use as carriers of ophthalmic drugs