Electroactive 4D Porous Scaffold Based on Conducting Polymer as a Responsive and Dynamic In Vitro Cell Culture Platform.

In vivo, cells reside in a 3D porous and dynamic microenvironment. It provides biochemical and biophysical cues that regulate cell behavior in physiological and pathological processes. In the context of fundamental cell biology research, tissue engineering, and cell-based drug screening systems, a challenge is to develop relevant in vitro models that could integrate the dynamic properties of the cell microenvironment. Taking advantage of the promising high internal phase emulsion templating, we here designed a polyHIPE scaffold with a wide interconnected porosity and functionalized its internal 3D surface with a thin layer of electroactive conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) to turn it into a 4D electroresponsive scaffold. The resulting scaffold was cytocompatible with fibroblasts, supported cellular infiltration, and hosted cells, which display a 3D spreading morphology. It demonstrated robust actuation in ion- and protein-rich complex culture media, and its electroresponsiveness was not altered by fibroblast colonization. Thanks to customized electrochemical stimulation setups, the electromechanical response of the polyHIPE/PEDOT scaffolds was characterized in situ under a confocal microscope and showed 10% reversible volume variations. Finally, the setups were used to monitor in real time and in situ fibroblasts cultured into the polyHIPE/PEDOT scaffold during several cycles of electromechanical stimuli. Thus, we demonstrated the proof of concept of this tunable scaffold as a tool for future 4D cell culture and mechanobiology studies.

Recent Advances in Polymers as Matrices for Drug Delivery Applications.

Polymeric-based drug delivery systems have become versatile and valuable candidates in sectors such as pharmaceuticals, health, medicine, etc [...].

Natural Ageing of PLA Filaments, Can It Be Frozen?

The physical ageing of polylactic acid (PLA) is a phenomenon that changes the material's properties over time. This ageing process is highly dependent on ambient variables, such as temperature and humidity. For PLA, the ageing is noticeable even at room temperatures, a process commonly referred to as natural ageing. Stopping the ageing by freezing the material can be helpful to preserve the properties of the PLA and stabilise it at any time during its storage until it is required for testing. However, it is essential to demonstrate that the PLA's mechanical properties are not degraded after defrosting the samples. Four different methods for stopping the ageing (anti-ageing processes) are analysed in this paper-all based on freezing and defrosting the PLA samples. We determine the temperature and ambient water vapor influence during the freezing and defrosting process using desiccant and zip bags. The material form selected is PLA filaments (no bulk material or scaffold structures) printed at 190 °C with diameters between 400 and 550 µm and frozen at -24 °C in the presence or absence of a desiccant. The impact of the anti-ageing processes on PLA's ageing and mechanical integrity is studied regarding the thermal, mechanical and fractographical properties. In conclusion, an anti-ageing process is defined to successfully stop the natural ageing of the PLA for an indefinite length of time. This process does not affect the mechanical properties or the structural integrity of the PLA. As a result, large quantities of this material can be produced in a single batch and be safely stored to be later characterised under the same manufacturing and ageing conditions, which is currently a limiting factor from an experimental point of view as polymeric filament properties can show significant variety from batch to batch.

The Effect of Ca2+ and Mg2+ Ions Loaded at Degradable PLA Membranes on the Proliferation and Osteoinduction of MSCs.

Biodegradable membranes, including Polylactic acid (PLA)-based membranes, are commonly used in bone-tissue-related clinical procedures as biointerface to promote bone tissue regeneration. Calcium (Ca2+) and Magnesium (Mg2+) ions have been related to the promotion of osteogenesis, where the PLA membranes could be used as carrier and delivery substrate for them to provide osteogenic properties to this material. For this aim, a new ion delivery system based on biodegradable PLA membranes loaded with Mg and hydroxyapatite (HA) particles has been processed by the combination of tape casting and colloidal route. Materials characterization shows that the incorporation of Mg and HA particles changes the surface and hydrophobicity of the PLA membrane, and the in vitro degradation test shows Mg2+ and Ca2+ ion release and occasionally the precipitation of different ion species onto the membrane surface. Mouse and human Mesenchymal Stem Cells (MSC) were used to define the biocompatibility and bioactivity of these PLA membrane composites, and data indicated Mg2+ promotes cell proliferation and potentiates osteoinductive signals, while Ca2+ induces the expression of ALP osteogenic marker in human MSCs. Biodegradable PLA membranes loaded with Mg and HA particles is a promising new ion delivery system of Mg2+ and Ca2+ ions that provides osteogenic signals and works as functional biointerface interfaces with bone tissues.

The Mechanical, Thermal, and Chemical Properties of PLA-Mg Filaments Produced via a Colloidal Route for Fused-Filament Fabrication.

The effect of Mg particles on the thermal, chemical, physical, and primarily mechanical properties of 3D-printed PLA/Mg composites is studied in this paper. Recently, new colloidal processing has been proposed to introduce Mg particles into the PLA matrix, which ensures good dispersion of the particles and better thermal properties, allowing for thermal processing routes such as extrusion or 3D printing via fused-filament fabrication. The thermal and physical properties are here studied in 1D single-filament-printed PLA/Mg composites with 0 to 10 wt.% of Mg particles by Differential Scanning Calorimetry (DSC); we analyse the PLA chain modifications produced, the crystallinity fraction, and the different crystalline forms of the PLA after thermal processing. Fourier Transform Infrared Spectroscopy (FTIR) is used to confirm the influence of the PLA/Mg colloidal processing after printing. The mechanical properties are measured with a universal tensile test machine on the 1D single-printed filaments via fused-filament fabrication (FFF); the filaments were naturally aged to stable conditions. Filaments with and without a notch are studied to obtain the materials' tensile strength, elastic modulus, and fracture toughness. Different analytical models to explain the results of the PLA-Mg were studied, in which the minimum values for the interface strength of the PLA-Mg composites were calculated.

Thermal treatment of magnesium particles in polylactic acid polymer films elicits the expression of osteogenic differentiation markers and lipidome profile remodeling in human adipose stem cells.

The efficacy of polylactic acid (PLA)/Magnesium (Mg)-based materials for driving stem cells toward bone tissue engineering applications requires specific Mg surface properties to modulate the interface of stem cells with the film. Here, we have developed novel PLA/Mg-based composites and explored their osteogenic differentiation potential on human adipose stem cells (hASCs). Mg-particles/polymer interface was improved by two treatments: heating in oxidative atmosphere (TT) and surface modification with a compatibilizer (PEI). Different contents of Mg particles were dispersed in PLA and composite surface and bulk properties, protein adsorption, stem cell-PLA/Mg interactions, osteogenic markers expressions, and lipids composition profile were evaluated. Mg particles were uniformly distributed on the surface and in the bulk PLA polymer. Improved and modulated particle-polymer adhesion was observed in Mg particle-treated composites. After 21 days in canonical growth culture conditions, hASCs on PLA/MgTT displayed the highest expression of the general osteogenic markers, RUNX2, SSP1, and BGLAP genes, Alkaline Phosphatase, type I Collagen, Osteopontin, and Calcium deposits. Moreover, by LC/MS QTOF mass-spectrophotometry lipidomic analysis, we found in PLA/MgTT-cells, for the first time, a remodeling of the lipid classes composition associated with the osteogenic differentiation. We ascribed these results to MgTT characteristics, which improve Mg availability and composite osteoinductive performance.

Characterisation and Modelling of PLA Filaments and Evolution with Time.

The properties of polylactic acid (PLA) filaments have not yet been analysed in detail, and they are strongly affected by the extrusion process used in some additive manufacturing systems. Here we present the mechanical, thermal, physical, and fractographical properties of an extruded filament (not the bulk material or scaffolds), the basic building block of any PLA structure printed via material extrusion. This research aims to create a reference point for the modelisation of additively manufactured structures via extrusion processes, as the main building block is characterised in detail for a deep understanding. Furthermore, we investigated the natural ageing (up to one year), the effect of the printing (extruding) temperature (180 and 190 °C), and the effect of the crosshead speed during the tensile tests (10-1 to 102 mm/min) to provide a deeper analysis of the material. The results showed that the material extruded at 190 °C performed better than the material extruded at 180 °C. However, after one hundred days of natural ageing, both materials behaved similarly. This was related to the flow-induced molecular orientation during the extrusion. The crosshead rate produced a logarithmic increase of the mechanical properties, consistent with the Eyring model. Additionally, the ageing produced significant changes in both the elastic modulus and the yield strength: from 2.4 GPa and 40 MPa, in one-day-aged samples, up to 4 GPa and 62 MPa once entirely aged. Finally, it was observed that the glass transition and the enthalpic relaxation increased with ageing, agreeing with the Kohlraushch-William-Watts model.

Controlled SrR Delivery by the Incorporation of Mg Particles on Biodegradable PLA-Based Composites.

Among several ions playing a vital role in the body, Sr2+ and Mg2+ are involved in the mechanism of bone formation, making them especially useful for bone tissue engineering applications. Recently, polylactic acid (PLA)/Mg composites have emerged as a promising family of biomaterials due to their inherent biocompatibility and biodegradability properties. In these composites, polymer and bio-metal have a synergetic effect-while the PLA inhibits the Mg fast reactivity, Mg provides bioactivity to the inert polymer buffering the medium pH during degradation. Meanwhile, the typical form of administrating Sr2+ to patients is through the medication strontium ranelate (SrR), which increases the bone mineral density. Following this interesting research line, a new group of composites, which integrates Mg particles and SrR charged onto halloysite nanotubes (HNT) in a polymeric matrix, was proposed. PLA/Mg/SrR-HNT composites have been processed following a colloidal route, obtaining homogenous composites granulated and film-shaped. The drug delivery profile was evaluated in terms of in vitro lixiviation/dissolution paying special attention to the synergism of both ions release. The combination of two of the most reported ions involved in bone regeneration in the composite biomaterial may generate extra interest in bone healing applications.

Impact of PLA/Mg films degradation on surface physical properties and biofilm survival.

New biocompatible and bioabsorbable materials are currently being developed for bone regeneration. These serve as scaffolding for controlled drug release and prevent bacterial infections. Films of polylactic acid (PLA) polymers that are Mg-reinforced have demonstrated they have suitable properties and bioactive behavior for promoting the osseointegration process. However little attention has been paid to studying whether the degradation process can alter the adhesive physical properties of the biodegradable film and whether this can modify the biofilm formation capacity of pathogens. Moreover, considering that the concentration of Mg and other corrosion products may not be constant during the degradation process, the question that arises is whether these changes can have negative consequences in terms of the bacterial colonization of surfaces. Bacteria are able to react differently to the same compound, depending on its concentration in the medium and can even become stronger when threatened. In this context, physical surface parameters such as hydrophobicity, surface tension and zeta potential of PLA films reinforced with 10% Mg have been determined before and after degradation, as well as the biofilm formation capacity of Staphylococcus epidermidis. The addition of Mg to the films makes them less hydrophobic and the degradation also reduces the hydrophobicity and increases the negative charge of the surface, especially over long periods of time. Early biofilm formation at 8 h is consistent with the physical properties of the films, where we can observe a reduction in the bacterial biofilm formation. However, after 24 h of incubation, the biofilm formation increases significantly on the PLA/Mg films with respect to PLA control. The explosive release of Mg ions and other corrosion products within the first hours were not enough to prevent a greater biofilm formation after this initial time. Consequently, the Mg addition to the polymer matrix had a bacteriostatic effect but not a bactericidal one. Future works should aim to optimize the design and biofunctionality of these promising bioabsorbable composites for a degradation period suitable for the intended application.

In vitro degradation of a biodegradable polylactic acid/magnesium composite as potential bone augmentation material in the presence of titanium and PEEK dental implants.

OBJECTIVE: The aim of this study was to assess the degradation behavior by measuring the H2 release of a biodegradable composite consisting of a polylactic acid matrix reinforced with 30% wt. spherical magnesium microparticles (PLA/Mg) as potential bone augmentation material in combination with dental implants of either titanium or polyetheretherketone (PEEK) in order to evaluate the potential influence of the titanium dental implants on the corrosion behavior of the Mg particles within the PLA matrix. METHODS: Three PEEK dental implants and three titanium dental implants were put into a central perforation of six PLA/Mg-discs. These samples were incubated at 37°C for 30days in McCoy's 5A modified medium and the H2 release was evaluated. RESULTS: Between day 7 and day 16 the average H2 release per cm2 of the surface of the PLA/Mg-samples in combination with the titanium implants was significantly higher than that of the sample group combined with the implants of PEEK (3.1±0.4ml vs. 2.8±0.4ml). This significant difference disappeared afterwards, whereas the H2 release was highest at day 30 and amounted 3.5±0.7ml/cm2 for the group with the titanium implants and 3.2±0.8ml/cm2 for the group with the PEEK implants. SIGNIFICANCE: Regarding the similar values of the degradation depending H2 release of the two implant material groups, the co-implantation of a PLA/Mg composite is not only possible with new metal-free implant materials such as PEEK, but also with conventional implants of titanium.