Development of Biocompatible Digital Light Processing Resins for Additive Manufacturing Using Visible Light-Induced RAFT Polymerization.

Patients with bone diseases often experience increased bone fragility. When bone injuries exceed the body's natural healing capacity, they become significant obstacles. The global rise in the aging population and the escalating obesity pandemic are anticipated to lead to a notable increase in acute bone injuries in the coming years. Our research developed a novel DLP resin for 3D printing, utilizing poly(ethylene glycol diacrylate) (PEGDA) and various monomers through the PET-RAFT polymerization method. To enhance the performance of bone scaffolds, triply periodic minimal surfaces (TPMS) were incorporated into the printed structure, promoting porosity and pore interconnectivity without reducing the mechanical resistance of the printed piece. The gyroid TPMS structure was the one that showed the highest mechanical resistance (0.94 ± 0.117 and 1.66 ± 0.240 MPa) for both variants of resin composition. Additionally, bioactive particles were introduced to enhance the material's biocompatibility, showcasing the potential for incorporating active compounds for specific applications. The inclusion of bioceramic particles produces an increase of 13% in bioactivity signal for osteogenic differentiation (alkaline phosphatase essay) compared to that of control resins. Our findings highlight the substantial improvement in printing precision and resolution achieved by including the photoabsorber, Rose Bengal, in the synthesized resin. This enhancement allows for creating intricately detailed and accurately defined 3D-printed parts. Furthermore, the TPMS gyroid structure significantly enhances the material's mechanical resistance, while including bioactive compounds significantly boosts the polymeric resin's biocompatibility and bioactivity (osteogenic differentiation).

Smart Polymer Surfaces with Complex Wrinkled Patterns: Reversible, Non-Planar, Gradient, and Hierarchical Structures.

This review summarizes the relevant developments in preparing wrinkled structures with variable characteristics. These include the formation of smart interfaces with reversible wrinkle formation, the construction of wrinkles in non-planar supports, or, more interestingly, the development of complex hierarchically structured wrinkled patterns. Smart wrinkled surfaces obtained using light-responsive, pH-responsive, temperature-responsive, and electromagnetic-responsive polymers are thoroughly described. These systems control the formation of wrinkles in particular surface positions and the reversible construction of planar-wrinkled surfaces. This know-how of non-planar substrates has been recently extended to other structures, thus forming wrinkled patterns on solid, hollow spheres, cylinders, and cylindrical tubes. Finally, this bibliographic analysis also presents some illustrative examples of the potential of wrinkle formation to create more complex patterns, including gradient structures and hierarchically multiscale-ordered wrinkles. The orientation and the wrinkle characteristics (amplitude and period) can also be modulated according to the requested application.

Fabrication and Testing of Multi-Hierarchical Porous Scaffolds Designed for Bone Regeneration via Additive Manufacturing Processes.

Bone implants or replacements are very scarce due to the low donor availability and the high rate of body rejection. For this reason, tissue engineering strategies have been developed as alternative solutions to this problem. This research sought to create a cellular scaffold with an intricate and complex network of interconnected pores and microchannels using salt leaching and additive manufacturing (3D printing) methods that mimic the hierarchical internal structure of the bone. A biocompatible hydrogel film (based on poly-ethylene glycol) was used to cover the surface of different polymeric scaffolds. This thin film was then exposed to various stimuli to spontaneously form wrinkled micropatterns, with the aim of increasing the contact area and the material's biocompatibility. The main innovation of this study was to include these wrinkled micropatterns on the surface of the scaffold by taking advantage of thin polymer film surface instabilities. On the other hand, salt and nano-hydroxyapatite (nHA) particles were included in the polymeric matrix to create a modified filament for 3D printing. The printed part was leached to eliminate porogen particles, leaving homogenously distributed pores on the structure. The pores have a mean size of 26.4 ± 9.9 µm, resulting in a global scaffold porosity of ~42% (including pores and microchannels). The presence of nHA particles, which display a homogeneous distribution according to the FE-SEM and EDX results, have a slight influence on the mechanical resistance of the material, but incredibly, despite being a bioactive compound for bone cells, did not show a significant increase in cell viability on the scaffold surface. However, the synergistic effect between the presence of the hydrogel and the pores on the material does produce an increase in cell viability compared to the control sample and the bare PCL material.

Innovation in Additive Manufacturing Using Polymers: A Survey on the Technological and Material Developments.

This review summarizes the most recent advances from technological and physico-chemical perspectives to improve several remaining issues in polymeric materials' additive manufacturing (AM). Without a doubt, AM is experimenting with significant progress due to technological innovations that are currently advancing. In this context, the state-of-the-art considers both research areas as working separately and contributing to developing the different AM technologies. First, AM techniques' advantages and current limitations are analyzed and discussed. A detailed overview of the efforts made to improve the two most extensively employed techniques, i.e., material extrusion and VAT-photopolymerization, is presented. Aspects such as the part size, the possibility of producing parts in a continuous process, the improvement of the fabrication time, the reduction of the use of supports, and the fabrication of components using more than one material are analyzed. The last part of this review complements these technological advances with a general overview of the innovations made from a material perspective. The use of reinforced polymers, the preparation of adapted high-temperature materials, or even the fabrication of metallic and ceramic parts using polymers as supports are considered. Finally, the use of smart materials that enable the fabrication of shape-changing 3D objects and sustainable materials will also be explored.

Wrinkling on Stimuli-Responsive Functional Polymer Surfaces as a Promising Strategy for the Preparation of Effective Antibacterial/Antibiofouling Surfaces.

Biocompatible smart interfaces play a crucial role in biomedical or tissue engineering applications, where their ability to actively change their conformation or physico-chemical properties permits finely tuning their surface attributes. Polyelectrolytes, such as acrylic acid, are a particular type of smart polymers that present pH responsiveness. This work aims to fabricate stable hydrogel films with reversible pH responsiveness that could spontaneously form wrinkled surface patterns. For this purpose, the photosensitive reaction mixtures were deposited via spin-coating over functionalized glasses. Following vacuum, UV, or either plasma treatments, it is possible to spontaneously form wrinkles, which could increase cell adherence. The pH responsiveness of the material was evaluated, observing an abrupt variation in the film thickness as a function of the environmental pH. Moreover, the presence of the carboxylic acid functional groups at the interface was evidenced by analyzing the adsorption/desorption capacity using methylene blue as a cationic dye model. The results demonstrated that increasing the acrylic acid in the microwrinkled hydrogel effectively improved the adsorption and release capacity and the ability of the carboxylic groups to establish ionic interactions with methylene blue. Finally, the role of the acrylic acid groups and the surface topography (smooth or wrinkled) on the final antibacterial properties were investigated, demonstrating their efficacy against both gram-positive and gram-negative bacteria model strains (E. coli and S. Aureus). According to our findings, microwrinkled hydrogels presented excellent antibacterial properties improving the results obtained for planar (smooth) hydrogels.

Three-Dimensional Whole-Organ Characterization of the Regional Alveolar Morphology in Normal Murine Lungs.

Alveolar architecture plays a fundamental role in the processes of ventilation and perfusion in the lung. Alterations in the alveolar surface area and alveolar cavity volume constitute the pathophysiological basis of chronic respiratory diseases such as pulmonary emphysema. Previous studies based on micro-computed tomography (micro-CT) of lung samples have allowed the geometrical study of acinar units. However, our current knowledge is based on the study of a few tissue samples in random locations of the lung that do not give an account of the spatial distributions of the alveolar architecture in the whole lung. In this work, we combine micro-CT imaging and computational geometry algorithms to study the regional distribution of key morphological parameters throughout the whole lung. To this end, 3D whole-lung images of Sprague-Dawley rats are acquired using high-resolution micro-CT imaging and analyzed to estimate porosity, alveolar surface density, and surface-to-volume ratio. We assess the effect of current gold-standard dehydration methods in the preparation of lung samples and propose a fixation protocol that includes the application of a methanol-PBS solution before dehydration. Our results show that regional porosity, alveolar surface density, and surface-to-volume ratio have a uniform distribution in normal lungs, which do not seem to be affected by gravitational effects. We further show that sample fixation based on ethanol baths for dehydration introduces shrinking and affects the acinar architecture in the subpleural regions. In contrast, preparations based on the proposed dehydration protocol effectively preserve the alveolar morphology.

Biocompatible fluorinated wrinkled hydrogel films with antimicrobial activity.

Surface-modified hydrogel films were designed to control the bacterial colonization on their surface and to promote cell proliferation through the gradual insertion of highly hydrophobic functional monomers. These hydrogel films were deposited via spin-coating technique, using muscovite mica as a substrate. These samples were then exposed to different external stimuli to produce wrinkled patterns. The relationship between the monomers which compose the hydrogel, was varied to alter the hydrophobic/hydrophilic balance of the final composite. Contact angle and confocal Raman spectroscopy measurements were carried out to characterize the surface and the bulk of the hydrogel film. Cell proliferation and antimicrobial tests were performed using premyoblastic murine cells (C2C12-GFP) and RAW 264.7 (ATCC® TIB-71) macrophagic cell lines, and also for bacteria strains, Staphylococcus aureus and Escherichia coli. The results indicate that the inclusion of the TFPMA produces an increase in cell proliferation, together with a decrease in living bacterial colonies after 48 h, both for Gram-positive or Gram-negative species.

Mapping regional strain in anesthetised healthy subjects during spontaneous ventilation.

Introduction: Breathing produces a phenomenon of cyclic deformation throughout life. Biomechanically, deformation of the lung is measured as strain. Regional strain recently started to be recognised as a tool in the study of lung pathophysiology, but regional lung strain has not been studied in healthy subjects breathing spontaneously without voluntary or pharmacological control of ventilation. Our aim is to generate three-dimensional (3D) regional strain and heterogeneity maps of healthy rat lungs and describe their changes over time. Methods: Micro-CT and image-based biomechanical analysis by finite element approach were carried out in six anaesthetised rats under spontaneous breathing in two different states, at the beginning of the experiment and after 3 hours of observation. 3D regional strain maps were constructed and divided into 10 isovolumetric region-of-interest (ROI) in three directions (apex to base, dorsal to ventral and costal to mediastinal), allowing to regionally analyse the volumetric strain, the strain progression and the strain heterogeneity. To describe in depth these parameters, and systematise their report, we defined regional strain heterogeneity index [1+strain SD ROI(x)]/[1+strain mean ROI(x)] and regional strain progression index [ROI(x)-mean of final strain/ROI(x)-mean of initial strain]. Results: We were able to generate 3D regional strain maps of the lung in subjects without respiratory support, showing significant differences among the three analysed axes. We observed a significantly lower regional volumetric strain in the apex sector compared with the base, with no significant anatomical systematic differences in the other directions. This heterogeneity could not be identified with physiological or standard CT methods. There was no progression of the analysed regional volumetric strain when the two time-points were compared. Discussion: It is possible to map the regional volumetric strain in the lung for healthy subjects during spontaneous breathing. Regional strain heterogeneity and changes over time can be measured using a CT image-based numerical analysis applying a finite element approach. These results support that healthy lung might have significant regional strain and its spatial distribution is highly heterogeneous. This protocol for CT image acquisition and analysis could be a useful tool for helping to understand the mechanobiology of the lung in many diseases.

Microwrinkled pH-sensitive hydrogel films and their role on the cell adhesion/proliferation.

In this work, hydrogels based on HEMA and DMAEMA (pH-sensitive monomer) were used to form biocompatible films which present microwrinkled patterns in their surface, with the focus of exploring the role of chemical composition on cell adhesion and proliferation. Three different pH (5.4, 7.4, and 8.3) were employed to prepare these hydrogels. The pre-polymerized hydrogel mixtures were deposited via spin coating, then exposed to vacuum for deswelling the films and finally, to UV-light to spontaneously generate the wrinkled pattern. By following this procedure, is possible to form a thin rigid layer on the top of the soft and incompletely polymerized hydrogel film which generates, in turn, a wrinkled pattern due to strain mismatch in the interface. FE-SEM and AFM micrographs allowed us to characterize the wrinkled pattern dimensions. The results evidenced that chemical composition is directly related to the surface pattern morphologies obtained, not so in the case of pH variation, which does not generate relevant changes in the pattern morphology. Interestingly, these pH variations resulted in significant alterations on the interface-cell interactions. More precisely, a premyoblastic cell monolayer was cultured over the wrinkled pattern, showing an optimal cell proliferation at neutral pH. Also, the variation of DMAEMA amount on the monomer feed composition employed for the preparation of the wrinkle surfaces revealed that a certain amount is required to favor cell attachment and growth.

The role of three-dimensionality and alveolar pressure in the distribution and amplification of alveolar stresses.

Alveolar stresses are fundamental to enable the respiration process in mammalians and have recently gained increasing attention due to their mechanobiological role in the pathogenesis and development of respiratory diseases. Despite the fundamental physiological role of stresses in the alveolar wall, the determination of alveolar stresses remains challenging, and our current knowledge is largely drawn from 2D studies that idealize the alveolar septal wall as a spring or a planar continuum. Here we study the 3D stress distribution in alveolar walls of normal lungs by combining ex-vivo micro-computed tomography and 3D finite-element analysis. Our results show that alveolar walls are subject to a fully 3D state of stresses rather than to a pure axial stress state. To understand the contributions of the different components and deformation modes, we decompose the stress tensor field into hydrostatic and deviatoric components, which are associated with isotropic and distortional stresses, respectively. Stress concentrations arise in localized regions of the alveolar microstructure, with magnitudes that can be up to 27 times the applied alveolar pressure. Interestingly, we show that the stress amplification factor strongly depends on the level of alveolar pressure, i.e, stresses do not scale proportional to the applied alveolar pressure. In addition, we show that 2D techniques to assess alveolar stresses consistently overestimate the stress magnitude in alveolar walls, particularly for lungs under high transpulmonary pressure. These findings take particular relevance in the study of stress-induced remodeling of the emphysematous lung and in ventilator-induced lung injury, where the relation between transpulmonary pressure and alveolar wall stress is key to understand mechanotransduction processes in pneumocytes.

Antimicrobial Polymers for Additive Manufacturing.

Three-dimensional (3D) printing technologies can be widely used for producing detailed geometries based on individual and particular demands. Some applications are related to the production of personalized devices, implants (orthopedic and dental), drug dosage forms (antibacterial, immunosuppressive, anti-inflammatory, etc.), or 3D implants that contain active pharmaceutical treatments, which favor cellular proliferation and tissue regeneration. This review is focused on the generation of 3D printed polymer-based objects that present antibacterial properties. Two main different alternatives of obtaining these 3D printed objects are fully described, which employ different polymer sources. The first one uses natural polymers that, in some cases, already exhibit intrinsic antibacterial capacities. The second alternative involves the use of synthetic polymers, and thus takes advantage of polymers with antimicrobial functional groups, as well as alternative strategies based on the modification of the surface of polymers or the elaboration of composite materials through adding certain antibacterial agents or incorporating different drugs into the polymeric matrix.

Micrometric Wrinkled Patterns Spontaneously Formed on Hydrogel Thin Films via Argon Plasma Exposure.

The generation of microstructured patterns on the surface of a specific polymeric material could radically improve their performance in a particular application. Most of the interactions with the environment occur at the material interface; therefore, increasing the exposed active surface considerably improves their range of application. In this article, a simple and reliable protocol to form spontaneous wrinkled patterns using a hydrogel layer is reported. For this purpose, we took advantage of the doctor blade technique in order to generate homogenous films over solid substrates with controlled thickness and large coverage. The hydrogel wrinkle formation involves a prepolymerization step which produces oligomers leading to a solution with increased viscosity, enough for doctor blade deposition. Subsequently, the material was exposed to vacuum and plasma to trigger wrinkled pattern formation. Finally, a UV-polymerization treatment was applied to fix the undulations on top. Interestingly, the experimental parameters allowed us to finely tune the wrinkle characteristics (period, amplitude, and orientation). For this study, two main aspects were explored. The first one is related to the role of the substrate functionalization on the wrinkle formation. The second study correlates the deswelling time and its relationship with the dimensions and distribution of the wrinkle pattern. In the first batch, four different 3-(trimethoxysilyl)propyl methacrylate (TSM) concentrations were used to functionalize the substrate in order to enhance the adhesion between hydrogel film and the substrate. The wrinkles formed were characterized in terms of wrinkle amplitude, wavelength, pattern roughness, and surface Young modulus, by using AFM in imaging and force spectroscopy modes. Moreover, the chemical composition of the hydrogel film cross-section and the effect of the plasma treatment were analyzed with confocal Raman spectroscopy. These results demonstrated that an oxidized layer was formed on top of the hydrogel films due to the exposure to an argon plasma.

Design and fabrication of biocompatible wrinkled hydrogel films with selective antibiofouling properties.

In this article, we explored the selective antibiofouling capacity acquired by functional wrinkled hydrogel films via a fine tuning of their chemical structure through the gradual insertion of hydrophobic radical groups in their network. The hydrogel consists of three main components: hydroxyethyl methacrylate (HEMA, amphiphilic monomer), trifluoroethyl methacrylate (TFMA, hydrophobic monomer), and poly(ethylene glycol) diacrylate (PEGDA, hydrophilic crosslinking agent). Interestingly, the manipulation of the chemical composition affects both, surface morphology and physicochemical characteristics of the patterns, inducing transitions between different surface microstructures, i.e. from wrinkles to creases, to folds, and to crumples. Contact angle measurements show that the insertion of TFMA produces a slight decrease in surface wettability, remaining however highly hydrophilic. By using confocal Raman spectroscopy, important information about wrinkle formation mechanism could be obtained. The procedure presented in this article involves two consecutive thermal and photopolymerization steps, generating a "pseudo" two-layer system, which contracts at different extents when is exposed to external stimuli, leading to the formation of wrinkled surfaces. Finally, bacterial and cellular adhesion/proliferation studies were carried out, evidencing that the amount of TFMA included clearly reduce the bacterial adhesion while mammalian cells are able to still proliferate.

Smart pH-Responsive Antimicrobial Hydrogel Scaffolds Prepared by Additive Manufacturing.

We report on the fabrication of 3D printed pH-responsive and antimicrobial hydrogels with a micrometer-scale resolution achieved by stereolithography (SLA) 3D printing. The preparation of the hydrogels was optimized by selecting the most appropriate difunctional polyethylene glycol dimethacrylates (testing cross-linking agents with chain lengths ranging from 2 up to 14 units ethylene glycol) and introducing acrylic acid (AA) as a monofunctional monomer. As a result of the incorporation of AA, the hydrogels described are able to reversibly swell and shrink upon environmental changes on the pH, and the swelling extent is directly related to the amount of AA and can be thus finely tuned. More interestingly, upon optimization of the UV penetration depth employing a photoabsorber (Sudan I), a reliable procedure for the fabrication of 3D objects with a high model accuracy is shown. Finally, the antimicrobial properties of all of the hydrogels were demonstrated using Staphylococcus aureus as a bacterial model. We found that even those hydrogels with a low amount of AA monomeric units presented excellent antimicrobial properties against S. aureus.

Silylated oligomeric poly(ether-azomethine)s from monomers containing biphenyl moieties: synthesis and characterization.

In this study, four new silicon-containing poly(ether-azomethine)s with linear structures were prepared using original silicon and biphenyl moiety-containing monomers: two diamines and two dialdehydes. The oligomeric natures of the samples were established by GPC analysis, which showed chains containing 3 to 5 repetitive units. The monomers and the oligomeric samples were structurally characterized by NMR and FT-IR spectroscopy. The solubilities of the samples in common organic solvents and their thermal behavior enable improvement of their industrial and technological processability. The optical band gaps of the oligomeric samples were estimated from optical measurements (UV-vis), and their electrical behavior in films was determined using the four-point method. The surface arrangements and morphological characteristics of the films were determined via atomic force microscopy measurements. The roughness, area increase percentage and layer stiffness of the films were also measured using this technique.

Advances in the Fabrication of Antimicrobial Hydrogels for Biomedical Applications.

This review describes, in an organized manner, the recent developments in the elaboration of hydrogels that possess antimicrobial activity. The fabrication of antibacterial hydrogels for biomedical applications that permits cell adhesion and proliferation still remains as an interesting challenge, in particular for tissue engineering applications. In this context, a large number of studies has been carried out in the design of hydrogels that serve as support for antimicrobial agents (nanoparticles, antibiotics, etc.). Another interesting approach is to use polymers with inherent antimicrobial activity provided by functional groups contained in their structures, such as quaternary ammonium salt or hydrogels fabricated from antimicrobial peptides (AMPs) or natural polymers, such as chitosan. A summary of the different alternatives employed for this purpose is described in this review, considering their advantages and disadvantages. Finally, more recent methodologies that lead to more sophisticated hydrogels that are able to react to external stimuli are equally depicted in this review.

Thermal Response Analysis of Phospholipid Bilayers Using Ellipsometric Techniques.

Biomimetic planar artificial membranes have been widely studied due to their multiple applications in several research fields. Their humectation and thermal response are crucial for reaching stability; these characteristics are related to the molecular organization inside the bilayer, which is affected by the aliphatic chain length, saturations, and molecule polarity, among others. Bilayer stability becomes a fundamental factor when technological devices are developed-like biosensors-based on those systems. Thermal studies were performed for different types of phosphatidylcholine (PC) molecules: two pure PC bilayers and four binary PC mixtures. These analyses were carried out through the detection of slight changes in their optical and structural parameters via Ellipsometry and Surface Plasmon Resonance (SPR) techniques. Phospholipid bilayers were prepared by Langmuir-Blodgett technique and deposited over a hydrophilic silicon wafer. Their molecular inclination degree, mobility, and stability of the different phases were detected and analyzed through bilayer thickness changes and their optical phase-amplitude response. Results show that certain binary lipid mixtures-with differences in its aliphatic chain length-present a co-existence of two thermal responses due to non-ideal mixing.

Strategies to Fabricate Polypeptide-Based Structures via Ring-Opening Polymerization of N-Carboxyanhydrides.

In this review, we provide a general and clear overview about the different alternatives reported to fabricate a myriad of polypeptide architectures based on the ring-opening polymerization of N-carbonyanhydrides (ROP NCAs). First of all, the strategies for the preparation of NCA monomers directly from natural occurring or from modified amino acids are analyzed. The synthetic alternatives to prepare non-functionalized and functionalized NCAs are presented. Protection/deprotection protocols, as well as other functionalization chemistries are discussed in this section. Later on, the mechanisms involved in the ROP NCA polymerization, as well as the strategies developed to reduce the eventually occurring side reactions are presented. Finally, a general overview of the synthetic strategies described in the literature to fabricate different polypeptide architectures is provided. This part of the review is organized depending on the complexity of the macromolecular topology prepared. Therefore, linear homopolypeptides, random and block copolypeptides are described first. The next sections include cyclic and branched polymers such as star polypeptides, polymer brushes and highly branched structures including arborescent or dendrigraft structures.

Thin and ordered hydrogel films deposited through electrospinning technique; a simple and efficient support for organic bilayers.

Thermal behavior of Dipalmitoylphosphatidylcholine (DPPC) bilayers deposited over hydrogel fibers was examined. Thus, membrane stability, water absorption-release, phase transitions and phase transition temperatures were studied through different methods during heating cycles. Hydrogel films were realized using an oligomer mixture (HEMA-PEGDA575/photo-initiator) with adequate viscosity. Then, the fibers were deposited over silicon wafers (hydrophilic substrate) through electrospinning technique using four different voltages: 15, 20, 25 and 30 kV. The films were then exposed to UV light, favoring polymer chain crosslinking and interactions between hydrogel and substrate. For samples deposited at 20 and 25 kV, hierarchical wrinkle folds were observed at surface level, their arrangement distribution depends directly on thickness and associated point defects. DPPC bilayers were then placed over hydrogel scaffold using Langmuir-Blodgett technique. Field emission scanning electron microscopy (FE-SEM) analysis were used to investigate sample surface, micrographies show homogeneous layer formation with chain polymer order/disorder related to applied voltage during hydrogel deposition process, among other parameters. According to the results obtained, it is possible to conclude that the oligomer deposited at 20 kV produce thin homogenous films (~40 nm) with enhanced ability to absorb water and release it in a controlled way during heating cycles. These scaffold properties confer to DPPC membrane thermal stability, which allow an easy detection of phase(s) and phase transitions. Thermal behavior was also studied via Atomic Force Microscopy (roughness analysis). Contact angle measurements corroborate system wettability, supporting the theory that hydrogel thin films act as DPPC membrane enhancers for thermal stability against external stimuli.