Dual-Crosslinked Degradable Elastomeric Networks With Self-Healing Properties: Bringing Multi(catechol) Star-Block Copolymers into Play.

In the biomedical field, degradable chemically crosslinked elastomers are interesting materials for tissue engineering applications, since they present rubber-like mechanical properties matching those of soft tissues and are able to preserve their three-dimensional (3D) structure over degradation. Their use in biomedical applications requires surgical handling and implantation that can be a source of accidental damages responsible for the loss of properties. Therefore, their inability to be healed after damage or breaking can be a major drawback. In this work, biodegradable dual-crosslinked networks that exhibit fast and efficient self-healing properties at 37 °C are designed. Self-healable dual-crosslinked (chemically and physically) elastomeric networks are prepared by two methods. The first approach is based on the mix of hydrophobic poly(ethylene glycol)-poly(lactic acid) (PEG-PLA) star-shaped copolymers functionalized with either catechol or methacrylate moieties. In the second approach, hydrophobic bifunctional PEG-PLA star-shaped copolymers with both catechol and methacrylate on their structure are used. In the two systems, the supramolecular network is responsible for the self-healing properties, thanks to the dynamic dissociation/reassociation of the numerous hydrogen bonds between the catechol groups, whereas the covalent network ensures mechanical properties similar to pure methacrylate networks. The self-healable materials display mechanical properties that are compatible with soft tissues and exhibit linear degradation because of the chemical cross-links. The performances of networks from mixed copolymers versus bifunctional copolymers are compared and demonstrate the superiority of the latter. The biocompatibility of the materials is also demonstrated, confirming the potential of these degradable and self-healable elastomeric networks to be used for the design of temporary medical devices.

Electrospun microstructured PLA-based scaffolds featuring relevant anisotropic, mechanical and degradation characteristics for soft tissue engineering.

Electrospun scaffolds combine suitable structural characteristics that make them strong candidates for their use in tissue engineering. These features can be tailored to optimize other physiologically relevant attributes (e.g. mechanical anisotropy and cellular affinity) while ensuring adequate degradation rates of the biomaterial. Here, we present the fabrication of microstructured scaffolds by using a combination of micropatterned electrospinning collectors (honeycomb- or square-patterned) and poly(lactic acid) (PLA)-based copolymers (linear or star-shaped). The resulting materials showed appropriate macropore size and fiber alignment that were key parameters to enhance their anisotropic properties in protraction. Moreover, their elastic modulus, which was initially similar to that of soft tissues, gradually changed in hydrolytic conditions, matching the degradation profile in a 2- to 3-month period. Finally, honeycomb-structured scaffolds exhibited enhanced cellular proliferation compared to standard electrospun mats, while cell colonization was shown to be guided by the macropore contour. Taking together, these results provide new insight into the rational design of microstructured materials that can mimic the progressive evolution of properties in soft tissue regeneration.

Electrospun Biomaterials from Chitosan Blends Applied as Scaffold for Tissue Regeneration.

Our objective in this work was to summarize the main results obtained in processing pure chitosan and chitosan/hyaluronan complex in view of biomedical applications, taking advantage of their original properties. In addition, an electrospinning technique was selected to prepare nanofiber mats well adapted for tissue engineering in relation to the large porosity of the materials, allowing an exchange with the environment. The optimum conditions for preparation of purified and stable nanofibers in aqueous solution and phosphate buffer pH = 7.4 are described. Their mechanical properties and degree of swelling are given. Then, the prepared biomaterials are investigated to test their advantage for chondrocyte development after comparison of nanofiber mats and uniform films. For that purpose, the adhesion of cells is studied by atomic force microscopy (AFM) using single-cell force spectroscopy, showing the good adhesion of chondrocytes on chitosan. At the end, adhesion and proliferation of chondrocytes in vitro are examined and clearly show the interest of chitosan nanofiber mats compared to chitosan film for potential application in tissue engineering.

Anisotropic electrospun honeycomb polycaprolactone scaffolds: Elaboration, morphological and mechanical properties.

Tissue engineering technology requires porous scaffolds, based on biomaterials, which have to mimic as closely as possible the morphological and anisotropic mechanical properties of the native tissue to substitute. Anisotropic fibrous scaffolds fabricated by template-assisted electrospinning are investigated in this study. Fibers of electrospun Polycaprolactone (PCL) were successfully arranged spatially into honeycomb structures by using well-shaped 3D micro-architected metal collectors. Fibrous scaffolds present 2 × 4 mm2 wide elementary patterns with low and high fiber density areas. Distinct regions of the honeycomb patterns were analyzed through SEM images revealing different fiber diameters with specific fiber orientation depending on the regions of interest. Tensile test experiments were carried out with an optical observation of the local deformation at the pattern scale, allowing the determination and analysis, at small and large deformation, of the axial and transverse local strains. The honeycomb patterned mats showed significantly different mechanical properties along the two orthogonal directions probing an anisotropic ratio of 4.2. Stress relaxation test was performed on scaffolds at 15% of strain. This measurement pointed out the low contribution of the viscosity of about 20% in the mechanical response of the scaffold. An orthotropic linear elastic model was consequently proposed to characterize the anisotropic behavior of the produced patterned membranes. This new versatile method to produce architected porous materials, adjustable to several polymers and structures, will provide appealing benefits for soft regenerative medicine application and the development of custom-made scaffolds.

Production of Chitosan/Hyaluronan Complex Nanofibers. Characterization and Physical Properties as a Function of the Composition.

In this work, optimized conditions for preparation of chitosan and hyaluronan polyelectrolyte complex are proposed. The objective was to produce new biomaterials being biocompatible and bioresorbable in the body as well as approaching the extracellular matrix (ECM) structure. These materials will be tested for chondrocyte development in tissue engineering and wound healing applications. Nanofibers made of the polyelectrolyte complex (PEC) were successfully manufactured by electrospinning, and casted films were used as a model for properties comparison. To our knowledge, it is the first time that stable chitosan/hyaluronan fibers are produced, which were observed to be long-lasting in buffer at pH~7.4. The role of thermal treatment at 120 °C for 4 h is examined to control the degree of swelling by crosslinking of the two polysaccharides by H-bonds and amide bonds formation. The properties of the materials are tested for different PEC compositions at different pH values, based on swelling and solubility degrees, diameters of nanofibers and mechanical performances. The influence of the solvent (acidic potential and composition) utilized to process biomaterials is also examined. Acid formic/water 50/50 v/v is observed to be the more appropriated solvent for the carried-out procedures.

Biopharmaceutical Assessment of Dexamethasone Acetate-Based Hydrogels Combining Hydroxypropyl Cyclodextrins and Polysaccharides for Ocular Delivery.

We previously developed two optimized formulations of dexamethasone acetate (DXMa) hydrogels by means of special cubic mixture designs for topical ocular administration. These gels were elaborated with hydroxypropyl-ß-CD (HPßCD) and hydroxypropyl-γ-CD (HPγCD) and commercial hydrogels in order to enhance DXMa water solubility and finally DXMa's ocular bioavailability and transcorneal penetration. The main objective of this study was to characterize them and to evaluate in vitro, ex vivo, and in vivo their safety, biopermanence, and transcorneal permeation. Gels A and B are Newtonian fluids and display a viscosity of 13.2 mPa.s and 18.6 mPa.s, respectively, which increases their ocular retention, according to the in vivo biopermanence study by PET/CT. These hydrogels could act as corneal absorption promoters as they allow a higher transcorneal permeation of DXMa through porcine excised cornea, compared to DEXAFREE® and MAXIDEX®. Cytotoxicity assays showed no cytotoxic effects on human primary corneal epithelial cells (HCE). Furthermore, Gel B is clearly safe for the eye, but the effect of Gel A on the human eye cannot be predicted. Both gels were also stable 12 months at 25 °C after sterilization by filtration. These results demonstrate that the developed formulations present a high potential for the topical ocular administration of dexamethasone acetate.

Biomaterials Based on Electrospun Chitosan. Relation between Processing Conditions and Mechanical Properties.

In this paper, it is shown that pure chitosan nanofibers and films were prepared with success in 0.5 M acetic acid as solvent using poly (ethylene oxide) (PEO) at different yields, allowing electrospinning of the blends. After processing, a neutralization step of chitosan followed by water washing is performed, preserving the initial morphology of chitosan materials. The influence of the yield in PEO in the blend on the degree of swelling and hydrophilicity of films and nanofibers is demonstrated. Then, the mechanical behavior of blended nanofibers and films used as reference are determined for small stress applied in the linear domain by DMA and by uniaxial traction up to rupture. The dried and wet states are covered for the first time. It is shown that the mechanical properties are increased when electrospinning is performed in the presence of PEO up to a 70/30 chitosan/PEO weight ratio even after PEO extraction. This result can be explained by a better dispersion of the chitosan in the presence of PEO.

Preparation of Pure and Stable Chitosan Nanofibers by Electrospinning in the Presence of Poly(ethylene oxide).

Electrospinning was employed to obtain chitosan nanofibers from blends of chitosans (CS) and poly(ethylene oxide) (PEO). Blends of chitosan (MW (weight-average molecular weight) = 102 kg/mol) and PEO (M (molecular weight) = 1000 kg/mol) were selected to optimize the electrospinning process parameters. The PEO powder was solubilized into chitosan solution at different weight ratios in 0.5 M acetic acid. The physicochemical changes of the nanofibers were determined by scanning electron microscopy (SEM), swelling capacity, and nuclear magnetic resonance (NMR) spectroscopy. For stabilization, the produced nanofibers were neutralized with K2CO3 in water or 70% ethanol/30% water as solvent. Subsequently, repeated washings with pure water were performed to extract PEO, potassium acetate and carbonate salts formed in the course of chitosan nanofiber purification. The increase of PEO content in the blend from 20 to 40 w% exhibited bead-free fibers with average diameters 85 ± 19 and 147 ± 28 nm, respectively. Their NMR analysis proved that PEO and the salts were nearly completely removed from the nanostructure of chitosan, demonstrating that the adopted strategy is successful for producing pure chitosan nanofibers. In addition, the nanofibers obtained after neutralization in ethanol-aqueous solution has better structural stability, at least for six months in aqueous solutions (phosphate buffer (PBS) or water).

Carbohydrate-decorated PCL fibers for specific protein adhesion.

Ultrafine biocompatible fibers decorated with carbohydrates were prepared by electrospinning. Both bulk- and surface-modification approaches have been investigated and compared in terms of practicability and grafting density along the fibrous mats. On one hand, bulk-functionalized fibers were prepared by electrospinning of native and galactose-modified PCL polymers. The size and morphology of the resulting fibers was strongly influenced by the galactose-PCL content as observed by electron microscopy. Successful surface modification was evidenced by water contact angle measurements, but a rather low carbohydrate density was attained, as indicated by a colorimetric quantification. On the other hand, efficient and versatile surface-glycosylation was achieved after modification of azido-functionalized electrospun fibers by CuAAC click-chemistry. Homogeneous ultrafine PCL fibers, decorated with azide functions, have been made completely hydrophilic upon coupling with propargyl-α-d-mannoside and propargyl-ß-d-galactoside. Specific adhesion of lectins further attested good bioavailability of the surface carbohydrate residues, suggesting interesting perspectives of the latter approach in the development of bioactive materials for tissue engineering.

Electrospun azido-PCL nanofibers for enhanced surface functionalization by click chemistry.

This paper reports highly surface functionalized and "clickable" α,ω-azido-poly(ε-caprolactone) fibers (f-PCL-N(3)), obtained by classical electrospinning setup. Azide-functionalized PCL was obtained from a commercially available α,ω-poly(ε-caprolactone)-diol, PCL(2), and electrospun with a nonderivative high-molecular-weight PCL. Successful chemical modifications of PCL(2) were confirmed by NMR, FTIR and MALDI-TOF mass spectroscopy. The high content of surface azides, as a response to the high electric field applied, was characterized using a colorimetric assay. In addition, azide reduction to amines revealed a nondestructive route for highly amine-functionalized fibers. Fluorescence labeling of f-PCL-N(3) fibers with FITC-alkyne fluorophore proved that the azide groups are mainly surface-localized as well as highly available for click-chemistry coupling.

Characterization of the core-shell nanoparticles formed as soluble hydrogen-bonding interpolymer complexes at low pH.

The formation of soluble hydrogen-bonding interpolymer complexes between poly(acrylic acid) (PAA) and poly(acrylic acid-co-2-acrylamido-2-methyl-1-propane sulfonic acid)-graft-poly(N,N-dimethylacrylamide) (P(AA-co-AMPSA)-g-PDMAM) at pH=2.0 was studied. A viscometric study showed that in semidilute solution a physical gel is formed due to the interconnection of the anionic P(AA-co-AMPSA) backbone of the graft copolymer, in a transient network, by means of the complexes formed between the PDMAM side chains of the graft copolymer and PAA. Dynamic and static light scattering measurements, in conjunction with small-angle neutron scattering measurements, suggest the formation of core-shell colloidal nanoparticles in dilute solution, comprised by an insoluble PAA/PDMAM core surrounded by an anionic P(AA-co-AMPSA) corona. Even if larger clusters are formed in semidilute solution, the size of the insoluble core remains practically stable. Atomic force microscopy performed under ambient conditions reveal that the particles collapse and flatten upon deposition on a substrate, with dimensions close to the ones of the dry hydrophobic core.

pH-Tunable rheological properties of a telechelic cationic polyelectrolyte reversible hydrogel.

Steady shear properties and linear and nonlinear viscoelastic behaviors of a poly(methyl methacrylate)-poly(dimethyl amino ethyl methacrylate)-poly(methyl methacrylate) polymer, (PMMA-PDMAEMA-PMMA), telechelic polymers in salt-free aqueous solution have been investigated as a function of concentration and pH. Above a critical concentration, a transient physical network is formed through an association mechanism between hydrophobic end groups, leading to a gel-like behavior. The gel-like polymer solutions were shown to exhibit a peculiar flow behavior, associated with time fluctuation of the transient first normal stress difference, attributed to orientation effects of the stiff charged polymer chains. The viscoelastic behavior was shown to be governed by two pH dependent time scales: a short time scale controlled by the lifetime of the hydrophobic associative junctions and a long time scale corresponding to the network relaxation time. All rheological results show strong evidence that Coulomb interactions, which control both macromolecular chain rigidity and inter-chain interactions, lead to specific pH-tunable properties of great potential interest.