Ulvan-chitosan polyelectrolyte complexes as matrices for enzyme induced biomimetic mineralization

Polyelectrolyte complexes (PEC) of chitosan and ulvan were fabricated to study alkaline phosphatase (ALP) mediated formation of apatitic minerals. Scaffolds of the PEC were subjected to ALP and successful mineral formation was studied using SEM, Raman and XRD techniques. Investigation of the morphology via SEM shows globular structures of the deposited minerals, which promoted cell attachment, proliferation and extracellular matrix formation. The PEC and their successful calcium phosphate based mineralization offers a greener route of scaffold fabrication towards developing resorbable materials for tissue engineering

Phenolic plant extract enrichment of enzymatically mineralized hydrogels

Hydrogel mineralization with calcium phosphate (CaP) and antibacterial activity are desirable for applications in bone regeneration. Mineralization with CaP can be induced using the enzyme alkaline phosphatase (ALP), responsible for CaP formation in bone tissue. Incorporation of polyphenols, plant-derived bactericidal molecules, was hypothesized to provide antibacterial activity and enhance ALP-induced mineralization. Three phenolic rich plant extracts from: (i) green tea, rich in epigallocatechin gallate (EGCG) (herafter referred to as EGCG-rich extract); (ii) pine bark and (iii) rosemary were added to gellan gum (GG) hydrogels and subsequently mineralized using ALP. The phenolic composition of the three extracts used were analyzed by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MSn). EGCG-rich extract showed the highest content of phenolic compounds and promoted the highest CaP formation as corroborated by dry mass percentage meassurements and ICP-OES de-termination of mass of elemental Ca and P. All three extracts alone exhibited antibacterial activity in the following order EGCG-rich > PI > RO, respectively. However, extract-loaded and mineralized GG hydro-gels did not exhibit appreciable antibacterial activity by diffusion test. In conclusion, only the EGCG-rich extract promotes ALP-mediated mineralization

Raman Spectroscopy for Advanced Polymeric Biomaterials

In recent years, polymeric biomaterials have emerged as a potential candidate for therapeutic applications owing to their salient features. The characterization of these drugs and bioactive molecules loaded polymer based biomaterials is an important prerequisite for obtaining a deep understanding of their physicochemical behavior in order to ensure their successful clinical use. There are a variety of complementary characterization techniques available for the characterization of the biomaterials. This review highlights the potential of Raman spectroscopy including its importance for investigating the molecular structure of polymeric materials along with a brief description of its principles, instrumentation, and recent advances

Bio-based Polyethylene–Lignin Composites Containing a Pro-oxidant/Pro-degradant Additive: Preparation and Characterization

Formulations of low cost bio-based oxo-biodegradable polyethylene (PE)/Lignin hybrid polymeric composites were prepared by using ethylene/vinyl acetate (EVA) copolymer as compatibilizer and a transition metal salt as oxo-biodegradation promoter. The hybrid composites and relevant Lignin-free blends were formulated by following a statistical mixture design. The effect of Lignin, pro-degradant additive, EVA copolymer and their compatibility with the PE continuous matrix, was evaluated by means of structural features by attenuated total reflectance, morphological by scanning electron microscopy, thermal by differential scanning calorimetry and thermo-gravimetric analysis and mechanical properties by an Instron Machine. The results attained in this study, regarding especially the thermal and mechanical properties, suggest that bio-based oxo-biodegradable hybrid composites offer an interesting way to produce low cost bio-based materials with fairly enhanced properties. The moderate-low cost hybrid materials appear to be attractive for their potential in the mercantile area of commodities including: packaging, personal care products, agricultural mulch films and disposable items. This will constitute a novel added-value contribution aimed at mitigating the environmental burden caused by plastic waste items improperly abandoned in the environment.Italian Ministry of Industr

Microgel/silica hybrid colloids: Bioinspired synthesis and controlled release application

In the present work, we demonstrate that polymer based functional microgels can be used as autocatalytic template for the bioinspired deposition of silica nanoparticles inside the microgel network under ambient conditions. Temperature responsive poly(N-vinyl caprolactam) based microgels were synthesized by precipitation polymerization using glycidyl methacrylate as a comonomer. These microgels were further reacted with aminoethanthiol to develop amine functional groups inside the microgels. A water soluble silica precursor (PEGPEOS) was developed by modification of hyperbranched polyethoxysiloxane with polyethylene glycol monomethyl ether. Microgel/silica hybrid colloids were prepared by simultaneous PEGPEOS conversion and silica deposition in the microgels. TEM studies showed that silica nanoparticles of approximately 10 nm in size were deposited inside the microgel network owing to the strong acidbase interaction between the acidic silica and basic amine groups. DLS results indicated that the incorporation of silica nanoparticles reduced the thermal sensitivity of microgels. Molecular dynamics simulations were performed to investigate the interaction between silica precursor and microgels based on radial distribution function and interaction energy. The developed hybrid microgels were further explored for controlled release of aspirin used as a model drug. The preliminary results indicated that the presence of silica rich domains in the microgel network remarkably retarded aspirin release

Enzymatically biomineralized chitosan scaffolds for tissue-engineering applications

Porous biodegradable scaffolds represent promising candidates for tissue‐engineering applications because of their capability to be preseeded with cells. We report an uncrosslinked chitosan scaffold designed with the aim of inducing and supporting enzyme‐mediated formation of apatite minerals in the absence of osteogenic growth factors. To realize this, natural enzyme alkaline phosphatase (ALP) was incorporated into uncrosslinked chitosan scaffolds. The uncrosslinked chitosan makes available amine and alcohol functionalities to enhance the biomineralization process. The physicochemical findings revealed homogeneous mineralization, with the phase structure of the formed minerals resembling that of apatite at low mineral concentrations, and similar to dicalcium phosphate dihydrate (DCPD) with increasing ALP content. The MC3T3 cell activity clearly showed that the mineralization of the chitosan scaffolds was effective in improving cellular adhesion, proliferation and colonization

Endosomal Size and Membrane Leakiness Influence Proton Sponge-Based Rupture of Endosomal Vesicles

In gene therapy, endosomal escape represents a major bottleneck since nanoparticles often remain entrapped inside endosomes and are trafficked toward the lysosomes for degradation. A detailed understanding of the endosomal barrier would be beneficial for developing rational strategies to improve transfection and endosomal escape. By visualizing individual endosomal escape events in live cells, we obtain insight into mechanistic factors that influence proton sponge-based endosomal escape. In a comparative study, we found that HeLa cells treated with JetPEI/pDNA polyplexes have a 3.5-fold increased endosomal escape frequency compared to ARPE-19 cells. We found that endosomal size has a major impact on the escape capacity. The smaller HeLa endosomes are more easily ruptured by the proton sponge effect than the larger ARPE-19 endosomes, a finding supported by a mathematical model based on the underlying physical principles. Still, it remains intriguing that even in the small HeLa endosomes, <10% of the polyplex-containing endosomes show endosomal escape. Further experiments revealed that the membrane of polyplex-containing endosomes becomes leaky to small compounds, preventing effective buildup of osmotic pressure, which in turn prevents endosomal rupture. Analysis of H1299 and A549 cells revealed that endosomal size determines endosomal escape efficiency when cells have comparable membrane leakiness. However, at high levels of membrane leakiness, buildup of osmotic pressure is no longer possible, regardless of endosomal size. Based on our findings that both endosomal size and membrane leakiness have a high impact on proton sponge-based endosomal rupture, we provide important clues toward further improvement of this escape strategy