Grafting of Crown Ether and Cryptand Macrocycles on Large Pore Stellate Mesoporous Silica for Sodium Cation Extraction.

Regulation of the sodium cations level in the case of renal failure diseases is a very challenging task for clinicians, and new pollutant extractors based on nanomaterials are emerging as potential treatments. In this work, we report different strategies for the chemical functionalization of biocompatible large pore mesoporous silica, denoted stellate mesoporous silica (STMS), with chelating ligands able to selectively capture sodium. We address efficient methods to covalently graft highly chelating macrocycles onto STMS NPs such as crown ethers (CE) and cryptands (C221) through complementary carbodiimidation reactions. Regarding sodium capture in water, C221 cryptand-grafted STMS showed better capture efficiency than CE-STMS due to higher sodium atom chelation in the cryptand cage (Na+ coverage of 15.5% vs. 3.7%). The sodium selectivity was hence tested with C221 cryptand-grafted STMS in a multi-element aqueous solution (metallic cations with the same concentration) and in a solution mimicking peritoneal dialysis solution. Results obtained indicate that C221 cryptand-grafted STMS are relevant nanomaterials to extract sodium cations in such media and allow us to regulate their levels.

Glycan-Functionalized Collagen Hydrogels Modulate the Glycoenvironment of a Neuronal Primary Culture.

Glycans play a central role in the development and homeostasis of the central nervous system (CNS), so changes in the glycosylation profile of the cell surface and extracellular matrix (ECM) components are evident in CNS disorders. Regenerative medicine-based strategies using biomaterial platforms are being increasingly used to target these diseases and to study the glyco-signature of the physiological and pathological conditions, particularly through the use of biomaterials able to recapitulate natural components and physical organization of the ECM. Collagen hydrogels have shown potential as a vehicle for the delivery of cells into the brain, and their therapeutic effect can be expanded by functionalizing them to address the glycosylation patterns altered with specific brain disorders. The goal of this study is to develop and optimize a glycan-functionalized tridimensional collagen-based hydrogel that will make contact with and modulate the differentiation of a primary neuronal culture. The developed system would provide more information on how a glyco-engineered material can influence the native glyco-signature profile during the process of cell differentiation by a differential modulation of glycan expression at the tissue level. To this purpose, collagen polymers underwent one step reductive amination with maltose (Glc(α1-4)α-Glc) and lactose (Gal(ß1-4)ß-Glc) so that the pyranosidic structure of the reducing sugar could be sacrificed (acting as a linker) and the α-Glc and ß-Gal residues exposed, respectively. The glycoconjugate biopolymers were used to formulate hydrogels that were chemically and biologically characterized, and the glyco-signature profile of a neuronal culture after hydrogel treatment was analyzed by lectin staining. The hydrogel conjugated with glucose limited the astrocytic proliferation for up to 2 weeks and promoted the increase in sialylation while decreasing fucosylation by 2-fold. These results indicate the differential influence of glycan residues present in the matrix on cellular sugar expression and thus have potential to enhance cell delivery systems in the central nervous system.

Protein sustained release from isobutyramide-grafted stellate mesoporous silica nanoparticles.

Proteins are great therapeutic candidates as endogenous biomolecules providing a wide range of applications. However, their delivery suffers from some limitations and specifically designed delivery systems having an efficient protein anchoring and delivery strategy are still needed. In this work, we propose to combine large pore stellate mesoporous silica (STMS) with isobutyramide (IBAM), as a "glue" molecule which has been shown promising for immobilization of various biomacromolecules at silica surface. We address here for the first time the ability of such IBAM-modified NPs to sustainably deliver proteins over a prolonged time. In this work, a quantitative loading study of proteins (serum albumin (HSA), peroxidase (HRP), immunoglobulin (IgG) and polylysine (PLL)) on STMS@IBAM is first presented using three complementary detection techniques to ensure precision and avoid protein quantification issues. The results demonstrated a high loading capacity for HSA and HRP (≥ ca. 350 µg.mg-1) but a moderate one for IgG and PLL. After evaluating the physicochemical properties of the loaded particles and their stability over scaling-up and washings, the ability of STMS@IBAM to release proteins over prolonged time was evaluated in equilibrium (static) and flow mimicking (dynamic) conditions and at different temperatures (25, 37, 45 °C). Results show not only the potential of such "glue" functionalized STMS to release proteins in a sustained way, but also the retention of the biological activity of immobilized and released HRP, used as an enzyme model. Finally, an AFM-force spectroscopy study was conducted to decipher the interactions between IBAM and proteins, showing the involvement of different interactions in the adsorption and release processes.

Design and applications of protein delivery systems in nanomedicine and tissue engineering.

Proteins are biological macromolecules involved in a wide range of biological functions, which makes them very appealing as therapeutics agents. Indeed, compared to small molecule drugs, their endogenous nature ensures their biocompatibility and biodegradability, they can be used in a large range of applications and present a higher specificity and activity. However, they suffer from unfolding, enzymatic degradation, short half-life and poor membrane permeability. To overcome such drawbacks, the development of protein delivery systems to protect, carry and deliver them in a controlled way have emerged importantly these last years. In this review, the formulation of a wide panel of protein delivery systems either in the form of polymer or inorganic nanoengineered colloids and scaffolds are presented and the protein loading and release mechanisms are addressed. A section is also dedicated to the detection of proteins and the characterization methods of their release. Then, the main protein delivery systems developed these last three years for anticancer, tissue engineering or diabetes applications are presented, as well as the major in vivo models used to test them. The last part of this review aims at presenting the perspectives of the field such as the use of protein-rich material or the sequestration of proteins. This part will also deal with less common applications and gene therapy as an indirect method to deliver protein.

Biomaterial based strategies to reconstruct the nigrostriatal pathway in organotypic slice co-cultures.

Protection or repair of the nigrostriatal pathway represents a principal disease-modifying therapeutic strategy for Parkinson's disease (PD). Glial cell line-derived neurotrophic factor (GDNF) holds great therapeutic potential for PD, but its efficacious delivery remains difficult. The aim of this study was to evaluate the potential of different biomaterials (hydrogels, microspheres, cryogels and microcontact printed surfaces) for reconstructing the nigrostriatal pathway in organotypic co-culture of ventral mesencephalon and dorsal striatum. The biomaterials (either alone or loaded with GDNF) were locally applied onto the brain co-slices and fiber growth between the co-slices was evaluated after three weeks in culture based on staining for tyrosine hydroxylase (TH). Collagen hydrogels loaded with GDNF slightly promoted the TH+ nerve fiber growth towards the dorsal striatum, while GDNF loaded microspheres embedded within the hydrogels did not provide an improvement. Cryogels alone or loaded with GDNF also enhanced TH+ fiber growth. Lines of GDNF immobilized onto the membrane inserts via microcontact printing also significantly improved TH+ fiber growth. In conclusion, this study shows that various biomaterials and tissue engineering techniques can be employed to regenerate the nigrostriatal pathway in organotypic brain slices. This comparison of techniques highlights the relative merits of different technologies that researchers can use/develop for neuronal regeneration strategies.

Albumin-driven disassembly of lipidic nanoparticles: the specific case of the squalene-adenosine nanodrug.

In the field of nanomedicine, nanostructured nanoparticles (NPs) made of self-assembling prodrugs emerged in the recent years with promising properties. In particular, squalene-based drug nanoparticles have already shown their efficiency through in vivo experiments. However, a complete pattern of their stability and interactions in the blood stream is still lacking. In this work we assess the behavior of squalene-adenosine (SQAd) nanoparticles - whose neuroprotective effect has already been demonstrated in murine models - in the presence of fetal bovine serum (FBS) and of bovine serum albumin (BSA), the main protein of blood plasma. Extensive physicochemical characterizations were performed using Small Angle Neutron Scattering (SANS), cryogenic transmission electron microscopy (Cryo-TEM), circular dichroism (CD), steady-state fluorescence spectroscopy (SSFS) and isothermal titration calorimetry (ITC) as well as in silico by means of ensemble docking simulations with human serum albumin (HSA). Significant changes in the colloidal stability of the nanoparticles in the presence of serum albumin were observed. SANS, CD and SSFS analyses demonstrated an interaction between SQAd and BSA, with a partial disassembly of the nanoparticles in the presence of BSA and the formation of a complex between SQAd and BSA. The interaction free energy of SQAd nanoparticles with BSA derived from ITC experiments, is about -8 kcal mol-1 which is further supported in silico by ensemble docking simulations. Overall, our results show that serum albumin partially disassembles SQAd nanoparticles by extracting individual SQAd monomers from them. As a consequence, the SQAd nanoparticles would act as a circulating reservoir in the blood stream. The approach developed in this study could be extended to other soft organic nanoparticles.

Synthesis and characterization of hyaluronic acid coated manganese dioxide microparticles that act as ROS scavengers.

Atherosclerosis is a chronic inflammatory disease of the arterial wall that leads to cardiovascular diseases which are the major cause of deaths worldwide. There is currently no treatment that can stop or reverse the disease. However, the use of microparticles with anti-inflammatory properties could represent a promising treatment. Herein, spherical microparticles with a core-shell structure and an average diameter of 1µm were synthesized. The microparticles were comprised of a MnCO3 and MnO2 core and a 4-arm PEG-amine cross-linked shell of hyaluronic acid. The HA-Mn-SM microparticles were loaded with D-α-tocopherol (vitamin-E) (TOC), to fabricate a targeted biocompatible delivery platform for the treatment of atherosclerotic inflamed cells. Loading and release studies of TOC demonstrated a lactic acid concentration dependant controlled release profile of the HA-Mn-SM mimicking the atherosclerotic environment where lactic acid is over-produced. The microparticles exhibited a high scavenging ability towards H2O2 in addition to the controlled generation of O2. The optimal results were obtained for 250µg/mL microparticles which in the presence of 1000µM H2O2 resulted in the scavenging of almost all the H2O2. Our results demonstrate that 50µg/mL of microparticles scavenged continuously produced H2O2 up to a concentration of 1000µM, a characteristic that demonstrates the sustained therapeutic effect of the HA-Mn-SM microparticles in an environment that mimics that of inflamed tissues. Our results indicate the potential use of HA-Mn-SM as a novel platform for the treatment of atherosclerosis. In vitro studies confirmed that the microparticles are not cytotoxic at concentrations up to 250µg/mL and for 72h. These preliminary results indicate the potential use of HA-Mn-SM as a novel drug delivery system for atherosclerotic tissues.