Chemical insights into bioinks for 3D printing

International audienc

Chemical cross-linking methods for cell encapsulation in hydrogels

International audienceCell-encapsulating hydrogels are of tremendous interest in regenerative medicine. Tissue engineering relies on biomaterials able to act as artificial extracellular matrices to guide cells towards the development of new tissues. Therefore, considerable efforts have been made to design biomaterials which mimic cells’ native environment, thus encouraging natural behavior. The choice of biomaterial in which cells are embedded is crucial for their survival, proliferation and differentiation. Being more stable, chemical hydrogels are preferred over physical hydrogels as cell-laden substrates. When designing chemical hydrogels, scientists must choose not only the nature of the network (synthetic and/or bio-polymers) but also the type of cross-link bridging hydrogel constituents. For that purpose, numerous chemistries have been used (i) to introduce reactive functions on the hydrogel precursors and (ii) to form covalent bonds in the presence of living cells. The review will discuss the advantages and limitations of each strategy

Inorganic polymerization: an attractive route to biocompatible hybrid hydrogels

International audienceAs an intermediate state between liquid and solid materials, hydrogels display unique properties, opening a wide scope of applications, especially in the biomedical field. Organic hydrogels are composed of an organic network cross-linked via chemical or physical reticulation nodes. In contrast, hybrid hydrogels are defined by the coexistence of organic and inorganic moieties in water. Inorganic polymerization, i.e. sol-gel process, is one of the main techniques leading to hybrid hydrogels. The chemoselectivity of this method proceeds through hydrolysis and condensation reactions of metal oxide moieties. In addition, the mild reaction conditions make this process very promising for the preparation of water-containing materials and their bio-applications

Peptide synthesis: ball-milling, in solution, or on solid support, what is the best strategy?

International audienceWhile presenting particularly interesting advantages, peptide synthesis by ball-milling was never compared to the two traditional strategies, namely peptide syntheses in solution and on solid support (solid-phase peptide synthesis, SPPS). In this study, the challenging VVIA tetrapeptide was synthesized by ball-milling, in solution, and on solid support. The three strategies were then compared in terms of yield, purity, reaction time and environmental impact. The results obtained enabled to draw some strengths and weaknesses of each strategy, and to foresee what will have to be implemented to build more efficient and sustainable peptide syntheses in the near future

Self-mineralization and assembly of a bis-silylated Phe–Phe pseudodipeptide to a structured bioorganic–inorganic material

International audienceSelf-mineralization of trialkoxysilyl hybrid peptide yield in a single step a nanostructured hybrid material. A bis-silylated pseudodipeptide inspired from the Phe-Phe dipeptide was used to program the assembly by sol-gel polymerization in heterogeneous conditions, in water at pH 1.5 without any structure-directing agent. A mechanism deciphering the hybrid material assembly was proposed thanks to 1H NMR spectroscopy. First, water-insoluble hybrid building blocks were hydrolysed into their soluble silanol counterparts. Then, these transitional species, thanks to hydrogen bonding and π-π stacking, self-assembled in solution. Last, the proximity of silanol moieties favoured their polycondensation into growing siloxane oligomers, which spontaneously precipitated to produce an ordered hybrid material

Solid-Phase Synthesis of Arylpiperazine Derivatives and Implementation of the Distributed Drug Discovery (D3) Project in the Search for CNS Agents

We have successfully implemented the concept of Distributed Drug Discovery (D3) in the search for CNS agents. Herein, we demonstrate, for the first time, student engagement from different sites around the globe in the development of new biologically active compounds. As an outcome we have synthesized a 24-membered library of arylpiperazine derivatives targeted to 5-HT1A and 5-HT2A receptors. The synthesis was simultaneously performed on BAL-MBHA-PS resin in Poland and the United States, and on BAL-PS-SynPhase Lanterns in France. The D3 project strategy opens the possibility of obtaining potent 5-HT1A/5-HT2A agents in a distributed fashion. While the biological testing is still centralized, this combination of distributed synthesis with screening will enable a D3 network of students world-wide to participate, as part of their education, in the synthesis and testing of this class of biologically active compounds

Conception et synthèse de nouveaux outils chimiques pour l'étude des phosphoprotéines et la caractérisation de la liaison d'un ligand à son récepteur par spectrométrie de masse MALDI-TOF

Depuis l'avènement des techniques d'ionisation douces comme le MALDI ou l'ESI, la spectrométrie de masse est devenue un outil incontournable pour l'étude des biomolécules, et en particulier, des protéines. Le mécanisme d'ionisation spécifique des sources MALDI introduit un phénomène de discrimination spectrale généralement considéré comme un frein. Le greffage d'un motif a-cyano 4-hydroxy cinnamique ou HCCA sur un peptide permet d'en tirer parti en créant une discrimination spectrale favorable au composé marqué. Deux applications du marquage HCCA ont été développées au cours de ce travail de thèse.Une des limitations des études phosphoprotéomiques réside dans la faible ionisation des peptides phosphorylés. De nombreuses méthodes de purification ont été proposées pour contourner ce problème. Nous avons étudié une approche alternative qui consiste à greffer spécifiquement un motif HCCA sur une position phosphorylée afin d'amplifier les signaux des peptides d'intérêt. Nous avons développé en parallèle une méthodologie pour l'étude de la liaison d'un ligand peptidique à son récepteur ne nécessitant pas de radioactivité. Le greffage covalent d'HCCA sur un ligand nous a permis de le détecter et de le quantifier dans une expérience de déplacement. Cette méthodologie a notamment permis de déterminer l'affinité d'un ligand de référence pour le récepteur V1A à la vasopressine.Since the advent of soft ionization techniques such as MALDI or ESI, mass spectrometry has become an indispensable tool for the study of biomolecules, and particularly proteins. The specific ionization mechanism of MALDI leads to a spectral discrimination phenomenon generally regarded as a restriction. The a-cyano 4-hydroxy cinnamic acid (HCCA) pattern grafting on a peptide can take advantage by introducing a spectral discrimination in favor of the labeled compound. Two applications have been developed during this thesis, based on HCCA tagging.A limitation of phosphoproteomic studies is the low ionization of phosphorylated peptides. Many purification methods have been reported to circumvent this problem. We studied an alternative approach which consists in specifically grafting a HCCA moiety on a phosphorylated position in order to amplify the signal of interest peptide.At the same time, we have developed a methodology for the study of ligand peptidic binding to its receptor which does not require radioactivity. The covalent HCCA tagged ligand has allowed us to detect and quantify it, in a binding displacement assay. More particularly, this methodology allowed us to determine the affinity of a reference ligand for the V1A vasopressin receptor.MONTPELLIER-BU Sciences (341722106) / SudocSudocFranceF

Microwave-assisted Solid Phase Peptide Synthesis on High Loaded Resins

International audienc

Silicone grafted bioactive peptides and their applications

International audienc

Chemical cross-linkers for protein structure studies by mass spectrometry

International audienceThe cross-linking approach combined with MS for protein structure determination is one of the most striking examples of multidisciplinary success. Indeed, it has become clear that the bottleneck of the method was the detection and the identification of low-abundance cross-linked peptides in complex mixtures. Sample treatment or chromatography separation partially addresses these issues. However, the main problem comes from over-represented unmodified peptides, which do not yield any structural information. A real breakthrough was provided by high mass accuracy measurement, because of the outstanding technical developments in MS. This improvement greatly simplified the identification of cross-linked peptides, reducing the possible combinations matching with an observed m/z value. In addition, the huge amount of data collected has to be processed with dedicated software whose role is to propose distance constraints or ideally a structural model of the protein. In addition to instrumentation and algorithms efficiency, significant efforts have been made to design new cross-linkers matching all the requirements in terms of reactivity and selectivity but also displaying probes or reactive systems facilitating the isolation, the detection of cross-links, or the interpretation of MS data. These chemical features are reviewed and commented on in the light of the more recent strategies