Engineering, on-demand manufacturing, and scaling-up ofpolymeric nanocapsules

Polymeric nanocapsules are versatile delivery systems with the capacity to load lipophilic drugs in their oily nucleus and hydrophilic drugs in their polymeric shell. The objective of this work was to expand the technological possibilities to prepare customized nanocapsules. First, we adapted the solvent displacement technique to modulate the particle size of the resulting nanocapsules in the 50–500 nm range. We also produced nanosystems with a shell made of one or multiple polymer layers i.e. chitosan, dextran sulphate, hyaluronate, chondroitin sulphate, and alginate. In addition, we identified the conditions to translate the process into a miniaturized high-throughput tailor-made fabrication that enables massive screening of formulations. Finally, the production of the nanocapsules was scaled-up both in a batch production, and also using microfluidics. The versatility of the properties of these nanocapsules and their fabrication technologies is expected to propel their advance from bench to clinicThis work was funded by CDTI (Spanish Ministry of Science and Innovation) and (Fondo Tecnológico Xunta de Galicia, FEDER funds) COLIVAC-FEDER INNTERCONECTA-2012-CE277 and Grupos de referencia competitiva (ED431C 2017/09, Xunta de Galicia)S

Engineering Anisotropic Meniscus: Zonal Functionality and Spatiotemporal Drug Delivery

This document is the preprint manuscript version of a published work that appeared in final form in Tissue Engineering Part B: Reviews, Copyright 2020, Mary Ann Liebert, Inc., publishers after peer review and technical editing by the publisher. To access the final edited and published work see: https://doi.org/10.1089/ten.teb.2020.0096Human meniscus is a fibrocartilaginous structure that is crucial for an adequate performance of the human knee joint. Degeneration of the meniscus is often followed by partial or total meniscectomy, which enhances the risk of developing knee osteoarthritis. The lack of a satisfactory treatment for this condition has triggered a major interest in drug delivery (DD) and tissue engineering (TE) strategies intended to restore a bioactive and fully functional meniscal tissue. The aim of this review is to critically discuss the most relevant studies on spatiotemporal DD and TE, aiming for a multizonal meniscal reconstruction. Indeed, the development of meniscal tissue implants should involve a provision for adequate active molecules and scaffold features that take into account the anisotropic ultrastructure of human meniscus. This zonal differentiation is reflected in the meniscus biochemical composition, collagen fiber arrangement, and cell distribution. In this sense, it is expected that a proper combination of advanced DD and zonal TE strategies will play a key role in the future trends in meniscus regenerationThis project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 814444 (MEFISTO); and from Xunta de Galicia’s Grupos de referencia competitiva (grant number ED431C 2017/09

New scaffolds encapsulating TGF-β3/BMP-7 combinations driving strong chondrogenic differentiation

The regeneration of articular cartilage remains an unresolved question despite the current access to a variety of tissue scaffolds activated with growth factors relevant to this application. Further advances might result from combining more than one of these factors; here, we propose a scaffold composition optimized for the dual delivery of BMP-7 and TGF-β3, two proteins with described chondrogenic activity. First, we tested in a mesenchymal stem cell micromass culture with TGF-β3 whether the exposure to microspheres loaded with BMP-7 would improve cartilage formation. Histology and qRT-PCR data confirmed that the sustained release of BMP-7 cooperates with TGF-β3 towards the generation of chondrogenic differentiation. Then, we optimized a scaffold prototype for tissue culture and dual encapsulation of BMP-7 and TGF-β3. The scaffolds were prepared from poly(lactic-co-glycolic acid), and BMP-7/TGF-β3 were loaded as nanocomplexes with heparin and Tetronic 1107. The scaffolds showed the sustained release of both proteins over four weeks, with minimal burst effect. We finally cultured human mesenchymal stems cells on these scaffolds, in the absence of exogenous chondrogenic factor supplementation. The cells cultured on the scaffolds loaded with BMP-7 and TGF-β3 showed clear signs of cartilage formation macroscopically and histologically. RT-PCR studies confirmed a clear upregulation of cartilage markers SOX9 and Aggrecan. In summary, scaffolds encapsulating BMP-7 and TGF-β3 can efficiently deliver a cooperative growth factor combination that drives efficient cartilage formation in human mesenchymal stem cell cultures. These results open attractive perspectives towards in vivo translation of this technology in cartilage regeneration experimentsFundación Ramón Areces (CIVP16A1832), Xunta de Galicia (Proxectos de Investigación Desenvolvidos por Investigadores Emerxentes, EM2013/042), Fundación BBVA, Proyectos de Investigación en Biomedicina (2014- PO0110) y Ministerio de Economía y Competitividad (SAF2014-58189-R)S

Studies on the base catalyzed synthesis of acetophenone enolcarbamate enhanced by microwaves

The 10th International Electronic Conference on Synthetic Organic Chemistry session Symposium on Microwave Assisted SynthesisMicrowave assisted organic synthesis (MAOS) of N,N-diisopropyl enol carbamate of acetophenone is achieved in good yield when acetophenone, N,N-diisopropylcarbamoyl chloride and Hüning's base, DABCO or TMEDA were irradiated with microwaves. It is remarkable that reaction times are reduced from six days with conventional heating, to twenty minutesXUNTA DE GALICIA for financial support: PGIDIT05PXIB26201PR, PR405 A 098/59-0 . USC for a predoctoral fellowship to JCC

Microwave-assisted synthesis of 3-arylcoumarins

The 12th International Electronic Conference on Synthetic Organic Chemistry session Symposium on Microwave Assisted Synthesis3-arylcoumarins are prepared in good yields by microwave assisted solvent-free condensation of phenylacetic acids and phenylacetic esters, using potassium tert-butoxide as base.We acknowledge XUNTA DE GALICIA for financial support: PGIDIT05PXIB26201PR, PR405 A098/59-0. USC for a predoctoral fellowship to J. C.-C. This work was undertaken as part of the EC sponsored programs D32 COST Program (Chemistry in High-Energy Microenvironments, WG10)

Polysaccharide nanoparticles can efficiently modulate the immune response against anHIV peptide antigen

The development of an effective HIV vaccine continues to be a major health challenge since, so far, only the RV144 trial has demonstrated a modest clinical efficacy. Recently, the targeting of the 12 highly conserved protease cleavage sites (PCS1–12) has been presented as a strategy seeking to hamper the maturation and infectivity of HIV. To pursue this line of research, and because peptide antigens have low immunogenicity, we have included these peptides in engineered nanoparticles, aiming at overcoming this limitation. More specifically, we investigated whether the covalent attachment of a PCS peptide (PCS5) to polysaccharide-based nanoparticles, and their coadministration with polyinosinic:polycytidylic acid (poly(I:C)), improved the generated immune response. To this end, PCS5 was first conjugated to two different polysaccharides (chitosan and hyaluronic acid) through either a stable or a cleavable bond and then associated with an oppositely charged polymer (dextran sulfate and chitosan) and poly(I:C) to form the nanoparticles. Nanoparticles associating PCS5 by ionic interactions were used in this study as the control formulation. In vivo, all nanosystems elicited high anti-PCS5 antibodies. Nanoparticles containing PCS5 conjugated and poly(I:C) seemed to induce the strongest activation of antigen-presenting cells. Interestingly, T cell activation presented different kinetics depending on the prototype. These findings show that both the nanoparticle composition and the conjugation of the HIV peptide antigen may play an important role in the generation of humoral and cellular responsesThis work was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health (Award No. R01AI111805, Subaward No. 41795-02) and by the European Union’s Horizon 2020 research program (NanoPilot project, Grant Agreement No. 646142). T.G.D. acknowledges a predoctoral FPU grant from the Spanish Ministry of Education, Culture and Sports (Grant No. FPU14/05866)S

Microwave assisted synthesis of tripodal triazines from 1,3,5-tris(2- hydroxyethyl)-1,3,5-triazinane-2,4,6-trione and fatty acids

The 13th International Electronic Conference on Synthetic Organic Chemistry session Symposium on Microwave Assisted SynthesisA new fast and direct synthesis for tripodal fatty acid esters of tris-hydroxyethyl isocyanurate 1 under microwave irradiation is developed. The method is valid for saturated and unsaturated acids, but the last produced a mixture of esters cis and transXUNTA DE GALICIA for financial support: PGIDIT05PXIB26201PR and USC for a predoctoral fellowship to JC

Microwave assisted synthesis of 1,3,5-tris(2-hydroxyethyl)isocyanurate carbamates with C3 symmetry

The 14th International Electronic Conference on Synthetic Organic Chemistry session Microwave Assisted SynthesisMicrowave assisted synthesis of 1,3,5-tris(2-hydroxyethyl)isocyanurate carbamates under solventless conditions in short times and good yields is describedXUNTA DE GALICIA for financial support: INCITE09 262346P

Solvent Free Microwave Assisted Synthesis of 1,3,5-tris(2-aryloxyethyl)isocyanurates

The 14th International Electronic Conference on Synthetic Organic Chemistry session Microwave Assisted SynthesisSynthesis of 1,3,5-tris(2-aryloxyethyl)isocyanurates by esterification of 1,3,5-tris(2-hydroxyethyl)isocyanurate under microwave irradiation in a direct and efficient way in solvent free conditionsXUNTA DE GALICIA for financial support: INCITE09 262346P

Microwave assisted synthesis of triskelium compounds from 1,3,5-triazines

The 11th International Electronic Conference on Synthetic Organic Chemistry session Symposium on Microwave Assisted SynthesisMicrowave assisted synthesis of 2,4,6-trishydrazino-1,3,5-triazine from hydrazine and cyanuric chloride is achieved in good yield. Its reaction with acetone and benzaldehyde yields the corresponding trishydrazones in good yields and short reaction times using MAOSXUNTA DE GALICIA for financial support: PGIDIT05PXIB26201PR, PR405 A 098/59-0. USC for a predoctoral fellowship to JC