Engineered nascent living human tissues with unit programmability.

Leveraging human cells as materials precursors is a promising approach for fabricating living materials with tissue-like functionalities and cellular programmability. Here we describe a set of cellular units with metabolically engineered glycoproteins that allow cells to tether together to function as macrotissue building blocks and bioeffectors. The generated human living materials, termed as Cellgels, can be rapidly assembled in a wide variety of programmable three-dimensional configurations with physiologically relevant cell densities (up to 108 cells per cm3), tunable mechanical properties and handleability. Cellgels inherit the ability of living cells to sense and respond to their environment, showing autonomous tissue-integrative behaviour, mechanical maturation, biological self-healing, biospecific adhesion and capacity to promote wound healing. These living features also enable the modular bottom-up assembly of multiscale constructs, which are reminiscent of human tissue interfaces with heterogeneous composition. This technology can potentially be extended to any human cell type, unlocking the possibility for fabricating living materials that harness the intrinsic biofunctionalities of biological systems.

Cell Surface Engineering Tools for Programming Living Assemblies.

Breakthroughs in precision cell surface engineering tools are supporting the rapid development of programmable living assemblies with valuable features for tackling complex biological problems. Herein, the authors overview the most recent technological advances in chemically- and biologically-driven toolboxes for engineering mammalian cell surfaces and triggering their assembly into living architectures. A particular focus is given to surface engineering technologies for enabling biomimetic cell-cell social interactions and multicellular cell-sorting events. Further advancements in cell surface modification technologies may expand the currently available bioengineering toolset and unlock a new generation of personalized cell therapeutics with clinically relevant biofunctionalities. The combination of state-of-the-art cell surface modifications with advanced biofabrication technologies is envisioned to contribute toward generating living materials with increasing tissue/organ-mimetic bioactivities and therapeutic potential.

Engineering mammalian living materials towards clinically relevant therapeutics.

Engineered living materials represent a new generation of human-made biotherapeutics that are highly attractive for a myriad of medical applications. In essence, such cell-rich platforms provide encodable bioactivities with extended lifetimes and environmental multi-adaptability currently unattainable in conventional biomaterial platforms. Emerging cell bioengineering tools are herein discussed from the perspective of materializing living cells as cooperative building blocks that drive the assembly of multiscale living materials. Owing to their living character, pristine cellular units can also be imparted with additional therapeutically-relevant biofunctionalities. On this focus, the most recent advances on the engineering of mammalian living materials and their biomedical applications are herein outlined, alongside with a critical perspective on major roadblocks hindering their realistic clinical translation. All in all, transposing the concept of leveraging living materials as autologous tissue-building entities and/or self-regulated biotherapeutics opens new realms for improving precision and personalized medicine strategies in the foreseeable future.

Coordination Compounds As Multi-Delivery Systems for Osteoporosis.

Osteoporosis therapies leveraging bisphosphonates and mineral components (e.g., magnesium, calcium, and strontium) have been raising attention because of their potential for managing this ever-growing disease. The administration of multicomponent therapeutics (combined therapy) in elderly patients is complex and suffers from low patient adherence. Herein, we report an all-in-one combination of four antiosteoporotic components into a new family of coordination complexes: [M2(H4alen)4(H2O)2]·1.5H2O [where M2+ = Mg2+ (1), (Mg0.535Ca0.465)2+ (2) and (Mg0.505Ca0.450Sr0.045)2+ (3)]. These solid-state complexes were prepared, for the first time, through microwave-assisted synthesis. It is demonstrated that the compounds are capable of releasing their antiosteoporotic components, both in conditions that mimic the path along the gastrointestinal tract and in long periods under physiological conditions (pH ∼7.4). More importantly, when administered in low concentrations, the compounds did not elicit a cytotoxic effect toward liver, kidney, and osteoblast-like cell lines. Besides, it is important to highlight the unique coordination complex with four bone therapeutic components, [(Mg0.505Ca0.450Sr0.045)2(H4alen)4(H2O)2]·1.5H2O (3), which significantly promoted osteoblast metabolic activity up to ca. 1.4-fold versus the control group. These findings bring this type of compounds one-step closer to be considered as an all-in-one and more effective treatment for managing chronic bone diseases, prompting further research on their therapeutic properties.

Double network laminarin-boronic/alginate dynamic bioink for 3D bioprinting cell-laden constructs.

The design of dynamically crosslinked hydrogel bioinks for three-dimensional (3D) bioprinting is emerging as a valuable strategy to advance the fabrication of mechanically tuneable cell-laden constructs for 3Din vitrodisease modelling and tissue engineering applications. Herein, a dynamic bioink comprising boronic acid-functionalised laminarin and alginate is explored for bioprinting 3D constructs under physiologically relevant conditions. The formulated bioink takes advantage of a double crosslinked network that combines covalent but reversible boronate ester bonds and ionic gelation via divalent cations. Moreover, it exhibits suitable rheological properties and improved mechanical features owing to its modular crosslinking chemistry, yielding stable constructs with user-programmable architecture. We explored such dynamic bioink as a supporting matrix for different cell classes, namely osteoblast precursors, fibroblasts and breast cancer cells. The resulting cell-laden bioprinted hydrogels display a homogeneous cell distribution post-printing and exceptional cell viability (>90%) that can be maintained for prolonged time periods in culture (14 days) for all cell lines. This simple and chemically versatile approach is envisaged to accelerate the development of multifunctional bioinks and contribute towards the fabrication of biomimetic 3D scaffolds with applicability in a wide range of predictive or exploratory biomedical platforms.

Gelatin Methacryloyl (GelMA) Nanocomposite Hydrogels Embedding Bioactive Naringin Liposomes.

The development of nanocomposite hydrogels that take advantage of hierarchic building blocks is gaining increased attention due to their added functionality and numerous biomedical applications. Gathering on the unique properties of these platforms, herein we report the synthesis of bioactive nanocomposite hydrogels comprising naringin-loaded salmon-derived lecithin nanosized liposomal building blocks and gelatin methacryloyl (GelMA) macro-sized hydrogels for their embedding. This platform takes advantage of liposomes' significant drug loading capacity and their role in hydrogel network reinforcement, as well as of the injectability and light-mediated crosslinking of bioderived gelatin-based biomaterials. First, the physicochemical properties, as well as the encapsulation efficiency, release profile, and cytotoxicity of naringin-loaded nanoliposomes (LipoN) were characterized. Then, the effect of embedding LipoN in the GelMA matrix were characterized by studying the release behavior, swelling ratio, and hydrophilic character, as well as the rheological and mechanical properties of GelMA and GelMA-LipoN functionalized hydrogels. Finally, the dispersion of nanoliposomes encapsulating a model fluorescent probe in the GelMA matrix was visualized. The formulation of naringin-loaded liposomes via an optimized procedure yielded nanosized (114 nm) negatively charged particles with a high encapsulation efficiency (~99%). Naringin-loaded nanoliposomes administration to human adipose-derived stem cells confirmed their suitable cytocompatibility. Moreover, in addition to significantly extending the release of naringin from the hydrogel, the nanoliposomes inclusion in the GelMA matrix significantly increased its elastic and compressive moduli and decreased its swelling ratio, while showing an excellent dispersion in the hydrogel network. Overall, salmon-derived nanoliposomes enabled the inclusion and controlled release of pro-osteogenic bioactive molecules, as well as improved the hydrogel matrix properties, which suggests that these soft nanoparticles can play an important role in bioengineering bioactive nanocomposites for bone tissue engineering in the foreseeable future.

Self-Assembled Bioactive Colloidal Gels as Injectable Multiparticle Shedding Platforms.

Self-assembled colloidal gels are highly versatile 3D nanocluster platforms with potential to overcome the rapid clearing issues associated with standard free nanotherapeutics administration. However, the development of nanoassembled colloidal gels exhibiting autonomous multiparticle release from the bulk particle network remains elusive. Herein, we generated multiparticle colloidal gels from two nanosized building blocks: cationic poly(d,l-lactide-co-glycolide)-polyethylenimine (PLGA-PEI) nanoparticles and anionic zein-hyaluronan (HA) nanogels that assemble into macrosized 3D constructs via attractive electrostatic forces. The resulting colloidal gels exhibited high stability in complex culture medium as well as fit-to-shape moldable properties and injectability. Moreover, nanoassembled colloidal gels encapsulated bioactive quercetin flavonoids with high loading efficacy and presented remarkable anti-inflammatory activities, reducing key proinflammatory biomarkers in inflammation-activated macrophages. More importantly, because of their rationally selected building blocks zein-HA/PLGA-PEI, self-assembled colloidal platforms displayed autonomous multiparticle shedding. Both positive and negative particles released from the colloidal system were efficiently internalized by macrophages along time as evidenced by quantitative particle uptake analysis. Overall, the generated nanostructured gels represent an implantable versatile platform for focalized multiparticle delivery. In addition, the possibility to combine a higher number of particle species with different properties or stimuli-responsiveness enables the manufacturing of combinatorial nanostructured gels for numerous biomedical applications.

Mechanochemical Patternable ECM-Mimetic Hydrogels for Programmed Cell Orientation.

Native human tissues are supported by a viscoelastic extracellular matrix (ECM) that can adapt its intricate network to dynamic mechanical stimuli. To recapitulate the unique ECM biofunctionality, hydrogel design is shifting from typical covalent crosslinks toward covalently adaptable networks. To pursue such properties, herein hybrid polysaccharide-polypeptide networks are designed based on dynamic covalent assembly inspired by natural ECM crosslinking processes. This is achieved through the synthesis of an amine-reactive oxidized-laminarin biopolymer that can readily crosslink with gelatin (oxLAM-Gelatin) and simultaneously allow cell encapsulation. Interestingly, the rational design of oxLAM-Gelatin hydrogels with varying aldehyde-to-amine ratios enables a refined control over crosslinking kinetics, viscoelastic properties, and degradability profile. The mechanochemical features of these hydrogels post-crosslinking offer an alternative route for imprinting any intended nano- or microtopography in ECM-mimetic matrices bearing inherent cell-adhesive motifs. Different patterns are easily paved in oxLAM-Gelatin under physiological conditions and complex topographical configurations are retained along time. Human adipose-derived mesenchymal stem cells contacting mechanically sculpted oxLAM-Gelatin hydrogels sense the underlying surface nanotopography and align parallel to the anisotropic nanoridge/nanogroove intercalating array. These findings demonstrate that covalently adaptable features in ECM-mimetic networks can be leveraged to combine surface topography and cell-adhesive motifs as they appear in natural matrices.

Advanced Bottom-Up Engineering of Living Architectures.

Bottom-up tissue engineering is a promising approach for designing modular biomimetic structures that aim to recapitulate the intricate hierarchy and biofunctionality of native human tissues. In recent years, this field has seen exciting progress driven by an increasing knowledge of biological systems and their rational deconstruction into key core components. Relevant advances in the bottom-up assembly of unitary living blocks toward the creation of higher order bioarchitectures based on multicellular-rich structures or multicomponent cell-biomaterial synergies are described. An up-to-date critical overview of long-term existing and rapidly emerging technologies for integrative bottom-up tissue engineering is provided, including discussion of their practical challenges and required advances. It is envisioned that a combination of cell-biomaterial constructs with bioadaptable features and biospecific 3D designs will contribute to the development of more robust and functional humanized tissues for therapies and disease models, as well as tools for fundamental biological studies.

Bioinstructive Naringin-Loaded Micelles for Guiding Stem Cell Osteodifferentiation.

Naringin is a naturally occurring flavanone with recognized neuroprotective, cardioprotective, anti-inflammatory, and antiosteoporotic properties. Herein, the delivery of Naringin-loaded methoxy-poly(ethylene glycol)-maleimide-thiol-poly(l-lactide) (mPEGMSPLA) diblock polymeric micelles to human adipose-derived stem cells (hASCs) with the aim to augment its pro-osteogenic effect in these cells is reported for the first time. The synthesis of the diblock copolymer is performed via Michael-type addition reaction between hydrophilic methoxy-poly(ethylene glycol)-maleimide (mPEGMAL) and hydrophobic thiol-poly(l-lactide) (PLASH) and confirmed by 1 H NMR and attenuated total reflectance Fourier transformed infrared (ATR-FTIR) spectroscopy. The resulting mPEGMSPLA copolymer self-assembles into monodispersed polymeric micelles (≈84.4 ± 2 nm) and presents a high Naringin encapsulation efficiency (87.8 ± 4%), with a sustained release profile at physiological pH. Alongside, in vitro data reveal that upon internalization into hASC 2D cultures, Naringin nanomicellar formulations attain a higher pro-osteogenic effect than that of free drug. Notably, these bioactive carriers also induce superior osteopontin expression and increase matrix mineralization in these cells over free drug administration. Overall, such findings support for the first time the use of polymeric nanomicelles for Naringin delivery into hASCs as a valid approach for modulating stem cell osteogenic differentiation.

Bioinspired bone therapies using naringin: applications and advances.

The use of natural compounds for treating chronic bone diseases holds remarkable potential. Among these therapeutics, naringin, a flavanone glycoside, represents one of the most promising candidates owing to its multifaceted effect on bone tissues. This review provides an up-to-date overview on naringin applications in the treatment of bone disorders, such as osteoporosis and osteoarthritis, and further highlights its potential for stem cell pro-osteogenic differentiation therapies. A critical perspective on naringin clinical translation is also provided. The topic is discussed in light of recently developed biomaterial-based approaches that potentiate its bioavailability and bioactivity. Overall, the reported pro-osteogenic, antiresorptive and antiadipogenic properties establish this flavanone as an exciting candidate for application in bone tissue engineering and regenerative medicine.

Stimuli-responsive nanocarriers for delivery of bone therapeutics - Barriers and progresses.

The development of stimuli-responsive nanomedicines with tunable cargo release is gathering an increased applicability in bone regeneration and precision biomedicine. Yet, the formulation of nanocarriers that explore skeletal-specific stimuli remains remarkably challenging to materialize due to several endogenous and disease-specific barriers that must be considered during particle design stages. Such anatomo-physiological constrains ultimately hinder nanocarriers bioavailability in target bone tissues and impact the overall therapeutic outcome. This review aims to showcase and critically discuss the hurdles encountered upon responsive nanocarriers delivery in the context of skeletal diseases or tissue regeneration scenarios. Such focus is complemented with an in-depth and up-to-date analysis of advances in the development of stimuli-responsive, bone-focused delivery systems. In a holistic perspective, a deeper knowledge of human osteology combined with advances in materials functionalization via simple precision-chemistry is envisioned to incite the manufacture of stimuli-triggered nanomedicines with more realistic potential for clinical translation.

Molybdenum(0) tricarbonyl and tetracarbonyl complexes with a cationic pyrazolylpyridine ligand: synthesis, crystal structures and catalytic performance in olefin epoxidation.

The synthesis of molybdenum(0) tricarbonyl and tetracarbonyl complexes of the form [Mo(CO)3(ptapzpy)Br] (1) and cis-[Mo(CO)4(ptapzpy)]Br (2) is reported, where ptapzpy = 2-(1-propyltrimethylammonium-3-pyrazolyl)pyridine. Preparation of these derivatives was accomplished either through thermal replacement of CO in Mo(CO)6 (for 1) or substitution under milder conditions of piperidine ligands in the precursor cis-[Mo(CO)4(pip)2] (for 2). The crystal structures of the ligand [ptapzpy]Br and complexes 1 and 2 were determined. Thermal treatment of 2 at 125-150 °C leads to mono decarbonylation and formation of 1. On the other hand, oxidative decarbonylation of 1 and 2 by reaction with tert-butylhydroperoxide (TBHP, 10 equiv.) gives a molybdenum oxide hybrid material formulated as [Mo3O9([ptapzpy]Br)2]·nH2O (3), which was characterised by FT-IR and Raman spectroscopy, thermogravimetric analysis, and 13C{1H} CP MAS NMR spectroscopy. Compounds 1-3 were effective (pre)catalysts for the epoxidation of cis-cyclooctene at 55 °C with aqueous H2O2 or TBHP (slightly better results were obtained with the former). The characterisation of the Mo-containing solids isolated after the catalytic reaction showed that poorly soluble ß-octamolybdate salts, (L) x [Mo8O26], were formed from 1-3 with TBHP and from 1 with H2O2, while soluble oxoperoxo species were formed from 3 with H2O2. These findings helped to explain the different catalytic performances obtained.

Nonlinear compensation with DBP aided by a memory polynomial.

Using digital backpropagation (DBP) based on the split step Fourier method (SSFM) aided by a memory polynomial (MP) model, we demonstrate an improved DBP approach for fiber nonlinearity compensation. The proposed technique (DBP-SSFM&MP) is numerically validated and its performance and complexity are compared against the benchmark DBP-SSFM, considering a single-channel 336 Gb/s PM-64QAM transmission system. We demonstrate that the proposed technique allows to maintain the performance achieved by DBP-SSFM, while decreasing the required number of iterations, by over 60%. For a transmission length of 1600 km we obtain a complexity reduction gain of 50.7% in terms of real multiplications in comparison with the standard DBP-SSFM.