Three-Dimensional Bioprinted Controlled Release Scaffold Containing Mesenchymal Stem/Stromal Lyosecretome for Bone Regeneration: Sterile Manufacturing and In Vitro Biological Efficacy

Recently, 3D-printed scaffolds for the controlled release of mesenchymal stem cell (MSC) freeze-dried secretome (Lyosecretome) have been proposed to enhance scaffold osteoinduction and osteoconduction; coprinting of poly(ε-caprolactone) (PCL) with alginate hydrogels allows adequate mechanical strength to be combined with the modulable kinetics of the active principle release. This study represents the feasibility study for the sterile production of coprinted scaffolds and the proof of concept for their in vitro biological efficacy. Sterile scaffolds were obtained, and Lyosecretome enhanced their colonization by MSCs, sustaining differentiation towards the bone line in an osteogenic medium. Indeed, after 14 days, the amount of mineralized matrix detected by alizarin red was significantly higher for the Lyosecretome scaffolds. The amount of osteocalcin, a specific bone matrix protein, was significantly higher at all the times considered (14 and 28 days) for the Lyosecretome scaffolds. Confocal microscopy further confirmed such results, demonstrating improved osteogenesis with the Lyosecretome scaffolds after 14 and 28 days. Overall, these results prove the role of MSC secretome, coprinted in PCL/alginate scaffolds, in inducing bone regeneration; sterile scaffolds containing MSC secretome are now available for in vivo pre-clinical tests of bone regeneration

A self-organized synthetic morphogenic liposome responds with shape changes to local light cues

Reconstituting artificial proto-cells capable of transducing extracellular signals into cytoskeletal changes can reveal fundamental principles of how non-equilibrium phenomena in cellular signal transduction affect morphogenesis. Here, we generated a Synthetic Morphogenic Membrane System (SynMMS) by encapsulating a dynamic microtubule (MT) aster and a light-inducible signaling system driven by GTP/ATP chemical potential into cell-sized liposomes. Responding to light cues in analogy to morphogens, this biomimetic design embodies basic principles of localized Rho-GTPase signal transduction that generate an intracellular MT-regulator signaling gradient. Light-induced signaling promotes membrane-deforming growth of MT-filaments by dynamically elevating the membrane-proximal tubulin concentration. The resulting membrane deformations enable recursive coupling of the MT-aster with the signaling system, which generates global self-organized morphologies that reorganize towards local external cues in dependence on prior shape. SynMMS thereby signifies a step towards bio-inspired engineering of self-organized cellular morphogenesis

Farmers-to-consumers: An example of sustainable soilless horticulture in Urban and Peri-Urban areas

As urban populations rise, new strategies to address nutritional requirements are needed. People moving from rural areas often end up in the suburbs, facing severe poverty and unbalanced nutrition. Improving urban agricultural production is constrained by limited access to fertile land, but the introduction of low cost hydroponic gardens may be a sustainable way to launch horticultural production on roofs, terraces and courtyards. Gardens also perform a social function, restoring the associative mentality lost when emigrants leave their communities. In this study, we propose the adoption of community hydroponic gardens as a tool to overtake poverty, improve nutrition and increase associative attitude. A democratic organization of women farming groups, together with the role of a technical assistance centre is described. Agricultural production is managed by the groups, where both intercropping patterns for home consumption and commercial leafy vegetable production are conducted. Fresh produce for the market is washed and packaged and sold to supermarkets and restaurants in the city. The technical assistance centre provides tutoring, agricultural inputs and manages commercial relations. Its sustainability is assured by a percent of the vegetable marketing income. It also offers training to students of local agricultural schools. A socioeconomic overview of the advantages created by the installation of community gardens is considered

Experimental characterization and computational modeling of hydrogel cross-linking for bioprinting applications

Alginate-based hydrogels are extensively used to create bioinks for bioprinting, due to their biocompatibility, low toxicity, low costs, and slight gelling. Modeling of bioprinting process can boost experimental design reducing trial-and-error tests. To this aim, the cross-linking kinetics for the chemical gelation of sodium alginate hydrogels via calcium chloride diffusion is analyzed. Experimental measurements on the absorbed volume of calcium chloride in the hydrogel are obtained at different times. Moreover, a reaction-diffusion model is developed, accounting for the dependence of diffusive properties on the gelation degree. The coupled chemical system is solved using finite element discretizations which include the inhomogeneous evolution of hydrogel state in time and space. Experimental results are fitted within the proposed modeling framework, which is thereby calibrated and validated. Moreover, the importance of accounting for cross-linking-dependent diffusive properties is highlighted, showing that, if a constant diffusivity property is employed, the model does not properly capture the experimental evidence. Since the analyzed mechanisms highly affect the evolution of the front of the solidified gel in the final bioprinted structure, the present study is a step towards the development of reliable computational tools for the in silico optimization of protocols and post-printing treatments for bioprinting applications