Enhanced osteogenic differentiation of human mesenchymal stem cells by direct delivery of Cbfß protein.

Core binding factor ß (Cbfß) is a non-DNA binding cofactor of Runx2 that potentiates DNA binding. Previously, it has been reported that Cbfß plays an essential role in osteogenic differentiation and skeletal development by inhibition adipogenesis. Here, we delivered the recombinant Cbfß protein into human mesenchymal stem cells (MSCs) and triggered osteogenic lineage commitment. The efficient delivery of Cbfß was achieved by fusing 30Kc19 protein, which is a cell-penetrating protein derived from the silkworm. After the production of the recombinant Cbfß-30Kc19 protein in the Escherichia coli expression system, and confirmation of its intracellular delivery, MSCs were treated with the Cbfß-30Kc19 once or twice up to 300 µg/ml. By investigating the upregulation of osteoblast-specific genes and phenotypical changes, such as calcium mineralization, we demonstrated that Cbfß-30Kc19 efficiently induced osteogenic differentiation in MSCs. At the same time, Cbfß-30Kc19 suppressed adipocyte formation and downregulated the expression of adipocyte-specific genes. Our results demonstrate that the intracellularly delivered Cbfß-30Kc19 enhances osteogenesis in MSCs, whereas it suppresses adipogenesis by altering the transcriptional regulatory network involved in osteoblast-adipocyte lineage commitment. Cbfß-30Kc19 holds great potential for the treatment of bone-related diseases, such as osteoporosis, by allowing transcriptional regulation in MSCs, and overcoming the limitations of current therapies.

Derivation of Cell-Engineered Nanovesicles from Human Induced Pluripotent Stem Cells and Their Protective Effect on the Senescence of Dermal Fibroblasts.

Stem cells secrete numerous paracrine factors, such as cytokines, growth factors, and extracellular vesicles. As a kind of extracellular vesicle (EV), exosomes produced in the endosomal compartment of eukaryotic cells have recently emerged as a biomedical material for regenerative medicine, because they contain many valuable contents that are derived from the host cells, and can stably deliver those contents to other recipient cells. Although we have previously demonstrated the beneficial effects of human induced potent stem cell-derived exosomes (iPSC-Exo) on the aging of skin fibroblasts, low production yield has remained an obstacle for clinical applications. In this study, we generated cell-engineered nanovesicles (CENVs) by serial extrusion of human iPSCs through membrane filters with diminishing pore sizes, and explored whether the iPSC-CENV ameliorates physiological alterations of human dermal fibroblasts (HDFs) that occur by natural senescence. The iPSC-CENV exhibited similar characteristics to the iPSC-Exo, while the production yield was drastically increased compared to that of iPSC-derived EVs, including exosomes. The proliferation and migration of both young and senescent HDFs were stimulated by the treatment with iPSC-CENVs. In addition, it was revealed that the iPSC-CNEV restored senescence-related alterations of gene expression. Treatment with iPSC-CENVs significantly reduced the activity of senescence-associated-ß-galactosidase (SA-ß-Gal) in senescent HDFs, as well as suppressing the elevated expression of p53 and p21, key factors involved in cell cycle arrest, apoptosis, and cellular senescence signaling pathways. Taken together, these results suggest that iPSC-CENV could provide an excellent alternative to iPSC-exo, and be exploited as a resource for the treatment of signs of skin aging.

Institutional decoupling and the limited implementation of certified environmental technologies.

New environmental policies and initiatives increasingly drive firms to develop sustainable technologies, yet it is unclear why firms often decide not to capture value from the technologies that they create. We argue that the under-implementation of environmental technologies can be explained by the concept of institutional decoupling, which suggests that organizations may engage in symbolic actions without necessarily taking the risk of implementing a new technology in a less established market when there is no decisive evidence of its commercial viability. Focusing on the Korean Green Certification Program, a government-initiated review system for environmental technologies, we find that the timing of certification is a key source of variation in the implementation of certified technologies. Our findings also show that the main effect is conditioned by performance feedback and niche density-the factors that shape a firm's risk perceptions. Implications for organization studies, sustainable innovation, and environmental policies are discussed.

Nano-structure of vitronectin/heparin on cell membrane for stimulating single cell in iPSC-derived embryoid body.

Individual cell environment stimulating single cell is a suitable strategy for the generation of sophisticated multicellular aggregates with localized biochemical signaling. However, such strategy for induced pluripotent stem cell (iPSC)-derived embryoid bodies (EBs) is limited because the presence of external stimulation can inhibit spontaneous cellular communication, resulting in misdirection in the maturation and differentiation of EBs. In this study, a facile method of engineering the iPSC membrane to stimulate the inner cell of EBs while maintaining cellular activities is reported. We coated the iPSC surface with nanoscale extracellular matrix fabricated by self-assembly between vitronectin and heparin. This nano-coating allowed iPSC to retain its in vitro properties including adhesion capability, proliferation, and pluripotency during its aggregation. More importantly, the nano-coating did not induce lineage-specific differentiation but increased E-cadherin expression, resulting in promotion of development of EB. This study provides a foundation for future production of sophisticated patient-specific multicellular aggregates by modification of living cell membranes.

Effects of the Anode Diffusion Layer on the Performance of a Nonenzymatic Electrochemical Glucose Fuel Cell with a Proton Exchange Membrane.

It is necessary to apply a nonenzymatic glucose fuel cell using a proton exchange membrane for an implantable biomedical device that operates at low power. The permeability of glucose with high viscosity and a large molecular weight in the porous medium of the diffusion layer was investigated for use in fuel cells. Carbon paper was prepared as an anode diffusion layer, and it was analyzed with a diffusion layer treated with polytetrafluoroethylene (PTFE) and a microporous layer (MPL). When untreated carbon paper was applied, the peak power density (PPD) and open-circuit voltage (OCV) increased as the glucose concentration and flow rate increased. On this occasion, the highest PPD of 17.81 µW cm-2 was achieved at 3 mM and a 2.0 mL min-1 glucose aqueous solution (at atmospheric pressure and 36.5 °C). The diffusion layer, which became more hydrophobic through PTFE treatment, adversely affected glucose permeability. In addition, the addition of an MPL decreased OCV and PPD with increasing glucose concentrations and flow rates. Compared with untreated carbon paper, the PPD was six times lower approximately. Consequently, it was confirmed that the properties of carbon paper, such as low hydrophobicity, high porosity, and thin thickness, would be advantageous for nonenzymatic glucose fuel cells.

Stem Cell-Derived Exosomes and Nanovesicles: Promotion of Cell Proliferation, Migration, and Anti-Senescence for Treatment of Wound Damage and Skin Ageing.

Extracellular vesicles (EVs), such as exosomes, are nano-sized vesicles derived from endocytic membranes and contain biomolecules such as proteins, lipids, RNAs, and DNAs for the transfer of signals to recipient cells, playing significant roles in cell-to-cell communication. Discovery of exosomes has attracted attention for possible use as next generation therapies in clinical applications; however, several studies suggest that cells secrete exosomes that perform as mediators in the tumor niche and play several roles in tumorigenesis, angiogenesis, and metastasis. Recently, stem cell-derived exosomes have been suggested as a desirable source for regenerative medicine due to their roles in the promotion of angiogenesis via migratory and proliferative mechanisms. This review is aimed at demonstrating the present knowledge of stem cell-derived exosomes and cell-engineered nanovesicles (CNVs) as proliferative, migratory, and anti-senescent therapeutic biomaterial for use in tissue regeneration; wound healing and anti-ageing are explained. We conclude this review by discussing the future perspectives of stem cell-derived exosomes and CNVs as a platform in therapeutic strategies for treatment of wound damage and skin aging.