A cell-free system toward deciphering the post-translational modification barcodes of Oct4 in different cellular contexts

AbstractThe octamer-binding transcription factor 4 (Oct4) is essential for maintaining the self-renewal and pluripotency of embryonic stem cells (ESCs). Post-translational modifications (PTMs) of Oct4 critically control its structure, function and intracellular localization. However, determination of Oct4 PTM profiles has largely been restricted by the quantity and purity of the Oct4 protein samples required for mass spectrometric analyses. In this study, by incubating the Escherichia coli-derived His-tagged Oct4 proteins with the whole cell lysates of a variety of human cells followed by retrieving the reacted Oct4 proteins with the Ni–NTA beads, we developed a labor- and cost-effective in vitro PTM method that allowed for mass spectrometric determination of the phosphorylation profiles of Oct4 proteins exposed to various cell-free systems. A number of Oct4 phosphorylation sites that were commonly present in all the cell-free systems or specifically present in a particular cellular context were identified, indicating that Oct4 is controlled by both common and distinct PTM regulatory pathways. Our work provided a proof-of-concept that such a cell-free system-based in vitro PTM approach can be applied to systematically map out the physiologically-relevant PTM sites in Oct4 proteins, which opened up an avenue to fully decipher the Oct4 PTM barcodes in various cellular contexts

Graphene reinforced polyether ether ketone nanocomposites for bone repair applications

To improve the performance of polyether ether ketone matrix (PEEK) in hard tissue repair and replacement applications, we fabricated graphene nanoplatelet (G) reinforced PEEK with different filler weight concentrations (0.1%–5%) through injection moulding. The mechanical properties, surface morphology, chemical composition and thermal stability of the composites have been characterized. The biocompatibility has been assessed in vitro and the bone repair function of the composite implants have been assessed in vivo using a rabbit mandibular bone defect model. Mechanical testing results suggest that the composite samples have compressive moduli similar to that of the natural bone. Although addition of G into PEEK does not significantly influence the composite tensile, flexural or compressive moduli, it can significantly enhance the ductility and toughness of the material. On the other hand, all G-reinforced PEEK implants demonstrated enhanced adhesion and differentiation of rat bone marrow stromal cells (BMSCs), with 5% G-PEEK showing the highest bioactivity among all samples. The in vivo osseointegration data further revealed that 5% G-PEEK has the best effect in promoting osseointegration and bone regeneration, in both early stage and late stage bone re-growth. Study shows that our G-reinforced PEEK-based implants provides a promising strategy for enhancing the performance of future regenerative bone implants

Graphene Reinforced Polyether Ether Ketone Nanocomposites for Bone Repair Applications

To improve the performance of polyether ether ketone matrix (PEEK) in hard tissue repair and replacement applications, we fabricated graphene (G) reinforced PEEK with graded G concentrations (0.1%-5%) through injection molding. The mechanical properties, surface morphology, chemical composition and thermal stability of the composites have been characterized through universal mechanical testing, scanning electron microscopy, contact-angle measurement, transmission electron microscope, X-ray photoelectron spectroscopy, X-ray diffraction and thermal analysis system. The biocompatibility has been assessed in vitro and the bone repair function of the composite implant have been assessed in vivo using a rabbit mandibular bone defect model. Mechanical testing results suggest that the composite samples have compressive moduli similar to that of the natural bone. Although addition of G into PEEK does not significantly influence the composite tensile, flexural or compressive moduli, it can significantly enhance the ductility and toughness of the material. On the other hand, all G-reinforced PEEK implants demonstrated enhanced adhesion and differentiation of rat bone marrow stromal cells (BMSCs), with 5% G-PEEK showing the highest bioactivity among all samples. The in vivo osseointegration data further revealed that 5% G-PEEK has the best effect in promoting osseointegration and bone regeneration, in both early stage and late stage bone re-growth. Study shows that our G-reinforced PEEK-based implants provides a promising strategy for enhancing the performance of future regenerative bone implants.<br /

A Fast UV-Curable PU-PAAm Hydrogel with Mechanical Flexibility and Self-Adhesion for Wound Healing

Hydrogels demonstrate superior properties that favor wound healing and have been widely used in clinical settings for wound dressing applications. However, commercial hydrogel dressings often lack flexibility/adhesiveness, and do not conform well to the irregular skin surfaces of complex wounds and/or wounds near joints. As a result, the wound is likely to be exposed to potential bacterial invasion. Herein, we designed and developed a mechanically flexible and self-adhesive polyurethane-poly(acrylamide) (PU-PAAm) hydrogel for wound healing applications. The hydrogel can be cured from a novel waterborne emulsion within 90 s under UV irradiation. The PU component within the PU-PAAm hydrogel plays a “bridging” role that accelerates the formation of an interpenetrating polymer network (IPN), which consists of a physically crosslinked PU network trapped within a chemically crosslinked PAAm network. The unique IPN structure endowes the hydrogel with superior stretchability and ductility. The hydrogen bonding formation and electrostatic interaction between the hydrogel and skin ensure strong adhesion without causing irritation to skin upon dressing removal. Animal studies further confirmed the PU-PAAm hydrogel's remarkable skin regeneration capability. This work shows our new hydrogel holds a promising prospect for treatment of complicated or challenging wounds such as burns and chronic wounds

Atmospheric Pressure Microplasma for Antibacterial Silver Nanoparticle/Chitosan Nanocomposites With Tailored Properties

Room temperature atmospheric pressure microplasma (APM) was deployed for the first time for the in situ synthesis of anti-bacterial silver nanoparticle / chitosan (AgNP/CS) nanocomposites. The plasma induced liquid chemistry plays a role in the in situ formation of AgNP, the size distribution of which depends on the silver salt precursor concentration. The microplasma process has also simultaneously tailored the physical properties of the composites, rendering more crosslinked chitosan polymer network with shorter molecular chains. The formation of AgNP within the in situ modified chitosan has led to nanocomposites with overall improved mechanical properties and better stability in simulated body fluid. Our plasma synthesized AgNP/CS nanocomposites also demonstrate effective antibacterial properties against E. Coli and S. Aureus bacterial strains, showing their promise in potential antimicrobial applications.</p

Atmospheric pressure microplasma for antibacterial silver nanoparticle/chitosan nanocomposites with tailored properties

Room temperature atmospheric pressure microplasma (APM) was deployed for the first time for the in situ synthesis of antibacterial silver nanoparticle/chitosan (AgNP/CS) nanocomposites. The plasma induced liquid chemistry plays a role in the in situ formation of AgNP, the size distribution of which depends on the silver salt precursor concentration. The microplasma process has also simultaneously tailored the physical properties of the composites, through molecular chain scission and formation of physically crosslinked polymer network. The formation of AgNP within the in situ modified chitosan has led to nanocomposites with overall improved mechanical properties and better stability in simulated body fluid. Our plasma synthesized AgNP/CS nanocomposites also demonstrate effective antibacterial properties against E. coli and S. aureus bacterial strains, showing their promise in potential antimicrobial applications