Alanine aminotransferase electrochemical sensor based on graphene@MXene composite nanomaterials.

Graphene@MXene composite nanomaterials were utilized to construct an electrochemical sensor for alanine aminotransferase (ALT) detection. The combination of graphene nanosheets with MXene avoids the self-stacking of MXene and graphene, and broadens the charge transfer channel. In addition, the composite nanomaterial provides increased loading sites for pyruvate oxidase. The principle of ALT detection is a two-step enzymatic reaction. L-Alanine was initially transferred to pyruvate catalyzed by ALT. The formed pyruvate was then oxidized by pyruvate oxidase, generating H2O2. Through the detection of the generated H2O2, ALT activity was measured. The linear range of the sensor to ALT was from 5 to 400 U·L-1 with a detection limit of 0.16 U·L-1 (S/N = 3). For real sample analysis, the spiked recovery test results of ALT in serum samples were between 96.89 and 103.93% with RSD < 5%, confirming the reliability of the sensor testing results and potential clinical application of the sensor.

Electrochemical sensor using cobalt oxide-modified porous carbon for uric acid determination.

An electrochemical sensor for the detection of uric acid was constructed using cobalt oxide-modified porous carbon@multi-walled carbon nanotube (MWCNTs) composite material for the modification of the electrode. Firstly, ZIF-67 is generated on carbon nanotubes using the surfactant cetylammonium bromide (CTAB) as template. The vesicles generated by CTAB act as nucleation sites for the in situ growth of ZIF-67. Then, cobalt oxide-modified porous carbon was obtained after high-temperature carbonization of ZIF-67, leading to the formation of cobalt oxide-modified porous carbon@MWCNT composite materials. Co-N and Co-O active sites on the composite material can improve the oxidation of uric acid on the electrode surface, leading to enhanced sensitivity and selectivity for uric acid detection. The sensor has a good linear range from 1 to 40 µM for uric acid detection with a detection limit of 0.09 µM. The sensor was utilized for determination of uric acid in actual serum samples.

[Technical Difficulties in the Implementation of the Continuous Glucose Monitoring System].

The continuous glucose monitoring system (CGMS) has been clinically applied to monitor the dynamic change of the subcutaneous interstitial glucose concentration which is a function of the blood glucose level by glucose sensors. It can track blood glucose levels all day along, and thus provide comprehensive and reliable information about blood glucose dynamics. The clinical application of CGMS enables monitoring of blood glucose fluctuations and the discovery of hidden hyperglycemia and hypoglycemia that are difficult to be detected by traditional methods. As a CGMS needs to work subcutaneously for a long time, a series of factors such as biocompatibility, enzyme inactivation, oxygen deficiency, foreign body reaction, implant size, electrode flexibility, error correction, comfort, device toxicity, electrical safety, et al. should be considered beforehand. The study focused on the difficulties in the technology, and compared the products of Abbott, Medtronic and DexCom, then summarized their cutting-edge. Finally, this study expounded some key technologies in dynamic blood glucose monitoring and therefore can be utilized as a reference for the development of CGMS.

Applications of oxidized alginate in regenerative medicine.

Because of its ideal degradation rate and features, oxidized alginate (OA) is selected as an appropriate substitute and has been introduced into hydrogels, microspheres, 3D-printed/composite scaffolds, membranes, and electrospinning and coating materials. By taking advantage of OA, the OA-based materials can be easily functionalized and deliver drugs or growth factors to promote tissue regeneration. In 1928, it was first found that alginate could be oxidized using periodate, yielding OA. Since then, considerable progress has been made in the research on the modification and application of alginate after oxidation. In this article, we summarize the key properties and existing applications of OA and various OA-based materials and discuss their prospects in regenerative medicine.

Immobilization of BMP-2-derived peptides on 3D-printed porous scaffolds for enhanced osteogenesis.

Three-dimensional (3D) printing technologies open up new perspectives for customizing the external shape and internal architecture of bone scaffolds. In this study, an oligopeptide (SSVPT, Ser-Ser-Val-Pro-Thr) derived from bone morphogenetic protein 2 was conjugated with a dopamine coating on a 3D-printed poly(lactic acid) (PLA) scaffold to enhance osteogenesis. Cell experiments in vitro showed that the scaffold was highly osteoconductive to the adhesion and proliferation of rat marrow mesenchymal stem cells (MSCs). In addition, RT-PCR analysis showed that the scaffold was able to promote the expression of osteogenesis-related genes, such as alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) and osteopontin (OPN). Images of the micro-CT 3D reconstruction from the rat cranial bone defect model showed that bone regeneration patterns occurred from one side edge towards the center of the area implanted with the prepared biomimetic peptide hydrogels, demonstrating significantly accelerated bone regeneration. This work will provide a basis to explore the application potential of bioactive scaffolds further.

pH-Triggered Conformational Change of Antp-Based Drug Delivery Platform for Tumor Treatment with Combined Photothermal Therapy and Chemotherapy.

Poor cellular uptake and low therapeutic efficacy of small-molecule antitumor drugs limit the application of drug delivery systems (DDSs) in cancer therapy. A conformational change of the Antp mimetic peptide (AMP) in tumor microenvironments can greatly increase the cellular uptake as well as control drug release from a DDS. In this study, AMP-based nanoparticles (AMP-NPs) conjugated with tyroserleutide (YSL), an immunologically therapeutic tripeptide, are designed to encapsulate doxorubicin (Dox) and indocyanine green (ICG) to improve cellular uptake and cancer therapeutic efficacy by combining chemotherapy with photothermal therapy. In vitro studies verify that AMP-NPs can control the release of Dox and YSL at different pH values. Cell experiments show that AMP-NPs can promote the cellular uptake of Dox, and YSL can promote hepatocarcinoma cell (H22) apoptosis through downregulating Bcl-2 and cyclin D1 expression. In a mouse xenograft model using H22 cells, tumors are ablated when Dox- and ICG-loaded AMP-NPs are injected with the combination of hyperthermia effect induced by near-infrared (NIR) laser irradiation and chemotherapy from Dox and YSL. The pH-, photothermal-, and glutathione-responsive AMP-NPs with a conformational transition strategy can be utilized to synergistically enhance the cancer therapeutic efficacy with few side effects upon NIR laser irradiation.

A Biomimicking Polymeric Cryogel Scaffold for Repair of Critical-Sized Cranial Defect in a Rat Model.

Mineralized polymeric cryogels with interconnective macroporous structure have demonstrated their potential as promising scaffolding material in bone tissue engineering. However, their capability in inducing osteogenic differentiation of mesenchymal stem cells (MSCs) in vitro and osteogenesis in vivo has not been explored yet. In this work, the roles of the mineralized cryogel on osteogenesis are systematically studied. Mineralized macroporous poly(ethylene glycol)-co-2-hydroxyethyl methacrylate cryogel promotes osteogenic differentiation of rat MSCs, particularly in upregulating the activity of alkaline phosphatase (ALP, ∼5.7-folds) and expression of related osteogenic gene markers (ALP ∼16-folds, osteocalcin ∼133-folds) at 14 days. In vivo implantation reveals that mineralized cryogels could promote fast osteogenesis and angiogenesis in critical-sized cranial bone defect of a Sprague-Dawley rat model in 4 weeks. The adsorption, entrapment, and concentration of osteogenic growth factors (bone morphogenetic protein 2) and angiogenesis growth factor (vascular endothelial growth factor [VEGF]) in the matrices in vivo may possibly participate in the process of osteogenesis and angiogenesis. Notably, the adsorption of larger amount of VEGF in nonmineralized cryogels facilitates obvious angiogenesis and comparable osteogenesis in bone defect in 8 weeks. Graphical abstract [Figure: see text] Impact Statement The current work reported the fabrication and characterization of a biomimicking mineralized polymeric cryogel as scaffolding material in bone regeneration. In addition to its three dimensional porous structure and the osteogenic potential, this biomimicking scaffold was also found to enhance the adsorption of biochemical cues, which in turn greatly promoted the angiogenesis as well as the tissue regeneration.

Bioactive 3D scaffolds self-assembled from phosphorylated mimicking peptide amphiphiles to enhance osteogenesis.

Being an active scaffold in bone tissue engineering, hydrogel self-assembled from biomimetic peptide amphiphile (PA) has excellent ability to induce osteogenic differentiation and osteogenesis. Here, a multifunctional scaffold based on bone morphogenetic protein-2 (BMP-2) mimicking peptide, RGDS, and phosphoserine has been developed to enhance osteogenesis. Cell experiments in vitro displayed that the hydrogel could effectively promote rat messenchymal stem cells (rMSCs) proliferation and induce them differentiation into oesteblasts. The up-regulated RNA expression of osteogenic marker genes, like BMP-2, osteopontin (OPN), osteocalcin (OCN) and runt-related transcription factor 2 (Runx2) revealed that the scaffold could accelerate rMSCs differentiation at RNA level. Further studies on rat skull defect model demonstrated that the multifunctional scaffold exhibited excellent repair ability due to a potential synergistic effect of biomimetic peptide and phosphoserine. Histochemical/immunohistochemical staining results showed that expressions of alkaline phosphatase (ALP) and OCN was significantly up-regulated, indicating that the hydrogel could accelerate maturation of osteoblast precursors during the whole repairing process and be a promising bioactive scaffold for bone repairing.

Bioactive gel self-assembled from phosphorylate biomimetic peptide: A potential scaffold for enhanced osteogenesis.

Bone morphogenetic protein-2 biomimetic peptide (BMPBP) is a potent osteoinductive cytokine and plays a critical role during bone regeneration. Efforts to prepare hydrogels with surface modification or physical absorption of bioactive molecules do not provide sufficient bioactivity to meet the requirements of clinical application. The goal of this study was to form a three-dimensional hydrogel comprised of BMP-2 core sequence oligopeptide, phosphoserine, a synthetic cell adhesion peptide (RGDS), and polyaspartic acid to synergistically promote osteogenesis. Experiments performed in vitro revealed that the peptide gel was conducive to adhesion and proliferation of rat marrow mesenchymal stem cells (rMSCs). In addition, RT-PCR analysis indicated that rMSCs allowed better expression of osteogenesis-related genes such as BMP-2, runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). Use of the rat cranial bone defects model with micro-CT 3D reconstruction showed that bone regeneration patterns occurred from one side edge toward the center of the area implanted with the prepared biomimetic peptide hydrogels, demonstrating significantly accelerated bone regeneration. This work will provide a basis to explore the further application potential of this bioactive scaffold.

Biological Activity of an Injectable Biphasic Calcium Phosphate/PMMA Bone Cement for Induced Osteogensis in Rabbit Model.

Polymethylmethacrylate (PMMA) bone cement is widely used in repair of vertebral fracture because of its good biomechanical properties and fast curing. However, the bioinertness of PMMA cement may cause interfacial loosening, fatigue, fracture, and ultimate failure. In this study, biphasic calcium phosphate (BCP) is introduced into PMMA cement to prepare an injectable composite bone cement (BCPx /PMMA) and the content of BCP is optimized to achieve appropriate rate of absorption that matches the bone regeneration. The compressive strength of BCPx /PMMA bone cement is found to comply with the International Standardization Organization standard 5833, and can promote biomineralization as well as adhesion, proliferation, and osteogenic differentiation of Sprague-Dawley rat bone marrow mesenchymal stem cells in vitro. Furthermore, in vivo test performed on a rabbit radius defect model demonstrates that the presence of BCP can significantly improve the osteogenic efficacy of PMMA cement. Therefore, it is anticipated that BCPx /PMMA bone cement, as a promising injectable biomaterial, is of great potential in bone tissue regeneration.

pH triggered re-assembly of nanosphere to nanofiber: The role of peptide conformational change for enhanced cancer therapy.

pH-triggered conformational change and subsequent re-assembly of nanostructures provide a new strategy in nanomedicine for controlled drug release and enhanced therapy. Here, we reported the development of a novel pH-responsive nano-assembly as a drug carrier from peptide amphiphile (PA) consisting of mimicking peptide and stearic acid moieties. The mimicking peptide is a basic 17-amino acid peptide derived from antennapedia homeodomain, and undergoes a conformational transition of the secondary structure from ß-sheet at pH7.4 to α-helix at pH5.0. Such transition therefore leads to simultaneous evolution of the self-assembled structure of PA from nanosphere to nanofiber, promotes assemblies retention and then release drugs in the cytoplasm of tumor cell. In vitro studies showed that the doxorubicin (Dox)-loaded PA nanoparticle (PA@Dox) could be uptaken efficiently by the cell due to the membrane penetrating capability of the mimicking peptide and subsequently the released Dox further induce apoptosis of murine colon carcinoma CT26 (MCCC) cell. In a mouse xenograft model of MCCC, administration of PA@Dox via lateral tail vein injection could remarkably retard the tumor growth. The overall results suggested that the PA-based nanocarriers adopting the novel strategy of pH-triggered secondary structural change could enhance therapeutic efficacy and be used as a promising platform for potential development of new generation of drug carriers for cancer therapy.

Metal-Free Poly-Cycloaddition of Activated Azide and Alkynes toward Multifunctional Polytriazoles: Aggregation-Induced Emission, Explosive Detection, Fluorescent Patterning, and Light Refraction.

The metal-free click polymerization (MFCP) of activated alkynes and azides or activated azide and alkynes have been developed into powerful techniques for the construction of polytriazoles without the obsession of metallic catalyst residues problem. However, the MFCP of activated azides and alkynes is rarely applied in preparation of functional polytriazoles. In this paper, soluble multifunctional polytriazoles (PIa and PIb) with high weight-average molecular weights (Mw up to 32 000) are prepared via the developed metal-free poly-cycloaddition of activated azide and alkynes in high yields (up to 90%). The resultant PIa and PIb are thermally stable, and show aggregation-induced emission characteristics, enabling their aggregates to detect explosives with superamplification effect. Moreover, thanks to their containing aromatic rings and polar moieties, PIa and PIb exhibit high refractive indices. In addition, they can also be cross-linked upon UV irradiation to generate 2D fluorescent patterning due to their remaining azide groups and containing ester groups. Thus, these multifunctional polytriazoles are potentially applicable in the optoelectronic and sensing fields.

pH-responsive nanoparticle assembly from peptide amphiphiles for tumor targeting drug delivery.

In this paper, the peptide amphiphiles (PA) which consists of RGDSEEEEEEEEEEK as pH-sensitive segment and stearic acid as hydrophobic segment named RGDS-E10-Lys(C18) was successfully synthesized. TEM images showed that uniformly dispersed nanoparticles could be formed by PA molecules in pH 7.4 medium, however, disintegrated in pH 5.0 medium. Circular dichroism (CD) spectrum indicated that polypeptide adopted a random-coil conformation in neutral medium (pH 7.4). The CD signal was significantly attenuate for decreased solubility of PA in medium with pH 5.0. As expected, the prepared RGDS-E10-Lys(C18) assembly showed high pH-sensitive property which demonstrated a much more rapid drug release from micelles in tumor tissue (acidic environment) than in physiological environment (neutral environment). After DOX-loaded micelles incubated with tumor cells, the cytotoxicity of the micelles against Hela cells was increased obviously, indicating the great potential of micelles developed here as promising vehicle for targeted pH-responsive drug delivery.

Poly(γ-glutamic acid) modulates the properties of poly(ethylene glycol) hydrogel for biomedical applications.

In this paper, a series of copolymer hydrogels were fabricated from methacrylated poly(γ-glutamic acid) (mPGA) and poly(ethylene glycol) diacrylate (PEGDA). The effect of ionic strength and pH on the swelling behavior and mechanical properties of these hydrogels were studied in detail. Release of Rhodamine B as a model drug from the hydrogel was evaluated under varied pH. In vitro photoencapsulation of bovine cartilage chondrocytes was performed to assess the cytotoxicity of this copolymer hydrogel. The results revealed that the copolymer hydrogel is ionic- and pH-sensitive, and does not exhibit acute cytotoxicity; this copolymer hydrogel may have promising application as matrix for controlled drug release and scaffolding material in tissue engineering.

Effect of modification degree of nanohydroxyapatite on biocompatibility and mechanical property of injectable poly(methyl methacrylate)-based bone cement.

The objective of this study is to prepare a biocompatible nanohydroxyapatite/poly(methyl methacrylate) (HA/PMMA) composite bone cement, which has good mechanical property and can be used for vertebroplasty. Up to 40 wt % of nanohydroxyapatite (nano-HA) in the power, which was surface modified with poly(methylmethacrylate-co-γ-methacryloxypropyl timethoxysilane) [P(MMA-co-MPS)] copolymer, was incorporated into the composite bone cement. The content of P(MMA-co-MPS) on the surface of nano-HA (18.7%, 22.8%, and 26%) was determined through thermogravimetric analysis (TGA). The morphology of biomineralized surface of composite bone cement was observed under scanning electron microscope (SEM). The mechanical measurements of the composite cements implied that the interfacial interaction between the HA and PMMA matrix may be greatly enhanced after surface modification of HA. Biochemical assays indicated that the HA/PMMA bone cement had no cytotoxicity and induced no hemolysis. The cell adhesion and alkaline phosphatase (ALP) activity assays indicated that the biocompatibility of HA/PMMA bone cement could be promoted, demonstrating that it can be used as an ideal weight-bearing bone repair materials on clinical application.

Using the isotope effect to probe an aggregation induced emission mechanism: theoretical prediction and experimental validation.

Aggregation-induced emission (AIE) has become a hot topic for a variety of potential applications, but the understanding of its working mechanism is still under scrutiny. Herein, we proposed the use of the isotope effect (IE) to identify the AIE mechanism: under the restriction of an internal motion mechanism, the IE is pronouncedly different in excited-state decay rates when contrasting AIE luminogens (AIEgens) and non-AIEgens in theoretical calculations. For the complete deuteration of AIEgens, the IE of nonradiative decay rate in solution (<-10%) is much weaker than that (-65% to -95%) in aggregate, because the former stems from the overall results of competitive vibronic coupling and the severe mixing of low-frequency modes while the latter mainly comes from the vibronic coupling only. The experimental results confirm the isotopic "jump" behaviors in AIEgens well. However, non-AIEgens exhibit equivalent IEs (-40% to -90%) in both solution and solid phases. Further partial deuteration schemes for the 6-ring AIE analogues show positional dependence.

Alendronate-decorated biodegradable polymeric micelles for potential bone-targeted delivery of vancomycin.

Osteomyelitis is a bone infection disease which is caused by bacteria or other germs, and could cause serious impact on the health and working capacity of the patients. Alendronate (ALN) can chelate strongly with the calcium ion of hydroxyapatite (HA) which is commonly used to treat osteoporosis. Nanomedicine has attracted a lot of attention in that the nano-sized carrier can deliver drug molecules to specific site of interest with the aid of targeting moiety and achieve sustained release, resulting in improved therapeutic effect and reduced side effect. In this study, micelles self-assembled from poly(lactic acid-co-glycolic acid)-block-poly(ethylene glycol)-alendronate (PLGA-PEG-ALN) copolymer were prepared for bone-targeted delivery of vancomycin (Van). The chemical structure of PLGA-PEG-ALN was confirmed by proton nuclear magnetic resonance ((1)H-NMR) spectroscopy. The formation of the nanoparticles was characterized by dynamic light scattering, transmission electronic microscopy as well as the critical micelle concentration measurement. Release profiles from the micelles revealed that the conjugation of ALN to the surface of micelle did not pose adverse effect on the drug-loading capacity and release behaviors. The cytotoxicity of Van-loaded PLGA-PEG-ALN micelles as well as the blank micelles was evaluated via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay toward rat bone marrow stromal cells (rBMSCs) and human embryonic hepatocytes (L02 cells), and results showed that this Van-loaded micelle possesses appropriate cytotoxicity and is safe in the potential treatment of osteomyelitis. The in vitro affinity of PLGA-PEG-ALN micelles to the HA was also confirmed in vitro. The antibacterial effect of Van-loaded PLGA-PEG-ALN micelles was tested against Staphylococcus aureus (SA) which is the main pathogenic bacteria in osteomyelitis, and the results showed that the Van-loaded micelles can effectively inhibit the growth of SA. These results demonstrated that the PLGA-PEG-ALN micelles may be potentially used for the bone targeted delivery of Van.

Synergistic effect of HA and BMP-2 mimicking peptide on the bioactivity of HA/PMMA bone cement.

HA and bone morphogenetic protein-2 (BMP-2) possess superior osteoinductive and osteoconductive activities in the repair of bone defects. In this study, a BMP-2 mimicking oligopeptide (serine-serine-valine-proline-threonine, SSVPT) was introduced to the surface of nano-sized HA as the reinforcement phase of a poly(methyl methacrylate) (PMMA) cement, and the synergistic action of HA and oligopeptide on the bioactivity of HA/PMMA bone cement was investigated. Incorporated with 3-trimethoxysilyl propyl methacrylate (MPS)-modified HA, the compressive strength, flexural strength, and flexural modulus of HA/PMMA complied with the international standardization organization 5833 standard. Adherence measurement demonstrated that the introduction of HA and SSVPT could promote the adhesion and proliferation of human fetal osteoblast (hFOB) on the surface of HA/PMMA cement. The increased alkaline phosphatase (ALP) activity indicated that HA/PMMA could promote osteogenic differentiation of hFOB. All of the results illuminated that the HA/PMMA cement could be used as a bioactive material for regeneration and reconstruction of load-bearing bone.

Modulation of aggregation-induced emission and electroluminescence of silole derivatives by a covalent bonding pattern.

The deciphering of structure-property relationships is of high importance to rational design of functional molecules and to explore their potential applications. In this work, a series of silole derivatives substituted with benzo[b]thiophene (BT) at the 2,5-positions of the silole ring are synthesized and characterized. The experimental investigation reveals that the covalent bonding through the 2-position of BT (2-BT) with silole ring allows a better conjugation of the backbone than that achieved though the 5-position of BT (5-BT), and results in totally different emission behaviors. The silole derivatives with 5-BT groups are weakly fluorescent in solutions, but are induced to emit intensely in aggregates, presenting excellent aggregation-induced emission (AIE) characteristics. Those with 2-BT groups can fluoresce more strongly in solutions, but no obvious emission enhancements are found in aggregates, suggesting they are not AIE-active. Theoretical calculations disclose that the good conjugation lowers the rotational motions of BT groups, which enables the molecules to emit more efficiently in solutions. But the well-conjugated planar backbone is prone to form strong intermoelcular interactions in aggregates, which decreases the emission efficiency. Non-doped organic light-emitting diodes (OLEDs) are fabricated by using these siloles as emitters. AIE-active silole derivatives show much better elecroluminescence properties than those without the AIE characterisic, demonstrating the advantage of AIE-active emitters in OLED applications.