Weiqun Tian

Construction of nerve guide conduits from cellulose/soy protein composite membranes combined with Schwann cells and pyrroloquinoline quinone for the repair of peripheral nerve defect.

Regeneration and functional reconstruction of peripheral nerve defects remained a significant clinical challenge. Nerve guide conduits, with seed cells or neurotrophic factors (NTFs), had been widely used to improve the repair and regeneration of injured peripheral nerve. Pyrroloquinoline quinone (PQQ) was an antioxidant that can stimulate nerve growth factors (NGFs) synthesis and accelerate the Schwann cells (SCs) proliferation and growth. In present study, three kinds of nerve guide conduits were constructed: one from cellulose/SPI hollow tube (CSC), another from CSC combined with SCs (CSSC), and the third one from CSSC combined with PQQ (CSSPC), respectively. And then they were applied to bridge and repair the sciatic nerve defect in rats, using autograft as control. Effects of different nerve guide conduits on the nerve regeneration were comparatively evaluated by general analysis, sciatic function index (SFI) and histological analysis (HE and TEM). Newly-formed regenerative nerve fibers were observed and running through the transparent nerve guide conduits 12 weeks after surgery. SFI results indicated that the reconstruction of motor function in CSSPC group was better than that in CSSC and CSC groups. HE images from the cross-sections and longitudinal-sections of the harvested regenerative nerve indicated that regenerative nerve fibers had been formed and accompanied with new blood vessels and matrix materials in the conduits. TEM images also showed that lots of fresh myelinated and non-myelinated nerve fibers had been formed. Parts of vacuolar, swollen and abnormal axons occurred in CSC and CSSC groups, while the vacuolization and swell of axons was the least serious in CSSPC group. These results indicated that CSSPC group had the most ability to repair and reconstruct the nerve structure and functions due to the comprehensive contributions from hollow CSC tube, SCs and PQQ. As a result, the CSSPC may have the potential for the applications as nerve guide conduits in the field of nerve tissue engineering.

Improvement in physical and biological properties of chitosan/soy protein films by surface grafted heparin

A series of chitosan/soy protein isolate (SPI) composite films (CS-n, n=0, 10 and 30, corresponding to SPI content in the composites) were prepared. Heparin was grafted onto the surface of CS-n to fabricate a series of heparinized films (HCS-n). CS-n and HCS-n were characterized by ATR-Fourier transform infrared spectroscopy and water contact angle. The surface heparin density was measured by toluidine blue assay. The results showed that heparin has been successfully grafted onto the surface of CS-n. Heparin evenly distributed on the surface of the films and the heparin content increased with the increase of SPI content, and the hydrophilicity of the films was enhanced due to the grafted heparin. The cytocompatibility and hemocompatibility of CS-n and HCS-n were evaluated by cell culture (MTT assay, live/dead assay, cell morphology and cell density observation), platelet adhesion test, plasma recalcification time (PRT) measurement, hemolysis assay and thrombus formation test. HCS-n showed higher cell adhesion rate and improved cytocompatibility compared to the corresponding CS-n. HCS-n also exhibited lower platelet adhesion, longer PRT, higher blood anticoagulant indexes (BCI) and lower hemolysis rate than the corresponding CS-n. The improved cytocompatibility and hemocompatibility of HCS-n would shed light on the potential applications of chitosan/soy protein-based biomaterials that may come into contact with blood.

In vitro hemocompatibility and cytotoxicity evaluation of halloysite nanotubes for biomedical application

Halloysite nanotubes (HNTs), due to their unique structures and properties, may play an important role in biomedical applications. In vitro test is usually conducted as a preliminary screening evaluation of the hemocompatibility and cytotoxicity of HNTs for its short term consuming, convenience, and less expense. In this work, HNTs were processed with anticoagulated rabbit blood to detect its blood compatibility. The result of hemolysis test shows that the hemolysis ratios are below 0.5%, indicating nonhemolysis of HNTs. Plasma recalcification time suggests that HNTs are dose-dependently contributing to blood coagulation in platelet poor plasma (PPP). The effect of platelet activation caused by HNTs was also examined by scanning electron microscopy (SEM). Meanwhile, HNTs were labeled with fluorescein isothiocyanate (FITC) to observe its intracellular distribution in A549 cells under confocal microscopy. CCK-8 test and TUNEL test of HNTs at different concentration levels were performed in vitro, respectively. Therefore, the potential usage of HNTs in medicine may be very meaningful in oral dosing, dermal application, dental uses, or medical implants.

Long-term antibacterial protected cotton fabric coating by controlled release of chlorhexidine gluconate from halloysite nanotubes

In this work, an environmentally friendly antibacterial composite was prepared by loading natural halloysite nanotubes (HNTs) with chlorhexidine gluconate (CG). Fourier transform-infrared spectroscopy and X-ray diffraction demonstrated that CG could be successfully loaded on HNTs without changing the crystalline structure of HNTs. In vitro drug release assay showed that CG could be released from HNTs in a sustainable manner and could last as long as about ten days. Inhibition zone test suggested that the new composite displayed significant bacterial killing activity. In addition, the composite was deposited on the surface of cotton fabric by dip-coat technique. Our results showed that the HNTs/CG composite could be distributed on the cotton fabric and possessed better thermal stability and hydrophilicity properties than raw cotton fabric. HNTs/CG-coated cotton fabric exhibited over 98% bacterial reduction of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, and still showed over 90% antibacterial activity even after being washed 20 times. MTT test and animal skin irritation tests indicate that HNTs/CG-coated cotton was nontoxic to mouse fibroblast cells (L929) and gave no irritation to the rabbits skin. Thus, HNTs/CG composite is a safe and promising antibacterial agent and has great potential application in the textile field.

Cellulose/soy protein composite-based nerve guidance conduits with designed microstructure for peripheral nerve regeneration

OBJECTIVE The objective of this work was to develop nerve guidance conduits from natural polymers, cellulose and soy protein isolate (SPI), by evaluating the effects of cellulose/SPI film-based conduit (CSFC) and cellulose/SPI sponge-based conduit (CSSC) on regeneration of nerve defects in rats. APPROACH CSFC and CSSC with the same chemical components were fabricated from cellulose and SPI. Effects of CSSC and CSFC on regeneration of the defective nerve were comparatively investigated in rats with a 10 mm long gap in sciatic nerve. The outcomes of peripheral nerve repair were evaluated by a combination of electrophysiological assessment, Fluoro-Gold retrograde tracing, double NF200/S100 immunofluorescence analysis, toluidine blue staining, and electron microscopy. The probable molecular mechanism was investigated using quantitative real-time PCR (qPCR) analysis. MAIN RESULTS Compared with CSFC, CSSC had 2.69 times higher porosity and 5.07 times higher water absorption, thus ensuring much higher permeability. The nerve defects were successfully bridged and repaired by CSSC and CSFC. Three months after surgery, the CSSC group had a higher compound muscle action potential amplitude ratio, a higher percentage of positive NF200 and S100 staining, and a higher axon diameter and myelin sheath thickness than the CSFC group, showing the repair efficiency of CSSC was higher than that of CSFC. qPCR analysis indicated the mRNA levels of nerve growth factor, IL-10, IL-6, and growth-associated protein 43 (GAP-43) were higher in the CSSC group. This also indicated that there was better nerve repair with CSSC due to the higher porosity and permeability of CSSC providing a more favourable microenvironment for nerve regeneration than CSFC. SIGNIFICANCE A promising nerve guidance conduit was developed from cellulose/SPI sponge that showed potential for application in the repair of nerve defect. This work also suggests that nerve guidance conduits with better repair efficiency could be developed through structure design and processing optimization.

Intracellular pathway of halloysite nanotubes: potential application for antitumor drug delivery

Natural halloysite nanotubes (HNTs), with nanotubular structure, are attracting considerable attention in recent years. The hollow tubular structure allows HNTs to play an important role in drug delivery system as drug carriers. However, the wide applications of HNTs in biomedicine have been hampered by the lack of sufficient intracellular researches so far. In this study, we systemically investigated the transport mechanisms of HNTs in A549 living cells. The colocalization and inhibition experiments illustrated FITC-labeled HNTs were readily internalized into cells by both clathrin- and caveolae-dependent endocytosis, and the transport pathway of HNTs is an actin- and microtubule-associated process via Golgi apparatus and lysosome. Meanwhile, the cell cycle assay clarified that HNTs can prompt the intracellular transportation of gemcitabine and enhance the gemcitabine concentration in A549 tumor cells. Such elucidation of intracellular transport pathway of HNTs offers insights into the site-specific delivery and cellular internalization of HNTs, which provide a reasonable guidance for the design of novel drug delivery system.

Strategies on Nanodiagnostics and Nanotherapies of the Three Common Cancers

The emergence of nanomedicine has enriched the knowledge and strategies of treating diseases, and especially some incurable diseases, such as cancers, acquired immune deficiency syndrome (AIDS), and neurodegenerative diseases. The application of nanoparticles in medicine is in the core of nanomedicine. Nanoparticles can be used in drug delivery for improving the uptake of poorly soluble drugs, targeted delivery to a specific site, and drug bioavailability. Early diagnosis and targeted therapies for cancers can significantly improve patients’ quality of life and extend patients’ lives. The advantages of nanoparticles have given them a progressively important role in the nanodiagnosis and nanotherapy of common cancers. To provide a reference for the further application of nanoparticles, this review focuses on the recent development and application of nanoparticles in the early diagnosis and treatment of the three common cancers (lung cancer, liver cancer, and breast cancer) by using quantum dots, magnetic nanoparticles, and gold nanoparticles.

Electrodeposition to construct mechanically robust chitosan-based multi-channel conduits

A series of electrodeposited chitosan-based multi-channel conduits (ECMC) with potential for peripheral nerve tissue engineering were constructed using a novel electrodeposition method combined with homemade molds. The structural and mechanical properties of the ECMC were characterized by scanning electron microscopy, Fourier-transformed infrared spectroscopy, X-ray diffraction patterns and mechanical testing. The results showed that the electrodeposition process did not change the chemical structure of the chitosan molecules, but endowed the ECMC with high levels of flexibility and elasticity. Hemocompatibility and cytocompatibility of the ECMC were evaluated by hemolysis assay, MTT assay and live/dead assay. The results indicated that the ECMC had a low hemolysis rate, and can promote cell proliferation and support cell adhesion. This work provides a safe and feasible electrodeposition method to construct chitosan-based conduits with potential applications for peripheral nerve tissue engineering.