Haifeng Chen

A functional biphasic biomaterial homing mesenchymal stem cells for in vivo cartilage regeneration.

Cartilage regeneration after trauma is still a great challenge for clinicians and researchers due to many reasons, such as joint load-bearing, synovial movement and the paucity of endogenous repair cells. To overcome these limitations, we constructed a functional biomaterial using a biphasic scaffold platform and a bone-derived mesenchymal stem cells (BMSCs)-specific affinity peptide. The biphasic scaffold platform retains more cells homogeneously within the sol-gel transition of chitosan and provides sufficient solid matrix strength. This biphasic scaffold platform is functionalized with an affinity peptide targeting a cell source of interest, BMSCs. The presence of conjugated peptide gives this system a biological functionality towards BMSC-specific homing both in vitro and in vivo. The functional biomaterial can stimulate stem cell proliferation and chondrogenic differentiation during in vitro culture. Six months after in vivo implantation, compared with routine surgery or control scaffolds, the functional biomaterials induced superior cartilage repair without complications, as indicated by histological observations, magnetic resonance imaging and biomechanical properties. Beyond cartilage repair, this functional biphasic scaffold may provide a biomaterial framework for one-step tissue engineering strategy by homing endogenous cells to stimulate tissue regeneration.

Investigation on the structure and upconversion fluorescence of Yb3+/Ho3+ co-doped fluorapatite crystals for potential biomedical applications

Rare-earth Yb3+ and Ho3+ co-doped fluorapatite (FA:Yb3+/Ho3+) crystals were prepared by hydrothermal synthesis, and their structure, upconversion properties, cell proliferation and imaging were investigated. The synthesized crystals, with a size of 16 by 286u2005nm, have a hexagonal crystal structure of classic FA and a Ca/Yb/Ho molar ratio of 100/16/2.1. Several reasonable Yb3+/Ho3+ -embedding lattice models along the fluorine channel of the FA crystal cell are proposed for the first time, such as models for (Ca7YbHo©)(PO4)6F2 and (Ca6YbHoNa2)(PO4)6F2. The activated FA:Yb3+/Ho3+ crystals were found to exhibit distinct upconversion fluorescence. The 543- and 654-nm signals in the emission spectra could be assigned, respectively, to the 5F4 (5S2) - 5I8 and 5F5 - 5I8 transitions of holmium via 980-nm near-infrared excitation and the energy transfer of ytterbium. After the surfaces were grafted with hydrophilic dextran, the crystals displayed clear fluorescent cell imaging. Thus, the prepared novel FA:Yb3+/Ho3+ upconversion fluorescent crystals have potential applications in the biomedical field.

Directing chondrogenic differentiation of mesenchymal stem cells with a solid-supported chitosan thermogel for cartilage tissue engineering.

Hydrogels are attractive for cartilage tissue engineering because of their high plasticity and similarity with the native cartilage matrix. However, one critical drawback of hydrogels for osteochondral repair is their inadequate mechanical strength. To address this limitation, we constructed a solid-supported thermogel comprising a chitosan hydrogel system and demineralized bone matrix. Scanning electron microscopy, the equilibrium scanning ratio, the biodegradation rate, biomechanical tests, biochemical assays, metabolic activity tests, immunostaining and cartilage-specific gene expression analysis were used to evaluate the solid-supported thermogel. Compared with pure hydrogel or demineralized matrix, the hybrid biomaterial showed superior porosity, equilibrium swelling and degradation rate. The hybrid scaffolds exhibited an increased mechanical strength: 75% and 30% higher compared with pure hydrogels and demineralized matrix, respectively. After three days culture, bone-derived mesenchymal stem cells (BMSCs) maintained viability above 90% in all three materials; however, the cell retention of the hybrid scaffolds was more efficient and uniform than the other materials. Matrix production and chondrogenic differentiation of BMSCs in the hybrid scaffolds were superior to its precursors, based on glycosaminoglycan quantification and hyaline cartilage marker expression after three weeks in culture. Its easy preparation, favourable biophysical properties and chondrogenic capacity indicated that this solid-supported thermogel could be an attractive biomaterial framework for cartilage tissue engineering.

Evaluation of the morphology and osteogenic potential of titania-based electrospun nanofibers

Submicron-scale titania-based ceramic fibers with various compositions have been prepared by electrospinning. The as-prepared nanofibers were heat-treated at 700°C for 3 h to obtain pure inorganic fiber meshes. The results show that the diameter and morphology of the nanofibers are affected by starting polymer concentration and sol-gel composition. The titania and titaniasilica nanofibers had the average diameter about 100-300 nm. The crystal phase varied from high-crystallized rutile-anatase mixed crystal to low-crystallized anatase with adding the silica addition. The morphology and crystal phase were evaluated by SEM and XRD. Bone-marrow-derived mesenchymal stem cells were seeded on titania-silica 50/50 fiber meshes. Cell number and early differentiation marker expressions were analyzed, and the results indicated osteogenic potential of the titania-silica 50/50 fiber meshes.

Dextran-coated fluorapatite crystals doped with Yb3+/Ho3+ for labeling and tracking chondrogenic differentiation of bone marrow mesenchymal stem cells in vitro and in vivo

Upconversion fluorescent nanoparticles are becoming more widely used as imaging contrast agents, owing to their high resolution and penetration depth, and avoidance of tissue auto-fluorescence and photodamage to cells. Here, we synthesized upconversion fluorescent crystals from rare-earth Yb3+ and Ho3+ co-doped fluorapatite (FA:Yb3+/Ho3+) suitable for long-term tracking and monitoring cartilage development (chondrogenesis) in bone marrow mesenchymal stem cells (BMSCs) in vitro and in vivo. We initially determined the structure, morphology and luminescence of the products using X-ray powder diffraction, transmission electron microscopy and two-photon confocal microscopy. When excited at 980 nm, FA:Yb3+/Ho3+ crystals exhibited distinct upconversion fluorescence peaks at 543 nm and 654 nm. We then conjugated FA:Yb3+/Ho3+ crystals with dextran to enhance hydrophilicity, biocompatibility and cell penetration. Next, we employed the dextran-coated FA:Yb3+/Ho3+ crystals in labeling and tracking chondrogenic differentiation processes in BMSCs stably expressing green fluorescent protein (BMSCsGFP) in vitro and in vivo. Labeled BMSCsGFP were shown to reproducibly exhibit chondrogenic differentiation potential in RT-PCR analysis, histological assessment and immunohistochemistry. We observed continuous luminescence from the FA:Yb3+/Ho3+ upconversion crystals at 4 weeks and 12 weeks post transplantation in BMSCsGFP, while GFP fluorescence in both control and crystal-treated groups significantly decreased at 12 weeks after BMSCsGFP transplantation. We therefore demonstrate the high biocompatibility and stability of FA:Yb3+/Ho3+ crystals in tracking and monitoring BMSCs chondrogenesis in vitro and in vivo, highlighting their excellent cell labeling potential in cartilage tissue engineering.

The potential of nanomaterials for drug delivery, cell tracking, and regenerative medicine

1Mawson Institute, University of South Australia, Mawson Lakes, SA 5095, Australia 2School of Advanced Manufacturing, University of South Australia, Mawson Lakes, SA 5095, Australia 3Department of Biomedical Engineering, College of Engineering, Peking University, Beijing 100871, China 4Institute of Translational Medicine, The University of Liverpool, Liverpool L69 3GE, UK 5Department of Min-Met-Materials Engineering & CHU de Quebec Research Center, Laval University, Quebec City, Canada

Synthesis of thermal polymerizable Alginate-GMA hydrogel for cell encapsulation

Alginate is a negative ionic polysaccharide that is found abundantly in nature. Calcium is usually used as a cross-linker for alginate. However, calcium cross-linked alginate is used only for in vitro culture. In the present work, alginate was modified with glycidyl methacrylate (GMA) to produce a thermal polymerizable alginate-GMA (AA-GMA) macromonomer. The molecular structure and methacrylation (%DM) of the macromonomer were determined by 1H NMR. After mixing with the correct amount of initiator, the AA-GMA aqueous solution can be polymerized at physiological temperature. The AA-GMA hydrogels exhibited a three-dimensional porous structure with an average pore size ranging from 50 to 200 µm, directly depending on the macromonomer concentration. Biocompatibility of the AA-GMA hydrogel was determined by in vivo muscle injection and cell encapsulation. Muscle injection in vivo showed that the AA-GMA solution mixed with initiator could form a hydrogel in situ and had a mild inflammatory effect. Human umbilical vein endothelial cells (HUVECs) were encapsulated in the AA-GMA hydrogels in situ at 37°C. Cell viability and proliferation were unaffected by macromonomer concentrations, which suggests that AA-GMA has a potential application in the field of tissue engineering, especially for myocardial repair.

Fabrication of conductive polypyrrole nanofibers by electrospinning

Electrospinning is employed to prepare conductive polypyrrole nanofibers with uniform morphology and good mechanical strength. Soluble PPy was synthesized with NaDEHS as dopant and then applied to electrospinning with or without PEO as carrier. The PEO contents had great influence on the morphology and conductivity of the electrospun material. The results of these experiments will allow us to have a better understanding of PPy electrospun nanofibers and will permit the design of effective electrodes in the BMIs fields.

Enhanced hydrophilicity and protein adsorption of titanium surface by sodium bicarbonate solution

The aim of this study was to investigate a novel and convenient method of chemical treatment to modify the hydrophilicity of titanium surfaces. Sand-blasted and acid-etched (SLA) titanium surfaces and machined titanium surfaces were treated with sodium bicarbonate (NaHCO3) solution. The wetting behavior of both kinds of surfaces was measured by water contact angle (WCA) test. The surface microstructure was assessed with scanning electron microscopy (SEM) and three-dimensional (3D) optical microscopy. The elemental compositions of the surfaces were analyzed by X-ray photoelectron spectroscopy (XPS). The protein adsorption analysis was performed with fibronectin. Results showed that, after 1M NaHCO3 treatment, the hydrophilicity of both SLA and machined surfaces was enhanced. No significant microstructural change presented on titanium surfaces after NaHCO3 treatment. The deprotonation and ion exchange activities might cause the enhanced hydrophilicity of titanium surfaces. The increased protein adsorption of NaHCO3-treated SLA surfaces might indicate their improved tissue-integration in clinical use.

Cellular Uptake and Delivery-Dependent Effects of Tb3+-Doped Hydroxyapatite Nanorods

With the increasing interest in hydroxyapatite (HA) nanostructures for use in biomedicine, the systematic evaluation of their potential effects on biological systems is becoming critically important. In this work, we report the in vitro cellular uptake, in vivo tissue distributions and toxicity of Tb3+-doped HA (HA-Tb) after short-, intermediate-, and long-term exposure. Transmission electron microscopy analysis indicated that HA-Tb was taken up by cells via vesicle endocytosis. Cell proliferation and cytotoxicity assay, combined with confocal laser scanning microscopy, indicated excellent cell viability with no changes in cell morphology at the examined doses. Three HA-Tb delivery methods (intraperitoneal, intragastric, and intravenous) resulted in similar time-dependent tissue distributions, while intraperitoneal injection produced the highest bioavailability. HA-Tb initially accumulated in livers and intestines of rats (4 h to one day after administration), then became increasingly distributed in the kidney and bladder (seven days), and finally decreased in all tissues after 30 to 90 days. No histopathological abnormalities or lesions related to treatment with HA-Tb were observed. These results suggest that HA-Tb has minimal in vitro and in vivo toxicity, regardless of the delivery mode, time, and dose. The findings provide a foundation for the design and development of HA for biological applications.