TMPyP-bound guanosine-borate supramolecular hydrogel as smart hemoperfusion device with real-time visualized/electrochemical bi-modal monitoring for selective blood lead elimination.

Blood lead poisoning is a universal and severe health problem that greatly threatens human health in various industries. Elimination of blood lead relying on chelating agents and combination with hemoperfusion adsorbents has achieved considerable progress, but it is still suffering from the compromised selectivity of adsorbents as well as in absence of real time monitoring during treatment. Herein, we proposed a selective blood lead adsorbent integrated with real-time visualized/electrochemical bi-modal monitoring based on TMPyP-bound guanosine-borate (GB) supramolecular hydrogel as potential smart hemoperfusion device. The GB hydrogel possessed stability in physiological environment, self-healing ability resistant to fluid shear, blood compatibility, selective adsorption of lead ions superior to conventional adsorbents, anti-fouling performance to blood components and renewability. Benefiting from binding with TMPyP and the intrinsic conductivity, GB hydrogel was endowed with the ability to qualitatively diagnose the presence of blood lead via simple color change and quantitatively reflect the amount of adsorbed lead from blood accurately through electrochemical technique. This work puts forward an integrated treatment/monitoring hemoperfusion device with high selectivity, simple fabrication and low-cost, providing a paradigm for next generation design of intelligent, monitorable theranostic hemopurification system, which is also an extensible platform for the other research fields such as environmental monitoring and remediation.

A quantitative study of MC3T3-E1 cell adhesion, morphology and biomechanics on chitosan-collagen blend films at single cell level.

The interaction between cells and biomaterials plays a key role in cell proliferation and differentiation in tissue engineering. However, a quantitative analysis of those interactions has been less well studied. The objective of this study was to quantitative recapitulate the difference of MC3T3-E1 cell adhesion, morphological and biomechanical properties on chitosan-collagen films in terms of chemical composition. Here, the unbinding force between MC3T3-E1 cell and a series of chitosan-collagen films was probed by a real-time and in situ atomic force microscopy-single cell force spectroscopy (AFM-SCFS). Meanwhile, changes in cell morphology and Young's modulus on different chitosan-collagen films were detected by AFM. The cell area and CCK-8 results showed that cell spreading and proliferation increased with increasing collagen content. AFM observations clearly showed cell height decreased and pseudopod fusion with the collagen content increased. Cell adhesive force increased from 0.76±0.17 nN to 1.70±0.19 nN. On the contrary, cells Young's modulus, which reflected biophysical changes of cells decreased from 11.94±3.19 kPa to 1.81±0.52 kPa, respectively. It suggested that stronger cell-substrate interactions benefit cell adhesion, and better cell flexibility improve cell spreading. The findings indicate that cell morphology, adhesive force and Young's modulus are significant affected by various chitosan-collagen substrates. Those methods and quantitative results have guiding significance for investigating the mechanism of chitosan and/or collagen based cell-targeting drug carrier and the preparation of chitosan-collagen composite biomaterials.

A novel resorbable strontium-containing α-calcium sulfate hemihydrate bone substitute: a preparation and preliminary study.

Distraction osteogenesis after aggrieved bone segment resections is promising in the treatment of bone tumors and osteomyelitis. However, there is ambiguity with regard to the optimal choice of bone substitute, with biodegradability and excellent bone repair performance constituting key requirements. The purpose of this study was to develop a novel resorbable strontium-containing α-calcium sulfate hemihydrate (Sr-CaS) bone substitute to provide an alternative option for surgeons that better meets these requirements. The Sr-CaS was prepared using co-precipitation and hydrothermal methods and analyzed using x-ray diffraction (XRD), Fourier transform infrared (FTIR) scanning and thermogravimetric differential scanning calorimeter (TG-DSC) patterns. Cytotoxicity by tetrazolium bromide (MTT), sub-acute toxicity and hemolysis tests were performed to assess the initial biocompatibility of the new bone substitute. Radiographic analysis, micro-CT measurements and histological observation were used to evaluate the bone repair ability in rat tibia bone defects. The XRD and FTIR patterns of Sr-CaS were both very similar to CaS and the product had comparable characteristics similar to α-CaS as demonstrated by TG-DSC. Cytotoxicity of the substitute was class 1 (no cytotoxicity) and hemolysis was 4.3% (no hemolysis). Sub-acute toxicity was not seen after a 14 day evaluation. The substitute was radio-opaque. The empty group exhibited the lowest levels of both bone mineral densities (BMD) and bone volume/total volume (BV/TV) of the defects when compared to all other groups. The two Sr-CaS groups resulted in significantly greater BMDs and BV/TV of the defect compared to the CaS only group. However, there was no significant difference between the 5% and 10% Sr-CaS groups. The Sr-CaS was resorbable with satisfactory biocompatibility. The doped strontium ions enhanced the bone repair performance of CaS in a rat model and the new substitute demonstrated promising results for clinical use.

Nano-composite of poly(L-lactide) and halloysite nanotubes surface-grafted with L-lactide oligomer under microwave irradiation.

In order to improve the bonding between halloysite nanotubes (HNTs) and poly(L-lactide) (PLLA), and hence to increase the mechanical properties of HNTs/PLLA nano-composite, HNTs were surface-grafted with PLLA under microwave irradiation and then blended with PLLA matrix. The optimal conditions for grafting polymerization are: irradiation time of 30 min, microwave power of 30 W and reaction temperature of 130 degrees C. The structure and properties of the surface-grafted HNTs (g-HNTs) were characterized by Fourier transformation infrared (FTIR), thermal gravimetric analysis (TGA), X-ray diffraction (XRD) and dynamic light scattering (DLS). Nano-composites of g-HNTs/PLLA and non-grafted HNTs/PLLA were subsequently evaluated in terms of crystallinity, dispersion, interfacial interaction, mechanical performance and cytocompatibility by polarized optical microscopy (POM), field scanning electron microscope (FESM), tensile testing and cell culture experiment. Results show that the grafted PLLA chains on the surfaces of HNTs, as inter-tying molecules, played an important role in improving the adhesive strength between the nanotubes and the polymer matrix. The enhanced interaction among g-HNTs and PLLA matrix resulted in a better tensile strength and modulus compared to the pristine PLLA and HNTs/PLLA. Cell culture results indicated that g-HNTs promoted both adhesion and proliferation of M3T3 fibroblasts on the g-HNTs/PLLA composite film.

[Preparation and in vitro drug release performance of morphine-loaded chitosan microspheres].

OBJECTIVE: To prepare morphine-loaded chitosan microspheres by emulsion ionic cross-linking and investigate the effect of initial morphine quantity and different cross-linking degrees on drug loading, encapsulation efficiency and in vitro drug release. METHODS: Chitosan (with a relative molecular mass of 50,000 and deacetylation degree no less than 90%) at 100 mg and morphine at 20, 30, 40, or 50 mg were dissolved by 2% acetate and dripped slowly into 15 ml soy-bean oil containing 0.75 ml Span80. After full emulsification at 35 degrees C; for 1.5 h, the mixture was dripped slowly into sodium tripolyphosphate (10 mg/ml) at the mass ratio of 5:1, 7:1, or 9:1 to allow cross-linking for 2 h. The drug loading, encapsulation efficiency and in vitro drug release of the preparations were measured. RESULTS: The drug loading in the microsphere increased while the encapsulation efficiency reduced with the increment of the initial morphine quantity. High cross-linking degree resulted in prolonged release time of the drug loaded in the preparations. CONCLUSION: The microspheres loaded with morphine allows sustained release of morphine.

[Role of enamel matrix proteins in inducing biomimetic mineralization of the enamel: a study with quartz crystal microbalance technique].

OBJECTIVE: To investigate the adsorption behavior of enamel matrix proteins (EMPs) on the enamel surface and study their effect on biomineralization of enamel using quartz crystal microbalance (QCM) technique. METHODS AND RESULTS: The EMPs were adsorbed on the enamel surface to form a protein film, which was soaked in simulated body fluid solutions. After 30 days of biomimetic mineralization, the hydroxyapatite nucleation, growth and aggregation occurred with hydroxyapatite crystal formation on the enamel surface. CONCLUSION: The EMPs play a key role in regulating enamel mineralization.

[The study on the enamel remineralization by enamel matrix proteins' inducing].

OBJECTIVE: To find the enamel matrix proteins on the impact of enamel mineralization through experiments. METHODS: A combination of protein and beneficial carboxyl groups was grafted on the surface of enamel defects of rats through UV radiation then put into the enamel matrix proteins of calcium phosphate agar acetate solution systems, through scanning enamel surface with the electron microscopy to observe the morphological changes of enamel then analyse the regulation that enamel matrix proteins have done to the white hydroxyapatite crystals on the composition and morphology. RESULTS: In the enamel matrix protein added gel system, we can see the growth of hydroxyapatite crystals, and crystal showed a good degree of crystallinity and contained a small amount of CO3(2-) substituted hydroxyapatite crystals. CONCLUSION: The temperature at 37 degrees C water bath, after adding the enamel matrix proteins to gel system, the new hydroxyapatite crystals were numerous which proved that enamel matrix proteins played an important role in nucleation and growth of hydroxyapatite crystal, so it could be indicated that enamel matrix proteins could induce the enamel remineralization.

[Grafting functional groups on enamel surface by a self-assembled monolayer technique].

OBJECTIVE: To study the influence of various active groups grafted on the enamel surface by means of self assembly on enamel biomineralization. METHODS: The enamel was prepared by immersing the bicuspid tooth into 1 mmol/L ethanolic solution of a omega-functionalized (omega=PO4H2, SO3H, COOH or OH) group and deionized water solution of HSCH2CH2SO3Na for 24 h at room temperature. The contact angles and infrared (IR) images were used to identify the morphological changes of the enamel with chemisorption of the functional groups. RESULTS: The contact angles and IR images showed that the omega-functionalized (omega=-PO4H2, -SO3H, -COOH, -OH or -CH3) group was chemisorbed on the enamel surface. CONCLUSION: Self assembled monolayers with omega-functionalized (omega=-PO4H2, -SO3H, -COOH, -OH or -CH3) group can be successfully formed on the enamel surface by hydrolyzation.

Study on machinable glass-ceramic containing fluorophlogopite for dental CAD/CAM system.

The glass-ceramic mainly containing fluorophlogopite is one of widely used dental ceramics. In the K2O-CaO-MgO-Al2O3-SiO2-F system, a new-type glass-ceramic containing fluorophlogopite Ca-mica has been synthesized. Its crystalline was studied by XRD and EDS. The fluorophlogopite whose formula postulated K(1 - X )Ca( X/2)Mg(3)AlSi(3)O(10)F(2) was its main crystalline. The microstructure of the glass-ceramic displayed typical machinable microstructure with lath like crystals isolated and interlocking with different aspect ratio. The material also showed better bending strength (228.11 +/- 7.55 MPa). It took less than 12 minutes to fabricate a whole crown by dental CAD/CAM system with the glass-ceramic.

Synthesis of novel liquid crystal compounds and their blood compatibility as anticoagulative materials.

The objective of this study was to synthesize new types of cholesteric liquid crystal compounds and study the anticoagulative properties of their composite membranes. Three kinds of cholesteric liquid crystal compounds were synthesized and characterized by infrared spectroscopy, differential scanning calorimetry and optical polarizing microscope. The polysiloxane, as a substrate, was blended with three liquid crystal compounds and was then used as membranes. The anticoagulative property of different polysiloxane liquid crystal composite membranes was identified by the blood compatibility tests. Three cholesteryl liquid crystals synthesized in this work contained hydrophilic soft chains and presented iridescent texture owned by cholesteric liquid crystals in the range of their liquid crystal state temperature, but only cholesteryl acryloyl oxytetraethylene glycol carbonate was in the liquid crystal state at body temperature. When liquid crystals were blended with polysiloxane to form polysiloxane/liquid crystal composite membranes, the haemocompatibility of these membranes could be improved to some extent. The blood compatibility of composite membranes whose hydrophilic property was the best was more excellent than that of other composite membranes, fewer platelets adhered and spread, and showed little distortion on the surface of materials.

Improving the cell affinity of a poly(D,L-lactide) film modified by grafting collagen via a plasma technique.

Poly(D,L-lactide) films were surface-modified by grafting collagen via NH(3) plasma to improve cell affinity. The modified films were characterized by IR analysis, contact angle measurement, SEM analysis and collagen quantity determination. It was demonstrated that -NH(2) and collagen were incorporated into the surface of PDLLA films. The hydrophilicity of the PDLLA film increased after NH(3) plasma treatment, but decreased with further collagen modification. More collagen was incorporated into the PDLLA films by a grating method as compared to that with an anchorage treatment. L929 fibroblast cells were used to evaluate the cell affinity of the modified films and control. It was shown that PDLLA films surface-modified by grafting collagen via NH(3) plasma more efficiently enhanced the cells attachment and proliferation than those films modified by collagen anchorage or only NH(3) plasma treatment.

Development and potential of a biomimetic chitosan/type II collagen scaffold for cartilage tissue engineering.

BACKGROUND: Damaged articular cartilage has very limited capacity for spontaneous healing. Tissue engineering provides a new hope for functional cartilage repair. Creation of an appropriate cell carrier is one of the critical steps for successful tissue engineering. With the supposition that a biomimetic construct might promise to generate better effects, we developed a novel composite scaffold and investigated its potential for cartilage tissue engineering. METHODS: Chitosan of 88% deacetylation was prepared via a modified base reaction procedure. A freeze-drying process was employed to fabricate a three-dimensional composite scaffold consisting of chitosan and type II collagen. The scaffold was treated with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide. Ultrastructure and tensile strength of the matrix were carried out to assess its physico-chemical properties. After subcutaneous implantation in rabbits, its in vivo biocompatibility and degradability of the scaffold were determined. Its capacity to sustain chondrocyte growth and biosynthesis was evaluated through cell-scaffold co-culture in vitro. RESULTS: The fabricated composite matrix was porous and sponge-like with interconnected pores measuring from 100-250 microm in diameter. After cross-linking, the scaffold displayed enhanced tensile strength. Subcutaneous implantation results indicated the composite matrix was biocompatible and biodegradable. In intro cell-scaffold culture showed the scaffold sustained chondrocyte proliferation and differentiation, and maintained the spheric chondrocytic phenotype. As indicated by immunohistochemical staining, the chondrocytes synthesized type II collagen. CONCLUSIONS: Chitosan and type II collagen can be well blended and developed into a porous 3-D biomimetic matrix. Results of physico-chemical and biological tests suggest the composite matrix satisfies the constraints specified for a tissue-engineered construct and may be used as a chondrocyte carrier for cartilage tissue engineering.

[Biocompatibility evaluation of chitosan-g-polyvinylpyrrolidone].

OBJECTIVE: To evaluate the biocompatibility of chitosan-g-polyvinylpyrrolidone as a new scaffold material. METHODS: The material was tested and measured for water absorption and contact angle, followed by evaluation of the biocompatibility by implantation into rabbits and in vitro cultured with the corneal epithelial cells. RESULTS: The water absorption rate of the material reached 1 100% with contact angle of 83-86. The results of implantation revealed partial degradation of the material 3 months after implantation, and much collagen and numerous corneal stromal cells appeared on the material without obvious inflammation reactions. In vitro coculture with epithelial cells showed good adhesion of the cells to the material which induced no obvious cytotoxicity. CONCLUSION: The novel chitosan derivative has excellent biocompatibility and can be used as a tissue scaffold material.

Preparation of polylactic acid/chitin composite material and its safety evaluation by animal experiments.

OBJECTIVE: To prepare a scaffold material with good biocompatibility and biodegradability by compounding polylactic acid (PLA) and chitin. METHODS: After preparation of PLA from lactic acid, the compounding of PLA and chitin was carried out by dissolving these 2 materials in one solution for reaction. The composite material was obtained and molded after the solvent was evaporized, and the safety tests of this resultant material were conducted in guinea pigs and New Zealand rabbits, respectively. RESULTS: In allergic test, the guinea pigs responded to the digestion solution of the composite material in almost the same manner as to normal saline (the latter serving as negative control), and no obvious allergic reaction was observed in the animals except those in positive control group. Pyrogenic test by injecting the digestion solution of the composite material in 6 rabbits found a raise in the body temperature less than 0.2 degrees Celsius, with the total increase (adding up the individual temperature raise) less than 1.0 degrees Celsius, to meet the accepted criteria for the pyrogenic test. In subsequent toxicity test, the rabbits showed no signs of agitation or inanimate behavior after injection. CONCLUSIONS: PLA/chitin composite material conforms to the ISO10993-1, and can be used as a basic scaffold material in tissue engineering.

Preparation and biocompatibility of tissue-engineered scaffold materials based on collagen.

OBJECTIVE: To prepare a tissue-engineered scaffold material using collagen as the matrices and to study the blood compatibility and tissue biocompatibility of this material. METHODS: Physical, chemical and physical/chemical methods were used for the crosslinking of the collagen. RESULTS: Dynamic blood clotting tests indicated that the blood clotting index (BCI) of the crosslinked collagen materials prepared by different means decreased as their contact with the blood was prolonged, and the collagen material obtained after crosslink through 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide method showed the highest BCI after contact with the blood within certain length of time. Hemolysis ratios of all the crosslinked collagen materials were shown to be much lower than 5%, well conforming to the requirement of biomaterials. Scanning electron microscopy showed that the platelets attached to the surface of the crosslinked collagen materials, having a fairly small number, were not morphologically distorted. CONCLUSION: The collagen materials obtained by the 3 crosslink methods have good blood compatibility. The cells grow well on the surfaces of the materials, indicating their good biocompatibility.