Ximing Pu

Preparation of PLLA/PLGA microparticles using solution enhanced dispersion by supercritical fluids (SEDS).

In this work, poly(L-lactic acid)/poly(lactide-co-glycolide) (PLLA/PLGA) microparticles were prepared using the technique of solution-enhanced dispersion by supercritical fluids (SEDS). For comparison, separate PLLA and PLGA microparticles were also produced by the same SEDS process. The produced microparticles were characterized by scanning electron microscopy, laser particle size analyzer, X-ray diffraction, differential scanning calorimetry, Fourier transform infrared spectroscopy, and gas chromatography. Results indicate that PLLA/PLGA microparticles possess sphere-like shapes with smooth surfaces. The mean particle size of PLLA/PLGA microparticles ranges from 1.76 to 2.15 microm, depending on the feeding ratio of PLLA to PLGA used in the SEDS process. The crystallinity of PLLA/PLGA microparticles decreases after the SEDS processing, so that the produced microparticles are in an amorphous state. Pure PLGA was hard to precipitate in small, fine microparticle form without the presence of PLLA. A model drug, paclitaxel, was encapsulated into PLLA/PLGA microparticles by the same SEDS process, and the in vitro release rate of paclitaxel from these PLLA/PLGA composites could be modulated by variation of the mixing ratio PLLA:PLGA. The prepared microparticles have negligible residual organic solvent. Drug-loaded PLLA/PLGA microparticles produced by SEDS have potential as an advanced colloidal suspension for pharmaceutical applications.

Preparation, characterization and in vitro cytotoxicity of indomethacin-loaded PLLA/PLGA microparticles using supercritical CO2 technique.

In this work, indomethacin-loaded poly(l-lactic acid)/poly(lactide-co-glycolide) (IDMC-PLLA/PLGA) microparticles were prepared using solution-enhanced dispersion by supercritical fluids (SEDS) technique in an effort to obtain alternative IDMC formulation for drug delivery system. Surface morphology, particle size and particle size distribution, drug encapsulation efficiency, drug release kinetics, in vitro cytotoxicity and the cellular uptake of drug-loaded microparticles were investigated. The drug-loaded microparticles exhibited sphere-like shape and small particle size with narrow particle size distribution. IDMC was amorphously dispersed within the PLLA/PLGA matrix after the SEDS process. In vitro release studies revealed that the drug-loaded microparticles substantially enhanced the dissolution rate of IDMC compared to the free IDMC, and demonstrated a biphasic drug release profile. In vitro cytotoxicity assays indicated that drug-loaded microparticles possessed longer sustained inhibition activity on proliferation of the non-small-cell lung cancer A549 cell lines than did free IDMC. Fluorescence microscopy and transmission electron microscopy identified the phagocytosis of drug-loaded microparticles into the A549 cells and characteristic morphology of cell apoptosis such as the nuclear aberrations, condensation of chromatin, and swelling damage in mitochondria. These results collectively suggested that IDMC-PLLA/PLGA microparticles prepared using SEDS would have potentials in anti-tumor applications as a controlled drug release dosage form without harmful organic solvent residue.

Localized delivery of growth factors for angiogenesis and bone formation in tissue engineering

Angiogenesis is a key component of bone formation. Delivery of growth factors for both angiogenesis and osteogenesis is about to gain important potential as a future therapeutic tool. This review focuses on these growth factors that have dual functions in angiogenesis and osteogenesis, and their localized application. A major hurdle in the clinical development of growth factor therapy so far is how to assure safe and efficacious therapeutic use of such factors and avoid unwanted side effects and toxicity. It is now firmly established from the available information that the type, dose, combinations and delivery kinetics of growth factors all play a decisive role for the success of growth factor therapy. All of these parameters have to be adapted and optimized for each animal model or clinical case. In this review we discuss some important parameters associated with growth factor therapy and present an overview of selected preclinical studies, followed by a conceptual description of both established and proposed delivery strategies meeting therapeutic needs.

Development of Fe3O4-poly(L-lactide) magnetic microparticles in supercritical CO2.

The Fe(3)O(4)-poly(L-lactide) (Fe(3)O(4)-PLLA) magnetic microparticles were successfully prepared in a process of solution-enhanced dispersion by supercritical CO(2) (SEDS), and their morphology, particle size, magnetic mass content, surface atom distribution and magnetic properties were characterized. Indomethacin (Indo) was used as a drug model to produce drug-polymer magnetic composite microparticles. The resulting Fe(3)O(4)-PLLA microparticles with mean size of 803 nm had good magnetic property and a saturation magnetization of 24.99 emu/g. The X-ray photoelectron spectroscopy (XPS) test indicated that most of the Fe(3)O(4) were encapsulated by PLLA, which indicated that the Fe(3)O(4)-PLLA magnetic microparticles had a core-shell structure. After further loading with drug, the Indo-Fe(3)O(4)-PLLA microparticles had a bigger mean size of 901 nm, and the Fourier transform infrared spectrometer (FTIR) analysis demonstrated that the SEDS process was a typical physical coating process to produce drug-polymer magnetic composite microparticles, which is favorable for drugs since there is no change in chemistry. The in vitro cytotoxicity test showed that the Fe(3)O(4)-PLLA magnetic microparticles had no cytotoxicity and were biocompatible, which means there is potential for biomedical application.

Synergistic and sequential effects of BMP-2, bFGF and VEGF on osteogenic differentiation of rat osteoblasts

In the present study, the effects of bone morphogenetic protein-2 (BMP-2), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) on regulation of rat osteoblast (ROB) maturation in vitro were investigated. It was found that the proliferation, differentiation and mineralization of ROBs were all dose-dependently increased at particular times in the case of treatment with only one growth factor. To investigate the effects of combined treatment, ROBs were treated with either a single application of a relatively high dose of each growth factor, or binary/triple combined applications of relatively low doses of the growth factors. Osteogenic differentiation was significantly promoted in the triple combination treatment of BMP-2, VEGF and bFGF compared with the single or binary combination treatments. The optimal timing of the triple combination to enhance osteogenesis was also tested. When bFGF and VEGF were added in the early stage, and BMP-2 and VEGF were added in the late stage, osteogenic differentiation of ROBs could be enhanced more effectively. These results could be used to construct bone tissue engineering scaffolds that release growth factors sequentially.

Fabrication of aligned, porous and conductive fibers and their effects on cell adhesion and guidance.

The potential applications of aligned, conductive electrospun fibers have been widely studied in anisotropic tissue regeneration. In this study, aligned porous poly L-lactic acid fibers were obtained with electrospinning, then polypyrrole nanoparticles (PPy NPs) were coated onto the porous fibers with oxidation polymerization to prepare electrically conductive fibers with about 1.24 μm of diameter, and their surface conductivity was about 50 mS. The results of L929 cell test showed that more than 55% of cells grew along the aligned porous fiber axis, confirming that the cell guidance of aligned porous fibers was better than that of non-porous fibers. The results of differentiated PC12 cells on porous fibers showed that the alignment degree of neurite outgrowth and average neurite length of the cells were 84% and 111 μm, respectively, which were larger than those on the non-porous fibers. A primary mechanism was proposed to explain effect of these pores on cell/neurite adhesion and orientation along the aligned porous fibers.

Amino-functionalized mesostructured cellular foams as carriers of glucose oxidase

The mesostructured cellular foams (MCFs) were synthesized in aqueous hydrochloric acid by using dilute Pluronic P123 solutions in the presence of 1,3,5-trimethylbenzene (TMB) as organic cosolvent. And the amino-functionalized MCFs (NH₂-MCFs) were prepared from primary MCFs by post-synthesis method using 3-aminopropyl-trimethoxysilane (APTMS) as the chemical modifier. The SEM and TEM observations showed the similar morphologies and pore structures of both MCFs and NH₂-MCFs, indicating that the surface modification had little effect on the morphologies and pore structures. Glucose oxidase (GOD) was physically adsorbed on MCFs and NH₂-MCFs at different pH. The maximum immobilized amount of GOD on NH₂-MCFs (487 mg g⁻¹) was much higher than that of MCFs (216 mg g⁻¹) at pH 5.0. The larger loading capacity of NH₂-MCFs suggested that the electrostatic interaction was the dominant force for GOD adsorption. Furthermore, the immobilized GOD exhibited improved thermal and storable stabilities. The GOD immobilized on NH₂-MCFs (NH₂-MCFs-GOD) still maintained 80% of initial activity after incubation at 60°C for 1 h, whereas the free GOD and the GOD immobilized on MCFs (MCFs-GOD) remained only 40% and 60%, respectively. Moreover, after stored at 4°C for 30 days, the free GOD, the MCFs-GOD and the NH₂-MCFs-GOD retained 37%, 52% and 73% of initial activities, respectively. Based on these results, possible mechanisms were also discussed.

Enhancement of neurite adhesion, alignment and elongation on conductive polypyrrole-poly(lactide acid) fibers with cell-derived extracellular matrix

Extracellular matrix (ECM) can promote peripheral nerve repair. In this study, a conductive fiber-film (CFF) with core-sheath structure and conductivity of ∼10Scm-1 was prepared by electrospinning of aligned poly(l-lactide acid) (PLLA) fibers and electrochemical deposition of polypyrole (PPy) nanoparticles. Then the multiple components of ECM, including laminin, fibronectin and collagen, were coated on the surface of CFF by culturing and lysing L929 cells to fabricate the bioactive scaffold of ECM-linked CFF (ECM-CFF). The electrical stimulation (ES) of 100mV/cm for 14days and 2h per day did not significantly decrease the conductivity of ECM-CFF. The results of PC12 cells test indicated that, cells adhesion rate, neurite-bearing cell rate and neurite alignment rate on ECM-CFF were ∼95%, ∼77%, ∼70%, respectively, significantly larger than the corresponding values on bare CFF (17%, 29% and 14%, respectively). The neurites length on ECM-CFF (∼79mm) was also larger than that on bare CFF (∼25mm). ES of 100mV/cm onto PC12 cells through ECM-CFF could significantly promote neurite extension in first 3days of the neurite growth. These results indicated that, the combination of ECM-CFF with ES could improve the nerve regeneration by encouraging neural-cell adhesion, neurite growth and extension.

Biopanning and characterization of peptides with Fe3O4 nanoparticles-binding capability via phage display random peptide library technique.

Functionalization of inorganic nanoparticles (NPs) play an important role in biomedical applications. A proper functionalization of NPs can improve biocompatibility, avoid a loss of bioactivity, and further endow NPs with unique performances. Modification with vairous specific binding biomolecules from random biological libraries has been explored. In this work, two 7-mer peptides with sequences of HYIDFRW and TVNFKLY were selected from a phage display random peptide library by using ferromagnetic NPs as targets, and were verified to display strong binding affinity to Fe3O4 NPs. Fourier transform infrared spectrometry, fluorescence microscopy, thermal analysis and X-ray photoelectron spectroscopy confirmed the presence of peptides on the surface of Fe3O4 NPs. Sequence analyses revealed that the probable binding mechanism between the peptide and Fe3O4 NPs might be driven by Pearson hard acid-hard base specific interaction and hydrogen bonds, accompanied with hydrophilic interactions and non-specific electrostatic attractions. The cell viability assay indicated a good cytocompatibility of peptide-bound Fe3O4 NPs. Furthermore, TVNFKLY peptide and an ovarian tumor cell A2780 specific binding peptide (QQTNWSL) were conjugated to afford a liner 14-mer peptide (QQTNWSLTVNFKLY). The binding and targeting studies showed that 14-mer peptide was able to retain both the strong binding ability to Fe3O4 NPs and the specific binding ability to A2780 cells. The results suggested that the Fe3O4-binding peptides would be of great potential in the functionalization of Fe3O4 NPs for the tumor-targeted drug delivery and magnetic hyperthermia.

Facile synthesis of a α-MoO3 nanoplate/TiO2 nanotube composite for high electrochemical performance

A novel α-MoO3/TiO2 composite electrode material for high performance supercapacitor applications was synthesized using a facile electrodeposition technique. The surface morphology, microstructure, chemical composition and chemical states of the electrode material were analyzed using scanning electron microscopy, X-ray diffraction, energy dispersive X-ray spectroscopy, and X-ray photoelectron spectroscopy. Cyclic voltammetry tests, galvanostatic charge–discharge measurements, and electrochemical impedance spectroscopy were employed to analyze electrochemical behavior. The results demonstrate that a very high capacitance (43.42 mF cm−2) can be achieved at a scan rate of 20 mV s−1 in 1 M aqueous Na2SO4 solution and good capacitance behavior even after 3000 continuous charge–discharge cycles, indicating potential applications as an electrode material for supercapacitors.