Xiumei Mo

Soft tissue adhesive composed of modified gelatin and polysaccharides

Although fibrin glue has been clinically used as a surgical adhesive, hemostatic agent, and sealant, it has the risk of virus infection because its components, fibrinogen and thrombin, are obtained from human blood. To circumvent this problem, we employed bioabsorbable gelatin and polysaccharides to prepare a safer hemostatic glue. Gelatin was modified with ethylenediamine using water-soluble carbodiimide to introduce additional amino groups into the original gelatin, while dextran and hydroxyethyl-starch were oxidized by sodium periodate to convert 1,2-hydroxyl groups into dialdehyde groups. Upon mixing of the two polymer components in aqueous solution, Schiff base was formed between the amino groups in the modified gelatin and the aldehyde groups in the modified polysaccharides, which thus resulted in intermolecular cross-linking and gel formation. The fastest gel formation took place within 2 s, and its bonding strength to porcine skin was about 225 gfcm-2 when 20 wt% of an amino-gelatin (55% amino) and 10 wt% of aldehyde-HES (>84% dialdehyde) aqueous solutions were mixed. In contrast, the gelation time and bonding strength of fibrin glue was 5 s and 120 gfcm-2, respectively.

Fabrication of silk fibroin blended P(LLA‐CL) nanofibrous scaffolds for tissue engineering

Electrospinning using natural proteins and synthetic polymers offers an attractive technique for producing fibrous scaffolds with potential for tissue regeneration and repair. Nanofibrous scaffolds of silk fibroin (SF) and poly(L-lactic acid-co-epsilon-caprolactone) (P(LLA-CL)) blends were fabricated using 1,1,1,3,3,3-hexafluoro-2-propanol as a solvent via electrospinning. The average nanofibrous diameter increased with increasing polymer concentration and decreasing the blend ratio of SF to P(LLA-CL). Characterizations of XPS and (13)C NMR clarified the presence of SF on their surfaces and no obvious chemical bond reaction between SF with P(LLA-CL) and SF in SF/P(LLA-CL) nanofibers was present in a random coil conformation, SF conformation transformed from random coil to beta-sheet when treated with water vapor. Whereas water contact angle measurements conformed greater hydrophilicity than P(LLA-CL). Both the tensile strength and elongation at break increased with the content increasing of P(LLA-CL). Cell viability studies with pig iliac endothelial cells demonstrated that SF/P(LLA-CL) blended nanofibrous scaffolds significantly promoted cell growth in comparison with P(LLA-CL), especially when the weight ratio of SF to P(LLA-CL) was 25:75. These results suggested that SF/P(LLA-CL) blended nanofibrous scaffolds might be potential candidates for vascular tissue engineering.

Genipin-crosslinked silk fibroin/hydroxybutyl chitosan nanofibrous scaffolds for tissue-engineering application

To improve water-resistant ability and mechanical properties of silk fibroin (SF)/hydroxybutyl chitosan (HBC) nanofibrous scaffolds for tissue-engineering applications, genipin, glutaraldehyde (GTA), and ethanol were used to crosslink electrospun nanofibers, respectively. The mechanical properties of nanofibrous scaffolds were obviously improved after 24 h of crosslinking with genipin and were superior to those crosslinked with GTA and ethanol for 24 h. SEM indicated that crosslinked nanofibers with genipin and GTA vapor had good water-resistant ability. Characterization of the microstructure (porosity and pore structure) demonstrated crosslinked nanofibrous scaffolds with genipin and GTA vapor had lager porosities and mean diameters than those with ethanol. Characterization of FTIR-ATR and (13)C NMR clarified both genipin and GTA acted as crosslinking agents for SF and HBC. Furthermore, genipin could induce SF conformation from random coil or α-helix to β-sheet. Although GTA could also successfully crosslink SF/HBC nanofibrous scaffolds, in long run, genipin maybe a better method due to lower cytotoxicity than GTA. Cell viability studies and wound-healing test in rats clarified that the genipin-crosslinked SF/HBC nanofibrous scaffolds had a good biocompatibility both in vitro and in vivo. These results suggested that genipin-crosslinked SF/HBC nanofibrous scaffolds might be potential candidates for wound dressing and tissue-engineering scaffolds.

Electrospun chitosan-P(LLA-CL) nanofibers for biomimetic extracellular matrix

Chitosan-poly(L-lactic acid-co-ε-caprolactone)(50:50) (P(LLA-CL)) (CS/P(LLA-CL)) blends were electrospun into nanofibers using 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) and trifluoroacetic acid (TFA) as solvents. Chitosan, which is difficult to electrospin into nanofibers, could be easily electrospun into nanofibers with addition of a small portion of P(LLA-CL). The fiber diameter depended on both the polymer concentration and the blend ratio of chitosan to P(LLA-CL). The average fiber diameter increased with increasing polymer concentration and decreasing the blend ratio of chitosan to P(LLA-CL). X-ray diffractometry (XRD) and Fourier-transform infrared (FT-IR) spectra were measured to characterize blended nanofibers. The porosity of CS/P(LLA-CL) nanofiber mats increased with increasing the weight ratio of chitosan to P(LLA-CL), while both the tensile strength and the ultimate strain increased with increasing P(LLA-CL) ratio. Fibroblast cell growth on nanofiber mats were investigated with MTT assay and scanning electron microscope (SEM) observation. The highest cell proliferation was observed on the nanofiber mats when the weight ratio of chitosan to P(LLA-CL) was 1:2. As SEM images shown, fibroblast cells showed a polygonal shape on blend nanofiber mats and migrated into the nanofiber mats.

Europium-doped amorphous calcium phosphate porous nanospheres: preparation and application as luminescent drug carriers.

Calcium phosphate is the most important inorganic constituent of biological tissues, and synthetic calcium phosphate has been widely used as biomaterials. In this study, a facile method has been developed for the fabrication of amorphous calcium phosphate (ACP)/polylactide-block-monomethoxy(polyethyleneglycol) hybrid nanoparticles and ACP porous nanospheres. Europium-doping is performed to enable photoluminescence (PL) function of ACP porous nanospheres. A high specific surface area of the europium-doped ACP (Eu3+:ACP) porous nanospheres is achieved (126.7 m2/g). PL properties of Eu3+:ACP porous nanospheres are investigated, and the most intense peak at 612 nm is observed at 5 mol% Eu3+ doping. In vitro cytotoxicity experiments indicate that the as-prepared Eu3+:ACP porous nanospheres are biocompatible. In vitro drug release experiments indicate that the ibuprofen-loaded Eu3+:ACP porous nanospheres show a slow and sustained drug release in simulated body fluid. We have found that the cumulative amount of released drug has a linear relationship with the natural logarithm of release time (ln(t)). The Eu3+:ACP porous nanospheres are bioactive, and can transform to hydroxyapatite during drug release. The PL properties of drug-loaded nanocarriers before and after drug release are also investigated.

Preparation of core-shell biodegradable microfibers for long-term drug delivery

A coaxial electrospun technique to fabricate core-shell microfibers (MFs) for drug delivery application is described. In one-step, Paclitaxel (PTX)-loaded poly(L-lactic acid-co-epsilon-caprolactone) (75:25) (P(LLA-CL)(core/shell)) was electrospun into MFs using 2,2,2-trifluoroethanol as the solvent. The physical and chemical properties of electrospun fibers were characterized by various techniques, such as scanning electron microscopy, transmission electron microscopy, X-ray diffractometry, and Fourier-transform infrared. The fiber diameter depended on both the polymer concentration and the flow ratio of PTX to P(LLA-CL). The encapsulation efficiency and in vitro release profile were measured using high performance liquid chromatography methods. PTX released from the MFs in a short burst over 24 h followed by very slow release over the following 60 days. In addition, the cytotoxicity of PTX-loaded P(LLA-CL) MFs was evaluated using 3-[4,5-dimehyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide assay on HeLa cell lines. These results indicate that PTX could be released from P(LLA-CL) fibers in a steady manner and effectively inhibit the activity of HeLa cells.

Cross-Linking of Gelatin and Chitosan Complex Nanofibers for Tissue-Engineering Scaffolds

The aim of this study is to investigate cross-linked gelatin–chitosan nanofibers produced by means of electrospinning. Gelatin and chitosan nanofibers were electrospun and then cross-linked by glutaraldehyde (GTA) vapor at room temperature. Scanning electron microscopy (SEM) images showed that the cross-linked mats could keep their nanofibrous structure after being soaked in deionized water at 37° C. The cross-linking mechanism was discussed based on FT-IR results. The two main mechanisms of cross-linking for chitosan and gelatin–chitosan complex are Schiff base reaction and acetalization reaction. For gelatin, the mechanism of cross-linking was Schiff base reaction. The mechanical properties of nanofibrous mats were improved after cross-linking. The biocompatibility of electrospun nanofibrous mats after cross-linking was investigated by the viability of porcine iliac endothelial cells (PIECs). The morphologies of PIECs on the cross-linked nanofibrous mats were observed by SEM. In addition, proliferation of PIECs was tested with the method of methylthiazol tetrazolium (MTT) assay. The results indicate that gelatin–chitosan nanofibrous mats could be a promising candidate for tissue-engineering scaffolds.

Electrospun scaffolds from silk fibroin and their cellular compatibility.

Electrospinning offers an attractive opportunity for producing silk fibroin (SF) nano/micro fibrous scaffolds with potential for tissue regeneration and repair. Electrospun scaffolds of silk fibroin were fabricated as a biomimetic scaffold for tissue engineering. The morphology of the electrospun scaffolds was investigated with SEM and AFM. The SEM images indicated that electrospun SF fibers were ribbon-shaped and the average width increased with increasing SF concentrations. The AFM images revealed that, after treated with methanol, there was a groove on the surface of fiber, which is conducive to cell attachment. The structure of electrospun SF fibers was characterized by NMR, WAXD, and DSC. The results displayed that SF in electrospun fibers was present in a random coil conformation, SF conformation transformed from random coil to beta-sheet when treated with methanol. Cell attachment and proliferation studies with pig iliac endothelial cells (PIECs) demonstrated that electrospun SF scaffolds significantly promoted cell attachment and proliferation in comparison with cast SF films. These results suggest electrospun SF scaffolds may be potential candidates for cardiovascular tissue engineering.

Biocompatibility, Alignment Degree and Mechanical Properties of an Electrospun Chitosan–P(LLA-CL) Fibrous Scaffold

Chitosan–poly(L-lactide-co-ε-caprolactone) (P(LLA-CL)) complex fibers, fibrous mats and a tubular scaffold have been obtained through electrospinning. Due to their high porosity, there were more porcine iliac artery endothelial cells (PIECs) attached to fiber mats than to tissue-culture plate (TCP) and coverslips. The cells could grow and spread well on nanofiber mats. There were many of native extracellular matrix (ECM)-like colloids above and under the surface of fibrous mats after cell culturing. The two-dimensional fast Fourier transform (2-D FFT) approach was used to analysis alignment degree of fibers collected on a rotary mandrel. The relations among mechanical properties, alignment degree, fiber diameter and rotary speed are discussed. Aligned fibers with various alignment degrees could be found through adjusting rotary speed. Fiber alignment was the variable most closely associated with the regulation of stress and strain. In this study, we show a feasible approach for producing scaffold with controllable mechanical property for soft tissue engineering through electrospinning.

Electrospinning Thermoplastic Polyurethane-Contained Collagen Nanofibers for Tissue-Engineering Applications

Electrospinning is a new method used in tissue engineering. It can spin fibers in nanoscale by electrostatic force. A series of thermoplastic polyurethane (TPU)/collagen blend nanofibrous membranes was prepared with different weight ratios and concentrations via electrospinning. The two biopolymers used 1,1,1,3,3,3,-hexafluoro-2-propanol (HFP) as solvent. The electrospun TPU-contained collagen nanofibers were characterized using scanning electron microscopy (SEM), XPS spectroscopy, atomic force microscopy, apparent density and porosity measurement, contact-angle measurement, mechanical tensile testing and viability of pig iliac endothelial cells (PIECs) on blended nanofiber mats. Our data indicate that fiber diameter was influenced by both polymer concentration and blend weight ratio of collagen to TPU. The average diameter of nanofibers gradually decreases with increasing collagen content in the blend. XPS analysis indicates that collagen is found to be present at the surface of blended nanofiber. The results of porosity and contact-angle measurement suggest that with the collagen content in the blend system, the porosity and hydrophilicity of the nanofiber mats is greatly improved. We have also characterized the molecular interactions in TPU/collagen complex by Fourier transform infrared spectroscopy (FT-IR). The result could demonstrate that there were no intermolecular bonds between the molecules of TPU and collagen. The ultimate tensile stress and strain were carried out and the data confirmed the FT-IR results. The TPU/collagen blend nanofibrous mats were further investigated as promising scaffold for PIEC culture. The cell proliferation and SEM morphology observations showed that the cells could not only favorably grow well on the surface of blend nanofibrous mats, but also able to migrate inside the scaffold within 24 h of culture. These results suggest that the blend nanofibers of TPU/collagen are designed to mimic the native extracellular matrix for tissue engineering and develop functional biomaterials.