There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechan...There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechanical properties of SF materials. In this study, SF was crosslinked effectively with genipin which is a naturally occurring iridoid glucoside and the crosslinking mechanism was investigated through FTIR and amino acid analysis. The results showed that genipin could react with the -NH2 groups on the side chains of SF macromolecules and to form inter- and intra-molecular covalent bonds, and improved the stability of SF materials significantly. In vitro, the performances of genipin-crosslinked SF films were assessed by seeding L929 cells and compared with ethanol-processed SF films, glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked ones. The genipin-crosslinked SF films showed a similar affinity to cells as ethanol-processed ones, and a higher bioactivity in promoting cell growth and proliferation, inhibition of cell apoptosis, and maintenance of normal cell cycle compared with glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked SF films. These features, combined with the decrease of brittleness of SF films crosslinked with chemical methods, substantiated genipin as an effective and biocompatible agent for the manufacturing of bioactive SF materials which used as tissue engineering scaffolds and drug delivery carriers.展开更多
To improve the toughness of silk fibroin( SF) films,poly( ethylene glycol-glycerin)( PEGG) was synthesized with ethylene glycol and epichlorohydrin. The SF / PEGG blend films were prepared by casting aqueous solution ...To improve the toughness of silk fibroin( SF) films,poly( ethylene glycol-glycerin)( PEGG) was synthesized with ethylene glycol and epichlorohydrin. The SF / PEGG blend films were prepared by casting aqueous solution and their structures were characterized. The PEGG was in liquid state at room temperature so it will not be a single phrase at blend film. It crosslinked with SF and made it insolubility in water. The results of X-ray diffraction( XRD) indicated that the crystallinity of the SF in the blend films decreased with the content of PEGG increasing. The tensile strength and elongation at break of blend films were measured using an instron tensile tester. The results showed that the tensile strength and elongation at break of blend films were high enough for application.After the blend films were stored at room temperature for 100 d,the crystallinity, the tensile strength and elongation at wet state increased. The blend films are superior to SF films in providing excellent flexibility and mechanical properties in both dry and wet states. Based on the fact that SF has good biocompatibility,the SF /PEGG blend film will offer new options in many different biomedical applications.展开更多
Objective： To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods： The effects of...Objective： To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods： The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor （VEGF） expression of ECV304 cells, and VEGF, angiopoietin-1 （Ang-1）, platelet-derived growth factor （PDGF） and fibroblast growth factor 2 （FGF2） expression of WI-38 cells were assessed by 3-（4,5）-dimethylthiahiazo （-z-yl）-3,5-di-phenytetrazoliumromide （MTT） assay, viable cell counting, flow cytometry （FCM）, and enzyme-linked immunosorbant assay （ELISA）. Results： We showed that the regenerated silk fibroin film was not cytotoxic to L929 cells and had no adverse influence on their adhesion, cell cycle or apoptosis; it had no adverse influence on the growth and VEGF secretion of ECV304 cells and no effect on the secretion of VEGF, Ang-1, PDGF and FGF2 by WI-38 cells. Conclusion： The regenerated silk fibroin film should be an excellent biomaterial with good cytocompatibility, providing a framework for reparation after trauma in clinical applications.展开更多
In this study, the outstanding biocompatibility of silk fibroin （SF） and the highly efficient anti-bacterial effect of nano silver （NS） were utilized to prepare SF/NS composite film with anti- bacterial property. ...In this study, the outstanding biocompatibility of silk fibroin （SF） and the highly efficient anti-bacterial effect of nano silver （NS） were utilized to prepare SF/NS composite film with anti- bacterial property. The structure and property of the film were characterized. The results showed that the structure of SF in the film was mainly silk I. SF in the film was almost insoluble in water. The tensile strength of film with NS was significantly lower than that of films without NS. When the addition of NS was within the range of 0%-0.6%, the elongation at break had no significant difference. The antibacterial rate of the film on staphylococcus aurens and escherichia coil increased with the amount of NS. The minimum amount of NS in the fdm was O. 1% and the maximum amount was 0.5%.展开更多
The objective of this research was to prepare Eri silk fibroin solution for preparing silk film loaded chlorhexidine drug as model for hydrophilic drug release. The Eri silk cocoons were boiled in 0.5%NaCO3 solution a...The objective of this research was to prepare Eri silk fibroin solution for preparing silk film loaded chlorhexidine drug as model for hydrophilic drug release. The Eri silk cocoons were boiled in 0.5%NaCO3 solution at 90℃, and then left in air dried at room temperature. The fibroin was dissolved in 9M (Ca(NO3)2) with ethanol (2 by mole) and heated at 70℃. The silk fibroin (SF) solution was then dialyzed to exclude salt in phosphate buffer. The SF and gelatin (G) solutions were mixed for preparation of films in both with and without chlorhexidine. The films were observed their morphology under scanning electron microscope. The results found that all of films were rough of their surfaces, homogeneous texture without phase separation. The native SF film composed of pores throughout the film area but did not observe in native G film. The results also showed that the SF and G can be good interacted to form hydrogen bonds. These were indicated from FTIR spectra and thermal analysis. The chlorhexidine drug has not affect on the changes of film properties. However, the releasing pattern of chlorhexidine from each film was varied. The highest rate of drug releasing was found in the native SF film while the native G film was the lowest. It might be suggested that the drug releasing rate was depended on polarity of each polymer components.展开更多
The blend film of silk fibroin (SF) and poly(ethylene glycol)400 (PEG400) with a blend ratio of 2/1 (wt/wt) wasprepared simply by dropping a little PEG400 into the SF solution and then casting the mixed aqueous soluti...The blend film of silk fibroin (SF) and poly(ethylene glycol)400 (PEG400) with a blend ratio of 2/1 (wt/wt) wasprepared simply by dropping a little PEG400 into the SF solution and then casting the mixed aqueous solution at 50℃. Theresulting film exhibited much better mechanical properties in the dry and wet state than SF itself, owing to theconformational change of SF in the blends from the random coil to the β-sheet structure and intermolecular hydrogen bondformation between SF and PEG400. Thermogravimetric analysis showed that the initial thermal decomposition temperatureof the blend film was 170℃, which was 80℃ lower than that of SF (250℃) and 20℃ higher than that of PEG400 (150℃),and indicated a Strong interaction between two components of the blend. No crystalline peaks were observed in the X-raydiffraction curve of the blend film. Cell culture test showed that SF/PEG400 was a suitable substrate for the growth of humanumbilical vein endothelial cells (HUVEC).展开更多
文摘There is an increasing demand for crosslinking methods of silk fibroin (SF) scaffolds in biomedical applications that could maintain the biocompatibility, bioactivity as well as improve the water resistance and mechanical properties of SF materials. In this study, SF was crosslinked effectively with genipin which is a naturally occurring iridoid glucoside and the crosslinking mechanism was investigated through FTIR and amino acid analysis. The results showed that genipin could react with the -NH2 groups on the side chains of SF macromolecules and to form inter- and intra-molecular covalent bonds, and improved the stability of SF materials significantly. In vitro, the performances of genipin-crosslinked SF films were assessed by seeding L929 cells and compared with ethanol-processed SF films, glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked ones. The genipin-crosslinked SF films showed a similar affinity to cells as ethanol-processed ones, and a higher bioactivity in promoting cell growth and proliferation, inhibition of cell apoptosis, and maintenance of normal cell cycle compared with glutaraldehyde and polyethylene glycol diglycidyl ether crosslinked SF films. These features, combined with the decrease of brittleness of SF films crosslinked with chemical methods, substantiated genipin as an effective and biocompatible agent for the manufacturing of bioactive SF materials which used as tissue engineering scaffolds and drug delivery carriers.
基金National Natural Science Foundations of China(Nos.51203107,51373114)the Priority Academic Program Development ofJiangsu Higher Education Institutions,ChinaNatural Science Foundation of Jiangsu Province,China(No.BK20131176)
文摘To improve the toughness of silk fibroin( SF) films,poly( ethylene glycol-glycerin)( PEGG) was synthesized with ethylene glycol and epichlorohydrin. The SF / PEGG blend films were prepared by casting aqueous solution and their structures were characterized. The PEGG was in liquid state at room temperature so it will not be a single phrase at blend film. It crosslinked with SF and made it insolubility in water. The results of X-ray diffraction( XRD) indicated that the crystallinity of the SF in the blend films decreased with the content of PEGG increasing. The tensile strength and elongation at break of blend films were measured using an instron tensile tester. The results showed that the tensile strength and elongation at break of blend films were high enough for application.After the blend films were stored at room temperature for 100 d,the crystallinity, the tensile strength and elongation at wet state increased. The blend films are superior to SF films in providing excellent flexibility and mechanical properties in both dry and wet states. Based on the fact that SF has good biocompatibility,the SF /PEGG blend film will offer new options in many different biomedical applications.
基金supported by the National Basic Research Program (973) of China (No.2005CB623906)the Medical Development Foundation of Soochow University (No.EE134702),China
文摘Objective： To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods： The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor （VEGF） expression of ECV304 cells, and VEGF, angiopoietin-1 （Ang-1）, platelet-derived growth factor （PDGF） and fibroblast growth factor 2 （FGF2） expression of WI-38 cells were assessed by 3-（4,5）-dimethylthiahiazo （-z-yl）-3,5-di-phenytetrazoliumromide （MTT） assay, viable cell counting, flow cytometry （FCM）, and enzyme-linked immunosorbant assay （ELISA）. Results： We showed that the regenerated silk fibroin film was not cytotoxic to L929 cells and had no adverse influence on their adhesion, cell cycle or apoptosis; it had no adverse influence on the growth and VEGF secretion of ECV304 cells and no effect on the secretion of VEGF, Ang-1, PDGF and FGF2 by WI-38 cells. Conclusion： The regenerated silk fibroin film should be an excellent biomaterial with good cytocompatibility, providing a framework for reparation after trauma in clinical applications.
基金National Natural Science Foundations of China，the Priority Academic Program Development of Jiangsu Higher Education Institutions,China
文摘In this study, the outstanding biocompatibility of silk fibroin （SF） and the highly efficient anti-bacterial effect of nano silver （NS） were utilized to prepare SF/NS composite film with anti- bacterial property. The structure and property of the film were characterized. The results showed that the structure of SF in the film was mainly silk I. SF in the film was almost insoluble in water. The tensile strength of film with NS was significantly lower than that of films without NS. When the addition of NS was within the range of 0%-0.6%, the elongation at break had no significant difference. The antibacterial rate of the film on staphylococcus aurens and escherichia coil increased with the amount of NS. The minimum amount of NS in the fdm was O. 1% and the maximum amount was 0.5%.
文摘The objective of this research was to prepare Eri silk fibroin solution for preparing silk film loaded chlorhexidine drug as model for hydrophilic drug release. The Eri silk cocoons were boiled in 0.5%NaCO3 solution at 90℃, and then left in air dried at room temperature. The fibroin was dissolved in 9M (Ca(NO3)2) with ethanol (2 by mole) and heated at 70℃. The silk fibroin (SF) solution was then dialyzed to exclude salt in phosphate buffer. The SF and gelatin (G) solutions were mixed for preparation of films in both with and without chlorhexidine. The films were observed their morphology under scanning electron microscope. The results found that all of films were rough of their surfaces, homogeneous texture without phase separation. The native SF film composed of pores throughout the film area but did not observe in native G film. The results also showed that the SF and G can be good interacted to form hydrogen bonds. These were indicated from FTIR spectra and thermal analysis. The chlorhexidine drug has not affect on the changes of film properties. However, the releasing pattern of chlorhexidine from each film was varied. The highest rate of drug releasing was found in the native SF film while the native G film was the lowest. It might be suggested that the drug releasing rate was depended on polarity of each polymer components.
基金This work was supported by the National Basic Science Research and Development Grants (973) of China (No. G1999064705).
文摘The blend film of silk fibroin (SF) and poly(ethylene glycol)400 (PEG400) with a blend ratio of 2/1 (wt/wt) wasprepared simply by dropping a little PEG400 into the SF solution and then casting the mixed aqueous solution at 50℃. Theresulting film exhibited much better mechanical properties in the dry and wet state than SF itself, owing to theconformational change of SF in the blends from the random coil to the β-sheet structure and intermolecular hydrogen bondformation between SF and PEG400. Thermogravimetric analysis showed that the initial thermal decomposition temperatureof the blend film was 170℃, which was 80℃ lower than that of SF (250℃) and 20℃ higher than that of PEG400 (150℃),and indicated a Strong interaction between two components of the blend. No crystalline peaks were observed in the X-raydiffraction curve of the blend film. Cell culture test showed that SF/PEG400 was a suitable substrate for the growth of humanumbilical vein endothelial cells (HUVEC).
