The Experts below are selected from a list of 4917 Experts worldwide ranked by ideXlab platform
Qingyou Xia - One of the best experts on this subject based on the ideXlab platform.
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Identification of Bombyx mori Sericin 4 protein as a new biological adhesive.
International journal of biological macromolecules, 2019Co-Authors: Zhaoming Dong, Kaiyu Guo, Xiaolu Zhang, Tong Zhang, Yan Zhang, Huaipu Chang, Muya Tang, Qingyou XiaAbstract:Abstract Sericins are large proteins with molecular weights >70 kDa. Three Sericin genes were reported in the silkworm, including Sericin 1, Sericin 2 and Sericin 3. In this study, we have identified a new Sericin gene and designated it as Sericin 4. The sequence, exon-intron structure, alternative splicing, and translation products of this gene have been described in this study. Quantitative RT-PCR analysis indicates that Sericin 4 is expressed in the middle silk gland. Immunofluorescence results show co-localization of Sericin 1 and Sericin 4 in the MSG. Western blot analysis revealed that Sericin 4 was found in the larval silk produced from the second instar to the fourth instar. Two protein bands at approximately 280 kDa and 260 kDa, were detected by western blot for Sericin 4. Two repetitive motifs that are rich in charged amino acids and glutamine have been identified, and they are likely to be responsible for the adhesiveness of Sericin 4. Overall, this study identifies a novel biological adhesive protein and provides new information for understanding how Sericins contribute to the adhesive properties of larval silks.
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fabrication of the fgf1 functionalized Sericin hydrogels with cell proliferation activity for biomedical application using genetically engineered bombyx mori b mori silk
Acta Biomaterialia, 2018Co-Authors: Feng Wang, Ping Zhao, Yuancheng Wang, David L Kaplan, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qingyou XiaAbstract:Abstract Sericin, as the major component of Bombyx mori silk, is a useful biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Here, we report the fabrication of a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. Sericin, together with FGF1, were simultaneously extracted from the silk fiber and then exposed to cold-induced hydrogel formation without additional crosslinking. The fabricated FGF1 Sericin hydrogels demonstrated injectability, useful mechanical properties and a porous microstructure, which contributed to cell adhesion and survival. In addition, FGF1 achieved long-term storage in the Sericin hydrogels over a wide range of temperatures. Further, the Sericin-FGF1 demonstrated sustained release to promote cell proliferation and wound healing. Furthermore, cellular inflammatory responses showed that the FGF1 Sericin hydrogels exhibited biocompatibility and no immunogenicity. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial to expand applications of FGF1 and Sericin in tissue and medical engineering. Further, we demonstrated a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale. Statement of Significance Sericin from the Bombyx mori silk, is regarded as a desirable biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Genetically engineering the Sericin with functional exogenous proteins would enhance its biofunctions and further expand its application in tissue engineering. In this study, we demonstrated a method to fabricate a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. The fabricated FGF1 Sericin hydrogels demonstrated injectability, porous microstructure, biocompatibility and no immunogenicity which contributed to cell adhesion and survival. Remarkably, FGF1 could achieve a long-term stability in the Sericin hydrogels over a wide range of temperatures and sustained release to promote cell proliferation and wound healing. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial in tissue and medical engineering application, and provided a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.
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in situ green synthesis and characterization of Sericin silver nanoparticle composite with effective antibacterial activity and good biocompatibility
Materials Science and Engineering: C, 2017Co-Authors: Gang Tao, Ping Zhao, Yejing Wang, Rui Cai, Pengchao Guo, Liqun Chen, Hua Zuo, Qingyou XiaAbstract:Silver nanoparticle has been widely applied to a variety of fields for its outstanding antimicrobial activity. However, the stability of silver nanoparticle limits its application under certain conditions. Thus, improving the stability of silver nanoparticle via biosynthesis is a promising shortcut to expand its application. Sericin from silkworm cocoon has good hydrophilicity, reaction activity, biocompatibility and biodegradability. In this study, we developed a novel, simple, one-step biosynthesis method to prepare Sericin-silver nanoparticle composite in situ in solution. Sericin served as the reductant of silver ion, the dispersant and stabilizer of the prepared Sericin-silver nanoparticle composite. Natural light was the only power source used to catalyze the synthesis of silver nanoparticle in situ in solution. The novel Sericin-silver nanoparticle composite was characterized by ultraviolet-visible and fluorescence spectroscopy, X-ray diffraction, transmission electron microscopy and fourier transform infrared spectroscopy. The results showed silver nanoparticle could be synthesized through the reduction of AgNO3 by the phenolic hydroxyl group of tyrosine residues of Sericin under the catalysis of natural light. The synthesized silver nanoparticle had good crystalline, size distribution and long-term stability at room temperature. Light irradiation was essential for the preparation of Sericin-silver nanoparticle composite. The antibacterial activity assay showed 25mg/L and 100mg/L were the minimum concentrations of Sericin-silver nanoparticle composite required to inhibit the growth of Staphylococcus aureus and kill this bacterium, respectively. The cytotoxicity assay showed cell viability and cell growth were almost not affected by Sericin-silver nanoparticle composite under the concentration of 25mg/L. Our study suggested the preparation of Sericin-silver nanoparticle composite was environmentally friendly and energy conservation, and the prepared Sericin-silver nanoparticle composite had long-term stability, effective antibacterial activity and good biocompatibility. This novel Sericin-silver nanoparticle composite has shown great potentials for biomedical application such as antibacterial agent and wound care.
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preparation and characterization of silk Sericin pva blend film with silver nanoparticles for potential antimicrobial application
International Journal of Biological Macromolecules, 2017Co-Authors: Rui Cai, Ping Zhao, Gang Tao, Yejing Wang, Pengchao Guo, Liqun Chen, Hua Zuo, Xinyu Liu, Qingyou XiaAbstract:Sericin has great potentials in biomedical applications for its good reactive activity, biocompatibility and biodegradability. However, the undesirable mechanical performance limits its application. Here, we developed a green, facile and economic approach to prepare Sericin/polyvinyl alcohol (PVA) blend film. Further, silver nanoparticles (AgNPs) were synthesized in situ on the surface of Sericin/PVA film via UV-assisted green synthesis method. Mechanical performance, swelling, mass losing and water retention tests showed the blend film had good mechanical performance, hygroscopicity, water retention capacity and low mass losing ratio. Scanning electron microscopy, fourier transfer infrared spectroscopy, X-ray diffractometry diffraction and X-ray photoelectron spectroscopy indicated the blending of PVA and Sericin promoted the formation of hydrogen bond network between Sericin and PVA, thus enhanced the mechanical performance and the stability of Sericin, as well as the hygroscopicity and water retention capacity. UV irradiation and AgNPs modification did not affect the inner crystalline structure of Sericin/PVA blend film. The inhibition zone and bacteria growth curve assay suggested AgNPs-Sericin/PVA film had good antibacterial activities against E. coli and S. aureus. This novel AgNPs-Sericin/PVA film shows great potentials in biomedical materials such as wound dressing and skin tissue engineering.
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advanced silk material spun by a transgenic silkworm promotes cell proliferation for biomedical application
Acta Biomaterialia, 2014Co-Authors: Feng Wang, Ping Zhao, Lin Yuan, Yuancheng Wang, Riyuan Wang, Huan Ding, Chunnuan Song, Zhixin Peng, Zhangchuan Peng, Qingyou XiaAbstract:Natural silk fiber spun by the silkworm Bombyx mori is widely used not only for textile materials, but also for biofunctional materials. In the present study, we genetically engineered an advanced silk material, named hSFSV, using a transgenic silkworm, in which the recombinant human acidic fibroblast growth factor (hFGF1) protein was specifically synthesized in the middle silk gland and secreted into the Sericin layer to surround the silk fiber using our previously optimized Sericin1 expression system. The content of the recombinant hFGF1 in the hSFSV silk was estimated to be approximate 0.07% of the cocoon shell weight. The mechanical properties of hSFSV raw silk fiber were enhanced slightly compared to those of the wild-type raw silk fiber, probably due to the presence of the recombinant of hFGF1 in the Sericin layer. Remarkably, the hSFSV raw silk significantly stimulated the cell growth and proliferation of NIH/3T3 mouse embryonic fibroblast cells, suggesting that the mitogenic activity of recombinant hFGF1 was well maintained and functioned in the Sericin layer of hSFSV raw silk. These results show that the genetically engineered raw silk hSFSV could be used directly as a fine biomedical material for mass application. In addition, the strategy whereby functional recombinant proteins are expressed in the Sericin layer of silk might be used to create more genetically engineered silks with various biofunctions and applications.
Ping Zhao - One of the best experts on this subject based on the ideXlab platform.
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transgenic pdgf bb Sericin hydrogel supports for cell proliferation and osteogenic differentiation
Biomaterials Science, 2020Co-Authors: Ping Zhao, Feng Wang, Yuancheng Wang, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qianqian Yang, Huijie ZhangAbstract:Sericin has been exploited as a biomaterial due to its biocompatibility, biodegradability, and low-immunogenicity as an isolated polymer and support for cell adhesion. In the present study, human platelet-derived growth factor (PDGF-BB)-functionalized Sericin hydrogels were generated using transgenic silkworms, where the as-spun silk incorporated engineered PDGF-BB (termed PDGFM) in the Sericin layers of the cocoons. Sericin and PDGFM were simultaneously extracted from the silk fibroin cocoon fibers, and the soluble extract was then formed into a hydrogel via thermal exposure. The PDGFM Sericin hydrogels exhibited increased β-sheet content and a compressive modulus of 74.91 ± 2.9 kPa comparable to chemically crosslinked Sericin hydrogels (1.68–55.53 kPa) and a porous microstructure, which contributed to cell adhesion and growth. A 13.1% of total extracted PDGFM from the initial silk fibers was incorporated and immobilized in the Sericin hydrogels during material processing, and 1.33% of PDGFM was released over 30 days from the hydrogels in vitro. The remaining PDGFM achieved long-term storage/stability in the Sericin hydrogels for more than 42 days at 37 °C. In addition, the PDGFM Sericin hydrogels were not immunogenic, were biocompatible and bioactive in promoting the support of cell proliferation. When combined with BMP-9, the PDGFM Sericin hydrogels provided synergy to support the osteoblastic differentiation of mesenchymal stem cells (hMSCs) in vitro and in vivo. This study demonstrates that genetically functionalized PDGFM Sericin hydrogels can provide useful biomaterials to support cell and tissue outcomes, here with a focus on osteogenesis.
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fabrication of the fgf1 functionalized Sericin hydrogels with cell proliferation activity for biomedical application using genetically engineered bombyx mori b mori silk
Acta Biomaterialia, 2018Co-Authors: Feng Wang, Ping Zhao, Yuancheng Wang, David L Kaplan, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qingyou XiaAbstract:Abstract Sericin, as the major component of Bombyx mori silk, is a useful biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Here, we report the fabrication of a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. Sericin, together with FGF1, were simultaneously extracted from the silk fiber and then exposed to cold-induced hydrogel formation without additional crosslinking. The fabricated FGF1 Sericin hydrogels demonstrated injectability, useful mechanical properties and a porous microstructure, which contributed to cell adhesion and survival. In addition, FGF1 achieved long-term storage in the Sericin hydrogels over a wide range of temperatures. Further, the Sericin-FGF1 demonstrated sustained release to promote cell proliferation and wound healing. Furthermore, cellular inflammatory responses showed that the FGF1 Sericin hydrogels exhibited biocompatibility and no immunogenicity. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial to expand applications of FGF1 and Sericin in tissue and medical engineering. Further, we demonstrated a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale. Statement of Significance Sericin from the Bombyx mori silk, is regarded as a desirable biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Genetically engineering the Sericin with functional exogenous proteins would enhance its biofunctions and further expand its application in tissue engineering. In this study, we demonstrated a method to fabricate a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. The fabricated FGF1 Sericin hydrogels demonstrated injectability, porous microstructure, biocompatibility and no immunogenicity which contributed to cell adhesion and survival. Remarkably, FGF1 could achieve a long-term stability in the Sericin hydrogels over a wide range of temperatures and sustained release to promote cell proliferation and wound healing. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial in tissue and medical engineering application, and provided a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.
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in situ green synthesis and characterization of Sericin silver nanoparticle composite with effective antibacterial activity and good biocompatibility
Materials Science and Engineering: C, 2017Co-Authors: Gang Tao, Ping Zhao, Yejing Wang, Rui Cai, Pengchao Guo, Liqun Chen, Hua Zuo, Qingyou XiaAbstract:Silver nanoparticle has been widely applied to a variety of fields for its outstanding antimicrobial activity. However, the stability of silver nanoparticle limits its application under certain conditions. Thus, improving the stability of silver nanoparticle via biosynthesis is a promising shortcut to expand its application. Sericin from silkworm cocoon has good hydrophilicity, reaction activity, biocompatibility and biodegradability. In this study, we developed a novel, simple, one-step biosynthesis method to prepare Sericin-silver nanoparticle composite in situ in solution. Sericin served as the reductant of silver ion, the dispersant and stabilizer of the prepared Sericin-silver nanoparticle composite. Natural light was the only power source used to catalyze the synthesis of silver nanoparticle in situ in solution. The novel Sericin-silver nanoparticle composite was characterized by ultraviolet-visible and fluorescence spectroscopy, X-ray diffraction, transmission electron microscopy and fourier transform infrared spectroscopy. The results showed silver nanoparticle could be synthesized through the reduction of AgNO3 by the phenolic hydroxyl group of tyrosine residues of Sericin under the catalysis of natural light. The synthesized silver nanoparticle had good crystalline, size distribution and long-term stability at room temperature. Light irradiation was essential for the preparation of Sericin-silver nanoparticle composite. The antibacterial activity assay showed 25mg/L and 100mg/L were the minimum concentrations of Sericin-silver nanoparticle composite required to inhibit the growth of Staphylococcus aureus and kill this bacterium, respectively. The cytotoxicity assay showed cell viability and cell growth were almost not affected by Sericin-silver nanoparticle composite under the concentration of 25mg/L. Our study suggested the preparation of Sericin-silver nanoparticle composite was environmentally friendly and energy conservation, and the prepared Sericin-silver nanoparticle composite had long-term stability, effective antibacterial activity and good biocompatibility. This novel Sericin-silver nanoparticle composite has shown great potentials for biomedical application such as antibacterial agent and wound care.
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preparation and characterization of silk Sericin pva blend film with silver nanoparticles for potential antimicrobial application
International Journal of Biological Macromolecules, 2017Co-Authors: Rui Cai, Ping Zhao, Gang Tao, Yejing Wang, Pengchao Guo, Liqun Chen, Hua Zuo, Xinyu Liu, Qingyou XiaAbstract:Sericin has great potentials in biomedical applications for its good reactive activity, biocompatibility and biodegradability. However, the undesirable mechanical performance limits its application. Here, we developed a green, facile and economic approach to prepare Sericin/polyvinyl alcohol (PVA) blend film. Further, silver nanoparticles (AgNPs) were synthesized in situ on the surface of Sericin/PVA film via UV-assisted green synthesis method. Mechanical performance, swelling, mass losing and water retention tests showed the blend film had good mechanical performance, hygroscopicity, water retention capacity and low mass losing ratio. Scanning electron microscopy, fourier transfer infrared spectroscopy, X-ray diffractometry diffraction and X-ray photoelectron spectroscopy indicated the blending of PVA and Sericin promoted the formation of hydrogen bond network between Sericin and PVA, thus enhanced the mechanical performance and the stability of Sericin, as well as the hygroscopicity and water retention capacity. UV irradiation and AgNPs modification did not affect the inner crystalline structure of Sericin/PVA blend film. The inhibition zone and bacteria growth curve assay suggested AgNPs-Sericin/PVA film had good antibacterial activities against E. coli and S. aureus. This novel AgNPs-Sericin/PVA film shows great potentials in biomedical materials such as wound dressing and skin tissue engineering.
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advanced silk material spun by a transgenic silkworm promotes cell proliferation for biomedical application
Acta Biomaterialia, 2014Co-Authors: Feng Wang, Ping Zhao, Lin Yuan, Yuancheng Wang, Riyuan Wang, Huan Ding, Chunnuan Song, Zhixin Peng, Zhangchuan Peng, Qingyou XiaAbstract:Natural silk fiber spun by the silkworm Bombyx mori is widely used not only for textile materials, but also for biofunctional materials. In the present study, we genetically engineered an advanced silk material, named hSFSV, using a transgenic silkworm, in which the recombinant human acidic fibroblast growth factor (hFGF1) protein was specifically synthesized in the middle silk gland and secreted into the Sericin layer to surround the silk fiber using our previously optimized Sericin1 expression system. The content of the recombinant hFGF1 in the hSFSV silk was estimated to be approximate 0.07% of the cocoon shell weight. The mechanical properties of hSFSV raw silk fiber were enhanced slightly compared to those of the wild-type raw silk fiber, probably due to the presence of the recombinant of hFGF1 in the Sericin layer. Remarkably, the hSFSV raw silk significantly stimulated the cell growth and proliferation of NIH/3T3 mouse embryonic fibroblast cells, suggesting that the mitogenic activity of recombinant hFGF1 was well maintained and functioned in the Sericin layer of hSFSV raw silk. These results show that the genetically engineered raw silk hSFSV could be used directly as a fine biomedical material for mass application. In addition, the strategy whereby functional recombinant proteins are expressed in the Sericin layer of silk might be used to create more genetically engineered silks with various biofunctions and applications.
Feng Wang - One of the best experts on this subject based on the ideXlab platform.
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transgenic pdgf bb Sericin hydrogel supports for cell proliferation and osteogenic differentiation
Biomaterials Science, 2020Co-Authors: Ping Zhao, Feng Wang, Yuancheng Wang, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qianqian Yang, Huijie ZhangAbstract:Sericin has been exploited as a biomaterial due to its biocompatibility, biodegradability, and low-immunogenicity as an isolated polymer and support for cell adhesion. In the present study, human platelet-derived growth factor (PDGF-BB)-functionalized Sericin hydrogels were generated using transgenic silkworms, where the as-spun silk incorporated engineered PDGF-BB (termed PDGFM) in the Sericin layers of the cocoons. Sericin and PDGFM were simultaneously extracted from the silk fibroin cocoon fibers, and the soluble extract was then formed into a hydrogel via thermal exposure. The PDGFM Sericin hydrogels exhibited increased β-sheet content and a compressive modulus of 74.91 ± 2.9 kPa comparable to chemically crosslinked Sericin hydrogels (1.68–55.53 kPa) and a porous microstructure, which contributed to cell adhesion and growth. A 13.1% of total extracted PDGFM from the initial silk fibers was incorporated and immobilized in the Sericin hydrogels during material processing, and 1.33% of PDGFM was released over 30 days from the hydrogels in vitro. The remaining PDGFM achieved long-term storage/stability in the Sericin hydrogels for more than 42 days at 37 °C. In addition, the PDGFM Sericin hydrogels were not immunogenic, were biocompatible and bioactive in promoting the support of cell proliferation. When combined with BMP-9, the PDGFM Sericin hydrogels provided synergy to support the osteoblastic differentiation of mesenchymal stem cells (hMSCs) in vitro and in vivo. This study demonstrates that genetically functionalized PDGFM Sericin hydrogels can provide useful biomaterials to support cell and tissue outcomes, here with a focus on osteogenesis.
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fabrication of the fgf1 functionalized Sericin hydrogels with cell proliferation activity for biomedical application using genetically engineered bombyx mori b mori silk
Acta Biomaterialia, 2018Co-Authors: Feng Wang, Ping Zhao, Yuancheng Wang, David L Kaplan, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qingyou XiaAbstract:Abstract Sericin, as the major component of Bombyx mori silk, is a useful biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Here, we report the fabrication of a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. Sericin, together with FGF1, were simultaneously extracted from the silk fiber and then exposed to cold-induced hydrogel formation without additional crosslinking. The fabricated FGF1 Sericin hydrogels demonstrated injectability, useful mechanical properties and a porous microstructure, which contributed to cell adhesion and survival. In addition, FGF1 achieved long-term storage in the Sericin hydrogels over a wide range of temperatures. Further, the Sericin-FGF1 demonstrated sustained release to promote cell proliferation and wound healing. Furthermore, cellular inflammatory responses showed that the FGF1 Sericin hydrogels exhibited biocompatibility and no immunogenicity. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial to expand applications of FGF1 and Sericin in tissue and medical engineering. Further, we demonstrated a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale. Statement of Significance Sericin from the Bombyx mori silk, is regarded as a desirable biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Genetically engineering the Sericin with functional exogenous proteins would enhance its biofunctions and further expand its application in tissue engineering. In this study, we demonstrated a method to fabricate a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. The fabricated FGF1 Sericin hydrogels demonstrated injectability, porous microstructure, biocompatibility and no immunogenicity which contributed to cell adhesion and survival. Remarkably, FGF1 could achieve a long-term stability in the Sericin hydrogels over a wide range of temperatures and sustained release to promote cell proliferation and wound healing. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial in tissue and medical engineering application, and provided a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.
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advanced silk material spun by a transgenic silkworm promotes cell proliferation for biomedical application
Acta Biomaterialia, 2014Co-Authors: Feng Wang, Ping Zhao, Lin Yuan, Yuancheng Wang, Riyuan Wang, Huan Ding, Chunnuan Song, Zhixin Peng, Zhangchuan Peng, Qingyou XiaAbstract:Natural silk fiber spun by the silkworm Bombyx mori is widely used not only for textile materials, but also for biofunctional materials. In the present study, we genetically engineered an advanced silk material, named hSFSV, using a transgenic silkworm, in which the recombinant human acidic fibroblast growth factor (hFGF1) protein was specifically synthesized in the middle silk gland and secreted into the Sericin layer to surround the silk fiber using our previously optimized Sericin1 expression system. The content of the recombinant hFGF1 in the hSFSV silk was estimated to be approximate 0.07% of the cocoon shell weight. The mechanical properties of hSFSV raw silk fiber were enhanced slightly compared to those of the wild-type raw silk fiber, probably due to the presence of the recombinant of hFGF1 in the Sericin layer. Remarkably, the hSFSV raw silk significantly stimulated the cell growth and proliferation of NIH/3T3 mouse embryonic fibroblast cells, suggesting that the mitogenic activity of recombinant hFGF1 was well maintained and functioned in the Sericin layer of hSFSV raw silk. These results show that the genetically engineered raw silk hSFSV could be used directly as a fine biomedical material for mass application. In addition, the strategy whereby functional recombinant proteins are expressed in the Sericin layer of silk might be used to create more genetically engineered silks with various biofunctions and applications.
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an optimized Sericin 1 expression system for mass producing recombinant proteins in the middle silk glands of transgenic silkworms
Transgenic Research, 2013Co-Authors: Feng Wang, Lin Yuan, Yuancheng Wang, Xiaoli Duan, Jianping Duan, Zhonghuai Xiang, Qingyou XiaAbstract:The middle silk gland (MSG) of silkworm is thought to be a potential host for mass-producing valuable recombinant proteins. Transgenic MSG expression systems based on the usage of promoter of Sericin1 gene (Sericin-1 expression system) have been established to produce various recombinant proteins in MSG. However, further modifying the activity of the Sericin-1 expression system to yield higher amounts of recombinant proteins is still necessary. In this study, we provide an alternative modification strategy to construct an efficient Sericin-1 expression system by using the hr3 enhancer (hr3 CQ) from a Chongqing strain of the Bombyx mori nuclear polyhedrosis virus (BmNPV) and the 3′UTRs of the fibroin heavy chain (Fib-HPA), the fibroin light chain (Fib-LPA), and Sericin1 (Ser1PA) genes. We first analyzed the effects of these DNA elements on expression of luciferase, and found that the combination of hr3 CQ and Ser1PA was most effective to increase the activity of luciferase. Then, hr3 CQ and Ser1PA were used to modify the Sericin1 expression system. Transgenic silkworms bearing these modified Sericin1 expression vectors were generated by a piggyBac transposon mediated genetic transformation method. Our results showed that mRNA level of DsRed reporter gene in transgenic silkworms containing hr3 CQ and Ser1PA significantly increased by 9 fold to approximately 83 % of that of endogenous Sericin1. As the results of that, the production of recombinant RFP increased by 16 fold to 9.5 % (w/w) of cocoon shell weight. We conclude that this modified Sericin-1 expression system is efficient and will contribute to the MSG as host to mass produce valuable recombinant proteins.
Subhas C. Kundu - One of the best experts on this subject based on the ideXlab platform.
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Non-Mulberry and Mulberry Silk Protein Sericins as Potential Media Supplement for Animal Cell Culture
Hindawi Limited, 2016Co-Authors: Neety Sahu, Shilpa Pal, Sunaina Sapru, Joydip Kundu, Sarmistha Talukdar, Ibotambi N. Singh, Juming Yao, Subhas C. KunduAbstract:Silk protein Sericins, in the recent years, find application in cosmetics and pharmaceuticals and as biomaterials. We investigate the potential of Sericin, extracted from both mulberry Bombyx mori and different non-mulberry sources, namely, tropical tasar, Antheraea mylitta; muga, Antheraea assama; and eri, Samia ricini, as growth supplement in serum-free culture medium. Sericin supplemented media containing different concentrations of Sericins from the different species are examined for attachment, growth, proliferation, and morphology of fibrosarcoma cells. The optimum Sericin supplementation seems to vary with the source of Sericins. The results indicate that all the Sericins promote the growth of L929 cells in serum-free culture media; however, S. ricini Sericin seems to promote better growth of cells amongst other non-mulberry Sericins
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the promotion of osseointegration of titanium surfaces by coating with silk protein Sericin
Biomaterials, 2013Co-Authors: Sunita Nayak, Tuli Dey, Deboki Naskar, Subhas C. KunduAbstract:A promising strategy to influence the osseointegration process around orthopaedic titanium implants is the immobilization of bioactive molecules. This recruits appropriate interaction between the surface and the tissue by directing cells adhesion, proliferation, differentiation and active matrix remodelling. In this study, we aimed to investigate the functionalization of metallic implant titanium with silk protein Sericin. Titanium surface was immobilized with non-mulberry Antheraea mylitta Sericin using glutaraldehyde as crosslinker. To analyse combinatorial effects the Sericin immobilized titanium was further conjugated with integrin binding peptide sequence Arg-Gly-Asp (RGD) using ethyl (dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling agents. The surface of Sericin immobilized titanium was characterized biophysically. Osteoblast-like cells were cultured on Sericin and Sericin/RGD functionalized titanium and found to be more viable than those on pristine titanium. The enhanced adhesion, proliferation, and differentiation of osteoblast cells were observed. RT-PCR analysis showed that mRNA expressions of bone sialoprotein, osteocalcin and alkaline phosphatase were upregulated in osteoblast cells cultured on Sericin and Sericin/RGD immobilized titanium substrates. Additionally, no significant amount of pro-inflammatory cytokines TNF-α, IL-1β and nitric oxide production were recorded when macrophages cells and osteoblast-macrophages co culture cells were grown on Sericin immobilized titanium. The findings demonstrate that the Sericin immobilized titanium surfaces are potentially useful bioactive coated materials for titanium-based medical implants.
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the influence of silkworm species on cellular interactions with novel pva silk Sericin hydrogels
Macromolecular Bioscience, 2012Co-Authors: Khoon S Lim, Subhas C. Kundu, Joydip Kundu, April Reeves, Laura A Poolewarren, Penny J MartensAbstract:Sericin peptides and PVA are chemically modified with methacrylate groups to produce a covalent PVA/Sericin hydrogel. Preservation of the Sericin bioactivity following methacrylation is confirmed, and PVA/Sericin hydrogels are fabricated for both B. mori and A. mylitta Sericin. Cell adhesion studies confirm the preservation of Sericin bioactivity post incorporation in PVA gels. PVA/A. mylitta gels are observed to facilitate cell adhesion to a significantly greater degree than PVA/B. mori gels. Overall, the incorporation of Sericin does not alter the physical properties of the PVA hydrogels but does result in significantly improved cellular interaction, particularly from A. mylitta gels.
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natural protective glue protein Sericin bioengineered by silkworms potential for biomedical and biotechnological applications
Progress in Polymer Science, 2008Co-Authors: Subhas C. Kundu, Biraja C Dash, Rupesh Dash, David L KaplanAbstract:Silk proteins consist of a fibrous core protein, fibroin, and glue proteins called Sericins, which envelop the fibroin fiber with successive sticky layers that help in the formation of a cocoon by cementing together the silk fibers. This system is essentially a protein–fiber composite system, with fibers of the high-molecular-weight fibroin proteins surrounded by the glue-like continuous phase of the Sericin. In mulberry silk produced by Bombyx mori, Ser1 and Ser2 are the two major genes encoding for Sericin. Ser1 is the most abundant silk Sericin of B. mori and consists of repeats of a 38-amino acid motif that contributes towards the hydrophilicity of Sericin. The protein has partially been characterized from non-mulberry silkworms like the tropical tasar Antheraea mylitta, and studied for various potential applications because of its unique biochemical and biophysical properties. These applications include the skincare and food industries, as coating material for biomedical applications like anticoagulants, in anticancer drugs, for drug delivery, and in tissue engineering. Sericin promotes proliferation of cells when used as a constituent of cell culture in serum-free media. Sericin also supports cell adhesion and proliferation when used in pure form and/or blended in matrices. Sericin films, 3D scaffolds, nanoparticles, composites, conjugated drugs, and recombinant Sericins offer potential future options for these needs.
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silk Sericin protein of tropical tasar silkworm inhibits uvb induced apoptosis in human skin keratinocytes
Molecular and Cellular Biochemistry, 2008Co-Authors: Rupesh Dash, Mahitosh Mandal, Sudip Kumar Ghosh, Subhas C. KunduAbstract:The silk protein Sericin has been identified as a potent antioxidant and photoprotective agent against ultraviolet B (UVB) irradiation in mouse skin model. In this study, we have investigated the anti-apoptotic effect of Sericin in UVB (30 mJ/cm2)-irradiated human keratinocytes. Flow cytometry analysis has shown that pre-treatment with Sericin inhibits UVB-induced apoptosis. The pre-treatment with Sericin suppresses bax expression, up-regulates the expression of bcl-2, prevents both the activation of caspase-3 and cleavage of Poly (ADP-ribose) polymerase. Generation of intracellular hydrogen peroxide in UVB-treated keratinocytes is inhibited through pre-treatment with Sericin suggesting that Sericin probably prevents mitochondrial damage.
Yuancheng Wang - One of the best experts on this subject based on the ideXlab platform.
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transgenic pdgf bb Sericin hydrogel supports for cell proliferation and osteogenic differentiation
Biomaterials Science, 2020Co-Authors: Ping Zhao, Feng Wang, Yuancheng Wang, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qianqian Yang, Huijie ZhangAbstract:Sericin has been exploited as a biomaterial due to its biocompatibility, biodegradability, and low-immunogenicity as an isolated polymer and support for cell adhesion. In the present study, human platelet-derived growth factor (PDGF-BB)-functionalized Sericin hydrogels were generated using transgenic silkworms, where the as-spun silk incorporated engineered PDGF-BB (termed PDGFM) in the Sericin layers of the cocoons. Sericin and PDGFM were simultaneously extracted from the silk fibroin cocoon fibers, and the soluble extract was then formed into a hydrogel via thermal exposure. The PDGFM Sericin hydrogels exhibited increased β-sheet content and a compressive modulus of 74.91 ± 2.9 kPa comparable to chemically crosslinked Sericin hydrogels (1.68–55.53 kPa) and a porous microstructure, which contributed to cell adhesion and growth. A 13.1% of total extracted PDGFM from the initial silk fibers was incorporated and immobilized in the Sericin hydrogels during material processing, and 1.33% of PDGFM was released over 30 days from the hydrogels in vitro. The remaining PDGFM achieved long-term storage/stability in the Sericin hydrogels for more than 42 days at 37 °C. In addition, the PDGFM Sericin hydrogels were not immunogenic, were biocompatible and bioactive in promoting the support of cell proliferation. When combined with BMP-9, the PDGFM Sericin hydrogels provided synergy to support the osteoblastic differentiation of mesenchymal stem cells (hMSCs) in vitro and in vivo. This study demonstrates that genetically functionalized PDGFM Sericin hydrogels can provide useful biomaterials to support cell and tissue outcomes, here with a focus on osteogenesis.
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fabrication of the fgf1 functionalized Sericin hydrogels with cell proliferation activity for biomedical application using genetically engineered bombyx mori b mori silk
Acta Biomaterialia, 2018Co-Authors: Feng Wang, Ping Zhao, Yuancheng Wang, David L Kaplan, Riyuan Wang, Chi Tian, Kai Hou, Wenjing Chen, Qingyou XiaAbstract:Abstract Sericin, as the major component of Bombyx mori silk, is a useful biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Here, we report the fabrication of a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. Sericin, together with FGF1, were simultaneously extracted from the silk fiber and then exposed to cold-induced hydrogel formation without additional crosslinking. The fabricated FGF1 Sericin hydrogels demonstrated injectability, useful mechanical properties and a porous microstructure, which contributed to cell adhesion and survival. In addition, FGF1 achieved long-term storage in the Sericin hydrogels over a wide range of temperatures. Further, the Sericin-FGF1 demonstrated sustained release to promote cell proliferation and wound healing. Furthermore, cellular inflammatory responses showed that the FGF1 Sericin hydrogels exhibited biocompatibility and no immunogenicity. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial to expand applications of FGF1 and Sericin in tissue and medical engineering. Further, we demonstrated a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale. Statement of Significance Sericin from the Bombyx mori silk, is regarded as a desirable biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Genetically engineering the Sericin with functional exogenous proteins would enhance its biofunctions and further expand its application in tissue engineering. In this study, we demonstrated a method to fabricate a human acidic fibroblast growth factor (FGF1)-functionalized Sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its Sericin layer. The fabricated FGF1 Sericin hydrogels demonstrated injectability, porous microstructure, biocompatibility and no immunogenicity which contributed to cell adhesion and survival. Remarkably, FGF1 could achieve a long-term stability in the Sericin hydrogels over a wide range of temperatures and sustained release to promote cell proliferation and wound healing. This study revealed the successful exploration of FGF1-functionalized Sericin hydrogels as a new protein-based biomaterial in tissue and medical engineering application, and provided a strategy for the predesign of exogenous protein-functionalized Sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.
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advanced silk material spun by a transgenic silkworm promotes cell proliferation for biomedical application
Acta Biomaterialia, 2014Co-Authors: Feng Wang, Ping Zhao, Lin Yuan, Yuancheng Wang, Riyuan Wang, Huan Ding, Chunnuan Song, Zhixin Peng, Zhangchuan Peng, Qingyou XiaAbstract:Natural silk fiber spun by the silkworm Bombyx mori is widely used not only for textile materials, but also for biofunctional materials. In the present study, we genetically engineered an advanced silk material, named hSFSV, using a transgenic silkworm, in which the recombinant human acidic fibroblast growth factor (hFGF1) protein was specifically synthesized in the middle silk gland and secreted into the Sericin layer to surround the silk fiber using our previously optimized Sericin1 expression system. The content of the recombinant hFGF1 in the hSFSV silk was estimated to be approximate 0.07% of the cocoon shell weight. The mechanical properties of hSFSV raw silk fiber were enhanced slightly compared to those of the wild-type raw silk fiber, probably due to the presence of the recombinant of hFGF1 in the Sericin layer. Remarkably, the hSFSV raw silk significantly stimulated the cell growth and proliferation of NIH/3T3 mouse embryonic fibroblast cells, suggesting that the mitogenic activity of recombinant hFGF1 was well maintained and functioned in the Sericin layer of hSFSV raw silk. These results show that the genetically engineered raw silk hSFSV could be used directly as a fine biomedical material for mass application. In addition, the strategy whereby functional recombinant proteins are expressed in the Sericin layer of silk might be used to create more genetically engineered silks with various biofunctions and applications.
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an optimized Sericin 1 expression system for mass producing recombinant proteins in the middle silk glands of transgenic silkworms
Transgenic Research, 2013Co-Authors: Feng Wang, Lin Yuan, Yuancheng Wang, Xiaoli Duan, Jianping Duan, Zhonghuai Xiang, Qingyou XiaAbstract:The middle silk gland (MSG) of silkworm is thought to be a potential host for mass-producing valuable recombinant proteins. Transgenic MSG expression systems based on the usage of promoter of Sericin1 gene (Sericin-1 expression system) have been established to produce various recombinant proteins in MSG. However, further modifying the activity of the Sericin-1 expression system to yield higher amounts of recombinant proteins is still necessary. In this study, we provide an alternative modification strategy to construct an efficient Sericin-1 expression system by using the hr3 enhancer (hr3 CQ) from a Chongqing strain of the Bombyx mori nuclear polyhedrosis virus (BmNPV) and the 3′UTRs of the fibroin heavy chain (Fib-HPA), the fibroin light chain (Fib-LPA), and Sericin1 (Ser1PA) genes. We first analyzed the effects of these DNA elements on expression of luciferase, and found that the combination of hr3 CQ and Ser1PA was most effective to increase the activity of luciferase. Then, hr3 CQ and Ser1PA were used to modify the Sericin1 expression system. Transgenic silkworms bearing these modified Sericin1 expression vectors were generated by a piggyBac transposon mediated genetic transformation method. Our results showed that mRNA level of DsRed reporter gene in transgenic silkworms containing hr3 CQ and Ser1PA significantly increased by 9 fold to approximately 83 % of that of endogenous Sericin1. As the results of that, the production of recombinant RFP increased by 16 fold to 9.5 % (w/w) of cocoon shell weight. We conclude that this modified Sericin-1 expression system is efficient and will contribute to the MSG as host to mass produce valuable recombinant proteins.
