• Journal of Sericultural and Entomological Science
• /
• v.52 no.2
• /
• pp.102-109
• /
• 2014

We constructed the fibroin H-chain expression system to produce enhanced yellow fluorescent proteins (EYFP) in the silk of transgenic silkworm. Fluorescent silk could be made by fusing EYFP cDNA to the heavy chain gene and injecting it into a silkworm. The EYFP fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the EYFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The yellow fluorescence proving that the fusion protein was present in the silk. Accordingly, we suggest that the EYFP fluorescence silk will enable the production of novel biomaterial based on the transgenic silk.

• Polymer(Korea)
• /
• v.35 no.1
• /
• pp.7-12
• /
• 2011

Poly(lactic-co-glycolic acid) (PLGA) is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. Also, it can be easily fabricated into many shapes. Silk fibroin contains powerful bioactive molecules. We fabricated natural/synthetic hybrid films using 0, 10, 20, 40 and 80 wt% of silk fibroin. Schwann cells (SCs) were seeded on PLGA/silk fibroin hybrid films and confirmed the effects of adhesion and proliferation on SCs according to the content of silk fibroin. In this study, we confirmed PLGA/silk fibroin hybrid film containing 40% and 80% of silk fibroin interrupted adhesion and proliferation of SCs. Films containing 10% and 20% of silk, however, provided suitable environment for growth and proliferation of SCs. These results suggest that silk fibroin provides suitables surface to neural cells and its proper content provides proper culture conditions to improve cell adhesion and proliferation.

• Journal of Sericultural and Entomological Science
• /
• v.51 no.2
• /
• pp.153-158
• /
• 2013

To express green fluorescent protein in the cocoon of silkworm, we constructed the fibroin H-chain expression system to produce enhanced green fluorescent protein (EGFP) in the cocoon of transgenic silkworms. The EGFP fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, was designed to be secreted into the lumen of the posterior silk glands. The expression of the EGFP/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3-driven DsRed2 cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. A mixture of the donor and helper vector was micro-injected into 1,200 eggs of bivoltin silkworms, Baegokjam. We obtained 8 broods. The cocoon displayed strong green fluorescence, proving that the fusion protein was present in the cocoon. Also, the presence of fusion proteins in cocoons was demonstrated by SDS-PAGE and immunoblotting. Accordingly, we suggest that the EGFP fluorescence silk will enable the production of the novel biomaterial based on the transgenic silk.

• Journal of Sericultural and Entomological Science
• /
• v.51 no.1
• /
• pp.68-72
• /
• 2013

Fibroin and sericin are natural proteins obtained from cocoon and one of the spotlight materials for medical device. Medical device made of these proteins also has the advantage that this material is biodegradable in to amino acid. In this study, we prepared silk sponges using fibroin, sericin and additives. The characterizations of the silk sponges such as morphology, stability, and blood absorbency were observed. The structural stability of the silk sponge decreased significantly by increasing sericin contents. The effect on the concentrations of ethanol to induce crystallization was observed to be superior to 70% ethanol. Structural stability of silk sponges containing additives was very lower than those not containing additives. The blood absorbency of the silk sponges was found to be excellent, regardless of the composition of sericin and fibroin. The resilient power of these sponges was also very good, in spite of the repeated soaking and drying. Therefore, we expect that the silk sponges can be used medical supplies such as plastic implants and hemostatic cotton.

• Journal of Sericultural and Entomological Science
• /
• v.52 no.1
• /
• pp.10-15
• /
• 2014

Regenerated Antheraea pernyi silk sponge was prepared using calcium nitrate 4 hydrate melt and examined the conformational changes treated with aqueous ethanol solution. The conformation of silk sponges was changed from random coil structure to ${\beta}$-sheet and ${\alpha}$-helix conformation with low ethanol concentration (50 ~ 70%). On the other hand, that of silk sponges with 80% ethanol treatment showed ${\beta}$-sheet ($700cm^{-1}$), ${\alpha}$-helix ($625cm^{-1}$), and random coil ($660cm^{-1}$) specific peaks. Wound healing effect in vitro was observed by cytoslective wound healing kit. Therefore, regenerated Antheraea pernyi silk sponges might be used as promising wound dressing materials.

• Journal of Sericultural and Entomological Science
• /
• v.50 no.1
• /
• pp.15-19
• /
• 2012

A number of researcher have studied biomaterials for cartilage regeneration and are now proceeding. Silk protein was attempted for use as biomedical materials by many researchers because it is natural polymer with biocompatibility and excellent mechanical strength. In this study, we want to know a possibility of silk protein on the cartilage regeneration. We isolated chondrocytes from nasal cartilage and confirmed optimal culture condition of the cells. To observe the effects of silk fibroin on chondrogenesis, we added silk fibroin solutions to the culture medium of chondrocyte and detected gene expression levels related chondrogenesis such as col2, col10. The chondrocytes showed optimal growth when they were cultured in DMEM medium supplemented with 10% FBS 100 ${\cdot}{\ddot{I}}$M ascorbic acid. The levels of col2 gene expression were increased in non-autoclaved silk fibroin, but decreased in autoclaved one. Also the gene expression levels of col10 were increased in silk fibroin, particulary at 3D culture. Based on the results of this study, we had seen the possibility of silk fibroin for cartilage regeneration. In future studies, we should know more clearly the relationship between cartilage regeneration and the silk protein.

• Proceedings of the Korean Fiber Society Conference
• /
• 2003.04a
• /
• pp.388-389
• /
• 2003

전기방사를 이용하여 나노섬유를 제조하려는 많은 연구가 진행되고 있다. 고분자 재료 측면에서 피브로인, 키토산, 콜라겐 등 천연고분자는 생분해성, 생체적합성, 환경친화성이 뛰어나 이들을 나노섬유소재로 개발할 경우 의료용소재 뿐만아니라 생명공학소재로서의 이용가능성이 크다. 특히, 실크피브로인은 생체적합성, 산소투과능, 세포부착능 둥 생체재료로서의 성능뿐만 아니라 우수한 물성과 기계적 성질을 가지고 있으므로 다양한 형태로 성형화시킴으로써 소재 성능을 높일 수 있다. (중략)

• Polymer(Korea)
• /
• v.36 no.5
• /
• pp.651-655
• /
• 2012

Considerable effort has been directed toward the use of silk fibroin as a biotechnological material in biomedical applications on account of its excellent biodegradability, biocompatibility, and unique mechanical properties. For use in tissue engineering, it is very important to design and control the pore architecture of polymeric scaffolds, which provide the vital framework for seeded cells to organize into functioning tissue. In the present study, a silk fibroin scaffold with controlled interconnectivity and pore size was prepared using an electrospinning method with poly(ethylene oxide).

• Proceedings of the Korean Fiber Society Conference
• /
• 2003.04a
• /
• pp.293-294
• /
• 2003

견 피브로인은 대표적인 섬유상 단백질의 하나로 생체적합성, 생분해성, 저독성 등의 유용한 특성을 가지므로 생체재료로 상당한 관심과 연구의 대상이 되어왔다. 우리는 최근의 연구에서 견 피브로인을 생사로부터 추출한 다음, 포름산을 용제로 하여 전기방사함으로써 나노섬유를 제조하고 이들의 각화세포에 대한 친화력을 확인한 바 있다. 본 연구에서는 견 피브로인의 구조체를 나노섬유 부직포, 필름, 마이크로 섬유로 구성된 직물 둥의 형태로 하여 그들의 2차 구조를 비교함과 동시에 구조적 특성이 각화세포와의 친화력에 어떠한 영향을 미치는 가를 비교ㆍ검토 하고자 하였다. (중략)

• Polymer Science and Technology
• /
• v.16 no.5
• /
• pp.556-567
• /
• 2005