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Previous studies indicate that BF-7 enhances learning and memory in normal and elderly individuals. Here, we evaluated whether milk containing BF-7 (BF-7 milk) could improve the brain function, with thirty normal university students . Two versions of the Paced Auditory Serial Addition Test were used under double-blinded conditions to measure the efficacy of BF-7 milk on learning and memory, especially working memory and attention, and on mathematical ability. As a result, BF-7 milk improved the accuracy of the task more than 3-fold. Furthermore, BF-7 milk protected cultured neuronal cells from 3-hydroxykynurenine, a normal endogenous brain stress agent. These results indicate that BF-7 milk enhances memory, attention and mathematical ability in normal persons.

The antioxidant activity of silkworm powder treated by proteolytic enzyme was investigated. Total protein content of silkworm power was assayed using BCA, Bradford assays and SDS-polyacrylamide gel electrophoresis (PAGE) with alkaline protease treatment conditions including temperature and pH. The optimum condition of alkaline protease treatment for silkworm powder was found to be 60℃ and pH 7. The alkaline protease treatment resulted in increased contents of free amino acids, total polyphenol and total flavonoid compared to control group. The silkworm hydrolysates showed excellent antioxidant activities in various in vitro models such as 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, 2,2 - azino-bis(3-ethylbenzthiazoline-6)-sulfonic acid (ABTS) radical scavenging activity. These results provide useful information for using silkworm powder as an ingredient in functional foods and for exploiting alkaline protease treatment to improve the extractability and bioactivity of a raw material.

Hydrogels are crosslinked hydrophilic matrices for a variety of biomedical applications. Silk fibroin (SF), one of typical natural biomaterials, has been explored as base material for hydrogel. Photocrosslinked SF hydrogel containing poly(ethylene glycol) (PEG) was formulated through visible light initiated thiol-acrylate photopolymerization. The morphological, structural and thermal properties of SF . PEG hydrogel was investigated through scanning electron microscopy, X-ray diffractometry, thermogravimetry, and differential scanning calorimetry. The morphology of SF hydrogel showed dot and uneven surface with network cross-section. X-ray diffraction curves showed that the specific diffraction peaks of PEG were not changed by the intensity of the peaks were affected by sonication. Thermo-degradation behavior of SF . PEG hydrogel sonicated was significantly affected and became complex pattern compared to unsonicated ones. However, the melting endothermic temperature of SF-PEG hydrogel was not changed but the crystalline enthalpy was decreased by gelation and sonication.

Silk fibroin (SF) and gelatin are natural polymers suitable for biomedical applications, including controlled protein release. SF offers high mechanical strength and slow enzymatic degradability, whereas gelatin contains bioactive motifs that can provide biomimicry to the resulting scaffolds. Owing to their complementary material properties, SF and gelatin are increasingly being used together to afford hybrid scaffolds with adjustable properties. Here, we report the use of in situ crosslinked SF/gelatin hydrogels as a platform for tunable growth factor sequestration and delivery. We demonstrate that the physical assembly of SF into insoluble networks could be accelerated by sonication even in the presence of gelatin. However, the processing conditions from which to prepare SF aqueous solution (e.g., heating duration and number of processing steps) drastically altered the resulting hydrogel physical properties. Furthermore, the stiffness of SF/gelatin hybrid gels displayed temperature dependency. Specifically, incorporation of gelatin increased gel stiffness at 25 °C but decreases hydrogel mechanical stability at 37 °C. The thermostability of SF/gelatin gels can be restored by using a low concentration of genipin, a naturally derived chemical crosslinker. We also incorporate heparin-conjugated gelatin (GH) into the hydrogels to create a hybrid matrix capable of sequestering growth factors, such as basic fibroblast growth factor (bFGF). Both sonicated SF (SSF) and hybrid SSF-GH gels exhibit moderate bFGF sequestration, but only SSF-GH gels afford slow release of bFGF. On the other hand, genipin-stabilized network exhibited the highest retention and sustained release of bFGF, suggesting the suitability of this particular formulation as a scaffold for tissue engineering applications.

Silk fibroin (SF) is a natural polymer widely used and studied for diverse applications in the biomedical field. Recently, genetically modified silks, particularly fluorescent SF fibers, were reported to have been produced from transgenic silkworms. However, they are currently limited to textile manufacturing. To expand the use of transgenic silkworms for biomedical applications, a solution form of fluorescent SF needed to be developed. Here, we describe a novel method of preparing a fluorescent SF solution and demonstrate long-term fluorescent function up to one year after subcutaneous insertion. We also show that fluorescent SF labeled p53 antibodies clearly identify HeLa cells, indicating the applicability of fluorescent SF to cancer detection and bio-imaging. Furthermore, we demonstrate the intraoperative use of fluorescent SF in an animal model to detect a small esophageal perforation (0.5 mm). This study suggests how fluorescent SF biomaterials can be applied in biotechnology and clinical medicine.

The structural transition of silk protein has been induced by various method including alcohol treatment. To know the effect of alcohol concentration on silk beads conformation, silk beads were prepared in different alcohol concentration (100%, 70%, and 50%) and then examined the infrared spectra of silk beads. Silk beads prepared in 100% alcohol showed at attributed conformation and did not showed a characteristic absorption peak from model drug. However, silk beads in 70% and 50% alcohol showed some peaks originated from model drug including , , and . These results means that the micro-environment of silk beads was affected by alcohol concentration.

There is no specific data on Entomogenous fungus (DongChungHaCho) in accordance with the silkworm varieties, despite of very high value industrial use as functional materials at domestic and abroad. In this study, we investigated culture condition and characteristics of Paecilomyces tenuipes in 3 silkworm cultivars and 2 specific silkworm breeds. Infection rate of P. tenuipes for silkworm varieties was the highest Kumokjam, followed by Baegokjam, Daeseongjam, Golden silk, Yeonnokjam in that order. The optimum culture conditions were as follows: dark condition, , and about in planting density per pupa. Also, we have developed annual production technologies of P. tenuipes. First, in order that the moisture content of the infection pupa became 3% or less in the natural condition. Second, the dried pupa must be vacuum-packed and was kept under . Finally, by restoring the moisture content of the dried pupa and culture in optimal condition, the technique for annual production of the P. tenuipes was achieved. Therefore, we expect that the use of Kumokjam and annual production technique will contribute significantly to farm income.

It is known that silk protein supports effectively proliferation of cell such as insect cell and hybridoma cell. Althoughthere are many varieties of Bombyx mori silkworm, the effect of silkworm varieties on cell proliferation has not beenconsidered in detail. We studied that characteristics of silk cocoon obtained from Baegokjam, Kumokjam, Daeseongjamsilkworm varieties and whether silk protein affected cell proliferation or not. Silk sericin was prepared under high tem-perature and high pressure condition. Silk fibroin was prepared using CaCl 2 :H 2 O : EtOH with different dissolutiontime. As a result, there are differences in silk cocoon from different silkworm varieties about cell proliferation. The pro-liferation was accelerated in the presence of Baegokjam silk sericin and Kumokjam silk fibroin with 5hr dissolutiontime. We expect that silk proteins could be a preferable culture medium supplement for stimulating the proliferation ofcell. Then, this results suggest silk as a new material for medium supplement replacing with fetal bovine serum.Key words : Silk fibroin, Silk sericin, Silkworm variety, Cell proliferation

Korean commercial Bombyx mori silkworm cocoon made by Kumokjam and Daesungjam was examined to characterize for application. Amino acid analysis showed that the main amino acid of Kumokjam and Daesungjam cocoon was glycine, alanine, serine, tyrosine, and aspartic acid. The content of three major amino acids was about 67% and the content of long chain amino acids was about 37%. Stress-strain curve showed that breaking stress of Kumokjam cocoon, 50 MPa, was higher than that of Daesungjam cocoon, 48 MPa. Breaking strain of Kumokjam cocoon, 37%, was also longer than that of Daesungjam cocoon, 30%. These results might be used as basic information for development of non-textile materials using Bombyx mori silkworm cocoon.

Enzymatic modification of proteins is often used to increase the biological activity of materials. Silkworm powder has been investigated as a functional food resource, but no study has been performed on its modification by commercial food enzyme. Therefore, this study aimed to determine the feasibility of such modification of silkworm powder by alkaline protease. The activity of the enzyme was confirmed using an azocasein assay. Subsequently the silkworm powder was hydrolyzed by enzymatic treatment. UV visible spectrometry showed that the supernatant of silkworm powder subjected to enzymatic treatment had a stronger absorption band than the untreated powder. SDS-PAGE electrophoresis showed that the molecular weight of silkworm powder decreased on enzymatic treatment. Thus the results indicate that commercial enzymes might be used to modify the characteristics of silkworm powder.

【Recently, many researchers have studied silk sericin because of its high water retention, good wound healing, good cyto-compatibility, and blood-glucose- and cholesterol-lowering effects. Although sericin film can be used in wound dressing and cosmetic packs, its poor mechanical properties have prevented its use in industrial fields. In the present study, sericin was obtained from different silkworm varieties, and the effect of silkworm variety on the structure and properties of sericin was examined. Except for a small difference in serine content, no significant difference in sericin was noted among the silkworm varieties. In addition, silkworm variety almost had no effect on solution viscosity, implying that it does not influence the molecular weight of sericin. Mechanical properties of sericin film were strongly affected by silkworm variety. Wonwon 126 showed the best mechanical properties, while N74 and Geumgwangju displayed the worst properties.】

This study aimed to investigate the effects of silk fibroin on bone metabolism in ovariectomized rats. A total of 30 Sprague-Dawley rats were randomized into sham-operated (SHAM), ovariectomized control (OVX), alendronate (OVX+ALEN, 10 mg/kg body weight/d), low silk fibroin (OVX+SF100, 100 mg/kg body weight/d), and high silk fibroin (OVX+SF300, 300 mg/kg body weight/d) groups. All the rats were fed by gavage for 12 wk. At the end of 12 wk, blood and urine were collected for analysis of bone turnover markers, and bone mineral density (BMD) was measured by micro-computed tomography. The results show that the OVX group (p < 0.05) displayed the highest mean body weight gain. Among the five groups, serum levels of bone alkaline phosphatase (ALP) and urine levels of deoxypyridinoline (DPD) were highest in the OVX group (p < 0.05). Bone ALP levels in the ALEN group were significantly lower than that of the silk-treated groups. On the other hand, DPD levels were not significantly different between the ALEN and silk-fibroin-treated groups ( p < 0.05). The trabecular BMD was significantly higher in the ALEN and silk-treated groups compared to the OVX group ( p < 0.05). In conclusion, this study showed that silk fibroin has similar effects as alendronate, which is used in osteoporosis medication. Therefore silk fibroin might be a new candidate for the prevention and treatment of osteoporosis in patients.

The objective of this study was to evaluate bone regeneration ability using different concentrations of tetracycline (TC) loaded into silk fibroin membranes (SFMs). Prior to animal experiments, MTT assays and alkaline phosphatase (ALP) assays were performed to evaluate the cellular response of each membrane. Critical sized bone defects (8-mm diameter) on rat calvaria were prepared and covered with SFM containing different concentrations of TC:1% TC (TC1), 5% TC (TC5), 10% TC (TC10), and 0% TC (SFM only). The bone regeneration was evaluated by micro-computerized tomography (μ-CT) and histomorphometric analysis at 4 weeks postoperatively. ALP activity was increased in a dose-dependent manner as the applied TC concentrations. By μ-CT analysis, newly formed bone volume in the TC5 group was significantly higher than that in the SFM only group (P 0.05). In conclusion, TC-loaded SFM showed more bone formation than SFM without TC, and the amount of new bone formation was dependent on TC concentrations.

In this study, functionality of products of silkworm hemolymph fermented by Bacillus species was studied such as cellviability, antioxidant effect, and inhibitory effect on tyrosinase activities. A matured silkworm hemolymph was degradedby fermentation with Bacillus subtilis 10854 and Bacillus amyloliquefaciens M27. Especially, proteins of matured silk-worm hemolymph were degraded to 3,000 Da by fermentation with B. amyloliquefaciens M27. Cell viability for MTTassay was higher than PBS in hemolymph and fermented hemolymph. A DPPH free radical scavenging activity of fer-mented hemolymph was as higher as Vitamin C and dependent on sample concentrations. Thus, these results suggestthat degraded hemolymph fermented by B. amyloliquefaciens M27 may have antioxidant properties as a material forcosmetics.Key word : Silkworm, Hemolymph, Fermentation, Bacillus 서론 우리나라에 해외기술이 도입되고 화장품 연구개발이 본격적으로 이루어지기 시작한 1960년대 이후 국내 화장품시장 및 연구수준은 비약적인 발전을 이루었다. 2013년국내 화장품 시장 규모는 17조 6,000억으로 특히 국산화장품의 시장 점유율이 60%에 이르는 것으로 조사되었으며, 이는 국내에서 생산되는 원료로 다양한 과학기술을이용하여 생산한 화장품의 경쟁력이 높아지고 있다는 것을 의미한다. 최근에는 미백, 보습, 자외선 차단, 주름개선과 같은 화장품의 트랜드를 반영하기 위하여, 천연물의기능성을 활용하는 화장품 연구개발이 활발하게 이루어지고 있다. 흑마늘은 일반 마늘에 비해 생리활성물질의 증가로 새로운 기능성 소재로 각광받고 있으며 흑마늘의 티로시나아제(tyrosinase)와 엘라스타아제(elasetase) 활성 억제능과 자외선으로 인한 멜라닌 내 과산화물의 억제 효과에 대해 보고되어 있다(Jung et al. 2010, Lee et al. 2010).오미자씨 오일은 높은 안정성과 DPPH 라디칼 소거능 및콜라겐 합성능력이 뛰어나 기능성 화장품 소재로의 가능성이 보고된 바 있다(Yang 2012). 또한, 이러한 천연물을단순추출 및 사용하는 것 뿐만 아니라 미생물 발효에 의해 생성된 생리활성물질의 기능성을 평가하는 연구도 보고되고 있다. 균주로는 유산균(락토바실러스, Lactobacillus)과 효모(사카로미세스속, Saccharomyces), 고초균(바실러스속, Bacillus)을 주로 사용하고 있으며, Shim et al.(2012)에따르면 유산균으로 발효한 더덕 발효추출물이 pH 및 점도변화에 안정하고 경피 수분증발량을 억제하여 보습성을 높인다고 보고하였다. 마디풀 속과에 속하는 호장근(Polygonumcuspidata SIEB. et ZUCC.)을 효모균(Saccharomycescerevisiae)으로 발효시켜 SOD(Superoxide dismutase),catalase 항산화 효소활성을 측정한 연구에서 호장근 발효추출물은 농도 의존적으로 항산화 효소 활성이 증가하였다고 보고되었다(Lim et al. 2010). 바실러스속 세균은 자연계에 널리 존재하는 그람 양성의 간균으로 호기성 또는 통성 혐기성 조건에서 생육하는 것으로 알려져 있고주로 전분과 단백질 분해효소를 생성하는 것으로 연구되

We analyzed rutin content using mulberry genetic resources. They were grown under the same environment and con-ditions. Mulberry leaves were collected and then freeze-dried and powdered for rutin test. As a result, mean content ofmulberry strains was 0.52 ± 0.25%, and the coefficient of variation (CV) was 47.9%. The variation between the strainswas greatly severe. Among the tested strains, ‘Buyoungsang’ was showed the highest content of 1.37%, whereas‘Cheongeunsipmunja’ and ‘Mujeonsipmunja’ were showed the lowest content of 0.06% respectively. The content ofrutin of 16 mulberry varieties for silkworm rearing were compared. ‘Cheongolppong’ was showed the highest contentof 0.69%, whereas ‘Cheongilppong’ was showed the lowest content of 0.14 %. Finally we selected rutin high-containing9 strains. They are as follows. ‘Buyoungsang’, ‘Yeulbon’, 'Dangsang 5’, ‘A8’, ‘Aja’, ‘Seongsu 5’, ‘Sawonppong 12’,‘Hwanyoupjosaengnosang’ and ‘Hwansipjosaeng’ which are more than twice of the overall average content. Key words : Mulberry leaf, Rutin, Genetic resource

Propionibacterium acnes (P. acnes) is a major contributing factor to the inflammatory component of acne. The many prescription medications for acne allow for a large number of potential combination treatments. However, several antibiotics, apart from their antibacterial effects, exert side‑effects, such as the suppression of host inflammatory responses. Purified bee venom (BV) is a natural toxin produced by honeybees (Apis mellifera L.). BV has been widely used as a traditional medicine for various diseases. In the present study, to investigate the therapeutic effects of BV against P. acnes-induced inflammatory skin disease, P. acnes was intradermally injected into the ears of mice. After the injection, BV was applied to the skin surface of the right ear. Histological observation revealed that P. acnes induced a considerable increase in the number of infiltrated inflammatory cells. However, treatment with BV markedly reduced these reactions compared with the P. acnes-injected mice not treated with BV. Moreover, the expression levels of tumor necrosis factor (TNF)-α, and interleukin (IL)-1β were significantly reduced in the BV-treated mice compared with the untreated P. acnes-injected mice. In addition, treatment with BV significantly inhibited Toll-like receptor (TLR)2 and CD14 expression in P. acnes-injected tissue. The binding activity of nuclear factor-κB (NF-κB) and activator protein (AP)-1 was markedly suppressed following treatment with BV. The results from our study, using an animal model, indicate that BV exerts an inhibitory effect on inflammatory skin diseases. In conclusion, our data indicate that BV has potential for use as an anti-acne agent and may be useful in the pharmaceutical and cosmetics industries.

This research was performed in order to investigate the origin, standard compound, and structural and physical properties of honeybee venom which used as natural antibiotic ingredients to animal. We compared the nucleotide sequence of mitochondrial cytochrome c oxidase subunit 1 gene (COI) of honeybees were collected from Gangwon, Gyeonggi, Chungnam, Gyeongbuk, Gyeongnam province and Suwon. As major constituent of honeybee venom, melittin was assayed by liquid chromatography. X-ray, differential scanning calorimetry (DSC) and fourier transform infrared spectroscopy (FT-IR) were utilized to examine the structural and physical properties of honeybee venom. Based on the 627bp sequence of COI, Apis mellifera ligustica was determinated honeybees collected from all six regions. Melittin content varied from 50.7 to 68.6 and averaged 59.8%. According to XRD analysis, honeybee venom showed regular crystal structure peaks at 2θ=8.5° and 21.5°. DSC showed that the maximum degration temperature of powder was around 230°C. Through FT-IR analysis, we could identify cross-linking by the presence of peptide peak at 1,500∼1,600 cm −1 . In conclusion, the origin of honeybee venom was Apis mellifera ligustica and effective ingredient standards was melittin content varied from 50.7 to 68.6 as natural antibiotic ingredients.

In order to the feasibility of silk materials as protein delivery system, silk beads incorporated with bovine serum albumin (BSA) were prepared by dropping silk fibroin extract into dope solution composed of ethanol and dichloromethane. Structural and morphological characteristics of silk beads were examined using scanning electron microscopy (SEM), infrared spectrometry, and X-ray diffractometry. Swelling ratio of silk beads was also measured. Release behavior of prototypical protein, BSA, was studied by observing the electropheretic phenomenon and release profile. SEM showed that silk beads are spherical with porous interior structure. Infrared spectrometry and X-ray diffraction confirm that the silk beads have a -sheet conformation. The swelling capability of silk beads increased with the incorporation of the protein. The protein was released from the beads with slow release following an initial burst release. Therefore, silk beads show promise as materials for encasing protein drugs to be delivered to targets in the human body.

Purpose: This study evaluated powdered burn wound dressing materials fr om wild silkworm fibroin in an animal model.Methods: Fifteen rats were used in this experiment. Full-thickness 2×2 cm burn wounds were created on the back of rats under anesthesia. In the two experimental groups, the woun ds were treated with two different dressing materials made from silkworm fibroin. In the Control Group, natural healing wi thout any dressing material was set as control. The wound surface area was measured at five days, seven days and 14 days. Wound healing was evaluated by histologic analysis.Results: By gross observation, there were no infections or severe infla mmations through 14 days post-injury. The differences among groups were statistically significant at seven days and 1 4 days, postoperatively (P＜0.037 and 0.001, respectively). By post hoc test, the defect size was significantly smaller in experimental Group 1 compared with the Control Group and experimental Group 2 at seven days postoperatively (P=0.022 and 0.029, respectively). The difference between Group 1 and Group 2 was statistically significant at 14 days postoperat ively (P＜0.001). Group 1 and control also differed significantly (P=0.002). Group 1 showed a smaller residual scar than the Contro l Group and Group 2 at 14 days post-injury. Histologic analysis showed more re-epithelization in Groups 1 and 2 than i n the Control Groups.Conclusion: Burn wound healing was accelerated with silk fibroin spun by w ild silkworm Antheraea pernyi. There was no atypical inflammation with silk dressing materials. In concl usion, silk dressing materials can be used for treatment of burn wound.Key words:Full thickness burn wound, Wound dressing, Silk fibroin, Antheraea pernyi, Rats

This study was for the investigation of the stability of purified bee venom (PBV) during the treatment in the pH rangefrom pH2 to pH9 for 24 hours, respectively. Changes of components and physiological functionalities in PBV were byevaluated silver staining, and melittin contents were measured by liquid chromatography. The antimicrobial activityagainst bacteria by minimum inhibitory concentration (MIC) and effect of the cell regeneration were measured by 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl tetrazolium bromide (MTT) assay using human dermal fibroblast (HDF) cell.The main proteins such as melittin and phospholipase A 2 showed no characteristic changes. The antimicrobial activityand effect of cell regeneration showed no difference from pH2 to pH9. From this study, we suggest that componentsand physiological functionalities of PBV against treated pH were kept stability at from pH2 to pH9.Key words : Apis mellifera, Bee venom, Component, pH 서론 봉독은 꿀벌(Apis mellifera L.)의 독낭에 들어있는 자기방어 물질로 봉침은 수천년 전부터 관절염, 통풍 등(Kimet al. 2002, Kim et al. 2003) 치료에 한방과 민간요법으로 이용되어 왔다(Kim 1992). 봉독은 40여 가지의 성분으로이루어져 있으며, peptide 성분으로는 melittin, apamin, mastcell degranulating(MCD) peptide 등이 있으며, 효소 성분으로는 phospholipase A

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 -sheet and -helix conformation with low ethanol concentration (50 ~ 70%). On the other hand, that of silk sponges with 80% ethanol treatment showed -sheet (), -helix (), and random coil () 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.

Breast cancer is one of the most common cancers among women worldwide. Recently anticancer agents have been developed using natural substances. To evaluate the anticancer effect of hydrolysates of silk fibroin (HSF), we investigated the effect of HSF on cell viability and apoptosis of a breast cancer cell line, MCF-7, induced through the mitochondrial pathway. The result showed that HSF decreased cell viability in MCF-7 cells in a dose- and time-dependent manner, resulting in an increase in the sub-G1 phase cell population. HSF increased the level of the pro-apoptotic Bax protein and decreased the levels of the anti-apoptotic Bcl-2 protein. In addition, HSF induced apoptosis in MCF-7 cells through a mitochondria-dependent pathway by increasing levels of cytochtome c, and cleavage of PARP. Taken together, these findings suggest that HSF inhibits the proliferation of MCF-7 breast cancer cells through a mitochondria and caspase dependent apoptotic pathway.

In this study, we compared the efficiency of osteoblast differentiation media (ODM) contain ing three distinct reagent combinations in osteoblastic differentiation of human bone marrowderived mesenchymal stem cells (hBMSCs) in monolayer culture. In addition, we analyzed growth and differentiation of hBMSCs on silk scaffolds and examined the bone-forming activity of a nanofibrous silk scaffold in a tibia diaphysis defect model of a rat hind limb with intramed ullary nailing. Although all three ODM increased alkaline phosphatase activity to a comparable extent, the ODM containing bone morphogenetic protein-2 (BMP-2) was found to be signifi cantly less effective in promoting mineral deposition than the others. Growth of hBMSCs on sponge-form silk scaffolds was faster than on nanofibrous ones, while osteoblastic differentia tion was apparent in the cells grown on either type of scaffold. By contrast, bone formation was observed only at the edge of the nanofibrous scaffold implanted in the tibia diaphysis defect, suggesting that use of the silk scaffold alone is not sufficient for the reconstitution of the long bone defect. Since silk scaffolds can support cell growth and differentiation in vitro, loading MSCs on scaffolds might be necessary to improve the bone-forming activity of the scaffold in the long bone defect model.

In this study, matured silkworm hemolymph was studied for inhibitory effect on melanin formation, tyrosinase activities.A matured silkworm hemolymph mass purification method was developed and some of suppressing melanin formationeffects were investigated, too. As a result, the advanced purification method was higher in yield rate than existing oneby 53%. A free radical scavenging activity of hemolymph was higher than sericin on the market(IC 50 , 202.9± 33.1) andlower than vitamin C(0.57± 0.02) by 6.4± 0.3 ㎍/㎖. An inhibitory effect on melanin synthesis was higher inhemolymph compared to Morus bark and arbutin by 9.15% and 11.56%, respectively. An inhibitory effect on tyrosinaseactivity was higher in hemolymph than Morus bark and lower than arbutin. Effect of hemolymph 45㎍/㎖ on sup-pressing of tyrosinase was almost 90%. Thus, these results suggest that hemolymph of matured silkworm may have ben-eficial properties as a material for cosmetics.Key words : Matured silkworm hemolymph, Antioxidant, Melanin synthesis, Tyrosinase activity

Silk fibroin is a natural biomaterial that has the biocompatibility and other many advantages. But as a silk fibroin mem-brane thickness increases, the transparency becomes more opaque. Because the transparency of membranes tissue suchas the cornea and dura mater are necessary, transparent membrane is required to replace these transparent membranes.In this study, we fabricated blending silk fibroin membranes that made by mixing the various inorganic salts or polymerin an aqueous solution of silk fibroin. The transparency of the membranes were analyzed. the transparency of thesemembranes is very different, depending on the mixed materials. Inorganic salts mixed silk membrane was more trans-parent than the polymer mixed one. Especially, the silk fibroin membrane with calcium chloride was very transparent.We showed the possibility of blending silk fibroin membrane, which can be used in perfect transparent membrane suchas the cornea. In the future, we expect that the transparent blending silk fibroin membrane can be used in various med-ical applications.Key words : Silk fibroin, Salt, Polymer, Transparency

In this study, bee venom (BV) isolated from honey bees Apis mellifera was assessed for its potential use as an antimicrobial agent against fish pathogenic bacteria. We used three bacterial isolates (Edwardsiella tarda, Vibrio ichthyoenteri, and Streptococcus iniae) that were originally obtained from the gastrointestinal tracts of moribund Olive Flounder Paralichthys olivaceus. Bee venom exhibited antibacterial activity against all three infectious fish pathogens. The minimum inhibitory concentration and minimum bactericidal concentration (mean ± SE) of BV were 17.6 ± 2.6 and 34.9 ± 3.4 μg/mL, respectively, against E. tarda; 1.76 ± 0.3 and 6.8 ± 2.6 μg/mL against V. ichthyoenteri; and 3.49 ± 0.9 and 11 ± 1.6 μg/mL against S. iniae. The postantibiotic effect of BV was 5 ± 0.6 h for E. tarda, 6 ± 0.3 h for V. ichthyoenteri, and 7 ± 0.6 h for S. iniae. In addition, the antimicrobial activity of BV was not pH dependent, as inhibition zones were identical at pH levels ranging from 2 to 11. These result...

Antheraea pernyi silkworm is one of typical wild silkworms, which spins a tawny color cocoon. The cocoon has been used as a resource for textile material due to strong chemical stability and good mechanical properties. In this study, to increase the solubility efficiency of A. pernyi silk fibroin, the composition of dissolution solvent were examined. Calcium chloride tertiary system, normally used for dissolution of Bombyx mori silk fibroin, does not act on A. pernyi silk fibroin. Calcium nitrate system dissolves A. pernyi silk fibroin, and calcium nitrate ethanol system do more easily than calcium nitrate system. Amino acid composition of A. pernyi silk fibroin obtained after dissolution is mainly composed of alanine, glycine, and serine. A. pernyi silk fibroin would be used for non-textile applications near future.

The propolis was collected from the whole part of Korea Peninsula, such as central, southern, and Jeju island, to analyze the general composition including total flavonoid and phenolic contents, heavy metals, colors, amino acids, and crude lipid. The total flavonoid and phenolic contents of middle region in Korea Peninsula (4.26% and 13.06 g/100 g gallic acid) were higher than southern region (2.53%, 10.9 g/100 g gallic acid) and Jeju island (0.03%, 7.22g/100 g gallic acid), respectively. The heavy metals contents showed that Zn contents were found in and a harmful heavy metal such as Cr, Cd, Pb, Ni, Cu, As, and Hg were not detected at all in our experiment. The colors of extracted propolis showed a various spectrum from yellow to red purple. Various amino acids were also detected as proline, valine, methionine and the average crude lipid contents of propolis were 42.4%. All data collected in South Korea were very first executed in our institution for the purposes of utilizing and developing the industrial insects, honeybees as well as silkworm.

Silk is a natural polymer that has the advantages of the biocompatibility, excellent mechanical strength, low immune rejection, and molding facility. But silk does not dissolve easily in water or general solvent. To investigate the characteristics of silk biological membranes according to dissolving condition of silk fibroin, we made the silk biological membranes using silk fibroin solutions with different amount and dissolving time of silk. The characterizations of the silk biological membranes such as morphology, structure, and mechanical strength were observed. Although each biological membrane has the same fibroin content, there was a significant difference in the thickness and transparency. But there was no significant change in the molecular weight of the silk fibroin solutions and morphology of silk biological membranes. We were established the manufacturing condition for silk fibroin biological membrane. So we expect that the conditions will help in the development of medical supplies in the future.

Heavy metals and residual agricultural chemicals in Bombyx mori silkworm cocoon for nontextile usages were examined. Proximate composition of silkworm cocoon was examined; contents of moisture, crude protein, crude fat, and ash were about 5 ~ 6%, 93 ~ 94%, 0.4 ~ 0.5%, and 0.3 ~ 0.4%, respectively. The Heavy metals were analyzed by ICP/MS after acid digestion. Arsenic, cadmium, and lead were not detected. The average contents of mercury, zinc, and cupric were 0.317, 3.99, and 2.06 mg/kg, respectively. Agricultural chemicals, such as dichlorvos and paraquart also were not detected. Silkworm cocoon may be safe used for nontextile applications.

Cordyceps and its allies fungi has been described as a secret medicine that gives eternal youth and a long life. Some species of Cordyceps are sources of biochemicals, such ascordycepin, with interesting biological and pharmacological properties. Hence, it has been studied to uncover its pharmacological effect. We attempted to study the formation of fruiting bodies and to develop means of mass production Korean isolate of Paecilomyces tenuipes has been inoculated into silkworms, where it reproduced using culture container, mini-kit successfully. Culture container, mini-kit is composed of a cylinder-shaped body and lid. The container is made of translucent polyethylene terephthalate. The size of the container is 82 × 75 mm, reduced by 10 times as compared with the conventional culture kit. The mini kit has many advantages - high culture amount, ability of maintaining optimal humidity, parasite-free cultivation and high-end appearance. With the kit, the optimal cultivation condition is under 22 o C, culture period of 53 days. And synnemata of P. tenuipes could be kept fresh for 14 days at the temperature of under 10 o C. Therefore, the Min-kit can be used in both ways as a culture container and a packing kit for end-user customers.

Background Silk cocoon is composed of multiple layers. The natural silk cocoon containing all layers was cut as a rectangular shape as defined as total group. The inner and outermost layers were removed from the total group and the remained mat was defined as the middle group. The objectives of this study was to compare the total group with the middle group as a barrier membrane for the guided bone regeneration. Methods The effects of these materials on the cellular proliferation and alkaline phosphatase (ALP) expression of MG63 cells were explored. For comparing bone regeneration ability, bilateral bone defects were created in calvarial areas in ten adult New Zealand white rabbits. The defects were covered with silk membranes of the middle group, with silk membrane of the total group used as the control on the contralateral side. The defects were allowed to heal for 4 and 8 weeks. Micro-computerized tomography (μCT) and histological examination were performed. Results The middle group exhibited a higher MTT value 48 and 72 h after treatment compared to the total group. ALP expression was also higher in the middle group. The results of μCT and histologic examination showed that new bone formation was significantly higher in the middle group compared to the total group 8 weeks postoperatively (P

The aim of this study was to evaluate the regenerative capacity of a newly developed nerve guidance conduit using electrospun silk fibroin (SFNC) implanted in a 10-mm defect of the sciatic nerve in rats. After evaluating the physical properties and cytocompatibility of SFNC in vitro, rats were randomly allocated into three groups: defect only, autograft and SFNC. To compare motor function and abnormal sensation among groups, ankle stance angle (ASA) and severity of autotomy were observed for 10 weeks after injury. Immunostaining with axonal neurofilament (NF) and myelin basic protein (MBP) antibodies were performed to investigate regenerated nerve fibres inside SFNC. ASA increased significantly in the SFNC group at 1, 7 and 10 weeks after injury compared to the defect only group (p

Bone disease can be associated with bone resorption by osteoclasts, and interest in the development of antiresorptive agents has recently increased. The hydrolysate of silk fibroin has been studied with respect to such biomedical applications. In a previous study, silk fibroin showed indirect inhibitory effects on the differentiation of osteoclasts. To further evaluate the effect of a hydrolysate of silk fibroin on osteoclasts, we investigated the direct effects of the silk fibroin hydrolysate on osteoclastogenesis and apoptosis of osteoclasts induced by receptor activation of nuclear factor κB ligand (RANKL). The silk fibroin hydrolysate inhibited RANKL-induced formation of tartrate-resistant acid phosphatase (TRAP) in RAW 264.7 cells. The inhibitory effect of the silk fibroin hydrolysate resulted in the decreased expression of osteoclast marker genes, such as matrix metalloproteinase-9 (MMP-9), cathepsin-K and calcitonin receptor (CTR). In addition, the silk fibroin hydrolysate blocked the signaling pathways of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) and expression of transcription factors, such as nuclear factor of activated T cells c1 (NFATc1) and NF-κB. Finally, the silk fibroin hydrolysate induced apoptosis signaling cascades. Taken together, the present results indicate that silk fibroin hydrolysate has antiresorptive activity by both inhibiting osteoclastogenesis and inducing osteoclast apoptosis.

Bee venom (Apis mellifera L., BV) possessing a rich source of pharmacologically active substances has the potential to be used as a cosmetic ingredient for antiaging, antiinflammatory and antibacterial functions. The aim of this study was to assess the skin sensitization of BV on experimental animals using the Buehler test.Guinea pigs were randomly allocated into three groups of BV-sensitization, positive control-sensitization, and ethyl alcohol-sensitization group for induction and challenge. On the other hand, two groups of rats were administered with BV at doses of 0 and 1500 mg/kg. Clinical signs, mortality and body weight changes were continually monitored during the study period.No treatment-related clinical signs or body weight changes were observed in both animal models. The average skin reaction evaluated by erythema and edema on the challenge sites, and sensitization rate in the BV-sensitization group of guinea pigs were substantially low compared with those in positive control group, representing a negligible sensitizing potential of BV.It was concluded that BV was well tolerated and exhibited no dermal irritation potential in guinea pigs and rats. Our findings may provide a developmental basis of BV for a cosmetic ingredient or external application for topical uses.

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 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.

In this study, silk gland powder of silkworm were investigated to see the possibility for baby powder cosmetics materials. To test possibility as a baby powder cosmetics, total content rate of amino acids, DPPH free radical scavenging assay, MTT assay, and clinical trial were done. According to the result of the analysis of the amino acids of silk gland powder, serin (26.77%) content was the highest and asparatic acid (15.47%), and glycine (9.62%) were followed. DPPH free radical scavenging activity of silk gland powder was lower than vitamin C by 82.3% and 97%, respectively, which is relatively good. Moisture effect were increased in silk gland powder compared to control cosmetics by 50%. Also, silk gland powder was classified as a practically non-irritating material based on the score 0.05 of primary irritation index. Thus, these results suggest that silk gland powder of silkworm may have beneficial properties as a material for baby powder cosmetics.

Sericultural and Agicultural Materials Division, National Academy of Agricultural Science, RDA, Suwon 441-100, KoreaReceived: November 10, 2011 / Revised: November 18, 2011 / Accepted: December 8, 2011Silk fibroin (SF) peptide has been traditionally used as atreatment for flatulence, spasms, and phlegm. In this study,we examined whether SF peptide enhanced the anti-inflammatory effect of PEP-1-FK506 binding protein(PEP-1-FK506BP) through comparing the anti-inflammatoryactivities of SF peptide and/or PEP-1-FK506BP. In thepresence or absence of SF peptide, transduction levels ofPEP-1-FK506BP into HaCaT cells and mice skin andanti-inflammatory activities of PEP-1-FK506BP wereidentified by Western blot and histological analyses. SFpeptide alone effectively reduced both mice ear edemaand the elevated levels of cyclooxygenase-2, interleukin-6and -1β, and tumor necrosis factor-α, showing similaranti-inflammatory effect to that of PEP-1-FK506BP.Furthermore, co-treatment with SF peptide and PEP-1-FK506BP exhibited more enhanced anti-inflammatoryeffects than the samples treated with SF peptides or PEP-1-FK506BP alone, suggesting the possibility that SFpeptide and PEP-1-FK506BP might interact with eachother. Moreover, the transduction data demonstrated thatSF peptide did not affect the transduction of PEP-1-FK506BP into HaCaT cells and mice skin, indicating thatthe improvement of anti-inflammatory effect of PEP-1-FK506BP was not caused by enhanced transduction ofPEP-1-FK506BP. Thus, these results suggest the possibilitythat co-treatment with SF peptide and PEP-1-FK506BP maybe exploited as a useful therapy for various inflammation-related diseases.Keywords: Inflammation, PEP-1-FK506BP, protein transduction,reactive oxygen species, silk fibroin peptide, TPADuring various cellular processes, reactive oxygen species(ROS), such as hydroxyl radicals, superoxide anion, andhydrogen peroxide, are generated and, consequently, can causelipid peroxidation, DNA damage, cell death, and variousdiseases, resulting in inflammation, cancer, Parkinson’s disease,and ischemia [1, 5, 19]. Treatment with lipopolysaccharides(LPS) or 12-O-tetradecanoylphorbol-13-actate (TPA), knownas inflammation inducers used in animal models [25, 30],can activate macrophages and secrete pro-inflammatorymediators, such as cyclooxygenase-2 (COX-2) and induciblenitric oxide synthase (iNOS), and pro-inflammatory cytokines,including interleukin-6 (IL-6), interleukin-1beta (IL-1β), andtumor necrosis factor-alpha (TNF-α). In addition, nuclearfactor-kappaB (NF- κB) and mitogen-activated protein kinase(MAPK) pathway are related to inflammatory responses[7, 8].FK506 binding proteins (FK506BPs) are members of animmunophilins family, which can bind immunosuppressivedrugs such as rapamycin and FK506, used in prevention ofgraft rejection following organ transplantation [2, 11, 21].In particular, FK506BP, a small peptide of 12 kDa, wasfirst associated with rapamycin. The resultant FK506BP-rapamycin complex binds to and then disrupts the formationof activated mammalian target of rapamycin complex 1(mTORC1), which can control protein synthesis, cellproliferation, and cell growth in response to nutrients andgrowth factors, leading to the suppression of proteinsynthesis and cell growth [4, 6, 16]. Furthermore, wepreviously reported that, through fusing FK506BP with a

A newly developed silkworm variety, GoldenSilk makes bright yellow cocoon. In this article, the basic characteristics of GoldenSilk cocoon were examined through color analysis, degumming characteristics and UV spectrophotometer. The colorant of GoldenSilk cocoon was not distributed evenly throughout the cocoon layer but existed in the outer layer of the cocoon. The colorant has not good resistance against sunlight. The degumming loss percentage of GoldenSilk cocoon was 29.2%, which is relatively higher than that of the authorized one. UV spectrophotometer showed specific absorption band around 400 to 500 nm, which is the specific yellow colorant of the GoldenSilk cocoon extracted to the degumming solution during the degumming process.

The effect of cultivation season on the cocoon filament quality of authorized silkworm variety, GoldenSilk was examined with cocoon filament character, reeling character, and fiber character during 3 years. In cocoon filament character, filament length of spring and autumn cocoon was 979 and 879 m, respectively. Cocoon filament weight of spring and autumn cocoon was 33.1 and 26.5 cg, which is lower than that of general bivoltine strain, BaeGokJam. Raw silk percentage of spring and autumn cocoon was 15.65 and 13.77%, respectively. In reeling character, reelability showed similar trend regardless of cultivation season and strain. However, the length and weight of non-broken filament of spring season were slightly higher than those of autumn cocoon. Cultivation season did not affect on the fiber characteristics such as clearness and neatness, lousiness, and degumming loss. In cocoon character, filament length, size, and weight, spring cocoon filament showed higher value than autumn cocoon, but lower than authorized general silkworm varieties. In reeling characteristics, non-broken length and weight showed the similar trends with the cocoon filament character. However, the reelability of GoldenSilk cocoon was not affected by cultivation season. The reelability of GoldenSilk cocoon was also similar with that of authorized general one. Fiber character including clearness and neatness, lousiness, and degumming loss was also similar regardless of the rearing season and different strain.

Bee venom (BV) has been used in medicine to treat a variety of diseases including arthritis, rheumatism, and various cancers. Recent reports indicate that BV has anti-angiogenic effects, but the precise molecular mechanism underlying the effects of BV against colorectal cancer remains to be elucidated. We examined the effects of BV and its major components (melittin and apamin) on tumor angiogenesis and found that BV significantly decreased protein levels of hypoxia-inducible factor- (HIF-), an important factor involved in angiogenesis and tumor progression, in human colorectal carcinoma HCT116 cells. BV also suppressed the transcription of HIF- under hypoxia, leading to a decrease in the expression of vascular endothelial growth factor (VEGF), a major target gene of HIF-. We also found that these effects were mainly elicited by apamin, but not melittin. BV specifically inhibited the phosphorylation of ERK1/2 without changing the total levels of this protein, but had no effect on kinases of p38/JNK and AKT. Our results suggest that BV may inhibit human colorectal cancer progression and angiogenesis by inhibiting HIF- and VEGF expression, thereby providing a novel potential mechanism for the anticancer action of BV.

The aim of this study was to access the irritant properties of bee venom (BV) after its application to skin and eye mucous membranes of the rabbit. The animals were also observed for clinical signs and mortality after the application of the test material. Six animals were used for the skin irritation test and nine rabbits for the eye irritation test. The acute BV application to the rabbit skin revealed no appreciable clinical signs throughout the observation period of 72 h and there was no mortality seen. In the eye irritation test, eye reactions were read and graded 24, 48, 72, 96 and 168 h after BV treatment. No changes in the cornea, iris or conjunctivae were observed at all time points of observations. Based on the present findings, it can be concluded that the irritation potential of BV is negligible.

Silk fibroin, a natural protein produced by silkworm, is a good biomaterial which has biodegradability and biocompatibility. To ascertain the effects of silk fibroin on cell growth, silk fibroin films were prepared using silk fibroin aqueous solutions of various concentrations. We investigated the attachment, proliferation, morphology of the cells and the expression levels of genes related to cell attachment and growth on the silk fibroin films. When the cells were cultured on the 0.1 and 1% silk fibroin film, the cell adhesion ability was very excellent. Particularly, overall cell growth on the 1% silk fibroin film was definitely superior to the others. Also, expression levels of genes related cell growth were increased on the 0.1 and 1% silk fibroin film. These results suggest silk as a material for medical applications.

Transparency and insolubility of eardrum patch against exudates are important for otolaryngological surgery. The author prepared silk fibroin (SF) films with various concentrations and temperature and then examined solubility and conformation of SF films. SF films were transparent regardless of the various preparation conditions. Although most SF films are soluble in 1X PBS solution at for 1 h, the SF film with 3.4% with was insoluble. Scanning electron microscopy (SEM) showed that the SF films have solid and smooth surface. Fourier transform infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC) results showed that the conformation of SF films was influenced by the preparation conditions including SF concentration and casting temperature. In conclusion, SF membrane with transparence and insolubility against exudates could be considered as eardrum patch resources.