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Medical/ Medicinal application

posted Mar 4, 2011, 7:16 AM by rajesh gk

Title

[A study of occupational asthma and specific IgE in sericulture workers]

[Article in Chinese]

Authors

Wang X, Zheng S, Zhang H.

Institute of Basic Medical Sciences, CAMS, Beijing.

Journal

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1994 Oct;16(5):323-7.

Abstract

Clinical investigation revealed that 75.6% (68/90) of sericulture workers examined had allergic respiratory symptoms. Of the 90 workers, 14 (15.6%) suffered from occupational asthma. Work-related allergens including silkworm cocoon (SC), silkworm urine (SU), moth excrete (ME) and moth scales (MS) were collected and prepared. Serum specific IgE levels were detected by ELISA. The results showed that the level of sIgE in asthmatic workers was significantly higher than that in the "common symptom" group, asymptomatic group and control group. No significant difference in total IgE was found between the workers and controls. The anti-SU-sIgE positive rate was significantly higher than those of sIgE to the other three allergens. These results indicate that the high levels of sIgE are responsible for occupational asthma in sericulture workers and that SU is an important allergen in these patients.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/7720120 PMID: 7720120 [PubMed - indexed for MEDLINE]

 

Title

A study of medico-social problems of the inmates of a leprosy colony in Mysore.

Authors

Vasundhra MK, Siddalingappa AS, Srinivasan BS.

Journal

Lepr India. 1983 Jul;55(3):553-9.

Abstract

A study of the inmates of a leprosy colony in Mysore revealed that there were more male cases and that the deformities were also more common among the male patients. There was no cases among inmates below 20 years of age group 31-50 years. 93.2% of the inmates were illiterate and 96.6% of the patients were ignorant of the cause, spread, prognosis of the disease and prevention of disabilities. This indicated an urgent need of health education among the leprosy patients and their families. All the inmates, including the apparently healthy relatives reported loss of income due to disease and disabilities. Rehabilitation of the inmates by raising poultry, breeding sheep and sericulture activities must be encouraged.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/6656213 PMID: 6656213 [PubMed - indexed for MEDLINE]

 

Title

[A case of hypersensitivity pneumonitis relation to sericulture (sericulturist's lung disease)]

[Article in Japanese]

Authors

Umegae Y, Matsui S, Irokawa M, Katakai S, Nakazawa T, Iizuka K, Miura S, Fueki R, Kobayashi S, Kitaichi M.

Journal

Nihon Kyobu Shikkan Gakkai Zasshi. 1986 Jul;24(7):804-9.

Abstract

http://www.ncbi.nlm.nih.gov/pubmed/3784179

Citation

PMID: 3784179 [PubMed - indexed for MEDLINE]

 

Title

[Antigenic substances of bronchial asthma related to sericulture. 2. Cross antigenicity between silkworms and ordinary moths]

[Article in Japanese]

Authors

Kobayashi S, Nakazawa T, Yoshida S.

Journal

Arerugi. 1971 Sep;20(9):694-9.

Abstract

http://www.ncbi.nlm.nih.gov/pubmed/5001755

Citation

PMID: 5001755 [PubMed - indexed for MEDLINE]

 

Title

Enhancing effects of sericin on corneal wound healing in rat debrided corneal epithelium. Free Article

Authors

Nagai N, Murao T, Ito Y, Okamoto N, Sasaki M.

School of Pharmacy, Kinki University, Osaka, Japan.

Journal

Biol Pharm Bull. 2009 May;32(5):933-6.

Abstract

The protein sericin is the main constituent of silk. We demonstrate the effects of sericin on corneal wound healing in rat debrided corneal epithelium. We also determined the effects of sericin on cell adhesion and proliferation in a human cornea epithelial cell line (HCE-T). Epithelium was removed from the corneas of rats with a BD Micro-Sharp, and wounded corneas were dyed with a 1% fluorescein solution. The corneal wounds were monitored using a fundus camera TRC-50X equipped with a digital camera. The corneal wound of rats instilled with saline was approximately 10% healing at 12 h, and approximately 65% healing at 24 h after corneal epithelial abrasion. The corneal wounds of rats instilled with saline showed almost complete healing by 36 h after corneal epithelial abrasion. On the other hand, the corneal healing rate of rats instilled with sericin solution was higher than that of rats instilled with saline, and the corneal healing rate constant increased with increasing sericin concentration. In addition, the adhesion and proliferation of HCE-T cells treated with 0.01-0.5% sericin solutions were enhanced, reaching a maximum at treatments with 0.2 and 0.1% sericin solutions, respectively. The present study demonstrates that the instillation of sericin solution has a potent effect in promoting wound healing and wound-size reduction in rats, probably caused by increasing cell movement and proliferation.

Citation

PMID: 19420767 [PubMed - indexed for MEDLINE]

 

Title

Silk film biomaterials for cornea tissue engineering.

Authors

Lawrence BD, Marchant JK, Pindrus MA, Omenetto FG, Kaplan DL.

Department of Biomedical Engineering, Tufts University, 4 Colby St., Medford, MA 02155, USA.

Journal

Biomaterials. 2009 Mar;30(7):1299-308. Epub 2008 Dec 6.

Abstract

Biomaterials for corneal tissue engineering must demonstrate several critical features for potential utility in vivo, including transparency, mechanical integrity, biocompatibility and slow biodegradation. Silk film biomaterials were designed and characterized to meet these functional requirements. Silk protein films were used in a biomimetic approach to replicate corneal stromal tissue architecture. The films were 2 microm thick to emulate corneal collagen lamellae dimensions, and were surface patterned to guide cell alignment. To enhance trans-lamellar diffusion of nutrients and to promote cell-cell interaction, pores with 0.5-5.0 microm diameters were introduced into the silk films. Human and rabbit corneal fibroblast proliferation, alignment and corneal extracellular matrix expression on these films in both 2D and 3D cultures were demonstrated. The mechanical properties, optical clarity and surface patterned features of these films, combined with their ability to support corneal cell functions suggest that this new biomaterial system offers important potential benefits for corneal tissue regeneration.

Citation

PMID: 19059642 [PubMed - indexed for MEDLINE]PMCID: PMC2670567 Free PMC Article

 

Title

Silk: a potential medium for tissue engineering.

Authors

Sobajo C, Behzad F, Yuan XF, Bayat A.

Plastic & Reconstructive Surgery Research, Manchester Interdisciplinary Biocentre, University of Manchester, Manchester, M1 7DN, UK.

Journal

Eplasty. 2008;8:e47. Epub 2008 Oct 10.

Abstract

OBJECTIVE: Human skin is a complex bilayered organ that serves as a protective barrier against the environment. The loss of integrity of skin by traumatic experiences such as burns and ulcers may result in considerable disability or ultimately death. Therefore, in skin injuries, adequate dermal substitutes are among primary care targets, aimed at replacing the structural and functional properties of native skin. To date, there are very few single application tissue-engineered dermal constructs fulfilling this criterion. Silk produced by the domestic silkworm, Bombyx mori, has a long history of use in medicine. It has recently been increasingly investigated as a promising biomaterial for dermal constructs. Silk contains 2 fibrous proteins, sericin and fibroin. Each one exhibits unique mechanical and biological properties. METHODS: Comprehensive review of randomized-controlled trials investigating current dermal constructs and the structures and properties of silk-based constructs on wound healing. RESULTS: This review revealed that silk-fibroin is regarded as the most promising biomaterial, providing options for the construction of tissue-engineered skin. CONCLUSION: The research available indicates that silk fibroin is a suitable biomaterial scaffold for the provision of adequate dermal constructs.

Citation

PMID: 18997857 [PubMed]PMCID: PMC2567119 Free PMC Article

 

Title

Protective effect of sericin peptide against alcohol-induced gastric injury in mice.

Authors

Li YG, Ji DF, Lin TB, Zhong S, Hu GY, Chen S.

Sericultural Research Institute, Zhejiang Academy of Agricultural Science, Hangzhou, Zhejiang 310021, China.

Journal

Chin Med J (Engl). 2008 Oct 20;121(20):2083-7.

Abstract

BACKGROUND: Sericin peptide (SP) has shown a powerful anti-oxidant property in a host of studies. The present study was designed to investigate the possible protective effects of SP against alcohol-induced gastric lesions in mice and to explore the potential mechanisms. METHODS: Animals were randomly divided into 5 groups: control, alcohol (56%, 14.2 ml/kg), SP-treated mice (0.2, 0.4, 0.8 g/kg). Mice were pretreated with SP before administering alcohol, the concentration of ethanol in serum and urine, the contents of malondialdehyde (MDA), glutathione (GSH) and the glutathione peroxidase (GSH-PX), catalase (CAT) and superoxide dismutase (SOD) activities in the gastric mucosa were measured, subsequently, the pathological evaluation of stomach was also observed. RESULTS: Of the animals pre-treated with SP (0.4, 0.8 g/kg), the concentration of ethanol in serum was significantly decreased, while increased in urine as compared to the alcohol-administered alone animals. Alcohol administration caused severe gastric damage as indicated by markedly increased MDA levels and decreased antioxidants, such as reduced GSH, GSH-PX and SOD in the gastric tissue while the CAT activity was not altered. On SP administration there was a reversal in these values towards normal. Histopathological studies confirmed the beneficial role of SP, which was in accordance with the biochemical parameters. CONCLUSIONS: SP could protect gastric mucosa from alcohol-induced mucosal injury. These gastroprotective effects might be due to increasing 'first-pass metabolism' in the stomach and hastening ethanol elimination directly through the urine. SP might also play an important role in the protection of the structure and function of gastric mitochondria, at least partly based on their anti-oxidant effect.

Citation

PMID: 19080280 [PubMed - indexed for MEDLINE] Free Article

 

Title

Preparation of gel film from Bombyx mori silk sericin and its characterization as a wound dressing.

Authors

Teramoto H, Kameda T, Tamada Y.

Division of Insect Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan. teramoto@affrc.go.jp

Journal

Biosci Biotechnol Biochem. 2008 Dec;72(12):3189-96. Epub 2008 Dec 7.

Abstract

Sericin is a highly hydrophilic protein family acting as the glue in Bombyx mori silk. In order to apply sericin as a wound dressing, a novel sericin film named gel film was prepared by a simple process without using any chemical modifications: sericin solution was gelled with ethanol into a sheet shape and then dried. Infrared analysis revealed that the sericin gel film contained water-stable beta-sheet networks formed in the gelation step. This structural feature rendered the gel film morphologically stable against swelling and gave it good handling properties in the wet state. The sericin gel film rapidly absorbed water, equilibrating at a water content of about 80%, and exhibited elastic deformation up to a strain of about 25% in the wet state. A culture of mouse fibroblasts on the gel film indicated that it had low cell adhesion properties and no cytotoxicity. These characteristics of sericin gel film suggest its potential as a wound dressing.

Citation

PMID: 19060395 [PubMed - indexed for MEDLINE] Free Article

 

Title

Peptide-based Biopolymers in Biomedicine and Biotechnology.

Authors

Chow D, Nunalee ML, Lim DW, Simnick AJ, Chilkoti A.

Department of Biomedical Engineering, Duke University, Box 90281, Durham, North Carolina 27708-0281.

Journal

Mater Sci Eng R Rep. 2008 Jan;62(4):125-155.

Abstract

Peptides are emerging as a new class of biomaterials due to their unique chemical, physical, and biological properties. The development of peptide-based biomaterials is driven by the convergence of protein engineering and macromolecular self-assembly. This review covers the basic principles, applications, and prospects of peptide-based biomaterials. We focus on both chemically synthesized and genetically encoded peptides, including poly-amino acids, elastin-like polypeptides, silk-like polymers and other biopolymers based on repetitive peptide motifs. Applications of these engineered biomolecules in protein purification, controlled drug delivery, tissue engineering, and biosurface engineering are discussed.

Citation

PMID: 19122836 [PubMed]PMCID: PMC2575411 Free PMC Article

 

Title

Biofunctionalized electrospun silk mats as a topical bioactive dressing for accelerated wound healing.

Authors

Schneider A, Wang XY, Kaplan DL, Garlick JA, Egles C.

Division of Cancer Biology and Tissue Engineering, Department of Oral and Maxillofacial Pathology, Tufts University, School of Dental Medicine, 55 Kneeland Street, Boston, MA 02111, USA.

Journal

Acta Biomater. 2009 Sep;5(7):2570-8. Epub 2008 Dec 31.

Abstract

Materials able to deliver topically bioactive molecules represent a new generation of biomaterials. In this article, we describe the use of silk mats, made of electrospun nanoscale silk fibers containing epidermal growth factor (EGF), for the promotion of wound healing processes. In our experiments, we demonstrated that EGF is incorporated into the silk mats and slowly released in a time-dependent manner (25% EGF release in 170h). We tested these materials using a new model of wounded human skin-equivalents displaying the same structure as human skin and able to heal using the same molecular and cellular mechanisms found in vivo. This human three-dimensional model allows us to demonstrate that the biofunctionalized silk mats, when placed on the wounds as a dressing, aid the healing by increasing the time of wound closure by the epidermal tongue by 90%. The preservation of the structure of the mats during the healing period as demonstrated by electronic microscopy, the biological action of the dressing, as well as the biocompatibility of the silk demonstrate that this biomaterial is a new and very promising material for medical applications, especially for patients suffering from chronic wounds.

Citation

PMID: 19162575 [PubMed - indexed for MEDLINE]PMCID: PMC2810481 Free PMC article

 

Title

Silk amino acids improve physical stamina and male reproductive function of mice.

Authors

Shin S, Yeon S, Park D, Oh J, Kang H, Kim S, Joo SS, Lim WT, Lee JY, Choi KC, Kim KY, Kim SU, Kim JC, Kim YB.

College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University, 410 Seongbongro (Gaeshin-dong), Cheongju, Chungbuk 361-763, Republic of Korea.

Journal

Biol Pharm Bull. 2010 Feb;33(2):273-8.

Abstract

The effects of a silk amino acid (SAA) preparation on the physical stamina and male reproductive function of mice were investigated. Eight-week-old male ICR mice (29-31 g) were orally administered SAA (50, 160 or 500 mg/kg) for 44 d during 30-min daily swimming exercise. The mice were subjected to a weight-loaded (5% of body weight) forced swimming on the 14th, 28th and 42nd day to determine maximum swimming time, and after a 2-d recovery period (treated with SAA without swimming exercise), parameters related to fatigue and reproductive function were analyzed from blood, muscles and reproductive organs. Repeated swimming exercise increased the maximum swimming time to some extent, in spite of a marked reduction in body weight gain, and SAA further enhanced the stamina in a dose-dependent manner. Forced swimming exercises increased blood parameters of tissue injury, but depleted blood glucose and tissue glycogen, which were substantially prevented by SAA treatment. In addition, SAA significantly reduced the muscular thiobarbituric acid-reactive substances and blood corticosterone content increased by forced swimming. Swimming exercise decreased the blood testosterone level, which was recovered by SAA, leading to enhanced sperm counts. These combined results indicate that SAA not only enhances physical stamina by minimizing damage to tissues, including muscles, as well as preventing energy depletion caused by swimming stress, but also improves male reproductive function by increasing testosterone and sperm counts.

Citation

PMID: 20118552 [PubMed - in process] Free Article

 

Title

Low molecular weight silk fibroin increases alkaline phosphatase and type I collagen expression in MG63 cells.

Authors

Kim JY, Choi JY, Jeong JH, Jang ES, Kim AS, Kim SG, Kweon HY, Jo YY, Yeo JH.

Department of Oral and Maxillofacial Surgery, Hallym University, Chuncheon 200-702, Korea.

Journal

BMB Rep. 2010 Jan;43(1):52-6.

Abstract

Silk fibroin, produced by the silkworm Bombyx mori, has been widely studied as a scaffold in tissue engineering. Although it has been shown to be slowly biodegradable, cellular responses to degraded silk fibroin fragments are largely unknown. In this study, silk fibroin was added to MG-63 cell cultures, and changes in gene expression in the MG-63 cells were screened by DNA microarray analysis. Genes showing a significant (2-fold) change were selected and their expression changes confirmed by quantitative RT-PCR and western blotting. DNA microarray results showed that alkaline phosphatase (ALP), collagen type-I alpha-1, fibronectin, and transforming growth factor-beta1 expressions significantly increased. The effect of degraded silk fibroin on osteoblastogenic gene expression was confirmed by observing up-regulation of ALP activity in MG-63 cells. The finding that small fragments of silk fibroin are able to increase the expression of osteoblastogenic genes suggests that controlled degradation of silk fibroin might accelerate new bone formation. [BMB reports 2010; 43(1): 52-56].

Citation

PMID: 20132736 [PubMed - indexed for MEDLINE] Free Article

 

Title

Biomedical applications of chemically-modified silk fibroin.

Authors

Murphy AR, Kaplan DL.

Department of Biomedical Engineering, 4 Colby Street, Tufts University, Medford, MA 02155. Email: david.kaplan@tufts.edu.

Journal

J Mater Chem. 2009 Jun 23;19(36):6443-6450.

Abstract

Silk proteins belong to a class of unique, high molecular weight, block copolymer-like proteins that have found widespread use in biomaterials and regenerative medicine. The useful features of these proteins, including self-assembly, robust mechanical properties, biocompatibility and biodegradability can be enhanced through a variety of chemical modifications. These modifications provide chemical handles for the attachment of growth factors, cell binding domains and other polymers to silk, expanding the range of cell and tissue engineering applications attainable. This review focuses on the chemical reactions that have been used to modify the amino acids in silk proteins, and describes their utility in biomedical applications.

Citation

PMID: 20161439 [PubMed]PMCID: PMC2790051 Free PMC Article

 

 

Title

Adenosine prevents kindled seizures--an effect as smooth as silk.

Authors

Stafstrom CE.

Journal

Epilepsy Curr. 2010 Mar;10(2):51-2.

Abstract

·         Exp Neurol. 2009 Sep;219(1):126-35.

Citation

PMID: 20231925 [PubMed]PMCID: PMC2836479Free PMC Article

 

Title

Silk-based materials for biomedical applications.

Authors

Leal-Egaña A, Scheibel T.

Lehrstuhl für Biomaterialien, Fakultät für Angewandte Naturwissenschaften, Universität Bayreuth, 95440 Bayreuth, Germany.

Journal

Biotechnol Appl Biochem. 2010 Mar 12;55(3):155-67.

Abstract

Since the beginning of civilization, humans have exploited nature as an extraordinary source of materials for medical applications. Most natural materials comprise biopolymers such as nucleic acids and protein-polysaccharides. For biomedical applications, proteins such as collagens have been traditionally employed. Other proteins are silk fibres produced by arthropods (e.g. silkworms and spiders), which provide interesting mechanical properties and the absence of toxicity. Silks present almost all characteristics desirable for biomedical applications, but the research on the underlying proteins has only recently commenced. In the present review, we summarize the current research related to silk being used as a material for cell culture and tissue engineering, particularly focusing on cell-surface adherence, mechanical and textural properties, toxicity, immunogenicity and biodegradability.

Citation

PMID: 20222871 PubMed - indexed for MEDLINE

 

Title

Sensitization to silk and childhood asthma in rural China.

Authors

Celedón JC, Palmer LJ, Xu X, Wang B, Fang Z, Weiss ST.

Channing Laboratory, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA. juan.celedon@channing.harvard.edu

Journal

Pediatrics. 2001 May;107(5):E80.

Abstract

Sensitization to perennial aeroallergens is associated with asthma in industrialized countries with a Western lifestyle. Because silk products are commonly used in Chinese society, we were interested in examining the relation between sensitization to silk and asthma. DESIGN: Cross-sectional study of 871 children in 503 families living in Anqing, a predominantly rural province of China. RESULTS: After adjustment for age, gender, familial correlations, and sensitization to other aeroallergens, skin test reactivity to silk was an independent predictor of asthma (odds ratio = 2.6; 95% confidence interval = 1.2-5.7). This association became stronger after inclusion of the eosinophil count and history of parasitic diseases of the participants in the multivariate model (odds ratio = 3.6; 95% confidence interval = 1.4-8.9). CONCLUSION: Because sericulture is an important activity in China and other countries throughout the world, sensitization to silk may influence the pathogenesis and severity of asthma in people living in these nations.

Citation

http://www.ncbi.nlm.nih.gov/pubmed/11331730 PMID: 11331730 [PubMed - indexed for MEDLINE]

 

posted Jul 5, 2010, 1:57 AM by rajesh gk

Title

Use of silkworms for identification of drug candidates having appropriate pharmacokinetics from plant sources. Free Article

Authors

Asami Y, Horie R, Hamamoto H, Sekimizu K.

Journal

BMC Pharmacol. 2010 Jun 11;10(1):7. [Epub ahead of print]

Abstract

ABSTRACT: BACKGROUND: We use silkworms to evaluate therapeutic effects of drug candidates. Our previous reports have revealed that there are common mechanisms of pharmacokinetics of chemicals in silkworms and mammals. In this report, we attempt to establish a method by using silkworms to identify chemicals from plant extracts which are absorbed from intestine and also stably exist in body fluids. RESULTS: Three compounds were detected in the silkworm hemolymph by HPLC analysis after midgut injection of acetone extracts of seihi, an herbal medicine obtained from orange peel. Analyses with MS and NMR revealed that the compounds were nobiletin, heptamethoxyflavone, and tangeretin. These compounds are reported to be stable in mammalian blood. The half-life of each of these compounds in the silkworm hemolymph was 18, 26 and 34 h, respectively. CONCLUSIONS: These findings suggest that silkworms can be used as a model animal to easily identify compounds with appropriate pharmacokinetic behavior.

Citation

PMID: 20537191 [PubMed - as supplied by publisher]

 

Title

Simple modular bioreactors for tissue engineering: a syst em for characterization of oxygen gradients, hMSC differentiation, and pre-vascularization.

Authors

Lovett M, Rockwood DN, Baryshyan A, Kaplan DL.

 

Tufts University, Biomedical Engineering, Medford, Massachusetts, United States; Michael.Lovett@tufts.edu.

Journal

Tissue Eng Part C Methods. 2010 Jun 9. [Epub ahead of print]

Abstract

Large-scale tissue engineering is limited by nutrient perfusion and mass transport limitations, especially oxygen diffusion, which restrict construct development to smaller than clinically relevant dimensions and limit the ability for in vivo integration. The goal of this work was to develop a modular approach to tissue engineering, where scaffold and tissue size, transport issues, and surgical implantation in vivo are considered from the outset. Human mesenchymal stem cells (hMSCs) were used as the model cell type, as their differentiation has been studied for several different cell lineages and often with conflicting results. Changes in the expression profiles of hMSCs differentiated under varied oxygen tensions are presented, demonstrating tissue-specific oxygen requirements for both adipogenic (20% O2) and chondrogenic (5% O2) differentiation. Oxygen and nutrient transport were enhanced by developing a bioreactor system for perfusing hMSC-seeded collagen gels using porous silk tubes, resulting in enhanced oxygen transport and cell viability within the gels. These systems are simple to use and scaled for versatility, to allow for the systematic study of relationships between cell content, oxygen and cell function. The data may be combined with oxygen transport modeling to derive minimally-sized modular units for construction of clinically-relevant tissue engineered constructs, a generic strategy that may be employed for vascularized target tissues.

Citation

PMID: 20528664 [PubMed - as supplied by publisher]

 

Title

An improvement in the attaching capability of cryopreserved human hepatocytes by a proteinaceous high molecule, Sericin, in the serum-free solution.

Authors

Miyamoto Y, Teramoto N, Hayashi S, Enosawa S.

Journal

Cell Transplant. 2010 Jun 3. [Epub ahead of print]

Abstract

The methodology of cryopreservation of human hepatocytes remains unsatisfactory. Even when the viability of thawed cells is tolerable, the cells often lose the attaching capability to a culture dish, resulting in the cells' inability to survive. Previously, we described the effectiveness of maltose on the attachment of hepatocytes. This communication demonstrates that a silk-derived high molecular protein, sericin, improves the cell-attaching capability in the serum-free freezing medium. When human hepatocytes (initial viability; 60.9+/-3.1% (mean+/-SD, n=3)) were frozen with serum-free Dulbecco modified Eagle medium (DMEM) containing 10% dimethylsulfoxide (DMSO), the viability was 29.4 +/- 3.2% and the cell-attaching capability 20.4+/-4.1%. On the other hand, DMEM containing 10%DMSO and 1% sericin increased the values to 45.0+/-0.8% and 26.2+/-3.2%. Moreover, the addition of 0.1mol/L maltose to the sericin-containing medium improved to 42.2+/-3.2% and 51.1 +/-1.0%, as we demonstrated in a previous report. The present results indicated that sericin combined with maltose is a novel additive in the serum-free freezing medium for human hepatocytes.

Citation

PMID: 20525438 [PubMed - as supplied by publisher]

 

Title

In vivo bioassay of recombinant human growth hormone synthesized in B. mori pupae. Free PMC Article

Authors

Lan H, Nie Z, Liu Y, Lv Z, Liu Y, Quan Y, Chen J, Zhen Q, Chen Q, Wang D, Sheng Q, Yu W, Chen J, Wu X, Zhang Y.

 

College of Life Sciences, Zhejiang University, Hangzhou, China.

Journal

J Biomed Biotechnol. 2010;2010:306462. Epub 2010 Mar 18.

Abstract

The human growth hormone (hGH) has been expressed in prokaryotic expression system with low bioactivity previously. Then the effective B. mori baculovirus system was employed to express hGH identical to mature hGH successfully in larvae, but the expression level was still limited. In this work, the hGH was expressed in B. mori pupae by baculovirus system. Quantification of recombinant hGH protein (BmrhGH) showed that the expression of BmrhGH reached the level of approximately 890 microg/mL pupae supernatant solution, which was five times more than the level using larvae. Furthermore, Animals were gavaged with BmrhGH at the dose of 4.5 mg/rat.day, and the body weight gain (BWG) of treated group had a significant difference (P < .01) compared with the control group. The other two parameters of liver weight and epiphyseal width were also found to be different between the two groups (P < .05). The results suggested that BmrhGH might be used as a protein drug by oral administration.

Citation

PMCID: PMC2842897, PMID: 20339512 [PubMed - indexed for MEDLINE]

 

Title

Enhancement of anti-tumor activity of gamma -irradiated silk fibroin via immunomodulatory effects.

Authors

Byun EB, Sung NY, Kim JH, Choi JI, Matsui T, Byun MW, Lee JW.

 

Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, 1266 Sinjeong-dong, Jeongeup, Republic of Korea.

Journal

Chem Biol Interact. 2010 Jun 7;186(1):90-5. Epub 2010 Mar 23.

Abstract

Silk fibers have proven to be effective in many clinical applications as well as for clothing. In addition to the substantial effect of silk fibers, the present study was conducted to explore its importance in a new dimension to reinforce the effects of its physiological function regarding anti-tumor activity and immune response with gamma-irradiated silk fibroin (GISF). The cytotoxicity results showed that pre-treatment of GISF in the mouse peritoneal macrophages (MPM) indicated a higher proliferative effect than that of non-irradiated silk fibroin (NISF) in a concentration-dependent manner. Based on the cytotoxicity result of MPM, GISF (50 and 150 kGy) was selected for an ex vivo study in an animal (C57BL6) system and evaluated about whether the non-specific immune response was also related to GISF. GISF (50 and 150 kGy) augmented immune responsiveness via activation of NK cells, T lymphocytes proliferation, NO production, and cytokine level, such as IL-6, IL-2, IL-12, IFN-gamma, TNF-alpha, as compared with NISF, which strongly suggested that GISF significantly augmented an important element of all aspects of the innate and adaptive immune system. Therefore, from these results, it seems likely that the GISF will play a potent role in eliciting the effect of the non-specific immune response and anti-tumor activity as a value-added product in the medical industry. Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.

Citation

PMID: 20338156 [PubMed - indexed for MEDLINE]

 

Title

Advancing towards a tissue-engineered tympanic membrane: silk fibroin as a substratum for growing human eardrum keratinocytes.

Authors

Ghassemifar R, Redmond S, Zainuddin, Chirila TV.

 

School of Surgery, Ear Sciences Centre, University of Western Australia, Nedlands, Western Australia, Australia.

Journal

J Biomater Appl. 2010 Mar;24(7):591-606. Epub 2009 Apr 22.

Abstract

Human tympanic membrane cells (hTMCs), harvested from tympanic membrane (TM) explants, were grown in culture and then seeded on membranes prepared from silkworm (Bombyx mori) silk fibroin (BMSF) and on tissue-culture plastic membranes (PET). Fibroin was isolated from silk cast into membranes with a thickness of 10-15 microm. The hTMCs were cultured on both materials for 15 days in a serum-containing culture medium. The cells grown on both substrata were subjected to nuclear staining (DAPI) and counted. Further, the cultures were immunostained for a number of protein markers related to the epithelial/keratinocyte phenotype and cell adhesion complexes. The BMSF membranes supported levels of hTMC growth higher than that observed on the PET membranes. The immunofluorochemical analysis indicated unequivocally that BMSF is a more suitable substratum than PET with respect to the growth patterns, proliferation, and cell-cell contact and adhesion. BMSF appear as a promising substratum in the tissue-engineered constructs for the replacement of TM in case of nonhealing perforations.

Citation

PMID: 20308345 [PubMed - indexed for MEDLINE]

 

Title

The synergistic effects of 3-D porous silk fibroin matrix scaffold properties and hydrodynamic environment in cartilage tissue regeneration.

Authors

Wang Y, Bella E, Lee CS, Migliaresi C, Pelcastre L, Schwartz Z, Boyan BD, Motta A.

 

Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332-0363, USA.

Journal

Biomaterials. 2010 Jun;31(17):4672-81. Epub 2010 Mar 19.

Abstract

Autologous cell-based tissue engineering using three-dimensional porous scaffolds has provided a good option for the repair of cartilage defects. Silk fibroin-based scaffolds are naturally degradable materials with excellent biocompatibility and robust mechanical properties, indicating potential applications in cartilage tissue engineering. In this study, silk fibroin scaffolds prepared by freeze-drying (FD) and salt-leaching (SL300 and SL500) were fully characterized and used to study the effects of silk fibroin scaffold properties on chondrocyte attachment, proliferation and differentiation. The synergistic effects of scaffold properties and hydrodynamic environment generated by in vitro rocking culture were also investigated using static cultures as control. FD scaffolds with small pore size and lower porosity increased cell attachment but inhibited cell penetration and limited cell proliferation and differentiation. In contrast, SL scaffolds displaying a bigger pore size, higher porosity and crystallinity resulted in homogenous cell distribution, increasing cell proliferation and advanced chondrocyte differentiation in terms of their spherical morphology, predominant chondrogenic gene expression and abundant cartilaginous extracellular matrix production. A hydrodynamic environment was beneficial to chondrocyte proliferation, differentiation, and integrin gene expression in a pore size dependent manner with superior cartilage matrix production but limited hypertrophic differentiation obtained using chondrocyte-seeded SL500 scaffolds. Integrin alpha5beta1 might mediate these effects. Chondrocyte/SL500 silk fibroin constructs obtained under in vitro rocking culture might serve as an excellent implant for in vivo cartilage defect reparation. Copyright 2010 Elsevier Ltd. All rights reserved.

Citation

PMID: 20303584 [PubMed - indexed for MEDLINE]

 

Title

Silk-based materials for biomedical applications.

Authors

Leal-Egaña A, Scheibel T.

 

Lehrstuhl für Biomaterialien, Fakultät für Angewandte Naturwissenschaften, Universität Bayreuth, 95440 Bayreuth, Germany.

Journal

Biotechnol Appl Biochem. 2010 Mar 12;55(3):155-67.

Abstract

Since the beginning of civilization, humans have exploited nature as an extraordinary source of materials for medical applications. Most natural materials comprise biopolymers such as nucleic acids and protein-polysaccharides. For biomedical applications, proteins such as collagens have been traditionally employed. Other proteins are silk fibres produced by arthropods (e.g. silkworms and spiders), which provide interesting mechanical properties and the absence of toxicity. Silks present almost all characteristics desirable for biomedical applications, but the research on the underlying proteins has only recently commenced. In the present review, we summarize the current research related to silk being used as a material for cell culture and tissue engineering, particularly focusing on cell-surface adherence, mechanical and textural properties, toxicity, immunogenicity and biodegradability.

Citation

PMID: 20222871 [PubMed - indexed for MEDLINE]

 

Title

Controlling silk fibroin particle features for drug delivery.

Authors

Lammel AS, Hu X, Park SH, Kaplan DL, Scheibel TR.

 

Lehrstuhl Zellbiophysik E27, James-Franck-Strasse, Technische Universität München, Garching, Germany.

Journal

Biomaterials. 2010 Jun;31(16):4583-91. Epub 2010 Mar 9.

Abstract

Silk proteins are a promising material for drug delivery due to their aqueous processability, biocompatibility, and biodegradability. A simple aqueous preparation method for silk fibroin particles with controllable size, secondary structure and zeta potential is reported. The particles were produced by salting out a silk fibroin solution with potassium phosphate. The effect of ionic strength and pH of potassium phosphate solution on the yield and morphology of the particles was determined. Secondary structure and zeta potential of the silk particles could be controlled by pH. Particles produced by salting out with 1.25 m potassium phosphate pH 6 showed a dominating silk II (crystalline) structure whereas particles produced at pH 9 were mainly composed of silk I (less crystalline). The results show that silk I-rich particles possess chemical and physical stability and secondary structure which remained unchanged during post treatments even upon exposure to 100% ethanol or methanol. A model is presented to explain the process of particle formation based on intra- and intermolecular interactions of the silk domains, influenced by pH and kosmotropic salts. The reported silk fibroin particles can be loaded with small molecule model drugs, such as alcian blue, rhodamine B, and crystal violet, by simple absorption based on electrostatic interactions. In vitro release of these compounds from the silk particles depends on charge-charge interactions between the compounds and the silk. With crystal violet we demonstrated that the release kinetics are dependent on the secondary structure of the particles. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Citation

PMCID: PMC2846964 [Available on 2011/6/1], PMID: 20219241 [PubMed - indexed for MEDLINE]

 

Title

A bFGF-releasing silk/PLGA-based biohybrid scaffold for ligament/tendon tissu e engineering using mesenchymal progenitor cells.

Authors

Sahoo S, Toh SL, Goh JC.

 

Department of Orthopaedic Surgery, National University of Singapore, Singapore.

Journal

Biomaterials. 2010 Apr;31(11):2990-8. Epub 2010 Jan 25.

Abstract

An ideal scaffold that provides a combination of suitable mechanical properties along with biological signals is required for successful ligament/tendon regeneration in mesenchymal stem cell-based tissue engineering strategies. Among the various fibre-based scaffolds that have been used, hybrid fibrous scaffolds comprising both microfibres and nanofibres have been recently shown to be particularly promising. This study developed a biohybrid fibrous scaffold system by coating bioactive bFGF-releasing ultrafine PLGA fibres over mechanically robust slowly-degrading degummed knitted microfibrous silk scaffolds. On the ECM-like biomimetic architecture of ultrafine fibres, sustained release of bFGF mimicked the ECM in function, initially stimulating mesenchymal progenitor cell (MPC) proliferation, and subsequently, their tenogeneic differentiation. The biohybrid scaffold system not only facilitated MPC attachment and promoted cell proliferation, with cells growing both on ultrafine PLGA fibres and silk microfibres, but also stimulated tenogeneic differentiation of seeded MPCs. Upregulated gene expression of ligament/tendon-specific ECM proteins and increased collagen production likely contributed to enhancing mechanical properties of the constructs, generating a ligament/tendon analogue that has the potential to be used to repair injured ligaments/tendons. Copyright 2010 Elsevier Ltd. All rights reserved.

Citation

PMID: 20089300 [PubMed - indexed for MEDLINE]

 

Title

Electrospun silk fibroin scaffolds with macropores for bone regeneration: an in vitro and in vivo study.

Authors

Park SY, Ki CS, Park YH, Jung HM, Woo KM, Kim HJ.

 

Department of Dental Anesthesiology and Dental Research Institute, Seoul National University, Seoul, Republic of Korea .

Journal

Tissue Eng Part A. 2010 Apr;16(4):1271-9.

Abstract

We developed three-dimensional electrospun silk fibroin (ESF) scaffolds with controllable pore size. The purpose of this study was to evaluate ESF scaffolds with pores (P-ESF) for bone regeneration via in vitro and in vivo studies, with a comparison to a commercially available porous three-dimensional polylactic acid (PLA) scaffold. P-ESF supported significantly higher proliferation and alkaline phosphatase activity of osteoblasts than PLA in vitro (p < 0.05). Moreover, higher expression levels of activated adhesion-related proteins, including focal adhesion kinase, were observed in the P-ESF than in PLA, as confirmed by western blot analyses. Microcomputed tomography revealed that 78.30% of the original bone volume was attained in the P-ESF implantation group at 7 weeks after critical bone defect formation in rat calvaria. Comparatively, the PLA implantation group showed only 49.31%. Histological evaluation also showed new bone tissue formation upon P-ESF implantation. Taken together, the P-ESF scaffold may be a good bone substitute for bone regeneration.

Citation

PMID: 19905876 [PubMed - indexed for MEDLINE]

 

Title

Antheraea pernyi silk fibroin maintains the immunosupressive properties of human bone marrow mesenchymal stem cells.

Authors

Huo GH, Li MZ, Lu SZ, Zhang XG.

 

Department of Immunology, Binzhou Medical college, Shandong Province, Yantai 264003, People's Republic of China. xiyingluan@yahoo.com.cn

Journal

Cell Biol Int. 2009 Nov;33(11):1127-34. Epub 2009 Aug 5.

Abstract

We reported previously that regenerated Antheraea pernyi silk fibroin (A. pernyi SF) could support the attachment and growth of human bone marrow mesenchymal stem cells (hBMSCs). In this work, the immunosupressive effects of hBMSCs cultured on the A. pernyi SF films on T-cells were investigated in vitro. The production of IL-6, CD80, CD86 and HLA-DR by the hBMSCs was also observed. The study showed that hBMSCs cultured on the regenerated A. pernyi SF films still kept their immunosupression on T-cell proliferation and IL-2 secretion. Moreover, regenerated A. pernyi SF like regenerated Bombyx mori SF and collagen did not elicit T-cell proliferation but it could support the expression of IL-6 and surface antigen of hBMSCs. Regenerated A. pernyi SF can maintain the function of hBMSCs in immunomodulation and cytokines production, which has the potential utility of hBMSCs combined with A. pernyi SF in tissue replacement and repair.

Citation

PMID: 19664716 [PubMed - indexed for MEDLINE]

 

Title

Mulberry fruit (Moris fructus) extracts induce human glioma cell death in vitro through ROS-dependent mitochondrial pathway and inhibits glioma tumor growth in vivo.

Authors

Jeong JC, Jang SW, Kim TH, Kwon CH, Kim YK.

 

Dongguk University, Kyung Ju, Korea.

Journal

Nutr Cancer. 2010 Apr;62(3):402-12.

Abstract

Mulberry has been reported to contain wide range of polyphenols and have chemopreventive activity. However, little has been known regarding the effect of mulberry fruit extracts on cell viability in vitro in human glioma cells and the anticancer efficacy in vivo. This study was undertaken to examine the effect of mulberry fruit (Moris fructus; MF) extracts on cell viability in vitro and anticancer efficacy in vivo. Cell viability and cell death were estimated by MTT assay and trypan blue exclusion assay, respectively. Reactive oxygen species (ROS) generation was measured using the fluorescence probe DCFH-DA. The mitochondrial transmembrane potential was measured with DiOC(6)(3). Bax expression and cytochrome c release were measured by Western blot analysis. Caspase activity was estimated using colorimetric kit. Cell migration was estimated using the scratched wound model. In vivo anticancer efficacy of MF extracts was evaluated using a subcutaneously injected mouse tumor model. Changes in proliferation and apoptosis were estimated by immunohistochemistric analysis. MF extracts resulted in apoptotic cell death in a dose- and time-dependent manner. MF extracts increased ROS generation, and the MF extract-induced cell death was also prevented by antioxidants, suggesting that ROS generation plays a critical role in the MF extract-induced cell death. Western blot analysis showed that treatment of MF extracts caused an increase in Bax expression, which was inhibited by the antioxidant N-acetylcysteine (NAC). MF extracts induced depolarization of mitochondrial membrane potential, and its effect was inhibited by the antioxidants NAC and catalase. MF extracts induced cytochrome c release, which was inhibited by NAC. Caspase activity was stimulated by MF extracts, and caspase inhibitors prevented the MF extract-induced cell death. Treatment of MF extracts inhibited cell migration. Oral MF extracts administration in animals with subcutaneous U87MG glioma cells reduced tumor volume. Subsequent tumor tissue analysis showed a decrease in PCNA-positive cells, an increase in TUNEL-positive cells, and caspase activation. From these data, we concluded that MF extracts reduce glioma tumor growth through inhibition of cell proliferation resulting from induction of apoptosis. These findings suggest that MF extracts result in human glioma cell death in vitro through ROS-dependent mitochondrial pathway and glioma tumor growth in vivo via reduction of tumor cell proliferation and induction of apoptosis.

Citation

PMID: 20358478 [PubMed - indexed for MEDLINE]

 

Title

Three new compounds from Morus nigra L.

Authors

Wang L, Yang Y, Liu C, Chen RY.

 

The Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education & Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Journal

J Asian Nat Prod Res. 2010 Jun;12(6):431-7.

Abstract

A new 2-arylbenzofuran derivative, mornigrol D (1), along with two new flavones, mornigrol G (2) and mornigrol H (3), and six known compounds, norartocarpetin (4), dihydrokaempferol (5), albanin A (6), albanin E (7), moracin M (8), and albafuran C (9), were isolated from the barks of Morus nigra. Their structures were elucidated by spectroscopic analysis. Compounds 1 and 9 showed antioxidative activities in vitro with inhibition ratios of 98 and 99% at the concentration of 10(-4) mol/l, and of 74 and 75% at the concentration of 10(-5) mol/l. In addition, compounds 1 and 4 showed potent anti-inflammatory activities (inhibition of release of beta-glucuronidase from rat polymorphonuclear leucocytes induced by platelet activating factor) with inhibitory ratios of 65.9% (P < 0.01) and 67.7% (P < 0.01) at a concentration of 10(-5) mol/l.

Citation

PMID: 20552479 [PubMed - in process]

 

Title

Chemical constituents of the stem bark of Morus cathayana.

Authors

Ni G, Zhang QJ, Wang YH, Chen RY, Zheng ZF, Yu DQ.

 

The Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Peking Union Medical College, Ministry of Education, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Journal

J Asian Nat Prod Res. 2010 Jun;12(6):505-15.

Abstract

Phytochemical investigation of the stem bark of Morus cathayana led to the isolation and identification of six new compounds, cathayanons F-J (1-5) and cathayanin A (6), and two known compounds, cathayanins B-C (7-8). Their structures were elucidated by spectroscopic methods. Compounds 1, 2, 3, 5, and 7 exhibited weak activities against five human cancer cell lines, with IC(50) values ranging from 4.7 to 9.8 microg/ml.

Citation

PMID: 20552491 [PubMed - in process]

 

Title

Hypolipidemic and antioxidant effects of mulberry (Morus alba L.) fruit in hyperlipidaemia rats.

Authors

Yang X, Yang L, Zheng H.

 

School of Life Science and Technology, Shanxi University, No. 92, Wucheng Road, Taiyuan 030006, Shanxi Province, PR China.

Journal

Food Chem Toxicol. 2010 Jun 1. [Epub ahead of print]

Abstract

The phytochemical constituents of a freeze-dried powder of mulberry (Morus alba L.) fruit (MFP) were determined. The hypolipidemic and antioxidant effects of the MFP as a dietary supplement were evaluated in rats who were fed 4 weeks of either a high-fat or a normal diet supplemented with 5% or 10% MFP. Administration of MFP to rats on a high-fat diet resulted in a significant decline in levels of serum and liver triglyceride, total cholesterol, serum low-density lipoprotein cholesterol, and a decrease in the atherogenic index, while the serum high-density lipoprotein cholesterol was significantly increased. In addition, the serum and liver content of thiobarbituric acid related substances, a lipid peroxidation product, significantly decreased, while the superoxide dismutase (SOD) of red blood cell and liver, as well blood glutathione peroxidase (GSH-Px) activities significantly increased. No significant changes in lipid profile in the serum and liver were observed in rats on a normal diet supplemented with MFP, but blood and liver antioxidant status improved, as measured by SOD and GSH-Px activity, and lipid peroxidation was reduced. These beneficial effects of MFP on hyperlipidaemia rats might be attributed to its dietary fiber, fatty acids, phenolics, flavonoids, anthocyanins, vitamins and trace elements content. Copyright © 2010 Elsevier Ltd. All rights reserved.

Citation

PMID: 20561945 [PubMed - as supplied by publisher]

 

Title

Mulberry fruit (Moris fructus) extracts induce human glioma cell death in vitro through ROS-dependent mitochondrial pathway and inhibits glioma tumor growth in vivo.

Authors

Jeong JC, Jang SW, Kim TH, Kwon CH, Kim YK.

 

Dongguk University, Kyung Ju, Korea.

Journal

Nutr Cancer. 2010 Apr;62(3):402-12.

Abstract

Mulberry has been reported to contain wide range of polyphenols and have chemopreventive activity. However, little has been known regarding the effect of mulberry fruit extracts on cell viability in vitro in human glioma cells and the anticancer efficacy in vivo. This study was undertaken to examine the effect of mulberry fruit (Moris fructus; MF) extracts on cell viability in vitro and anticancer efficacy in vivo. Cell viability and cell death were estimated by MTT assay and trypan blue exclusion assay, respectively. Reactive oxygen species (ROS) generation was measured using the fluorescence probe DCFH-DA. The mitochondrial transmembrane potential was measured with DiOC(6)(3). Bax expression and cytochrome c release were measured by Western blot analysis. Caspase activity was estimated using colorimetric kit. Cell migration was estimated using the scratched wound model. In vivo anticancer efficacy of MF extracts was evaluated using a subcutaneously injected mouse tumor model. Changes in proliferation and apoptosis were estimated by immunohistochemistric analysis. MF extracts resulted in apoptotic cell death in a dose- and time-dependent manner. MF extracts increased ROS generation, and the MF extract-induced cell death was also prevented by antioxidants, suggesting that ROS generation plays a critical role in the MF extract-induced cell death. Western blot analysis showed that treatment of MF extracts caused an increase in Bax expression, which was inhibited by the antioxidant N-acetylcysteine (NAC). MF extracts induced depolarization of mitochondrial membrane potential, and its effect was inhibited by the antioxidants NAC and catalase. MF extracts induced cytochrome c release, which was inhibited by NAC. Caspase activity was stimulated by MF extracts, and caspase inhibitors prevented the MF extract-induced cell death. Treatment of MF extracts inhibited cell migration. Oral MF extracts administration in animals with subcutaneous U87MG glioma cells reduced tumor volume. Subsequent tumor tissue analysis showed a decrease in PCNA-positive cells, an increase in TUNEL-positive cells, and caspase activation. From these data, we concluded that MF extracts reduce glioma tumor growth through inhibition of cell proliferation resulting from induction of apoptosis. These findings suggest that MF extracts result in human glioma cell death in vitro through ROS-dependent mitochondrial pathway and glioma tumor growth in vivo via reduction of tumor cell proliferation and induction of apoptosis.

Citation

PMID: 20358478 [PubMed - indexed for MEDLINE]

 

Title

Topical formulations of serratiopeptidase: development and pharmacodynamic evaluation.

Authors

Nirale NM, Menon MD.

 

Department of Pharmaceutics, Bombay College of Pharmacy, Kalina, Santacruz (East), Mumbai-400 098, India.

Journal

Indian J Pharm Sci. 2010 Jan;72(1):65-71.

Abstract

Serratiopeptidase, an enzyme derived from Serratia marcescences strain E-15 (ATCC 21074), present in the gut wall of the silk worm possesses anti-inflammatory properties, and can prove to be a suitable alternative to commonly used non steroidal antiinflammatory agents. Being sensitive to gastric degradation, serratiopeptidase is conventionally given orally in the form of enteric coated tablet formulations. Topical formulations of serratiopeptidase would be useful to treat local inflammations and may prove to be more effective compared to non steroidal antiinflammatory agents. The present study investigates the feasibility of developing topical preparations of serratiopeptidase in the form of ointments and gels. Excipient compatibility of serratiopeptidase with various excipients and polymers, formulation development, characterization and stability studies have been carried out. Stable formulation was evaluated for anti-inflammatory activity by oxazolone induced ear edema method in mice and allergenic potential by passive cutaneous anaphylaxis.

Citation

PMCID: PMC2883229, PMID: 20582192 [PubMed - in process]

 

Title

Efficient production of human Fas receptor extracellular domain-human IgG1 heavy chain Fc domain fusion protein using baculovirus/silkworm expression system.

Authors

Muraki M, Honda S.

 

Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1, Higashi, Tsukuba, Ibaraki 305-8566, Japan; Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1, Higashi, Tsukuba, Ibaraki 305-8566, Japan.

Journal

Protein Expr Purif. 2010 May 16. [Epub ahead of print]

Abstract

The fusion protein consisting of human Fas receptor extracellular domain and human IgG1 heavy chain Fc domain (hFasRECD-Fc) is a medically important protein that potentially has therapeutic uses. The fusion gene composed of a synthetic human Fas receptor extracellular domain gene and the cDNA encoding human IgG1 heavy chain Fc domain was investigated on the secretory expression using two baculovirus systems which employed either Spodoptera frugiperda 9 (Sf9) cell line or Bombyx mori (silkworm) larvae as the host organism. Both expression systems produced the functional hFasRECD-Fc as a dimer molecule linked by disulfide bridges. The secretion level per unit volume was much higher in the case of silkworm larvae as compared to Sf9 cell line, and was estimated to be more than 150 times. A substantially pure hFasRECD-Fc sample from silkworm larvae was obtained by single step Protein G-agarose affinity column chromatography. The affinity purified sample was further fractionated by anion-exchange chromatography with the final purification yield of 22.5mg from 26ml hemolymph. The hFasRECD-Fc from silkworm larvae and the tag-free human Fas ligand extracellular domain derivative from Pichia pastoris formed a stable complex in solution, which was verified by size-exclusion chromatography. This study demonstrated that the baculovirus/silkworm expression system provided the means for efficient production of highly pure hFasRECD-Fc with functional activity. Copyright © 2010. Published by Elsevier Inc.

Citation

PMID: 20576530 [PubMed - as supplied by publisher]

 

 

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