Regeneration of the femoral epicondyle on calcium-binding silk scaffolds developed using transgenic silk fibroin produced by transgenic silkworm

2011 ◽  
Vol 7 (3) ◽  
pp. 1192-1201 ◽  
Author(s):  
Aya Nagano ◽  
Yumi Tanioka ◽  
Nobuko Sakurai ◽  
Hideki Sezutsu ◽  
Noboru Kuboyama ◽  
...  
Keyword(s):  
Silk Fibroin ◽  
Calcium Binding ◽  
Transgenic Silkworm ◽  
Silk Scaffolds

Recombinant silk fibroin incorporated cell-adhesive sequences produced by transgenic silkworm as a possible candidate for use in vascular graft

2014 ◽  
Vol 2 (42) ◽  
pp. 7375-7383 ◽  
Author(s):  
Tetsuo Asakura ◽  
Makoto Isozaki ◽  
Toshiki Saotome ◽  
Ken-ichiro Tatematsu ◽  
Hideki Sezutsu ◽  
...  
Keyword(s):  
Silk Fibroin ◽  
Vascular Graft ◽  
Transgenic Silkworm ◽  
Cell Adhesive ◽  
Silk Fibroins

Transgenic silk fibroins that incorporated the laminin sequence were prepared. The adhesive activities tend to increase in the TG silk fibroins relative to WT silk fibroins.

scholarly journals Effect of Pore Size on the Biodegradation Rate of Silk Fibroin Scaffolds

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Zuwei Luo ◽  
Qin Zhang ◽  
Meijing Shi ◽  
Yang Zhang ◽  
Wei Tao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Weight Loss ◽  
Pore Size ◽  
Silk Fibroin ◽  
Degradation Rate ◽  
Structural Integrity ◽  
Pore Density ◽  
Tissue Engineering Scaffolds ◽  
Silk Scaffolds

Controlling the degradation rate of silk fibroin-based biomaterial is an important capability for the fabrication of silk-based tissue engineering scaffolds. In this study, scaffolds with different pore sizes were prepared by controlling the freezing temperature and the silk fibroin concentration.In vitrodegradation results showed that the internal pore walls of the scaffolds with a larger pore size collapsed upon exposure to collagenase IA for times ranging from 6 to 12 days, and the silk scaffolds exhibited a faster rate of weight loss. The morphological and structural features of the silk scaffolds with a smaller pore size maintained structural integrity after incubation in the protease solution for 18 days, and the rate of weight loss was relatively slow. Scaffolds with a smaller pore size or a higher pore density degraded more slowly than scaffolds with a larger pore size or lower pore density. These results demonstrate that the pore size of silk biomaterials is crucial in controlling the degradation rate of tissue engineering scaffolds.

scholarly journals Chondrogenic differentiation of Wharton’s Jelly mesenchymal stem cells on silk spidroin-fibroin mix scaffold supplemented with L-ascorbic acid and platelet rich plasma

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Anggraini Barlian ◽  
Hermawan Judawisastra ◽  
Ahmad Ridwan ◽  
Antonia Ratih Wahyuni ◽  
Meidiana Ebtayani Lingga
Keyword(s):  
Cell Proliferation ◽  
Compressive Strength ◽  
Ascorbic Acid ◽  
Silk Fibroin ◽  
Chondrogenic Differentiation ◽  
Platelet Rich Plasma ◽  
Blue Staining ◽  
Silk Fibroin Scaffold ◽  
Rgd Sequence ◽  
Silk Scaffolds

Abstract In this research, hWJ-MSCs were grown on silk scaffolds and induced towards chondrogenesis by supplementation with L-ascorbic acid (LAA) or platelet rich plasma (PRP). Silk scaffolds were fabricated with salt leaching method by mixing silk fibroin (SF) with silk spidroin (SS). The silk fibroin was obtained from Bombyx mori cocoon that had been degummed, and the silk spidroin was obtained from wild-type spider Argiope appensa. The effect of scaffold composition and inducer on cell proliferation was observed through MTT assay. The most optimal treatment then continued to be used to induce hWJ-MSC towards chondrogenic differentiation for 7 and 21 days. Scaffolds characterization showed that the scaffolds produced had 3D structure with interconnected pores, and all were biocompatible with hWJ-MSCs. Scaffold with the addition of 10% SS + 90% SF showed higher compressive strength and better pore interconnectivity in comparison to 100% silk fibroin scaffold. After 48 h, cells seeded on scaffold with spidroin and fibroin mix had flattened morphology in comparison to silk fibroin scaffold which appeared to be more rounded on the scaffold surface. Scaffold with 10% (w/w) of silk spidroin (SS) + 90% (w/w) of silk fibroin (SF) was the most optimal composition for cell proliferation. Immunocytochemistry of integrin β1 and RGD sequence, showed that scaffold with SS 10% provide better cell attachment with the presence of RGD sequence from the spidroin silk which could explain the higher cell proliferation than SF100% scaffold. Based on Alcian Blue staining and Collagen Type II immunocytochemistry (ICC), cells grown on 10% SS + 90% SF scaffold with 10% PRP supplementation were the most optimal to support chondrogenesis of hWJ-MSCs. These results showed that the addition of spidroin silk from A. appensa. had impact on scaffold compressive strength and chondrogenic differentiation of hWJ-MSC and had the potential for further development of bio-based material scaffold in cartilage tissue engineering.

Introduction of VEGF or RGD sequences improves revascularization properties of Bombyx mori silk fibroin produced by transgenic silkworm

2015 ◽  
Vol 3 (35) ◽  
pp. 7109-7116 ◽  
Author(s):  
Toshiki Saotome ◽  
Haruki Hayashi ◽  
Ryo Tanaka ◽  
Atsushi Kinugasa ◽  
Shouji Uesugi ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Silk Fibroin ◽  
Platelet Adhesion ◽  
Transgenic Silkworm ◽  
Silk Fibroins ◽  
Bombyx Mori Silk

Transgenic silk fibroins incorporated the VEGF and RGD were prepared. The VEGF SF showed lower platelet adhesion than the RGD SF and WT SF. An in vivo implantation study supported these in vitro results.

scholarly journals Characterization of Transgenic Silkworm Yielded Biomaterials with Calcium-Binding Activity

PLoS ONE ◽  
2016 ◽  
Vol 11 (7) ◽  
pp. e0159111 ◽  
Author(s):  
Shaohua Wang ◽  
Yuyu Zhang ◽  
Mingying Yang ◽  
Lupeng Ye ◽  
Lu Gong ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Binding Activity ◽  
Transgenic Silkworm

Immunogold localization of calmodulin in root caps of the maize Cultivar Merit

1990 ◽  
Vol 48 (3) ◽  
pp. 674-675
Author(s):  
P.T. Nguyen ◽  
C. Uphoff ◽  
C.L. Stinemetz
Keyword(s):  
Calcium Binding ◽  
Calcium Transport ◽  
Primary Role ◽  
Immunogold Localization ◽  
Root Tips ◽  
Electrical Currents ◽  
Considerable Evidence ◽  
Maize Cultivar ◽  
Gravity Response ◽  
Induced Changes

Considerable evidence suggest that the calcium-binding protein calmodulin (CaM) may mediate calcium action and/or transport important in the gravity response of plants. Calmodulin is present in both shoots and roots and is capable of regulating calcium transport in plant vesicles. In roots calmodulin is concentrated in the tip, the gravisensing region of the root; and is reported to be closely associated with amyloplasts, organelles suggested to play a primary role in gravi-perception. Inhibitors of CaM such as chlorpromazine, calmidazolium, and compound 48/80 interfere with the gravitropic response of both snoots and roots. The magnitude of the inhibition corresponded well with the extent to which the drug binds to endogenous CaM. Compound 48/80 and calmidazolium block gravi-induced changes in electrical currents across root tips, a phenomenon thought to be associated with the sensing of the gravity stimulus.In this study, we have investigated the subcellular distribution of CaM in graviresponsive and non-graviresponsive root caps of the maize cultivar Merit.

Immunogold labeling of the calcium binding protein synexin in isolated adrenal chromaffin granules and chromaffin cells

1990 ◽  
Vol 48 (3) ◽  
pp. 952-953
Author(s):  
Gemma A.J. Kuijpers ◽  
Harvey B. Pollard
Keyword(s):  
Adrenal Medulla ◽  
Calcium Binding ◽  
Chromaffin Cells ◽  
Cell Activation ◽  
Structural Information ◽  
Dependent Manner ◽  
Chromaffin Granules ◽  
Immuno Electron Microscopy ◽  
Ultrathin Sections ◽  
Adrenal Chromaffin

Exocytotic fusion of granules in the adrenal medulla chromaffin cell is triggered by a rise in the concentration of cytosolic Ca2+ upon cell activation. The protein synexin, annexin VII, was originally found in the adrenal medulla and has been shown to cause aggregation and to support fusion of chromaffin granules in a Ca2+-dependent manner. We have previously suggested that synexin may there fore play a role in the exocytotic fusion process. In order to obtain more structural information on synexin, we performed immuno-electron microscopy on frozen ultrathin sections of both isolated chromaffin granules and chromaffin cells.Chromaffin granules were isolated from bovine adrenal medulla, and synexin was isolated from bovine lung. Granules were incubated in the presence or absence of synexin (24 μg per mg granule protein) and Ca2+ (1 mM), which induces maximal granule aggregation, in 0.3M sucrose-40m MMES buffer(pH 6.0). Granules were pelleted, washed twice in buffer without synexin and fixed with 2% glutaraldehyde- 2% para formaldehyde in 0.1 M phosphate buffer (GA/PFA) for 30 min. Chromaffin cells were isolated and cultured for 3-5 days, and washed and incubated in Krebs solution with or without 20 uM nicotine. Cells were fixed 90 sec after on set of stimulation with GA/PFA for 30 min. Fixed granule or cell pellets were washed, infiltrated with 2.3 M sucrose in PBS, mounted and frozen in liquid N2.

Intracisternal BioGlue injection in the fetal lamb: a novel model for creation of obstructive congenital hydrocephalus without additional chemically induced neuroinflammation

2019 ◽  
Vol 24 (6) ◽  
pp. 652-662 ◽  
Author(s):  
Marc Oria ◽  
Soner Duru ◽  
Federico Scorletti ◽  
Fernando Vuletin ◽  
Jose L. Encinas ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Repeated Measures ◽  
Obstructive Hydrocephalus ◽  
Critical Threshold ◽  
Control Group ◽  
Chronic Obstructive ◽  
Fetal Sheep ◽  
Cisterna Magna ◽  
Lateral Ventricles ◽  
Fetal Lamb

OBJECTIVEThe authors hypothesized that new agents such as BioGlue would be as efficacious as kaolin in the induction of hydrocephalus in fetal sheep.METHODSThis study was performed in 34 fetal lambs randomly divided into 2 studies. In the first study, fetuses received kaolin, BioGlue (2.0 mL), or Onyx injected into the cisterna magna, or no injection (control group) between E85 and E90. In the second study, fetuses received 2.0-mL or 2.5-mL injections of BioGlue into the cisterna magna between E85 and E90. Fetuses were monitored using ultrasound to assess lateral ventricle size and progression of hydrocephalus. The fetuses were delivered (E120–E125) and euthanized for histological analysis. Selected brain sections were stained for ionized calcium binding adaptor 1 (Iba1) and glial fibrillary acidic protein (GFAP) to assess the presence and activation of microglia and astroglia, respectively. Statistical comparisons were performed with Student’s t-test for 2 determinations and ANOVA 1-way and 2-way repeated measures for multiple determinations.RESULTSAt 30 days after injection, the lateral ventricles were larger in all 3 groups that had undergone injection than in controls (mean diameter in controls 3.76 ± 0.05 mm, n = 5). However, dilatation was greater in the fetuses injected with 2 mL of BioGlue (11.34 ± 4.76 mm, n = 11) than in those injected with kaolin (6.4 ± 0.98 mm, n = 7) or Onyx (5.7 ± 0.31 mm, n = 6) (ANOVA, *p ≤ 0.0001). Fetuses injected with 2.0 mL or 2.5 mL of BioGlue showed the same ventricle dilatation but it appeared earlier (at 10 days postinjection) in those injected with 2.5 mL. The critical threshold of ventricle dilatation was 0.1 for all the groups, and only the BioGlue 2.0 mL and BioGlue 2.5 mL groups exceeded this critical value (at 30 days and 18 days after injection, respectively) (ANOVA, *p ≤ 0.0001). Moderate to severe hydrocephalus with corpus callosum disruption was observed in all experimental groups. All experimental groups showed ventriculomegaly with significant microgliosis and astrogliosis in the subventricular zone around the lateral ventricles. Only kaolin resulted in significant microgliosis in the fourth ventricle area (ANOVA, *p ≤ 0.005).CONCLUSIONSThe results of these studies demonstrate that BioGlue is more effective than Onyx or kaolin for inducing hydrocephalus in the fetal lamb and results in a volume-related response by obstructive space-occupancy without local neuroinflammatory reaction. This novel use of BioGlue generates a model with potential for new insights into hydrocephalus pathology and the development of therapeutics in obstructive hydrocephalus. In addition, this model allows for the study of acute and chronic obstructive hydrocephalus by using different BioGlue volumes for intracisternal injection.

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