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  1 / 1854 MEDLINE  
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[PMID]:29352308
[Au] Autor:Qian Q; You Z; Ye L; Che J; Wang Y; Wang S; Zhong B
[Ad] Endereço:College of Animal Sciences, Zhejiang University, Hangzhou, P. R. China.
[Ti] Título:High-efficiency production of human serum albumin in the posterior silk glands of transgenic silkworms, Bombyx mori L.
[So] Source:PLoS One;13(1):e0191507, 2018.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Human serum albumin (HSA) is an important biological preparation with a variety of biological functions in clinical applications. In this study, the mRNA of a fusion transposase derived from the pESNT-PBase plasmid and a pBHSA plasmid containing the HSA gene under the control of a fibroin light chain (FL) promoter were co-injected into fertilized eggs. Fifty-six transgenic silkworm pedigrees expressing theexogenous recombinant HSA (rHSA) in the posterior silk glands (PSGs) with stable inheritance were successfully obtained. The SDS-PAGE and Western blot results confirmed that the rHSA was secreted into the transgenic silkworm cocoon, and the rHSA could be easily extracted with phosphate-buffered saline (PBS). In our research, the isolated highest amount rHSA constituted up to 29.1% of the total soluble protein of the cocoon shell, indicating that the transgenic silkworm produced an average of 17.4 µg/mg of rHSA in the cocoon shell. The production of soluble rHSA in the PSGs by means of generating transgenic silkworms is a novel approach, whereby a large amount of virus-free and functional HSA can be produced through the simple rearing of silkworms.
[Mh] Termos MeSH primário: Bombyx/genética
Bombyx/metabolismo
Albumina Sérica Humana/biossíntese
Albumina Sérica Humana/genética
[Mh] Termos MeSH secundário: Animais
Animais Geneticamente Modificados
Bombyx/crescimento & desenvolvimento
Fibroínas/genética
Expressão Gênica
Genes de Insetos
Seres Humanos
Plasmídeos/genética
Regiões Promotoras Genéticas
Proteínas Recombinantes/biossíntese
Proteínas Recombinantes/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Recombinant Proteins); 9007-76-5 (Fibroins); ZIF514RVZR (Serum Albumin, Human)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180226
[Lr] Data última revisão:
180226
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180121
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0191507


  2 / 1854 MEDLINE  
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[PMID]:29025657
[Au] Autor:Johari N; Madaah Hosseini HR; Samadikuchaksaraei A
[Ad] Endereço:Department of Materials Science and Engineering, Sharif University of Technology, Tehran 1458889694, Iran.
[Ti] Título:Novel fluoridated silk fibroin/ TiO nanocomposite scaffolds for bone tissue engineering.
[So] Source:Mater Sci Eng C Mater Biol Appl;82:265-276, 2018 Jan 01.
[Is] ISSN:1873-0191
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:It is known that Fluoride ions strongly affect bone mineralization and formation. In the present study, the engineered bone tissue scaffolds are fabricated using silk fibroin (SF) and flouridated TiO nanoparticles. TiO nanoparticles are modified by fluoride ions, and different levels (0, 5, 10, 15 and 20wt%) of the fluoridated TiO nanoparticles (TiO -F) were subsequently added to the SF matrix through phase separation method to prepare silk fibroin/flouridated TiO nanocomposite scaffolds (SF/TiO -F). Phase structure, functional groups, morphology and mechanical properties of the obtained scaffolds were evaluated by X-ray diffraction method (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and compressive testing, respectively. In vitro degradation studies of scaffolds were performed by incubating the samples in phosphate buffered saline (PBS) at 37°C and pH7.4 for 30days. Additionally, the bioactivity of scaffolds was estimated in a simulated body fluid (SBF) buffered at 37°C and pH7.4 for 28days. Moreover, MTT assay was used to confirm the biocompatibility of the scaffolds using human like SaOS-2 osteoblast cell line for 1, 3 and 5days. The obtained results indicated that the mechanical properties of scaffolds have been improved by increasing the TiO -F amount up to 15wt%. However, a detrimental effect was observed by a further increase in the TiO -F content. The bioactivity of SF/TiO -F nanocomposite scaffolds was promoted by flouridation of TiO . Furthermore, cell cytotoxicity results demonstrated that the SF/TiO -F nanocomposite scaffolds are nontoxic to osteoblasts. The cell fixation results after 3days of incubation revealed that the cell attachment and spreading on SF/TiO -F nanocomposite scaffolds are improved with respect to SF/TiO nanocomposite scaffolds control sample.
[Mh] Termos MeSH primário: Materiais Biocompatíveis/química
Fibroínas/química
Nanocompostos/química
Titânio/química
[Mh] Termos MeSH secundário: Materiais Biocompatíveis/toxicidade
Linhagem Celular
Sobrevivência Celular/efeitos dos fármacos
Força Compressiva
Halogenação
Seres Humanos
Microscopia Eletrônica de Varredura
Porosidade
Seda/química
Espectroscopia de Infravermelho com Transformada de Fourier
Difração de Raios X
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biocompatible Materials); 0 (Silk); 15FIX9V2JP (titanium dioxide); 9007-76-5 (Fibroins); D1JT611TNE (Titanium)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180205
[Lr] Data última revisão:
180205
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171014
[St] Status:MEDLINE


  3 / 1854 MEDLINE  
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[PMID]:29037999
[Au] Autor:Aykac A; Karanlik B; Sehirli AO
[Ad] Endereço:Near East University, Faculty of Medicine, Department of Biophysics, Nicosia, Cyprus.
[Ti] Título:Protective effect of silk fibroin in burn injury in rat model.
[So] Source:Gene;641:287-291, 2018 Jan 30.
[Is] ISSN:1879-0038
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Activation of pro-inflamatuar pathways play major role in formation of major complications as a result of burns. This study was planned to investigate the protective effect of Silk Fibroin in lung injury caused by burn in the experimental rat model. After rinsing the skin of rats under ether anesthesia, the exposed back region, covers 30% of the total body, was kept in the 90°C water bath for 10s. The control rats were kept in the 25°C water bath for 10s. Immediately after burning process, silk fibroin was administered orally at a dose of 600mg/kg. After 24h following burning from all groups the levels of TNF-α, IL-1ß in blood samples and the MDA, GSH and the activity of MPO were determined from taken lung tissues. Moreover, the expression of Bcl-2/Bax, Caspase-3 and Caspase-9 were determined. Significant increase in TNF-α, IL-1ß, Casp-3 and Casp-9 levels were observed in the Silk Fibroin-treated burn group (p<0.05) whereas for ratio of Bcl-2/Bax, a significant reduction was observed compared to control group (p<0.05). Increased levels of TNF-α, IL-1ß, Caspase-3 and Caspase-9 in Silk Fibroin-treated burn groups were found to be reversed. Silk fibroin can be an effective biomaterial in diminishing burn injury in tissue and apoptosis.
[Mh] Termos MeSH primário: Bombyx/metabolismo
Queimaduras/tratamento farmacológico
Fibroínas/farmacologia
Lesão Pulmonar/tratamento farmacológico
Substâncias Protetoras/farmacologia
Seda/farmacologia
[Mh] Termos MeSH secundário: Animais
Apoptose/efeitos dos fármacos
Caspase 3/metabolismo
Caspase 9/metabolismo
Modelos Animais de Doenças
Feminino
Interferon-alfa/metabolismo
Interleucina-1beta/metabolismo
Lesão Pulmonar/metabolismo
Masculino
Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
Ratos
Ratos Wistar
Proteína X Associada a bcl-2/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Interferon-alpha); 0 (Interleukin-1beta); 0 (Protective Agents); 0 (Proto-Oncogene Proteins c-bcl-2); 0 (Silk); 0 (bcl-2-Associated X Protein); 0 (fibroin, silkworm); 9007-76-5 (Fibroins); EC 3.4.22.- (Caspase 3); EC 3.4.22.- (Caspase 9)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171018
[St] Status:MEDLINE


  4 / 1854 MEDLINE  
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[PMID]:28213972
[Au] Autor:Wei G; Wang L; Dong D; Teng Z; Shi Z; Wang K; An G; Guan Y; Han B; Yao M; Xian CJ
[Ad] Endereço:Department of Orthopaedics, The 1st Affiliated Hospital of Harbin Medical University, Harbin, China.
[Ti] Título:Promotion of cell growth and adhesion of a peptide hydrogel scaffold via mTOR/cadherin signaling.
[So] Source:J Cell Physiol;233(2):822-829, 2018 Feb.
[Is] ISSN:1097-4652
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Understanding neurite outgrowth, orientation, and migration is important for the design of biomaterials that interface with the neural tissue. However, the molecular signaling alternations have not been well elucidated to explain the impact of hydrogels on cell morphology. In our previous studies, a silk fibroin peptide (SF16) hydrogel was found to be an effective matrix for the viability, morphology, and proliferation of PC12 rat pheocrhomocytoma cells. We found that PC12 cells in the peptide hydrogel exhibited adhesive morphology compared to those cultured in agarose or collagen. Moreover, we identified that cell adhesion molecules (E- and N-cadherin) controlled by mTOR signaling were highly induced in PC12 cells cultured in the SF16 peptide hydrogel. Our findings suggest that the SF16 peptide might be suitable to be a cell-adhesion material in cell culture or tissue engineering, and mTOR/cadherin signaling is required for the cell adhesion in the SF16-peptide hydrogel.
[Mh] Termos MeSH primário: Caderinas/metabolismo
Adesão Celular
Proliferação Celular
Fibroínas/metabolismo
Proteínas do Tecido Nervoso/metabolismo
Neurônios/enzimologia
Peptídeos/metabolismo
Serina-Treonina Quinases TOR/metabolismo
Tecidos Suporte
[Mh] Termos MeSH secundário: Animais
Adesão Celular/efeitos dos fármacos
Proliferação Celular/efeitos dos fármacos
Forma Celular
Hidrogéis
Neurônios/efeitos dos fármacos
Células PC12
Inibidores de Proteínas Quinases/farmacologia
Ratos
Transdução de Sinais
Sirolimo/farmacologia
Serina-Treonina Quinases TOR/antagonistas & inibidores
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (CDH1 protein, rat); 0 (Cadherins); 0 (Hydrogels); 0 (N-cadherin, rat); 0 (Nerve Tissue Proteins); 0 (Peptides); 0 (Protein Kinase Inhibitors); 0 (fibroin, silkworm); 9007-76-5 (Fibroins); EC 2.7.1.1 (TOR Serine-Threonine Kinases); EC 2.7.1.1 (mTOR protein, rat); W36ZG6FT64 (Sirolimus)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170219
[St] Status:MEDLINE
[do] DOI:10.1002/jcp.25864


  5 / 1854 MEDLINE  
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[PMID]:28919426
[Au] Autor:Jung H; Kim YY; Kim B; Nam H; Suh JG
[Ad] Endereço:Department of Medical Genetics, College of Medicine, Hallym University, Chuncheon, Gangwon-do, 24252, Republic of Korea.
[Ti] Título:Improving glycemic control in model mice with type 2 diabetes by increasing superoxide dismutase (SOD) activity using silk fibroin hydrolysate (SFH).
[So] Source:Biochem Biophys Res Commun;493(1):115-119, 2017 Nov 04.
[Is] ISSN:1090-2104
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Islet cell dysfunction in type 2 diabetes is primarily attributed to the increased apoptosis of pancreatic beta cells. Silk fibroin hydrolysate (SFH) has an effect on blood in type 2 diabetes model mice (C57BL/KsJ-db/db). However, its exact mechanism is unknown. The type 2 diabetes model mice were randomly divided into non-diabetic mice (ND), diabetic mice (DB), and diabetic mice treated with silk fibroin hydrolysate (DB-SFH). The results showed that SFH significantly decreased fasting blood glucose and hemoglobin A1c (HbA1c). The oral glucose tolerance and insulin tolerance were significantly improved in the DB-SFH group. The DB-SFH group exhibited increased superoxide dismutase (SOD) activity in the plasma, as well as increased Mn-SOD and CuZn-SOD activities in the pancreatic islets. Furthermore, the pancreatic islet cells' death was decreased in the DB-SFH group. In the DB-SFH group, the protein expression of caspase-3 was significantly decreased compared with the DB group. The expression of the Nkx6.1 and Pdx1 proteins were increased in the DB-SFH group. The results suggest that SFH prevents the degeneration of pancreatic islets via increasing SOD while hyperglycemia is alleviated by maintaining beta cell mass in type 2 diabetes model mice.
[Mh] Termos MeSH primário: Diabetes Mellitus Tipo 2/tratamento farmacológico
Diabetes Mellitus Tipo 2/metabolismo
Fibroínas/administração & dosagem
Ilhotas Pancreáticas/metabolismo
Hidrolisados de Proteína/administração & dosagem
Superóxido Dismutase/metabolismo
[Mh] Termos MeSH secundário: Animais
Ativação Enzimática/efeitos dos fármacos
Insulina/sangue
Resistência à Insulina
Ilhotas Pancreáticas/efeitos dos fármacos
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Camundongos Transgênicos
Superóxido Dismutase/efeitos dos fármacos
Resultado do Tratamento
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Insulin); 0 (Protein Hydrolysates); 9007-76-5 (Fibroins); EC 1.15.1.1 (Superoxide Dismutase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171023
[Lr] Data última revisão:
171023
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170919
[St] Status:MEDLINE


  6 / 1854 MEDLINE  
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[PMID]:28832627
[Au] Autor:Malay AD; Arakawa K; Numata K
[Ad] Endereço:Enzyme Research Team, Center for Sustainable Resource Science, RIKEN, Wako-shi, Saitama, Japan.
[Ti] Título:Analysis of repetitive amino acid motifs reveals the essential features of spider dragline silk proteins.
[So] Source:PLoS One;12(8):e0183397, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The extraordinary mechanical properties of spider dragline silk are dependent on the highly repetitive sequences of the component proteins, major ampullate spidroin 1 and 2 (MaSp2 and MaSp2). MaSp sequences are dominated by repetitive modules composed of short amino acid motifs; however, the patterns of motif conservation through evolution and their relevance to silk characteristics are not well understood. We performed a systematic analysis of MaSp sequences encompassing infraorder Araneomorphae based on the conservation of explicitly defined motifs, with the aim of elucidating the essential elements of MaSp1 and MaSp2. The results show that the GGY motif is nearly ubiquitous in the two types of MaSp, while MaSp2 is invariably associated with GP and di-glutamine (QQ) motifs. Further analysis revealed an extended MaSp2 consensus sequence in family Araneidae, with implications for the classification of the archetypal spidroins ADF3 and ADF4. Additionally, the analysis of RNA-seq data showed the expression of a set of distinct MaSp-like variants in genus Tetragnatha. Finally, an apparent association was uncovered between web architecture and the abundance of GP, QQ, and GGY motifs in MaSp2, which suggests a co-expansion of these motifs in response to the evolution of spiders' prey capture strategy.
[Mh] Termos MeSH primário: Motivos de Aminoácidos
Fibroínas/química
Seda/química
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
Sequência Conservada
Aranhas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Silk); 0 (spidroin 1); 147883-43-0 (spidroin 2); 9007-76-5 (Fibroins)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171019
[Lr] Data última revisão:
171019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170824
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183397


  7 / 1854 MEDLINE  
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[PMID]:28817694
[Au] Autor:Duchi S; Piccinini F; Pierini M; Bevilacqua A; Torre ML; Lucarelli E; Santi S
[Ad] Endereço:Osteoarticolar Regeneration Laboratory, Rizzoli Orthopaedic Institute, Bologna, Italy.
[Ti] Título:A new holistic 3D non-invasive analysis of cellular distribution and motility on fibroin-alginate microcarriers using light sheet fluorescent microscopy.
[So] Source:PLoS One;12(8):e0183336, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading. In this work, we propose a non-invasive and non-destructive imaging analysis for investigating motility, viability and distribution of Mesenchymal Stem Cells (MSCs) on silk fibroin-based alginate microcarriers, to test the adhesion capacity of the fibroin coating onto alginate which is known to be unsuitable for cell adhesion. However, in depth characterization of the biomaterial is beyond the scope of this paper. Scaffold-loaded MSCs were stained with Calcein-AM and Ethidium homodimer-1 to detect live and dead cells, respectively, and counterstained with Hoechst to label cell nuclei. Time-lapse Light Sheet Fluorescent Microscopy (LSFM) was then used to produce three-dimensional images of the entire cells-loaded fibroin/alginate microcarriers. In order to quantitatively track the cell motility over time, we also developed an open source user friendly software tool called Fluorescent Cell Tracker in Three-Dimensions (F-Tracker3D). Combining LSFM with F-Tracker3D we were able for the first time to assess the distribution and motility of stem cells in a non-invasive, non-destructive, quantitative, and three-dimensional analysis of the entire surface of the cell-loaded scaffold. We therefore propose this imaging technique as an innovative holistic tool for monitoring cell-biomaterial interactions, and as a tool for the design, fabrication and functionalization of a scaffold as a medical device.
[Mh] Termos MeSH primário: Alginatos/química
Fibroínas/química
Microscopia de Fluorescência/métodos
[Mh] Termos MeSH secundário: Adesão Celular
Ácido Glucurônico/química
Ácidos Hexurônicos/química
Seres Humanos
Células Mesenquimais Estromais/citologia
Engenharia Tecidual/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Alginates); 0 (Hexuronic Acids); 8A5D83Q4RW (Glucuronic Acid); 8C3Z4148WZ (alginic acid); 9007-76-5 (Fibroins)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171016
[Lr] Data última revisão:
171016
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170818
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183336


  8 / 1854 MEDLINE  
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[PMID]:28687847
[Au] Autor:Vázquez N; Rodríguez-Barrientos CA; Aznar-Cervantes SD; Chacón M; Cenis JL; Riestra AC; Sánchez-Avila RM; Persinal M; Brea-Pastor A; Fernández-Vega Cueto L; Meana Á; Merayo-Lloves J
[Ad] Endereço:Instituto Universitario Fernández-Vega, Fundación de Investigación Oftalmológica, Universidad de Oviedo, Asturias, Spain.
[Ti] Título:Silk Fibroin Films for Corneal Endothelial Regeneration: Transplant in a Rabbit Descemet Membrane Endothelial Keratoplasty.
[So] Source:Invest Ophthalmol Vis Sci;58(9):3357-3365, 2017 Jul 01.
[Is] ISSN:1552-5783
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Purpose: Develop a silk fibroin (SF)-based artificial endothelial graft for its use in a rabbit Descemet membrane endothelial keratoplasty (DMEK). Methods: Human and rabbit artificial corneal endothelial grafts were developed through the culture of human and rabbit corneal endothelial cells (CECs) on SF films. Rabbit artificial SF endothelial grafts were transplanted in a DMEK surgery into a rabbit in vivo model. Results: SF artificial endothelial grafts showed the characteristic endothelial markers: zonula occludens (ZO-1) and Na+/K+ ATPase. In a rabbit model of DMEK surgery, SF artificial endothelial graft restored the corneal transparency and thickness at 6 week of follow-up. Anterior segment optical coherence tomography revealed the SF graft as a fully integrated component in the corneal tissue, displaying a similar corneal thickness and endothelial cell count when compared with its healthy contralateral cornea. Histologic analysis showed that the SF artificial endothelial graft was attached and integrated on the surface of the corneal stroma without a significant inflammatory reaction, and rabbit CECs consisted in a monolayer that showed their characteristic markers ZO-1 and Na+/K+ ATPase, suggesting proper intercellular junctions and cellular pump function. Conclusions: We have developed SF films with biological properties that supported the growth of rabbit and human CECs, which showed normal morphology and characteristic markers; and with mechanical properties that allowed its use in a DMEK surgery, proving its in vivo functionality in a rabbit model of endothelial dysfunction.
[Mh] Termos MeSH primário: Edema da Córnea/cirurgia
Ceratoplastia Endotelial com Remoção da Lâmina Limitante Posterior/métodos
Epitélio Posterior/transplante
Fibroínas
Membranas Artificiais
Seda
Engenharia Tecidual
[Mh] Termos MeSH secundário: Animais
Células Cultivadas
Edema da Córnea/patologia
Modelos Animais de Doenças
Células Endoteliais/citologia
Seres Humanos
Coelhos
Engenharia Tecidual/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Membranes, Artificial); 0 (Silk); 9007-76-5 (Fibroins)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170714
[Lr] Data última revisão:
170714
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170709
[St] Status:MEDLINE
[do] DOI:10.1167/iovs.17-21797


  9 / 1854 MEDLINE  
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[PMID]:28686698
[Au] Autor:Vetsch JR; Betts DC; Müller R; Hofmann S
[Ad] Endereço:Institute for Biomechanics, Swiss Federal Institute of Technology Zürich, Zurich, Switzerland.
[Ti] Título:Flow velocity-driven differentiation of human mesenchymal stromal cells in silk fibroin scaffolds: A combined experimental and computational approach.
[So] Source:PLoS One;12(7):e0180781, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Mechanical loading plays a major role in bone remodeling and fracture healing. Mimicking the concept of mechanical loading of bone has been widely studied in bone tissue engineering by perfusion cultures. Nevertheless, there is still debate regarding the in-vitro mechanical stimulation regime. This study aims at investigating the effect of two different flow rates (vlow = 0.001m/s and vhigh = 0.061m/s) on the growth of mineralized tissue produced by human mesenchymal stromal cells cultured on 3-D silk fibroin scaffolds. The flow rates applied were chosen to mimic the mechanical environment during early fracture healing or during bone remodeling, respectively. Scaffolds cultured under static conditions served as a control. Time-lapsed micro-computed tomography showed that mineralized extracellular matrix formation was completely inhibited at vlow compared to vhigh and the static group. Biochemical assays and histology confirmed these results and showed enhanced osteogenic differentiation at vhigh whereas the amount of DNA was increased at vlow. The biological response at vlow might correspond to the early stage of fracture healing, where cell proliferation and matrix production is prominent. Visual mapping of shear stresses, simulated by computational fluid dynamics, to 3-D micro-computed tomography data revealed that shear stresses up to 0.39mPa induced a higher DNA amount and shear stresses between 0.55mPa and 24mPa induced osteogenic differentiation. This study demonstrates the feasibility to drive cell behavior of human mesenchymal stromal cells by the flow velocity applied in agreement with mechanical loading mimicking early fracture healing (vlow) or bone remodeling (vhigh). These results can be used in the future to tightly control the behavior of human mesenchymal stromal cells towards proliferation or differentiation. Additionally, the combination of experiment and simulation presented is a strong tool to link biological responses to mechanical stimulation and can be applied to various in-vitro cultures to improve the understanding of the cause-effect relationship of mechanical loading.
[Mh] Termos MeSH primário: Calcificação Fisiológica
Fibroínas/farmacologia
Células Mesenquimais Estromais/citologia
Osteogênese
Engenharia Tecidual/métodos
[Mh] Termos MeSH secundário: Fenômenos Biomecânicos
Reatores Biológicos
Regeneração Óssea/fisiologia
Osso e Ossos/citologia
Osso e Ossos/fisiologia
Técnicas de Cultura de Células
Diferenciação Celular/efeitos dos fármacos
Proliferação Celular
Matriz Extracelular/metabolismo
Fibroínas/química
Seres Humanos
Peptídeos e Proteínas de Sinalização Intercelular/farmacologia
Células Mesenquimais Estromais/efeitos dos fármacos
Células Mesenquimais Estromais/fisiologia
Cultura Primária de Células
Reologia
Estresse Mecânico
Imagem com Lapso de Tempo
Tecidos Suporte
Microtomografia por Raio-X
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Intercellular Signaling Peptides and Proteins); 9007-76-5 (Fibroins)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170922
[Lr] Data última revisão:
170922
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170708
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0180781


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[PMID]:28542539
[Au] Autor:Lyda TA; Wagner EL; Bourg AX; Peng C; Tomaraei GN; Dean D; Kennedy MS; Marcotte WR
[Ad] Endereço:Department of Genetics and Biochemistry, Clemson University, Clemson, South Carolina, United States of America.
[Ti] Título:A Leishmania secretion system for the expression of major ampullate spidroin mimics.
[So] Source:PLoS One;12(5):e0178201, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Spider major ampullate silk fibers have been shown to display a unique combination of relatively high fracture strength and toughness compared to other fibers and show potential for tissue engineering scaffolds. While it is not possible to mass produce native spider silks, the potential ability to produce fibers from recombinant spider silk fibers could allow for an increased innovation rate within tissue engineering and regenerative medicine. In this pilot study, we improved upon a prior fabrication route by both changing the expression host and additives to the fiber pulling precursor solution to improve the performance of fibers. The new expression host for producing spidroin protein mimics, protozoan parasite Leishmania tarentolae, has numerous advantages including a relatively low cost of culture, rapid growth rate and a tractable secretion pathway. Tensile testing of hand pulled fibers produced from these spidroin-like proteins demonstrated that additives could significantly modify the fiber's mechanical and/or antimicrobial properties. Cross-linking the proteins with glutaraldehyde before fiber pulling resulted in a relative increase in tensile strength and decrease in ductility. The addition of ampicillin into the spinning solution resulted in the fibers being able to inhibit bacterial growth.
[Mh] Termos MeSH primário: Materiais Biomiméticos
Fibroínas/biossíntese
Leishmania/metabolismo
[Mh] Termos MeSH secundário: Ampicilina/farmacologia
Antibacterianos/farmacologia
Materiais Biomiméticos/farmacologia
Reatores Biológicos
Western Blotting
Reagentes para Ligações Cruzadas/química
Escherichia coli
Fibroínas/química
Fibroínas/farmacologia
Fibroínas/ultraestrutura
Glutaral/química
Leishmania/genética
Indústria Manufatureira
Teste de Materiais
Microscopia Eletrônica de Varredura
Projetos Piloto
Proteínas Recombinantes/biossíntese
Proteínas Recombinantes/química
Proteínas Recombinantes/farmacologia
Proteínas Recombinantes/ultraestrutura
Soluções
Resistência à Tração
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anti-Bacterial Agents); 0 (Cross-Linking Reagents); 0 (Recombinant Proteins); 0 (Solutions); 7C782967RD (Ampicillin); 9007-76-5 (Fibroins); T3C89M417N (Glutaral)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170918
[Lr] Data última revisão:
170918
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170526
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0178201



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