Construct Human Keratinocytes and Fibroblasts in Bovine Collagen Scaffold

2021 ◽  
Keyword(s):  
Collagen Scaffold ◽  
Human Keratinocytes ◽  
Bovine Collagen

scholarly journals Valorization of Cyprinus Carpio skin for biocompatible collagen hydrolysates with potential application in foods, cosmetics and pharmaceuticals

2021 ◽  
Author(s):  
Elena Danila ◽  
Raluca Stan ◽  
Mădălina ALBU KAYA ◽  
Georgeta VOICU ◽  
Maria Minodora MARIN ◽  
...  
Keyword(s):  
Cyprinus Carpio ◽  
Ph Value ◽  
Human Keratinocytes ◽  
Cd Spectroscopy ◽  
Random Coil ◽  
Collagen Hydrolysate ◽  
Bovine Collagen ◽  
Fish Collagen ◽  
Collagen Hydrolysates ◽  
Hydrolysis Of

Abstract Fish collagen is the safest source of collagen at present, the extraction being performed on secondary sources as skin, bones, scales or fins resulted after fish processing. The aim of the present study was to obtain biocompatible collagen hydrolysates from waste Cyprinus carpio skin, the main aquaculture species in Romania using an inexpensive and “green” neutral hydrolysis process. Neutral hydrolysis of pretreated fish skins performed for 6 hours at a temperature of 135°C and a pressure of 315 kPa produced collagen hydrolysates in 24.6-35.5% yields depending on the adopted pretreatment procedure. The extensive characterization of hydrolysate samples revealed a high purity degree (98% protein content, undetected ash content, pH value in the range 6-7), also confirmed by the absence of undesired aggregates in the characteristic fibril structure as determined by electronic microscopy. A specific collagen hydrolysate random coil structure and the absence of triple helix was determined by FTIR analysis and sustained by CD spectroscopy and X-ray diffraction. The biocompatibility assessment for the obtained fish collagen hydrolysates revealed no cytotoxic effect on Human keratinocytes, with an 80% cell viability, superior as compared to conventional bovine collagen hydrolysate.Neutral hydrolysis of waste Cyprinus carpio skin yielded collagen hydrolysates with determined characteristics and biocompatibility superior to bovine collagen, suitable for application in foods, cosmetics and pharmaceutical industry.

scholarly journals Model for human skin reconstructed in vitro composed of associated dermis and epidermis

2006 ◽  
Vol 124 (2) ◽  
pp. 71-76 ◽  
Author(s):  
Luís Ricardo Martinhão Souto ◽  
Jussara Rehder ◽  
José Vassallo ◽  
Maria Letícia Cintra ◽  
Maria Helena Stangler Kraemer ◽  
...  
Keyword(s):  
Human Skin ◽  
Collagen Type ◽  
Human Fibroblasts ◽  
Skin Grafting ◽  
Human Keratinocytes ◽  
Collagen Type I ◽  
Type I ◽  
Bovine Collagen

CONTEXT AND OBJECTIVE: The technique of obtaining human skin with dermis and epidermis reconstructed from cells isolated from patients can enable autologous skin grafting on patients with few donor sites. It also enables in vitro trials on chemicals and drugs. The objective of this work was to demonstrate a method for obtaining human skin composed of associated dermis and epidermis, reconstructed in vitro. DESIGN AND SETTING: Experimental laboratory study, in the Skin Cell Culture Laboratory of Faculdade de Ciências Médicas, Universidade Estadual de Campinas. METHODS: Cells from human fibroblast cultures are injected into bovine collagen type I matrix and kept immersed in specific culturing medium for fibroblasts. This enables human dermis reconstruction in vitro. On this, by culturing human keratinocytes and melanocytes, differentiated epidermis is formed, leading to the creation of human skin composed of associated dermis and epidermis, reconstructed in vitro. RESULTS: We showed that human skin composed of associated dermis and epidermis can be successfully reconstructed in vitro. It is histologically formed in the same way as human skin in vivo. Collagen tissue can be identified in the dermis, with cells and extracellular matrix organized in parallel to multilayer epidermis. CONCLUSIONS: It is possible to obtain completely differentiated human skin composed of associated dermis and epidermis, reconstructed in vitro, from injection of human fibroblasts into bovine collagen type I matrix and culturing of human keratinocytes and melanocytes on this matrix.

scholarly journals Biomechanical, Biochemical, and Cell Biological Evaluation of Different Collagen Scaffolds for Tendon Augmentation

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Carolin Gabler ◽  
Juliane Spohn ◽  
Thomas Tischer ◽  
Rainer Bader
Keyword(s):  
Biomechanical Testing ◽  
Cell Activity ◽  
Collagen Scaffold ◽  
Biomechanical Properties ◽  
Enzymatic Digestion ◽  
Biological Evaluation ◽  
Collagen Scaffolds ◽  
Beneficial Effects ◽  
Bovine Collagen

Tendon augmentation is increasingly clinically relevant due to rising amount of tendon ruptures because of the aging and more demanding population. Therefore, newly developed scaffolds based on bovine epoxide stabilized collagen maintaining the native fibril-like collagen structure were characterized and compared to two commercially available porcine collagen scaffolds. For biomechanical testing (ultimate load, ultimate stress, stiffness, and elastic modulus), bovine collagen scaffolds were hydrated and compared to reference products. Cell viability and proliferation were assessed by seeding human primary fibroblasts on each collagen-based scaffold and cultured over various time periods (3 d, 7 d, and 14 d). Live/dead staining was performed and metabolic cell activity (WST-1 assay) was measured. Biochemical degradability was investigated by enzymatic digestion. The bovine collagen scaffold showed significantly enhanced biomechanical properties. These persisted over different rehydration times. Cell biological tests revealed that the bovine collagen scaffolds support reproducible cell colonization and a significant increase in the number of viable cells during cultivation. The results are comparable with the viability and proliferation rate of cells grown on porcine reference materials. With regard to biochemical degradability, all tested materials showed comparable resistance to enzymatic degradation in vitro. Due to imitating the natural tendon structure the new scaffold material is supposed to provide beneficial effects in future clinical application.

Kalinin: A new epithelial basement membrane adhesion molecule

1991 ◽  
Vol 49 ◽  
pp. 178-179
Author(s):  
Douglas R. Keene ◽  
Gregory P. Lunstrum ◽  
Patricia Rousselle ◽  
Robert E. Burgeson
Keyword(s):  
Basement Membrane ◽  
Culture Media ◽  
Polyacrylamide Gel Electrophoresis ◽  
Human Keratinocytes ◽  
Positive Staining ◽  
Human Amnion ◽  
Epithelial Basement Membrane ◽  
Type Vii Collagen ◽  
Keratinocyte Cell ◽  
Epithelial Basement

A mouse monoclonal antibody produced from collagenase digests of human amnion was used by LM and TEM to study the distribution and ultrastructural features of an antigen present in epithelial tissues and in cultured human keratinocytes, and by immunoaffinity chromatography to partially purify the antigen from keratinocyte cell culture media.By immunofluorescence microscopy, the antigen displays a tissue distribution similar to type VII collagen; positive staining of the epithelial basement membrane is seen in skin, oral mucosa, trachea, esophagus, cornea, amnion and lung. Images from rotary shadowed preparations isolated by affinity chromatography demonstrate a population of rod-like molecules 107 nm in length, having pronounced globular domains at each end. Polyacrylamide gel electrophoresis suggests that the size of this molecule is approximately 440kDa, and that it is composed of three nonidentical chains disulfide bonded together.

Alterations of ultrastructure and elemental composition in cultured human epidermal keratinocytes after treatment with nitrofurazone and silver sulfadiazine

1987 ◽  
Vol 45 ◽  
pp. 676-677
Author(s):  
A. R. Crooker ◽  
M. C. Myers ◽  
T. L. Beard ◽  
E. S. Graham
Keyword(s):  
Electron Microscopy ◽  
Elemental Composition ◽  
Pyrolytic Carbon ◽  
Human Keratinocytes ◽  
Silver Sulfadiazine ◽  
Epidermal Keratinocytes ◽  
Human Epidermal Keratinocytes ◽  
Culture Systems ◽  
Cytotoxicity Endpoints

Cell culture systems have become increasingly popular as a means of screening toxic agents and studying toxic mechanisms of drugs and other chemicals at the cellular and subcellular levels. These in vitro tests can be conducted rapidly in a broad range of relevant mammalian culture systems; a variety of biological and biochemical cytotoxicity endpoints can be examined. The following study utilized human keratinocytes to evaluate the relative cytotoxicities of nitrofurazone (NF) and silver sulfadiazine (SS), the active ingredients of FURACIN(R) Topical Cream and SILVADENE(R) Cream, respectively. These compounds are anti-infectives used in the treatment of burn patients. Cell ultrastructure and elemental composition were utilized as cytotoxicity endpoints.Normal Human Epidermal Keratinocytes (HK) were prepared from the EpiPackTM culture system (Clonetics Corporation, Boulder, CO). For scanning electron microscopy (SEM) and transmission electron microscopy (TEM), cells were seeded on sterile 35 mm Falcon plastic dishes; for elemental microanalysis, cells were plated on polished pyrolytic carbon discs (E. Fullam, Latham, NY) placed in the culture dishes.

Non-cellulosic glucans as inductors for differentiation of human keratinocytes

Planta Medica ◽  
2013 ◽  
Vol 79 (13) ◽  
Author(s):  
D Zacharski ◽  
S Brandt ◽  
A Hensel
Keyword(s):  
Human Keratinocytes

Thrombin Stimulated Platelet Accumulation on Protein Coated Glass Capillaries: Role of Adhesive Platelet α-Granule Proteins

1989 ◽  
Vol 62 (03) ◽  
pp. 989-995 ◽  
Author(s):  
Juliette N Mulvihill ◽  
J Andrew Davies ◽  
Florence Toti ◽  
Jean-Marie Freyssinet ◽  
Jean-Pierre Cazenave
Keyword(s):  
Human Platelets ◽  
Capillary Perfusion ◽  
Coated Glass ◽  
Bovine Collagen ◽  
Glass Capillaries ◽  
Human Thrombin ◽  
Platelet Accumulation ◽  
Granule Proteins ◽  
Von Willebrand

SummaryThe generation of trace amounts of thrombin at artificial surfaces in contact with blood is likely to be a contributing factor in thrombosis on biomaterials. Using an in vitro capillary perfusion system, platelet accumulation on glass surfaces, uncoated or precoated with purified bovine collagen or human plasma proteins, was determined in the presence or absence of preadsorbed purified human thrombin. Static adsorption for 15 min at 22° C from solutions of thrombin 100 NIH units (33 μg)/ml gave surface concentrations in the range 0.019-0.101 μg/cm2. Protein coated capillaries, thrombin treated or untreated, were perfused for 2 min at 37° C with suspensions of washed 111In-labeled human platelets in Tyrode's-albumin buffer containing 40% washed red blood cells, under conditions of controlled, non pulsatile laminar flow (50 s−1 or 2,000 s−1). Platelet accumulation was increased in the presence of surface adsorbed thrombin on uncoated and albumin or fibrinogen coated glass but little affected on fibronectin or collagen coated glass. On von Willebrand factor (vWF) coated glass, thrombin enhancement was observed only at high shear forces. In experiments using antibodies against human platelet α-granule proteins, thrombin stimulated platelet deposition in uncoated glass capillaries was inhibited at 2,000 s−1 by anti-vWF and to a lesser extent by anti-fibrinogen but not by antithrombospondin antibodies.

Fetal Bovine Collagen Grafts for Repair of Tegmen Defects and Encephaloceles via Middle Cranial Fossa Approach

2020 ◽  
Author(s):  
Anand Patel ◽  
Matthew R. Bartindale ◽  
Jehad A. Zakaria ◽  
Anand V. Germanwala ◽  
Douglas E. Anderson ◽  
...  
Keyword(s):  
Middle Cranial Fossa ◽  
Middle Cranial Fossa Approach ◽  
Bovine Collagen ◽  
Cranial Fossa ◽  
Fetal Bovine
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