Comparison of Physicochemical Characteristics and Fibril Formation Ability of Collagens Extracted from the Skin of Farmed River Puffer (Takifugu obscurus) and Tiger Puffer (Takifugu rubripes)

Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) from the skin of river puffer (ASC-RP and PSC-RP) and tiger puffer (ASC-TP and PSC-TP) were extracted and physicochemically examined. Denaturation temperature (Td) for all the collagens was found to be 25.5–29.5 °C, which was lower than that of calf skin collagen (35.9 °C). Electrophoretic patterns indicated all four samples were type I collagen with molecular form of (α1)2α2. FTIR spectra confirmed the extracted collagens had a triple-helical structure, and that the degree of hydrogen bonding in ASC was higher than PSC. All the extracted collagens could aggregate into fibrils with D-periodicity. The fibril formation rate of ASC-RP and PSC-RP was slightly higher than ASC-TP and PSC-TP. Turbidity analysis revealed an increase in fibril formation rate when adding a low concentration of NaCl (less than 300 mM). The fibril formation ability was suppressed with further increasing of NaCl concentration, as illustrated by a reduction in the turbidity and formation degree. SEM analysis confirmed the well-formed interwoven structure of collagen fibrils after 24 h of incubation. Summarizing the experimental results suggested that the extracted collagens from the skin of river puffer and tiger puffer could be considered a viable substitute to mammalian-derived collagens for further use in biomaterial applications.

Download Full-text

Extraction and Partial Characterization of Pepsin-Soluble Collagens from the Skin of Amiurus Nebulosus

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.236-238.2926 ◽

2011 ◽

Vol 236-238 ◽

pp. 2926-2934 ◽

Cited By ~ 4

Author(s):

Li Li Chen ◽

Li Zhao ◽

Hua Liu ◽

Run Feng Wu

Keyword(s):

Type I Collagen ◽

Type I ◽

Alternative Source ◽

Soluble Collagen ◽

Acid Soluble Collagen ◽

Sds Page ◽

Skin Collagen ◽

Maximal Yield ◽

Pepsin Soluble Collagen

Pepsin-soluble collagen (PSC) was successfully extracted from the skin of Amiurus nebulosus. The skin of Amiurus nebulosus was immersed in 0.3 mol/L acetic acid (1: 20, m: V) for 6 h at 37°C, while pepsin was added, at a level of 5000U/g dosage of defatted skin. The maximal yield of the collagen was 97.44%, which was higher than that of acid-soluble collagen (ASC) at 62.05%. Some properties of pepsin-soluble collagens from the skin of Amiurus nebulosus were characterized. Amino acid composition and SDS-PAGE suggested that the collagen might be classified as type I collagen. Moreover, FTIR investigations showed the existence of helical arrangements in PSC of Amiurus nebulosus skin of collagen. There is a possibility to use Amiurus nebulosus skin collagen as an alternative source of collagen for industrial purposes and subsequently it may maximize the economical value of the fish.

Download Full-text

Preparation and Characterization of Thermally Stable Collagens from the Scales of Lizardfish (Synodus macrops)

Marine Drugs ◽

10.3390/md19110597 ◽

2021 ◽

Vol 19 (11) ◽

pp. 597

Author(s):

Junde Chen ◽

Guangyu Wang ◽

Yushuang Li

Keyword(s):

Type I Collagen ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Type I ◽

Salting Out ◽

Soluble Collagen ◽

Acid Soluble Collagen ◽

Skin Collagen ◽

Rat Tail

Marine collagen is gaining vast interest because of its high biocompatibility and lack of religious and social restrictions compared with collagen from terrestrial sources. In this study, lizardfish (Synodus macrops) scales were used to isolate acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC). Both ASC and PSC were identified as type I collagen with intact triple-helix structures by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and spectroscopy. The ASC and PSC had high amino acids of 237 residues/1000 residues and 236 residues/1000 residues, respectively. Thus, the maximum transition temperature (Tmax) of ASC (43.2 °C) was higher than that of PSC (42.5 °C). Interestingly, the Tmax of both ASC and PSC was higher than that of rat tail collagen (39.4 °C) and calf skin collagen (35.0 °C), the terrestrial collagen. Solubility tests showed that both ASC and PSC exhibited high solubility in the acidic pH ranges. ASC was less susceptible to the “salting out” effect compared with PSC. Both collagen types were nontoxic to HaCaT and MC3T3-E1 cells, and ASC was associated with a higher cell viability than PSC. These results indicated that ASC from lizardfish scales could be an alternative to terrestrial sources of collagen, with potential for biomedical applications.

Download Full-text

Physicochemical and Biocompatibility Properties of Type I Collagen from the Skin of Nile Tilapia (Oreochromis Niloticus) for Biomedical Applications

Marine Drugs ◽

10.3390/md17030137 ◽

2019 ◽

Vol 17 (3) ◽

pp. 137 ◽

Cited By ~ 13

Author(s):

Wen-Kui Song ◽

Dan Liu ◽

Lei-Lei Sun ◽

Ba-Fang Li ◽

Hu Hou

Keyword(s):

Nile Tilapia ◽

Biomedical Applications ◽

Type I Collagen ◽

Molecular Form ◽

Umbilical Vein ◽

Treated Group ◽

Systemic Toxicity ◽

Type I ◽

Sem Images ◽

Soluble Collagen

The aim of this study is to investigate the physicochemical properties, biosafety, and biocompatibility of the collagen extract from the skin of Nile tilapia, and evaluate its use as a potential material for biomedical applications. Two extraction methods were used to obtain acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) from tilapia skin. Amino acid composition, FTIR, and SDS-PAGE results showed that ASC and PSC were type I collagen. The molecular form of ASC and PSC is (α1)2α2. The FTIR spectra of ASC and PSC were similar, and the characteristic peaks corresponding to amide A, amide B, amide I, amide II, and amide III were 3323 cm−1, 2931 cm−1, 1677 cm−1, 1546 cm−1, and 1242 cm−1, respectively. Denaturation temperatures (Td) were 36.1 °C and 34.4 °C, respectively. SEM images showed the loose and porous structure of collagen, indicting its physical foundation for use in applications of biomedical materials. Negative results were obtained in an endotoxin test. Proliferation rates of osteoblastic (MC3T3E1) cells and fibroblast (L929) cells from mouse and human umbilical vein endothelial cells (HUVEC) were increased in the collagen-treated group compared with the controls. Furthermore, the acute systemic toxicity test showed no acute systemic toxicity of the ASC and PSC collagen sponges. These findings indicated that the collagen from Nile tilapia skin is highly biocompatible in nature and could be used as a suitable biomedical material.

Download Full-text

Evaluation of biocompatibility and immunogenicity of micro/nanofiber materials based on tilapia skin collagen

Journal of Biomaterials Applications ◽

10.1177/0885328218820180 ◽

2019 ◽

Vol 33 (8) ◽

pp. 1118-1127 ◽

Cited By ~ 4

Author(s):

Dongsheng Li ◽

Yonglin Gao ◽

Yunzhi Wang ◽

Xiaoping Yang ◽

Chuanglong He ◽

...

Keyword(s):

Type I Collagen ◽

Helical Structure ◽

Raw Material ◽

Type I ◽

Skin Collagen ◽

Fish Collagen ◽

Native Collagen ◽

Mouse Immunization ◽

Good Biocompatibility ◽

Self Assembled

Type I collagen, used as a raw material, plays a pivotal role in the development of medical devices and tissue engineering. Due to the risk of zoonotic transmission and religious constraints for mammalian collagen, fish collagen gains increased attention and is widely seen as an alternative. In this study, two collagen micro/nanofiber materials, self-assembled collagen nanofiber and electrospun collagen nanofiber, were prepared by tilapia skin collagen and their biocompatibility and immunogenicity was thoroughly investigated. The result revealed that the state of tilapia skin collagen in self-assembled collagen nanofiber and electrospun collagen nanofiber was different. The circular dichroism spectrum indicated that collagen in self-assembled collagen nanofiber retained the triple helical structure of the native collagen, while collagen in electrospun collagen nanofiber was denatured into gelatin. Nevertheless, the evaluation according to ISO10993, including tests of cytotoxicity, hemolysis, skin sensitization, acute systemic toxicity, mouse immunization and lymphocyte proliferation, demonstrated good biocompatibility and low immunogenicity for both self-assembled and electrospun collagen nanofiber materials. Overall, the present study highlighted that type I collagen from tilapia skin would be a promising biomaterial for the development of regenerate medical products.

Download Full-text

MOLECULAR CHARACTERISTICS OF ACID AND PEPSIN SOLUBLE COLLAGENS FROM THE SCALES OF GOLDEN CARP (PROBARBUS JULLIENI)

Emirates Journal of Food and Agriculture ◽

10.9755/ejfa.2016-09-1316 ◽

2017 ◽

pp. 450 ◽

Cited By ~ 8

Author(s):

Ali Muhammed Moula Ali ◽

Soottawat Benjakul ◽

Hideki Kishimura

Keyword(s):

Amino Acid ◽

Type I Collagen ◽

Helical Structure ◽

Dry Weight ◽

Total Amino Acid ◽

Molecular Characteristics ◽

Type I ◽

Scanning Calorimetry ◽

Soluble Collagen ◽

Golden Carp

Acid soluble collagen (ASC) and pepsin soluble collagen (PSC) were isolated from the scales of golden carp (Probarbus jullieni). Both ASC and PSC, identified as type I collagen, contained α- and β-chains as the dominant constituents. ASC and PSC had the yields of 0.42 and 1.16 g 100 g‒1 (dry weight basis), respectively. Amino acid composition revealed that glycine constituted 1/3 of total amino acid residues and no cysteine was found. ASC and PSC had imino acid contents of 197 and 202 residues/1000 residues, respectively. Fourier transform infrared spectroscopy (FTIR) and circular dichroism (CD) spectra indicated high integrity of the triple helical structure of both collagens. Based on differential scanning calorimetry (DSC), ASC and PSC had Tmax of 37.67 and 37.83 °C, respectively. Both collagens exhibited high solubility in acidic pH range (1‒3) and the decrease in solubility was found in the presence of NaCl at concentrations above 30 g L‒1. The overall results demonstrated that scales of golden carp could serve as another source of collagen.

Download Full-text

Isolation and properties of collagen extracted from mixed by-products obtained from different fish species

Biotecnia ◽

10.18633/biotecnia.v23i3.1463 ◽

2021 ◽

Vol 23 (3) ◽

pp. 109-116

Author(s):

Celia Olivia García-Sifuentes ◽

Julio Cesar Zamorano-Apodaca ◽

Marcel Martinez-Porchas ◽

Susana Maria Scheuren-Acevedo ◽

Miguel Angel Mazorra-Manzano

Keyword(s):

Fish Species ◽

Type I Collagen ◽

Helical Structure ◽

Extraction Process ◽

Type I ◽

Denaturation Temperature ◽

Soluble Collagen ◽

Acid Soluble Collagen ◽

Sds Page ◽

By Products

Fish by-products consisting of skin, bones, or scales are collagen sources. Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) mixed by-products derived from different fish species were extracted and evaluated. The properties evaluated for both collagens were chemical composition, amino acid- and SDS-PAGE- protein profiles, Fourier transform infrared spectroscopy (FTIR), denaturation temperature (Tmax), enthalpy (ΔH), and solubility. The ASC and PSC registered a protein content of 48.56 and 38.80 %, respectively. From the total amino acids detected, hydroxyproline accounted for 7 % and 6 % for ASC and PSC, respectively. The electrophoretic profile showed the presence of the type I collagen bands (α1, α2, β, and γ), whereas FTIR spectrum showed the presence of diverse collagen functional groups (Amide A, B, I, II, and III) for both extracted types, and demonstrated that the extraction process did not affect the collagen´s triple-helical structure. The Tmax of ASC and PSC were 38.27 and 38.07° C, respectively, whereas ΔH were 0.64 and 0.33 J g-1. The lowest solubility was registered at pH 5 for ASC and pH 9 for PSC. The caractheristics of the collagen extracted, indicated that a mixture of by-products from different species could be an alternative for their reutilization by the local markets.

Download Full-text

Comprehensive Assessment of Nile Tilapia Skin (Oreochromis niloticus) Collagen Hydrogels for Wound Dressings

Marine Drugs ◽

10.3390/md18040178 ◽

2020 ◽

Vol 18 (4) ◽

pp. 178 ◽

Cited By ~ 5

Author(s):

Baosheng Ge ◽

Haonan Wang ◽

Jie Li ◽

Hengheng Liu ◽

Yonghao Yin ◽

...

Keyword(s):

Wound Dressing ◽

Type I Collagen ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Wound Dressings ◽

Type I ◽

Rheological Characterization ◽

Scanning Calorimetry ◽

Soluble Collagen ◽

Collagen Hydrogels

Collagen plays an important role in the formation of extracellular matrix (ECM) and development/migration of cells and tissues. Here we report the preparation of collagen and collagen hydrogel from the skin of tilapia and an evaluation of their potential as a wound dressing for the treatment of refractory wounds. The acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) were extracted and characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), differential scanning calorimetry (DSC), circular dichroism (CD) and Fourier transform infrared spectroscopy (FTIR) analysis. Both ASC and PSC belong to type I collagen and have a complete triple helix structure, but PSC shows lower molecular weight and thermal stability, and has the inherent low antigenicity. Therefore, PSC was selected to prepare biomedical hydrogels using its self-aggregating properties. Rheological characterization showed that the mechanical strength of the hydrogels increased as the PSC content increased. Scanning electron microscope (SEM) analysis indicated that hydrogels could form a regular network structure at a suitable PSC content. Cytotoxicity experiments confirmed that hydrogels with different PSC content showed no significant toxicity to fibroblasts. Skin repair experiments and pathological analysis showed that the collagen hydrogels wound dressing could significantly accelerate the healing of deep second-degree burn wounds and the generation of new skin appendages, which can be used for treatment of various refractory wounds.

Download Full-text

Effect of hydroxylysine-O-glycosylation on the structure of type I collagen molecule: A computational study

Glycobiology ◽

10.1093/glycob/cwaa026 ◽

2020 ◽

Vol 30 (10) ◽

pp. 830-843

Author(s):

Ming Tang ◽

Xiaocong Wang ◽

Neha S Gandhi ◽

Bethany Lachele Foley ◽

Kevin Burrage ◽

...

Keyword(s):

Force Field ◽

Type I Collagen ◽

Computational Study ◽

Experimental Studies ◽

Helical Structure ◽

Atomic Level ◽

Collagen Molecule ◽

Type I ◽

Dynamic Simulations ◽

Force Field Parameters

Abstract Collagen undergoes many types of post-translational modifications (PTMs), including intracellular modifications and extracellular modifications. Among these PTMs, glycosylation of hydroxylysine (Hyl) is the most complicated. Experimental studies demonstrated that this PTM ceases once the collagen triple helix is formed and that Hyl-O-glycosylation modulates collagen fibrillogenesis. However, the underlying atomic-level mechanisms of these phenomena remain unclear. In this study, we first adapted the force field parameters for O-linkages between Hyl and carbohydrates and then investigated the influence of Hyl-O-glycosylation on the structure of type I collagen molecule, by performing comprehensive molecular dynamic simulations in explicit solvent of collagen molecule segment with and without the glycosylation of Hyl. Data analysis demonstrated that (i) collagen triple helices remain in a triple-helical structure upon glycosylation of Hyl; (ii) glycosylation of Hyl modulates the peptide backbone conformation and their solvation environment in the vicinity and (iii) the attached sugars are arranged such that their hydrophilic faces are well exposed to the solvent, while their hydrophobic faces point towards the hydrophobic portions of collagen. The adapted force field parameters for O-linkages between Hyl and carbohydrates will aid future computational studies on proteins with Hyl-O-glycosylation. In addition, this work, for the first time, presents the detailed effect of Hyl-O-glycosylation on the structure of human type I collagen at the atomic level, which may provide insights into the design and manufacture of collagenous biomaterials and the development of biomedical therapies for collagen-related diseases.

Download Full-text

A Newly Identified Platelet and Megakaryocyte Lysyl Oxidase-Adhesion to Collagen Axis in Human Primary Myelofibrosis

Blood ◽

10.1182/blood.v128.22.3133.3133 ◽

2016 ◽

Vol 128 (22) ◽

pp. 3133-3133

Author(s):

Alessandra Balduini ◽

Vittorio Abbonante ◽

Shinobu Matsuura ◽

Vittorio Rosti ◽

Katya Ravid

Keyword(s):

Type I Collagen ◽

Conflicts Of Interest ◽

Lysyl Oxidase ◽

Primary Myelofibrosis ◽

Collagen Type I ◽

Type I ◽

Soluble Collagen ◽

Peripheral Blood Cd34 ◽

Acid Soluble Collagen

Abstract Controlling platelet function is central to management of various pathologies, including Primary Myelofibrosis (PMF), which is associated with increased incidence of thrombosis and cardiovascular disease. In recent studies we showed that the matrix cross-linking enzyme, Lysyl Oxidase (LOX) is elevated in platelets and megakartocytes of myelofibrotic mice, and transgenic upregulation of LOX increases platelet and megakaryocyte adhesion to monomeric type I collagen (preferred by alpha2β1 collagen receptors), and augments propensity for in vivo thrombosis. Here, we examined the relevance of these findings to human disease, by first determining platelet LOX level, as well as platelet and megakaryocyte adhesion to collagen using samples derived from PMF patients and matching controls. In analyzing 10 PMF platelet samples (5 males and 5 females; 6 JAK2V617F; 4 CALR mutations; age range 30-55; PMF grade 1-3), we found a nearly 20 fold upregulation of LOX expression compared to matching healthy controls (p<0.001). Intriguingly, there was a significant increase in adhesion (plt/mm2) and spreading (pixel2) of PMF platelets relative to control on monomeric, pepsinated acid soluble collagen (PSCI) (p<0.05), while no differences were observed between the samples on native triple helical acid soluble collagen type I collagen (ASCI). To examine the role of LOX in this phenotype, we treated control and PMF-derived human megakaryocytes, differentiated from peripheral blood CD34+ cells, grown in presence or not of LOX inhibitor, β-aminopropionitrile (BAPN) from day 2 of culture. Our preliminary data, based on a cohort of 2 controls and 5 PMF samples, demonstrated that although on ASCI megakaryocyte adhesion is not altered by BAPN treatment both in CTRL and PMF derived megakaryocytes, on PSCI the adhesion of PMF derived megakaryocytes was reduced by about a 50% by BAPN treatment, while the adhesion of CTRL derived MKs was not significantly affected. Taken together, we identified LOX level to be upregulated in human PMF platelets and megakaryocytes, and LOX activity to be important for PMF cells adhesion to collagen. These newly identified properties are highly relevant to megakaryocyte adhesion to the niche, and to platelet activation in PMF. Disclosures No relevant conflicts of interest to declare.

Download Full-text