Fish Scale Collagen Functionalized Thermo-Responsive Nanofibres

Smart thermosensitive polymer such as poly (N-isopropyl acrylamide) (PNIPAM) and dominant fibrous protein of connective tissue such as collagen (CLG) possess great potential in biomedical and tissue engineering applications. The objectives of current work aim to explore potential of PNIPAM and collagen by (i) establish a stable procedure to extract collagen from fresh water Tilapia fish scale (TFS) and (ii) fabricate PNIPAM and hybrid PNIPAM-CLG nanofibrous scaffolds through electrospinning technique and investigate their material-process-structure behaviour. Type I collagen was derived through acid hydrolysis of TFS. Electrospinning of PNIPAM was carried out with 16, 18 and 20 wt% PNIPAM concentration in methanol (MeOH) while PNIPAM-CLG was prepared through blending measured quantity of PNIPAM dissolved in water with collagen dissolved in acetic acid. Material properties, viscosity, morphology and thermo-physical behaviors of the derived collagen, electrospun PNIPAM and PNIPAM-CLG scaffolds were characterized. Results from SDS-PAGE and FTIR confirmed that the isolated TFS collagen is of type I. EDX revealed that demineralization eliminated the aluminium, magnesium, silicon and phosphorus while significantly reduced the sulfur elements from raw TFS. SEM observation of the collagen morphology shown a fluffy and fibrillary lamellae structure. Electrospun scaffolds were successfully fabricated with 16 and 18 wt% PNIPAM in MeOH. Both homogeneity and average fibre diameter (Davg) were greater in the 18 wt% PNIPAM scaffold, in which the Davg for 16 and 18 wt% were ~110 and ~131.7 nm respectively. However, PNIPAM at 20 wt% failed to be electrospun owing to its excessively high viscosity. On the other hand, SEM observation revealed that the electrospun hybrid PNIPAM-CLG scaffold has Davg of ~105.5 nm amid the presence of numerous elongated beads.

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Maturation process and characterization of a novel thermostable and halotolerant subtilisin-like protease with high collagenolytic but low gelatinolytic activity

Applied and Environmental Microbiology ◽

10.1128/aem.02184-21 ◽

2021 ◽

Author(s):

Kui Zhang ◽

Qianqian Huang ◽

Yu Li ◽

Lanhua Liu ◽

Xiao-Feng Tang ◽

...

Keyword(s):

High Temperatures ◽

Type I Collagen ◽

Elevated Temperatures ◽

Catalytic Domain ◽

Ion Pairs ◽

Gelatinolytic Activity ◽

Type I ◽

Fish Scale ◽

Collagenolytic Proteases ◽

Collagenolytic Protease

Enzymatic degradation of collagen is of great industrial and environmental significance; however, little is known about thermophile-derived collagenolytic proteases. Here, we report a novel collagenolytic protease (TSS) from thermophilic Brevibacillus sp. WF146. The TSS precursor comprises a signal peptide, an N-terminal propeptide, a subtilisin-like catalytic domain, a β-jelly roll (βJR) domain, and a prepeptidase C-terminal (PPC) domain. The maturation of TSS involves a stepwise autoprocessing of the N-terminal propeptide and the PPC domain, and the βJR rather than the PPC domain is necessary for correct folding of the enzyme. Purified mature TSS displayed optimal activity at 70°C and pH 9.0, a half-life of 1.5 h at 75°C, and an increased thermostability with rising salinity up to 4 M. TSS possesses an increased number of surface acidic residues and ion pairs, as well as four Ca 2+ -binding sites, which contribute to its high thermostability and halotolerance. At high temperatures, TSS exhibited high activity toward insoluble type I collagen and azocoll, but showed a low gelatinolytic activity, with a strong preference for Arg and Gly at the P1 and P1’ positions, respectively. Both the βJR and PPC domains could bind but not swell collagen, and thus facilitate TSS-mediated collagenolysis via improving the accessibility of the enzyme to the substrate. Additionally, TSS has the ability to efficiently degrade fish scale collagen at high temperatures. IMPORTANCE Proteolytic degradation of collagen at high temperatures has the advantages of increasing degradation efficiency and minimizing the risk of microbial contamination. Reports on thermostable collagenolytic proteases are limited, and their maturation and catalytic mechanisms remain to be elucidated. Our results demonstrate that the thermophile-derived TSS matures in an autocatalytic manner, and represents one of the most thermostable collagenolytic proteases reported so far. At elevated temperatures, TSS prefers hydrolyzing insoluble heat-denatured collagen rather than gelatin, providing new insight into the mechanism of collagen degradation by thermostable collagenolytic proteases. Moreover, TSS has the potential to be used in recycling collagen-rich wastes such as fish scales.

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Curcumin loaded nano graphene oxide reinforced fish scale collagen – a 3D scaffold biomaterial for wound healing applications

RSC Advances ◽

10.1039/c5ra15726a ◽

2015 ◽

Vol 5 (119) ◽

pp. 98653-98665 ◽

Cited By ~ 30

Author(s):

Tapas Mitra ◽

Piyali Jana Manna ◽

S. T. K. Raja ◽

A. Gnanamani ◽

P. P. Kundu

Keyword(s):

Tissue Engineering ◽

Wound Healing ◽

Graphene Oxide ◽

Type I Collagen ◽

Type I ◽

Fish Scale ◽

3D Scaffold ◽

Engineering Research ◽

Nano Graphene ◽

Tissue Engineering Research

We prepare a highly stabilized nano graphene oxide functionalized with type I collagen to make a 3D scaffold as a novel platform for better tissue engineering research..

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Effect Collagen Concentration Tilapia Scales on the Quality of Burn Ointment

Asian Journal of Fisheries and Aquatic Research ◽

10.9734/ajfar/2021/v15i630353 ◽

2021 ◽

pp. 87-95

Author(s):

Muhammad Firham Ramadhan ◽

Junianto . ◽

Rusky Intan Pratama ◽

Iis Rostini

Keyword(s):

Type I Collagen ◽

Ph Value ◽

Quality Standard ◽

National Standard ◽

Control Treatment ◽

Type I ◽

Fish Scale ◽

Collagen Concentration ◽

Biomedical Material ◽

Skin Collagen

Collagen is one of the main connective tissue animal proteins and has been widely used as a biomedical material. Collagen is divided into XIX types. Type I collagen, among others, is obtained from bone, scales and skin. Collagen derived from type I can repair tissue or accelerate tissue regeneration to heal burns. The purpose of this research was to determine the addition of fish scale collagen extract to the characteristics of the burn ointment preparation in accordance with the Indonesian National Standard (SNI) and the best quality. The method used in this research is an experimental method of Completely Randomized Design (CRD) consisting of 4 collagen addition treatments: 0%, 2%, 4% and 6% repeated 5 times. Parameters in this method include physical-chemical parameters (pH, spreadability, shelf life and homogeneity) and organoleptic parameters (appearance, aroma, texture and color). Bayes test results, the concentration of the addition of tilapia scale collagen in the ointment preparation of 4% resulted in a value close to the control treatment. The addition of 4% collagen was the best treatment compared to 2% and 6% with a pH value of 6.12, dispersion of 3.22 cm, safe ointment preparation did not change at all during 28 days of storage. Based on the results of the organoleptic test parameters, the ointment at this concentration had a homogeneous appearance, slightly yellowish white color, a distinctive smell of collagen and a semi-solid texture, this was in accordance with the quality standard of the ointment and had the best quality characteristics.

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Elemental distribution analysis of type I collagen fibrils in tilapia fish scale with energy-filtered transmission electron microscope

Micron ◽

10.1016/j.micron.2009.04.001 ◽

2009 ◽

Vol 40 (5-6) ◽

pp. 665-668 ◽

Cited By ~ 19

Author(s):

Mitsuhiro Okuda ◽

Masaki Takeguchi ◽

Motohiro Tagaya ◽

Toru Tonegawa ◽

Ayako Hashimoto ◽

...

Keyword(s):

Electron Microscope ◽

Transmission Electron Microscope ◽

Type I Collagen ◽

Collagen Fibrils ◽

Elemental Distribution ◽

Type I ◽

Distribution Analysis ◽

Fish Scale ◽

Transmission Electron

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In Situ Localization of Two Fibrillar Collagens in Two Compact Connective Tissues by Immunoelectron Microscopy After Cryotechnical Processing

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215549704500115 ◽

1997 ◽

Vol 45 (1) ◽

pp. 119-128 ◽

Cited By ~ 13

Author(s):

G. Nicolas ◽

F. Gaill ◽

L. Zylberberg

Keyword(s):

Osmium Tetroxide ◽

Type I Collagen ◽

Freeze Substitution ◽

Immunogold Electron Microscopy ◽

Type I ◽

Fish Scale ◽

Electron Microscopic ◽

Lr White ◽

Fibrillar Collagens ◽

Freeze Fixation

Two fibrillar collagens, the worm cuticular collagen and the vertebrate Type I fish scale collagen, both organized in a compact tissue, were localized by immunogold electron microscopy in resin sections after freeze-fixation and freeze-substitution. Identification of these two fibrillar collagens failed with the-use of postembedding labeling after conventional electron microscopic processing. Positive labeling of the Type I collagen was observed in sections of fish scales freeze-fixed by either slam-freezing or high-pressure freezing, freeze-substituted in acetone with or without osmium tetroxide, and embedded in LR White. The worm cuticular collagen was detected in sections of cuticle that were freeze-fixed, freeze-substituted (necessarily with osmium tetroxide added to acetone), and embedded in either LR White or Epon. It was also detected in specimens pre-fixed by aldehydes before freeze-fixation. The Type I fish scale collagen appears to be more sensitive than the fibrillar cuticular collagen of worms to the procedures employed for postembedding immunoelectron microcopy. Our results have shown that freeze-fixation and freeze-substitution preserved the antigenicity of the fibrillar collagens organized in a compact three-dimensional network, whereas immunolabeling failed after conventional electron microscopic procedures. These cryostabilization techniques appear to be of value to improve the immunolocalization of collagens.

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Synthesis and Characterization of Wound Dressing Material from Bio-Wastes Impregnated with the Spider Web and the Ethanolic Weaves Extract of Mangifera indica (L.)

Biointerface Research in Applied Chemistry ◽

10.33263/briac122.19982012 ◽

2021 ◽

Vol 12 (2) ◽

pp. 1998-2012

Keyword(s):

Cost Effectiveness ◽

Wound Dressing ◽

Type I Collagen ◽

Mangifera Indica ◽

Biological Evaluation ◽

Spectroscopic Techniques ◽

Type I ◽

Fish Scale ◽

Good Efficacy ◽

Spider Web

Substantial progress in wound therapy has not snuffed out the passion in search of innovative wound dressing materials. This work is to analyze the physiochemical characterizations and biological evaluation of a wound dressing material. A wound dressing material had been synthesized from Physiologically Clotted Fibrin (PCF), Fish Scale Collagen (FSC). Also, the wound dressing material had impregnated with the folklore medicinal impact of the Spider Web (SW) and the Ethanolic Extract of Mangifera indica (L.)(EEMI). Infrared spectroscopic techniques confirmed the presence of Type I collagen. Surface morphology established the smooth, uniform, porous, biocompatible surface of the material. Water absorption studies, porosity measurements showed the required characteristics, the antibacterial activity favored the resistance, and the ash test supported the eco-friendly environment of the wound dressing material. Human erythrocytes had reviewed biocompatibility. The supernatant of the wound dressing material at different concentrations and incubation times had determined for percent hemolysis. Plots between percent hemolysis and concentration showed the non-hemolytic behavior of the wound dressing material. The synthesized biomaterial could propose as a wound dressing material with good efficacy, cost-effectiveness, and eco-friendly. The synthesized biomaterial could have been a better wound dressing material with good efficacy, cost-effectiveness, and eco-friendly.

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Biocompatibility of Novel Type I Collagen Purified from Tilapia Fish Scale: An In Vitro Comparative Study

BioMed Research International ◽

10.1155/2015/139476 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 17

Author(s):

Jia Tang ◽

Takashi Saito

Keyword(s):

Cellular Proliferation ◽

Type I Collagen ◽

Type I ◽

Fish Scale ◽

Porcine Skin ◽

Religious Issues ◽

Skin Collagen ◽

Potential Alternative ◽

Tilapia Scale

Type I collagen (COL-1) is the prevailing component of the extracellular matrix in a number of tissues including skin, ligament, cartilage, bone, and dentin. It is the most widely used tissue-derived natural polymer. Currently, mammalian animals, including pig, cow, and rat, are the three major sources for purification of COL-1. To reduce the risk of zoonotic infectious diseases transmission, minimize the possibility of immunogenic reaction, and avoid problems related to religious issues, exploration of new sources (other than mammalian animals) for the purification of type I collagen is highly desirable. Hence, the purpose of the current study was to investigate the in vitro responses of MDPC-23 to type I collagen isolated from tilapia scale in terms of cellular proliferation, differentiation, and mineralization. The results suggested that tilapia scale collagen exhibited comparable biocompatibility to porcine skin collagen, indicating it might be a potential alternative to type I collagen from mammals in the application for tissue regeneration in oral-maxillofacial area.

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Manufacturing micropatterned collagen scaffolds with chemical-crosslinking for development of biomimetic tissue-engineered oral mucosa

Scientific Reports ◽

10.1038/s41598-020-79114-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Ayako Suzuki ◽

Yoshihiro Kodama ◽

Keito Miwa ◽

Kazuma Kishimoto ◽

Emi Hoshikawa ◽

...

Keyword(s):

Oral Mucosa ◽

Type I Collagen ◽

Three Dimensional ◽

Collagen Scaffold ◽

Type I ◽

Chemical Crosslinking ◽

Graft Material ◽

Fish Scale ◽

Collagen Scaffolds ◽

Aspect Ratios

AbstractThe junction between the epithelium and the underlying connective tissue undulates, constituting of rete ridges, which lack currently available soft tissue constructs. In this study, using a micro electro mechanical systems process and soft lithography, fifteen negative molds, with different dimensions and aspect ratios in grid- and pillar-type configurations, were designed and fabricated to create three-dimensional micropatterns and replicated onto fish-scale type I collagen scaffolds treated with chemical crosslinking. Image analyses showed the micropatterns were well-transferred onto the scaffold surfaces, showing the versatility of our manufacturing system. With the help of rheological test, the collagen scaffold manufactured in this study was confirmed to be an ideal gel and have visco-elastic features. As compared with our previous study, its mechanical and handling properties were improved by chemical cross-linking, which is beneficial for grafting and suturing into the complex structures of oral cavity. Histologic evaluation of a tissue-engineered oral mucosa showed the topographical microstructures of grid-type were well-preserved, rather than pillar-type, a well-stratified epithelial layer was regenerated on all scaffolds and the epithelial rete ridge-like structure was developed. As this three-dimensional microstructure is valuable for maintaining epithelial integrity, our micropatterned collagen scaffolds can be used not only intraorally but extraorally as a graft material for human use.

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Collagen fibrillogenesis in vitro: interaction of types I and V collagen

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142670 ◽

1986 ◽

Vol 44 ◽

pp. 210-211

Author(s):

Arthur J. Wasserman ◽

Kathy C. Kloos ◽

David E. Birk

Keyword(s):

Type I Collagen ◽

Corneal Stroma ◽

Minor Component ◽

Homogeneous Distribution ◽

Type I ◽

Type V Collagen ◽

Fibril Morphology ◽

Type V ◽

In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

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