Juntao Zhang

Detection of type I collagen fibrils formation and dissociation by a fluorescence method based on thioflavin T.

In this study, fibrillogenesis and thermal dissociation of pepsin-soluble collagen (PSC), extracted from snakehead (Channa argus) skin, were monitored by fluorescence method based on thioflavin T (Th-T), where the accuracy and sensitivity were evaluated and compared with those of turbidity assay. The fluorescence method revealed the fibrillogenesis dynamics of collagen with better sensitivity, especially at nucleation and plateau stages. The melting temperature (Tm) of PSC was estimated to be 47°C by circular dichroism spectroscopy; below this temperature, the triple-helical structure should be intact. After that, the dynamic process of collagen dissociation was explored by the fluorescence method, and verified by morphological analysis of the fibrils and the proportion of retained fibrils. The thermal dissociation critical temperature (TDCT) of PSC fibrils was confirmed to be 39°C. The fluorescence intensity of fibril-incorporated Th-T gradually decreases in the dissociation process, and the decrease rate can be accelerated by increasing temperature. Finally, the thermal stability of triple-helical structures of free-, assembled- and dissociated-PSC was compared. Thus, we demonstrated the formation and thermal dissociation of collagen fibrils in vitro by a fluorescence method based on Th-T. This approach may advance the understanding of fibril formation and inverse dissociation of fish-sourced collagen in vitro.

Impact of Telopeptides on Self-Assembly Properties of Snakehead (Channa argus) Skin Collagen

Much work has been done to understand the self-assembly properties of collagens from mammals. However, there is little information about fish-sourced collagens that are advantageous in certain applications. In this study, the self-assembly dynamics of tropocollagen (with telopeptides) and atelocollagen (without telopeptides) extracted from snakehead (Channa argus) skins was studied with turbidity, dynamic rheology, scanning electron microscopy (SEM), and quantitative percentage of self-assembly. Turbidity results indicate that the self-assembly of fish-sourced collagens followed the nucleation-growth two-step model, while rheological results unveiled two growth stages in the development of collagen gels. Based on SEM, telopeptides promoted the formation of thicker fibrils and increased the density of network that provided the structural basis of increased turbidity and strengthened storage and loss moduli. The SEM data were supported by the increased percentage of self-assembled collagens by telopeptides. Findings from this work may facilitate the understanding of structures and functions of products containing fish-sourced collagens and their application.

Systematic modulation of gelation dynamics of snakehead (Channa argus) skin collagen by environmental parameters

AbstractGel matrices of mammalian collagen are routinely used as bioengineering materials and food thickening agents, for which the modulation of their mechanical properties is a key issue. However, little information is available regarding gel matrices based on fish-sourced collagen, which offer unique advantages for some applications. Here, a rheology assay, which is the most commonly used method for monitoring developing gels, was used to systematically reveal the influence of environmental parameters on the gelation dynamics and rheological properties of pepsin-soluble collagen (PSC) extracted from snakehead (Channa argus) skins. The gelation dynamics and equilibrium elastic moduli of PSC were affected by concentration, temperature, pH, buffer, and ion strength and type. SEM and TEM images of gels at different concentrations and incubation temperatures confirmed that the mechanical properties of PSC gels are directly related to the density, rather than size, of fibrils. Additionally, the relationship between the biological and mechanical properties of these collagen gels was also evaluated. The present study would facilitate a better understanding of the gelation of fish-sourced collagen and enable more precise control of the mechanical properties of these gel matrices.

Effect of Ionic Liquids on the Fibril-Formation and Gel Properties of Grass Carp ( Ctenopharyngodon idellus ) Skin Collagen

Self-assembled environment of collagen is one of the important factors for improving and regulating the properties of collagen-based biomaterials. This study aimed to investigate the effect of ionic liquids (ILs) on the fibril-formation and gel properties of grass carp (Ctenopharyngodon idellus) skin collagen. Fibrillogenic kinetics analysis showed that the collagen self-assembly can be suppressed by the introduction of ILs, and the inhibitory effect is influenced by concentration and types of ILs. Scanning electron microscopy test indicated that the assembled collagen fibrils in the presence of ILs had bigger diameters than that in the conventional buffer. Differential scanning calorimetry analysis revealed that the thermal stability of collagen fibrils can be significantly increased when self-assembly is performed in the presence of ILs. Moreover, the introduction of ILs enhanced the mechanical strength of collagen gels. Finding from this work provides a new idea for improving the performance of fish-sourced collagen biomaterials.

Graphene-Oxide-Based FRET Platform for Sensing Xenogeneic Collagen Coassembly

Xenogeneic collagen coassembly (XCCA) offers a new view for the design and performance regulation of novel collagen-based biomaterials. But there is still a lack of accurate and sensitive method for monitoring XCCA. In this study, a simple and efficient graphene-oxide (GO)-based fluorescence resonance energy transfer (FRET) platform has been developed to sense XCCA. We first designed a fluorescein isothiocyanate (FITC)-labeled porcine skin collagen (PSC) that adsorbed on the GO surface and effectively quenched its fluorescence. Upon the addition of grass carp skin collagen (GCSC), the XCCA between PSC and GCSC resulted in desorption of FITC-PSC from GO surface and thus caused an increase in fluorescence signal. Under the optimal conditions, the fluorescence signal linearly increased with the increase in the GCSC concentration in the range of 50-1000 μg/mL, with a sensitivity of 22 μg/mL (S/N = 3). Furthermore, the developed strategy also exhibited excellent specificity and anti-interference ability. More interestingly, the thermal stability of collagen fibrils formed by XCCA is linearly related to the GCSC concentration. These results open a facile, effective, and sensitive approach for sensing XCCA and provide a new strategy for arbitrarily regulating the thermal stability of collagen fibrils.

Modulation of the Self-Assembly of Collagen by Phytic Acid: An In Vitro Study

Phytic acid, containing a myoinositol ring coupled with six phosphate groups, can react with the amino groups of collagen to regulate their self-assembly behavior. The aim of this research is to evaluate the effects of phytic acid on the selfassembly behavior of collagen, the structures and properties of the resulting fibrils and hydrogels. Turbidity and chloramine T assay suggested that phytic acid could improve the self-assembly kinetics and degree of collagen, and the optimal ratio of phytic acid/collagen was 1/1 (w/w). Scanning electron microscopy (SEM) analysis indicated that co-fibrils of collagen with phytic acid are more slender than that of pure collagen, and transmission electron microscopy (TEM) reveals that the characteristic D-periodicity of collagen fibrils is not affected by phytic acid. Besides, differential scanning calorimetry (DSC) and rheology revealed that the thermal stability of collagen fibrils and the viscoelasticity of collagen hydrogels could be improved by phytic acid and the optimal ratio of phytic acid/collagen is 1/1 (w/w).

In situ photopolymerization of dimethacrylamide-based resins and composites

A dimethacrylamide was synthesized and used to formulate with the selected (meth)acrylates to form the in situ photocureable resins and composites. The effects of the selected comonomers with different functional groups on polymerization rate, degree of conversion, gel time, and compressive strength were investigated. The results show that in situ photopolymerization of the synthesized dimethacrylamide with comonomers having an electron-withdrawing and/or acrylate group dramatically increased the polymerization rate, degree of conversion, and compressive strength. On the other hand, an electron-donating group on either carbon-carbon double bond or ester linkage slowed down the polymerization. In contrast, the triethylene glycol dimethacrylate-based system did not show a clear pattern. The synergistic effect of the strong hydrogen-bonding between dimethacrylamide and organic acid groups may be responsible for higher compressive strengths. The formed composites showed the similar trend in compressive strength to the corresponding resins. Within the limitation of this study, it seems that in situ photopolymerization of dimethacrylamide or diacrylamide can be greatly accelerated by copolymerization with monomers having electron-withdrawing and/or acrylate groups. The monomers with methacrylate group can significantly reduce the polymerization rate and degree of conversion.

Effect of ultra-high pressure on molecular structure and properties of bullfrog skin collagen

Ultra-high pressure technology has attracted a great deal of attention in recent years, and has been widely used in food science, medicine, and other fields. This study aimed to determine the effect of ultra-high pressure on the structure and properties of collagen. Native collagen extracted from bullfrog skin was processed under different ultra-high pressure treatment conditions (300, 400, and 500MPa). Then systematic analysis of the molecular structures and properties of the samples after ultra-high pressure treatment were performed. It was found that the conformation of collagen molecules could be adjusted by ultra-high pressure treatment, and this regulation was closely related to the level of treatment pressure. A possible mechanism of the impact of ultra-high pressure on the collagen molecular structures was speculated according to the experimental results. At low pressure levels (300-400MPa), the pressure perpendicular to collagen axis dominates and leads to a tightening of the triple helix structure of collagen, while the pressure parallel to collagen axis is dominant and the triple helix tends to dissociate like a zipper at high pressure levels (>400MPa). These structural changes would simultaneously result in various changes to thermal stability, self-assembly properties, and antigenicity of collagen.

Effects of preparation processes on the structure and properties of collagen gel

ABSTRACT In this study, the effects of collagen self-assembly, drying methods, and reswelling on collagen gel structure and biological properties were determined. The results showed that the self-assembly of collagen improved the regularity and pore size of the collagen gel and promoted collagen gel thermostability and cell proliferation. After drying and reswelling, the main structure of the self-assembly gel was composed of lamellar gel walls, but the gel structure shrank to some extent and pore size was reduced. The structure of the self-assembled gel after drying and reswelling was closely related to that of the drying method.

Preparation of a low viscosity urethane-based composite for improved dental restoratives

Several new urethane-based dimethacrylates were synthesized, characterized and used to formulate the resin composites. Compressive strength (CS) was used as a screen tool to evaluate the mechanical property of the formed composites. Flexural strength, diametral tensile strength, water sorption, degree of conversion and shrinkage of the composites were also evaluated. The results show that most of the synthesized urethane-based dimethacrylates were solid, which are not suitable to dental filling restorations. However, it was found that liquid urethane-based dimethacrylates could be derivatized using asymmetrical methacrylate synthesis. Not only the newly synthesized urethane-based dimethacrylates showed lower viscosity values but also their constructed composites exhibited higher mechanical strengths. Without triethyleneglycol dimethacrylate (TEGDMA) addition, the new urethane-constructed composites showed significantly lower water sorption and shrinkage.