| Fish collagen and collagen-based nanofibre membranes are currently a hot research area and are utilized extensively in the cosmetic,food,and pharmaceutical industries.Therefore,in this work,double-spotted pufferfish skin collagen was prepared,and its structure,functional and rheological characteristics,and biocompatibility investigations were accomplished.This study prepared Col/SF composite nanofibre membranes by using double-spotted pufferfish skin collagen and silk fibroin,and then completed structure,thermal stability,mechanical properties,and biocompatibility to address the issues of poor mechanical properties and thermal stability of fish collagen nanofibre membranes.In the present study,in order to maximize the use of the skin resource of the double-spotted puffer fish skin and to offer new ideas for the development of high value-added collagen,and the main findings are as follows:(1)The collagen was prepared from the skin of double-spotted pufferfish.The extraction rate was 49.83%±1.85%(dry basis weight),higher than pig skin collagen(38.9%,dry basis weight),and the purity was 95.86%±0.66%,higher than the standard rat tail type I collagen(94.86%±0.51%).Thermal denaturation occurred at a temperature of 27.8°C,which was higher than that of deep-sea redfish and baltic cod.The foaming of double-spotted pufferfish skin collagen was superior was superior to chicken foot collagen and red stingray fish skin collagen.The foaming was ranged from 8.26±0.38%to 20.4±0.57%and emulsification was ranged from 44.36±0.001 m~2/g to 12.99±0.0014 m~2/g.The rheological characteristics demonstrate that the collagen has good viscoelasticity.Biocompatibility tests revealed that the relative growth rate of collagen was higher than 70%and it had good biocompatibility.Therefore,double-spotted pufferfish skin collagen is expected to become an alternative product to land-derived collagen.(2)The silk fibroin was created from degummed silk,and its structural and rheological and thermal stability were examined.SDS-PAGE showed that the relative molecular mass of the light chain of silk fibroin was 24.697 k Da.FTIR and secondary structure analysis showed that silk fibroin was mainlyα-helix,and it making up 50.49%of the secondary structure.XRD analysis showed that the silk fibroin had silk II and silk I.TGA showed that the maximum thermal decomposition temperature of silk fibroin was279.19℃and the residual amount was 7.62%.The rheological properties show that the constant viscosity of the silk fibroin at a certain shear frequency is helpful for spinning.Therefore,silk fibroin is a good material source for composite nanofibre membranes.(3)Collagen/silk fibroin(Col/SF)composite nanofibre membranes were prepared by using electrostatic spinning technology to investigate the effect of SF content on the morphology,structure and properties.SEM results showed that the average diameter of composite nanofibre membranes decreased as the SF content increased.The secondary structure analysis showed that the Col/SF composite nanofibre membrane consists mainly of alpha-helix,and the content of alpha-helix increased from 46.38%to 52.63%with the increase of SF content,and there was no random coils.The highest temperature was 303.14°C at Col:SF=60:40.Compared with collagen nanofibre membrane,the maximum thermal decomposition temperature of the Col/SF composite nanofibre membrane increased by 8.23°C and the residual amount increased from 28.13%to31.27%,the addition of SF improved the thermal stability.The mechanical properties showed that the elongation at break and the average tensile strength of the composite nanofibre film remained maximum at Col:SF=60:40,with 21.6±1.62%and 3.27±0.19Mpa respectively.The elongation at break and the average strength at break were both higher than those of the collagen nanofibre film.Biocompatibility experiments showed that the the rate of cellular viability of composite nanofibre membrane above 70%and they could be used as a non-toxic scaffold. |
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