Preparation Of Tilapia Collagen Powder At Low Temperature And Evaluation Of Its Promoting Wound Healing Properties

Collagen widely exists in human and mammalian cartilage,skin and other connective tissues.It is a natural biological macromolecule prepared by itself.At present,the common collagen in the market mainly comes from mammals such as cattle.However,due to the high risk of zoonosis,it also increases the potential risk of collagen in the field of medical materials.Therefore,marine collagen has become a new type of collagen,which is widely used in the field of wound hemostasis and healing.Therefore,in this paper,Tilapia Skin is used as raw material,rich collagen is extracted and further developed into a wound hemostatic excipient,so as to achieve the effect of wound hemostasis and healing.First,different extraction methods were carried to extract collagen from Tilapia Skin,and the highest extraction yield was obtained by enzymatic method through detecting the protein content,followed by single factor experiments to obtain approximate parameters suitable for the extraction of Tilapia Skin Collagen,and then analyzed by response surface method,the obtained results had a high fit with the box Behnken model,And it was concluded that the optimal conditions for the laboratory extraction of tilapia skin collagen were the feed to liquid ratio 1:21,enzyme addition 3.5%,and extraction time 14.5 h,and the final yield of fish collagen could reach 81.92%.Then,the powdered fish collagen prepared at low temperature was freeze pulverized by liquid nitrogen,and the performance physicochemical analysis,UV absorption spectra,and SDS-PAGE electropherograms showed that the extracted fish collagen conformed to the characteristics of type I collagen;CD analysis as well as FTIR infrared spectroscopy indicated a better degree of helix between the three peptide chains of Tilapia Skin Collagen,which did not denature;Differential calorimetric scanning results indicated that the thermal denaturation temperature of tilapia skin collagen was 57.5 ° C.Second,because the ultimate goal of this study is to develop a medical device product,biological tests on medical devices were conducted according to the standard of the series of biological evaluation tests of GB / T 16886 to judge the expected risks of the products in a cautionary manner.The collagen powder was shown to be noncytotoxic by the results of cytotoxicity tests;The results of the intradermal response test showed that none of them was irritant to the intradermal tissue.The results of acute systemic toxicity test and implantation test were negative,and the pyrogen test and hemolysis test met the standard requirements,indicating that fish collagen powder had good biocompatibility whether it was indicated by trauma or did not cause immune rejection of the body after implantation.Finally,as illustrated by the skin repair test and liver hemostasis experiment,the results indicated that tilapia skin collagen powder had a good effect on promoting wound healing and hemostasis of organs.