Preparation Of Salmon Collagen Film For Bone Repair And Study Of Its Properties

Objective:In order to make full use of salmon skin resources,we prepared a biodegradable collagen membrane material with excellent performance and biocompatibility,and investigated the effect of Salmon Collagen Membrane（SCM）in bone tissue repair.Methods:Firstly,the salmon skin was treated with different concentrations of Na OH and H₂O₂ solutions at different times to select the best pretreatment conditions to achieve the purpose of decolourisation and deliquescence of protein;and the effects of glutaraldehyde,genipin and EDC cross-linking agents on SCM were compared by examining the in vitro degradation rate,mechanical properties,denaturation temperature,cross-linking degree and microstructure to finally establish The optimum process conditions for SCM were established.Secondly,the physical and chemical properties of the final SCM were investigated to verify whether they met the basic requirements for medical devices,including suture tearing force,p H,moisture,total protein,hydroxyproline,fat,ash,heavy metals and exogenous DNA residues.Finally,the biological safety of SCM was verified by cytotoxicity test,cell proliferation test,cell adhesion test,pyrogen test,haemolysis test,systemic acute toxicity test in mice,partial implantation test,skin sensitization test,sub-chronic systemic toxicity test and intradermal reaction test according to the series of standards of“GB/T 16886 Biological Evaluation Test”.We also established a model of cranial defects in SD rats,observed bone repair in rats by micro X-ray tomography,calculated the rate of cranial healing in rats,and conducted a preliminary study on the effectiveness of SCM.Results:The best decolourisation and decontamination process for salmon skin was determined by treating the skin with 0.1 mol/L Na OH and 4%H₂O₂ for 6 h.The results of the SCM assay for different groups showed that the SCM obtained by cross-linking 10mmol/L carbodiimide had good in vitro degradation rate,mechanical properties,denaturation temperature,cross-linking degree and microstructure,which basically met the requirements.The physical and chemical properties of SCM were investigated as follows:suture tearing force of 3.33±0.4 N,acidity of 7.39±0.03,moisture content of 10.22±0.04%,total protein content of 90.69±0.02%,hydroxyproline content of 8.90±0.98%,fat content of0.96±0.60%,ash content of 0.95±0.33%and The biocompatibility of SCM was investigated as follows:the cytotoxicity test showed that SCM was not cytotoxic;the cell proliferation and cell adhesion test showed that mouse cranial anterior bone cells could adhere and proliferate on SCM.The pyrogen and haemolysis tests showed that SCM was not pyrogenic or haemolytic;the acute systemic toxicity test and the sub-chronic systemic toxicity test showed that SCM was not acutely or sub-chronicly systemic toxic.The local implantation test,intradermal reaction test and sensitization test also showed that SCM has good biocompatibility and safety,and basically meets the requirements of restorative materials.The results of the cranial bone repair study in rats showed that the biodegradable collagen membrane made from salmon skin had certain bone repair effects.