| Collagen is the most abundant protein in animals and constitutes about 25% of total proteins.They comprise the major structural element of all connective tissues and are also found in the interstitial tissue of virtually all organs,where they contribute to the stability of tissues and organs,and maintain their structural integrity.Due to its unique properties such as low antigenicity,high biodegradability and biocompatibility,collagen is widely used in food,biomedical and tissue engineering.To the best of our knowledge,however,the report has been quite limited concerning application of fish skin collagen in the biomedical material field.The aim of this article was to extract collagen from Paugusiushamiltoa fish skin and prepare guided tissue regeneration membrane for studying feasibility as biomedical material.It may provide experimental evidence for exploration and make good use of this fish skin collagen.The extraction collagen from fish skin and the physicochemical characteristics of collagen were measured.Different concentrations solution of sodium hydroxide and hydrogen peroxide treatment Paugusiushamiltoa skin,can also play a role on bleaching,removing non-collagenous protein and defatting.The skin were defatted with 5% H2O2 and 0.2%NaOH to achieve optimum effect.After a series of preliminary procedures,acid soluble collagen and pepsin soluble collagen were isolated from about 100 g with yields of 4.34 g and 5.62 g on a dry weight basis,respectively,this result suggested appropriate amount pepsin may can improve extracted capacity of collagen.The amino acid compositions showed collagen were rich in Gly,Pro,and Hyp,which were characteristic amino acid.Gly was the most abundant amino acid,and the collagen were found to contain no Try,Tyr and Met.There was imino acid(Pro and Hyp)content as relatively high as 26.40%.The denaturation temperature of collagen was 32.0 ?C by viscosity determined.The maximum ultraviolet absorption of collagen was 232 nm,which was accorded with the characteristic absorption peak at 226 nm.It was indicated by the FTIR spectra suggested that the collagen retained molecular order of nature collagen and existed the triple-helical structure.There was collagen consisted of two different type chains,which are normally designated as ?1 and ?2,having approximately the same molecular weight with approximately 130 kDa by the electrophoretic patterns.According to this,the collagen extracted from fish skin was type I collagen and existed triplehelical structure.Preparation of collagen membrane and the biochemical characteristics were analyzed.By using the technology of the vacuum freeze drying,the collagen membrane was prepared by crosslinking treatment at 110?C.The surface morphology was observed by SEM that membrane presented a porous network structure and bilayer structure including a dense layer on the one side and a loose layer on the other side.The porosity rate,water absorption rate and average tensile stress were 78%,4 times than itself and 18.82 MPa respectively,suggesting the collagen membrane had good mechanical strength and porosity.The collagen membrane was thoroughly degraded in PBS containing lysozyme in the two weeks.It did not produce sensibilization reaction to the rabbit skin,and does not have any provocation to the skin of rabbit.Taken together,the collagen membrane meet the basic demand of biomedical engineering material.The capacity of collagen membrane guiding new bone regeneration of rabbit and its degradability was evaluated.New Zealand rabbits were used to establish three 8 mm calvarial defect models which was covered by collagen membrane,Bio-Gide membrane and was not treated with any materials,the status of bone repairing were presented by naked eyes,imaging and morphoanatomical analysis of bone defect models.The effect assessed by micro-CT and HE staining observation in every phase,the region of the bone defect covered by collagen membrane and Bio-Gide membrane have different degrees of new bone accumulation in the bone defect.The speed of degradation of Bio-Gide membrane was faster than Bio-Gide membrane.The new bone regeneration effect of BioGide membrane was superior to collagen membrane.Bone defect which was not treated with any materials was covered with fibrous tissue and had no new bone formation.It can concluded that collagen membrane featured excellent biomedical material property.It can restrict the growth of fibrous connective tissue in the reparative process and promote new bone regeneration.It is recommended to apply collagen membrane as bone tissue repairing substitute.In conclusion,Paugusiushamiltoa fish skin is a good collagen source.Collagen membrane derived from the fish possess the basic need of biomedical materials,and has the ability to guide bone regeneration.It will be a kind of potential biomedical materials and have a broad application prospect. |
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