Growth factor-loaded collagen group composite material as well as preparation method and application thereof
A growth factor and composite material technology, applied in medical science, prosthesis, etc., can solve the problems of uncontrolled growth factor release direction, loss of growth factor activity, waste of growth factor, etc., to achieve good biocompatibility, preparation process Mild conditions and structurally stable effects
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[0025] Example 1: Preparation of two kinds of loose and dense collagen membranes
[0026] 1. Preparation of beef tendon type Ⅰ collagen swelling solution
[0027] Wash the commercially available fresh beef tendon thoroughly, remove the peritendinous tissue, tendon sheath and fascia, and freeze it in a freezer after washing. Take out the frozen beef tendon and cut it into slices with a thickness of about 1mm. Add ficin solution (concentration 0.05-0.25%) for treatment. After stirring, place it in a thermostat for enzymatic hydrolysis for 24 hours, and add excess H 2 O 2 Terminate the reaction, rinse with distilled water for 3-4 times, decant and add 0.3% malonic acid solution to swell for 24 hours. The swollen tendon slices are fully stirred for 6 hours, the viscosity is adjusted with 0.3% malonic acid solution, and the 80-120 mesh stainless steel mesh is filtered under positive pressure to remove impurities and unswelled materials, and then packaged.
[0028] 2. Preparation of air-d...
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[0038] Example 2: Preparation of chitosan-heparin-bFGF nanoparticles by ion cross-linking method
[0039] 1. Dissolve chitosan (molecular weight 10000, degree of deacetylation 85%) in 1% acetic acid solution, filter and dialyze for 4 days, freeze-dry to obtain purified chitosan.
[0040] 2. Weigh 0.2g chitosan and dissolve it in 5ml 1% acetic acid, 0.1mol·L -1 Adjust the pH to 6.0 with NaOH and dilute to 100ml to obtain a concentration of 2mg·ml -1 The chitosan solution is filtered to remove impurities.
[0041] 3. In 1mg·ml -1 Add the expected amount of bFGF to the heparin (molecular weight 12000) solution and mix overnight (8-12 hours). Mix 2ml 2mg·ml at 4℃ and 700rpm stirring -1 Chitosan solution drop by drop 5ml 1mg·ml -1 In the compound bFGF heparin, the dropping rate is 20 drops / min, and the chitosan-heparin nano-particles encapsulating bFGF are obtained.
[0042] The size of the prepared nanoparticles is relatively uniform when observed under transmission electric ytterbium, s...
Example Embodiment
[0043] Example 3: Preparation of collagen-based composite material loaded with bFGF
[0044] The cross-linked loose layer collagen membrane prepared in the above-mentioned embodiment was wetted with double distilled water and then spread flat, and Ch-He nanoparticles carrying bFGF were slowly dropped on the surface to cover the entire surface. Air-dry at 4°C until there is no mobile phase liquid on the surface of the CFF, then moisten the cross-linked dense layer collagen membrane and spread it on the surface of the cross-linked loose layer collagen membrane with a small amount of liquid, and dry it thoroughly at 4°C to obtain bFGF-loaded collagen Matrix composite material, in which the amount of bFGF is 100ng / cm -2 .
[0045] The cross section of the double layer material is sprayed with gold, and the field emission scanning electron microscope observation shows that ( image 3 ): The composite material is formed by a two-layer structure, the dense layer structure is compact and f...
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