Artificial bone made of recombinant collagen and fluorine-containing nano hydroxyapatite composite collagen
A technology of nano-hydroxyapatite and composite glue, which is applied in the direction of phosphorus compounds, nanotechnology, genetic engineering, etc., can solve the problem of lack of stability of FA coating, achieve excellent biocompatibility and biosafety, and slow down degradation Good speed and mechanical strength
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preparation example 1
[0082] Preparation Example 1 Preparation of Recombinant Collagen
[0083] Human-like collagen (composed of the amino acid sequence shown in SEQ ID NO: 1) was prepared according to the method described in Chinese patent application publication CN1371919A, and its molecular weight was confirmed to be about 90 kD. Here, this recombinant collagen is simply referred to as human-like collagen, and is used in the following examples.
Embodiment 1
[0085] Step 1: Prepare 0.5mol / L calcium nitrate solution, 0.3mol / L sodium hydrogen phosphate solution, and 0.5mol / L sodium fluoride solution, take 6mL, 6mL, and 0.3mL in sequence and add 5mL oleic acid and 15mL ethanol to mix To the solution, 0.5 g of polyvinylpyrrolidone K50 (dissolved in 5 mL of deionized water) was added thereto, and mixed well at room temperature.
[0086]Step 2: Transfer the mixed solution in step 1 to a 50 mL closed reactor with a polytetrafluoroethylene liner, react at 120° C. for 15 h, cool naturally to room temperature, and wash the precipitate to obtain the synthesized product. The product was identified by X-ray powder diffraction as fluorine nano-hydroxyapatite ( figure 1 ); The prepared product can be dispersed in dispersion liquids of different polarities simultaneously ( figure 2 ); Detecting by transmission electron microscopy shows that the prepared product is about 10nm in diameter, and a nanorod between about 150-300nm in length ( image ...
Embodiment 2
[0090] Step 1: Prepare 0.5mol / L calcium nitrate solution, 0.3mol / L sodium phosphate solution, and 0.5mol / L sodium fluoride solution, take 6mL, 6mL, and 0.6mL in sequence and add 5mL oleic acid and 15mL ethanol to the mixed solution , and then add 0.5 g of polyvinylpyrrolidone K50 (dissolved in 5 mL of deionized water), and mix well at room temperature.
[0091] Step 2: Transfer the mixed solution in step 1 to a 50 mL closed reactor with a polytetrafluoroethylene liner, react at 150° C. for 10 h, cool naturally to room temperature, and wash to obtain fluorine nano-hydroxyapatite.
[0092] Step 3: Mix 4 g of fluorine nano-hydroxyapatite in step 2 with 8 mL of 150 mg / mL human-like collagen aqueous solution, and add 180 μL of 10% (w / v) cross-linking agent genipin to it, and mix well , transferred to the mold, aged at 25°C for 36h, then placed in an ultra-low temperature refrigerator at -80°C for 24h, and then freeze-dried the frozen sample for 48h to obtain an artificial collagen ...
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