Injectable nano-composite hydrogel material loaded with growth factors as well as construction method and application of injectable nano-composite hydrogel material
A technology of growth factors and construction methods, applied in the field of injectable nanocomposite hydrogel materials and their construction, can solve the problems that growth factors cannot exist for a long time, and achieve the effect of ensuring long-term action and enhancing effective controlled release
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Embodiment 1
[0037] Step 1: Preparation of VEGF-loaded BPNSs (VEGF-BPNSs)
[0038] (1) Mix 0.6 μg VEGF with 200 μL of 300 ppm BPNSs dispersion;
[0039] (2) stirring at room temperature for 30 minutes;
[0040] (3) Incubate at 37° C. for 6 hours to obtain a VEGF-BPNSs dispersion. figure 1 Shown are the TEM pictures of the BPNSs used. figure 2 TEM pictures of VEGF used.
[0041] Step 2: Preparation of injectable nanocomposite hydrogel (VEGF-BP / DNA) loaded with VEGF
[0042] (1) 0.036g DNA polymer (deoxyribonucleic acid sodium salt derived from salmon testis, Sigma-Aldrich, molecular weight about 1.3×10 6 g / mol) was dissolved in 1mL DPBS, and stirred at 50°C until the DNA polymer was fully dissolved to obtain a DNA prepolymer;
[0043] (2) Put the DNA prepolymer in a high-temperature water bath at 100°C for 1 minute, then quickly transfer it to a constant temperature water bath at 40°C, add 200 μL of VEGF-BPNSs dispersion, and stir thoroughly to obtain a VEGF-BP / DNA prepolymer;
[004...
Embodiment 2
[0046] Step 1: Preparation of VEGF-loaded BPNSs (VEGF-BP NSs)
[0047] (1) Mix 2.4 μg VEGF with 200 μL of 600 ppm BPNSs dispersion;
[0048] (2) stirring at room temperature for 60 minutes;
[0049] (3) Incubate at 4° C. for 12 hours to obtain a VEGF-BPNSs dispersion. image 3 Shown is the TEM picture of the prepared VEGF-BPNSs.
[0050] Step 2: Preparation of injectable nanocomposite hydrogel (VEGF-BP / DNA) loaded with VEGF
[0051] (1) 0.06g DNA polymer (deoxyribonucleic acid sodium salt derived from salmon testis, Sigma-Aldrich, molecular weight about 1.3×10 6 g / mol) was dissolved in 1mL DPBS, and stirred at 50°C until the DNA polymer was fully dissolved to obtain a DNA prepolymer;
[0052] (2) Put the DNA prepolymer in a high-temperature water bath at 95°C for 5 minutes, then quickly transfer it to a constant temperature water bath at 40°C, add 200 μL of VEGF-BP NSs dispersion, and stir thoroughly to obtain a VEGF-BP / DNA prepolymer;
[0053] (3) Transfer the prepolymer...
Embodiment 3
[0058] Step 1: Preparation of VEGF-loaded BPNSs (VEGF-BP NSs)
[0059] (1) Mix 3.6 μg VEGF with 400 μL of 600 ppm BPNSs dispersion;
[0060] (2) stirring at room temperature for 120 minutes;
[0061] (3) Incubate at 25°C for 24 hours to obtain VEGF-BP NSs dispersion;
[0062] Step 2: Preparation of injectable nanocomposite hydrogel (VEGF-BP / DNA) loaded with VEGF
[0063] (1) 0.048g DNA polymer (deoxyribonucleic acid sodium salt derived from salmon testis, Sigma-Aldrich, molecular weight about 1.3×10 6 g / mol) was dissolved in 0.8mL DPBS, and stirred at 50°C until the DNA polymer was fully dissolved to obtain a DNA prepolymer;
[0064] (2) Put the DNA prepolymer in a high-temperature water bath at 90°C for 10 minutes, then quickly transfer it to a constant temperature water bath at 40°C, add 400 μL of VEGF-BP NSs dispersion, and stir thoroughly to obtain a VEGF-BP / DNA prepolymer;
[0065] (3) Transfer the prepolymer to the mold, stand at room temperature for cross-linking fo...
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