Collagen Matrix Cartilage Repair Membrane, Preparation Method and Application
A technology of collagen matrix and cartilage repair, applied in medical science, prosthesis, etc., can solve problems such as cartilage calcification, achieve the effect of inhibiting cartilage calcification, simple preparation process, and smooth surface
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Embodiment 1
[0027] A) strip the tendons from the leg of two-month-old healthy white pigs, remove fascia and fat, shred 120g, soak in 500mL of ethanol aqueous solution with a volume concentration of 75%, overnight at 4°C; B) 95g of the tendon fragments in step A Rinse 5 times with distilled water, add 0.5 mol / L acetic acid aqueous solution 1800mL, adjust pH to 1 with 6M sodium hydroxide aqueous solution, add pepsin (Sigma-ALDRICH) 9g, stand at 4°C for 3 days to obtain 1800mL collagen sol solution; C) Centrifuge the collagen sol solution obtained in step B horizontally at 4°C and 4000rpm for 25 minutes, discard the upper fat layer, dilute the precipitate a with 0.5mol / L acetic acid aqueous solution to 3 times the volume, and filter it with 4 layers of gauze in an ultra-clean bench, Mix the filtrate with 0.9mol / L NaCl aqueous solution at a volume ratio of 1:10, shake intermittently, overnight at 4°C, then centrifuge horizontally at 4°C and 4000rpm for 25 minutes, discard the supernatant and s...
Embodiment 2
[0029]The water-soluble type I collagen extracted by the method in Example 1 was dissolved in deionized water to prepare 10 mL of a collagen aqueous solution with a concentration of 10 mg / mL, and 40 mL of a 1M sodium bicarbonate solution was added for neutralization. Take out 4.3 mL of the neutralized type Ⅰ collagen solution and pour it into a cell culture dish (Corning) with a diameter of 6 cm. Place the culture dish in an oven with a temperature of 37°C and dry it for 48 hours to form a type I collagen film in the mold. Take out the dried type Ⅰ collagen film and cut it into 5mm×5mm size. Take 10 slices of cut membranes, place them in a cell culture dish (Corning) with a diameter of 10 cm, add 5 mL of parathyroid hormone-related peptide (Bachem) solution with a concentration of 100 ug / mL prepared in PBS (pH=7.4), Put it in the cell incubator and soak for 1 hour at 37°C to obtain the type Ⅰ collagen membrane material compounded with parathyroid hormone-related peptide, that...
Embodiment 3
[0031] The water-soluble type I collagen extracted by the method of Example 1 was dissolved in deionized water to prepare 10 mL of collagen aqueous solution with a concentration of 5 mg / mL, and 10 mL of sodium bicarbonate solution with a concentration of 1M was added for neutralization. Take out 4.3 mL of the neutralized type Ⅰ collagen solution and pour it into a cell culture dish (Corning) with a diameter of 6 cm. Place the culture dish in an oven with a temperature of 37°C and dry it for 48 hours to form a type I collagen film in the mold. Take out the dried type Ⅰ collagen film and cut it into 5mm×5mm size. Take 10 slices of cut membranes, place them in a cell culture dish (Corning) with a diameter of 10 cm, add 5 mL of parathyroid hormone-related peptide (Bachem) solution with a concentration of 100 ug / mL prepared in PBS (pH=7.4), Soak in a cell culture box at 37°C for 1 hour to obtain a type I collagen membrane material compounded with parathyroid hormone-related peptid...
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