II-type collagen and hyaluronic acid composite gel solution and preparation method and application thereof
A composite gel and hyaluronic acid technology, applied in the field of biomedicine, can solve the problems of limiting the research progress of type II collagen and HA composite agent, polymer precipitation, high production cost, etc., and achieve easy large-scale production, stable system and low cost. cheap effect
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[0052] Example 1: Preparation of Type II Collagen and Detection of Physical and Chemical Properties
[0053] 1. Pretreatment: hyaline cartilage was excised from pig joints, washed with normal saline, defatted with 75% ethanol for 30 minutes, washed with normal saline, and then encapsulated, and treated with Co. 60 disinfect. The remaining steps are performed under sterile conditions.
[0054] 2. Preparation of high-purity type II collagen solution: add an appropriate amount of pre-cooled 0.01M EDTA to the hyaline cartilage, mash it in a homogenizer, collect the precipitate by centrifugation, and stir overnight with 10 times the volume of 4M, pH 7.5 guanidine hydrochloride Remove the proteoglycan; fully wash with water, take the precipitate after centrifugation, add 0.5M acetic acid enzymatic hydrolysis solution, wherein the weight ratio of cartilage and pepsin is 50:1, this process is carried out below 4°C, and stirred for 24 hours. The supernatant was collected by high-spee...
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[0060] Example 2: Preparation of the composite gel solution of the present application (pH 6.5)
[0061] a. Preparation of balance substance (90ml): add NaH to deionized water 2 PO 4 0.402g, Na 2 HPO 4 0.474 g, NaCl 0.42 g, glucose 0.1 g, 1,3 propylene glycol 0.5 ml, Tween 80 0.1 ml, pH 6.5.
[0062] b. Measure 10ml of 10mg / ml type II collagen solution.
[0063] c. 10mg of HA is dissolved in 20ml of the balanced substance prepared in a, and the HA concentration is 2mg / ml.
[0064] D. mixing step a, b, c, obtain the composite gel liquid 100ml containing type II collagen and HA, the pH of this gel liquid is 6.5, NaH 2 PO 4 The concentration of Na is about 0.402% by weight; Na 2 HPO 4 The concentration of NaCl was about 0.474% by weight, the concentration of NaCl was about 0.42% by weight, the concentration of glucose was about 0.1% by weight, the concentration of 1,3 propylene glycol was 0.5 ml / ml, the concentration of Tween 80 was 0.1 ml / ml, II Collagen type was 1 mg / m...
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[0065] Example 3: Preparation of the composite gel solution of the present application (pH 7)
[0066] a. Preparation of balance substance (70ml): add NaH to deionized water 2 PO 4 0.322g, Na 2 HPO4 0.566 g, NaCl 0.44 g, glucose 5 g, 1,3 propylene glycol 1.5 ml, Tween 80 0.5 ml.
[0067] b. Measure 30ml of 10mg / ml type II collagen solution.
[0068] c. 200mg of HA was dissolved in 20ml of the equilibrium solution prepared in a, and the concentration of HA was 10mg / ml.
[0069] D. mixing steps a, b, c, obtain the composite gel solution 100ml containing type II collagen and hyaluronic acid, the pH of this gel solution is 7, NaH 2 The concentration of PO4 is about 0.322 wt%, Na 2 The concentration of HPO4 was about 0.566% by weight, the concentration of NaCl was about 0.44% by weight, the concentration of glucose was about 5% by weight, the concentration of 1,3-propylene glycol was 1.5ml / ml, and the concentration of Tween 80 was 0.5ml / ml. The concentration of hyaluronic acid ...
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