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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

Pending Publication Date: 2020-08-04
广州市红十字会医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the nature of these two substances, the research progress of type II collagen and HA compound is limited: HA exists in the form of acid in the body, and the existing extraction methods can only obtain HA salt products due to technological limitations. ; Collagen and HA are oppositely charged, and it is easy to form a polymer in a liquid state to produce precipitation
The methods reported in existing patents and literatures include storing the two separately before use, and then mixing them together during use, as described in CN201310099817.4; but this method still does not solve the problem of precipitation when the two are mixed
CN200910041033.X report adopts the method for electrochemical treatment to remove the sodium salt in HA, then the HA solution after dialysis is mixed with collagen solution, thereby obtains the solution that contains HA solution and collagen; But this method takes a long time, and the production cost is high

Method used

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  • II-type collagen and hyaluronic acid composite gel solution and preparation method and application thereof
  • II-type collagen and hyaluronic acid composite gel solution and preparation method and application thereof
  • II-type collagen and hyaluronic acid composite gel solution and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Preparation of Type II Collagen and Detection of Physicochemical Properties

[0053] 1. Pre-treatment: cut hyaline cartilage from pig joints, wash with normal saline, degrease with 75% ethanol for 30 minutes, wash with normal saline, encapsulate, and 60 disinfect. The remaining steps are performed under sterile conditions.

[0054] 2. Prepare high-purity type II collagen solution: Add appropriate amount of pre-cooled 0.01M EDTA to the hyaline cartilage, mash it in a homogenizer, collect the precipitate by centrifugation, and then stir overnight with 10 times the volume of 4M, pH 7.5 guanidine hydrochloride Remove proteoglycan; fully wash with water, take the precipitate after centrifugation, add 0.5M acetic acid enzymatic hydrolysis solution, wherein the weight ratio of cartilage to pepsin is 50:1, this process is carried out below 4°C, and stirred for 24 hours. The supernatant collected by high-speed centrifugation is a type II collagen solution, and this ...

Embodiment 2

[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. Dissolve 10mg of HA in 20ml of the balance substance prepared in a, and the concentration of HA is 2mg / ml.

[0064] d. Mix steps a, b, c to obtain 100ml of composite gel solution containing type II collagen and HA, the pH of the gel solution is 6.5, NaH 2 PO 4 The concentration is about 0.402% by weight; Na 2 HPO 4 The concentration of NaCl is about 0.474% by weight, the concentration of NaCl is about 0.42% by weight, the concentration of glucose is about 0.1% by weight, the concentration of 1,3 propanediol is 0.5 ml / ml, the concentration of Tween 80 is 0.1ml / ml, II The concentration of type collagen was 1 ...

Embodiment 3

[0065] Example 3: Preparation of the composite gel solution of the present application (pH 7)

[0066] a. Prepare 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. Dissolve 200mg of HA in 20ml of the balance solution prepared in a, and the concentration of HA is 10mg / ml.

[0069] d. mixing steps a, b, c to obtain 100ml of composite gel containing type II collagen and hyaluronic acid, the pH of the gel is 7, NaH 2 The concentration of PO4 is about 0.322% by weight, 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 propanediol was 1.5ml / ml, and the concentration of Tween 80 was 0.5ml / ml. The concentration of hyaluronic acid was 2 mg / ml, and the concentratio...

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Abstract

The invention provides a composite gel solution containing II-type collagen and hyaluronic acid, and is characterized in that the composite gel solution comprises II-type collagen, hyaluronic acid anda balance substance, wherein the balance substance comprises a NaH2PO4-Na2HPO4 system, a Tris-HCl system, a Tris-maleic acid system or an HEPES system, and comprises NaCl and / or KCl; and the pH valueof the composite gel solution is 6.5-7.5. The invention also provides a preparation method of the composite gel solution containing II-type collagen and hyaluronic acid. According to the composite gel solution provided by the invention, in a physiological pH value range, II-type collagen and hyaluronic acid can be mutually dissolved in any proportion, the system is stable, and precipitation or denaturation does not occur after long-time placement; and the osmotic pressure and the tension of the composite gel solution are suitable for a physiological system. The preparation method of the composite gel solution containing II-type collagen and hyaluronic acid provided by the invention is very simple, low in cost and easy for large-scale production.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a composite gel solution containing collagen and hyaluronic acid, a preparation method of the composite gel solution, and an application of the composite gel solution in preparing medicine for repairing cartilage damage. Background technique [0002] Osteoarthritis (OA) is the most common chronic chronic disease of middle-aged and elderly people in the world, with a high incidence rate, which tends to occur earlier and increases with age. The etiology of OA is complex and diverse, and general clinical treatment, drugs, and surgery are usually used, and there is no effective clinical control method at present. The main component of cartilage tissue is a rich extracellular matrix, which is mainly composed of type II collagen to form a solid fiber network, filled with liquid hyaluronic acid (HA), and the two-phase system forms a cartilage lacuna to protect the chondrocytes gro...

Claims

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Application Information

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IPC IPC(8): A61K38/39A61K31/728A61K9/06A61K9/52A61K47/02A61K47/36A61P19/02A61P29/00
CPCA61K38/39A61K31/728A61K36/537A61K9/5036A61K9/06A61K47/02A61P19/02A61P29/00A61K2300/00
Inventor 杨小红朱伟聪崔树良刘少杰
Owner 广州市红十字会医院
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