Preparation method of polysaccharide-active protein/polypeptide-based active hydrogel microspheres with high cell affinity

A technology of hydrogel microspheres and active proteins, which is applied in the field of preparation of polysaccharide-active protein/polypeptide-based active hydrogel microspheres with high cell affinity, and can solve the problem of insufficient cell affinity of polysaccharide materials, etc. problems, to achieve the effect of improving biological safety, avoiding toxic and side effects, and promoting normal tissue formation

Pending Publication Date: 2022-06-24
SHANGHAI QISHENG BIOLOGICAL PREPARATION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the problem of insufficient cell affinity of polysaccharide materials, the present invention provides a high cell affinity polysaccharide-active protein / polypeptide-based active hydrogel microsphere and its preparation method

Method used

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  • Preparation method of polysaccharide-active protein/polypeptide-based active hydrogel microspheres with high cell affinity
  • Preparation method of polysaccharide-active protein/polypeptide-based active hydrogel microspheres with high cell affinity
  • Preparation method of polysaccharide-active protein/polypeptide-based active hydrogel microspheres with high cell affinity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of Hyaluronic Acid-Lysozyme Hydrogel Microspheres

[0038] Solution 1: Accurately weigh 0.12g of sodium hyaluronate powder (Mw=40kDa) and dissolve it in 2.5g of phosphate buffer solution (pH=7.2), stir evenly to a clear and transparent solution.

[0039] Solution 2: Accurately weigh 0.24 g of lysozyme powder, dissolve it in 2.5 g of phosphate buffer (pH=7.2), and stir evenly to a clear and transparent solution.

[0040] After the solution 1 and the solution 2 were mixed and stirred to form a homogeneous phase, 0.02 g of DMTMM powder was added thereto, and the mixture was continued to be uniformly stirred to obtain a gel-forming solution 3.

[0041] Solution 4: Add 0.75g of surfactant Span-80 to 50g of dimethicone, and stir to a clear and transparent solution at room temperature.

[0042] At room temperature, solution 3 was added dropwise to solution 4, and the stirring speed was 400 rpm. After 24 hours of reaction at room temperature, the superna...

Embodiment 2

[0043] Example 2: Hyaluronic acid-lysozyme hydrogel microsphere particle size range test

[0044] The gel spheres in Example 1 were soaked in phosphate buffer (pH=7.2), and an appropriate amount was drawn to measure the particle size distribution of the gel spheres. The size of the gel spheres is 20-100 μm, and the particle size is normally distributed, as shown in the appendix. figure 2 shown.

Embodiment 3

[0045] Example 3: Observation of the growth of L929 cells on the surface of hyaluronic acid-lysozyme hydrogel microspheres

[0046] The gel spheres in Example 1 were sterilized with 75% ethanol solution, and fibroblasts were planted on the surface of the gel spheres. After 1 day, the cells adhered and grew on the surface of the material. image 3 As shown, the fibroblasts adhered to the surface of the gel spheres exhibited both spindle and round morphologies.

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Abstract

The invention relates to preparation of polysaccharide-active protein / polypeptide-based active hydrogel microspheres with high cell affinity. Specifically, the hydrogel active microspheres adopt a two-component system, namely (1) water-soluble polysaccharide with a plurality of carboxyl functional groups; and (2) a water-soluble protein or polypeptide chain segment with an arginine-glycine-asparaginic acid sequence (RGD peptide segment) and two or more lysine (amino functional groups) as a cross-linking agent. Through an emulsion method, according to different properties of a gel forming component solution, under the catalytic action of a condensing agent such as 4-(4, 6-dimethoxytriazine-2-yl)-4-methylmorpholine hydrochloride (DMTMM) or 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride / N-hydroxysuccinimide (EDC / NHS) and the like, the gel forming component solution is subjected to a condensation reaction in the presence of a catalyst, so that the gel forming component solution is prepared. The two components can be subjected to amidation covalent crosslinking (-(C = O)-N-) in an emulsion system to form hydrogel microspheres. The polysaccharide-active protein / polypeptide-based hydrogel disclosed by the invention has the functions of supporting cell growth, stimulating generation of collagen secreted by fiber cells, promoting generation of normal tissues and the like; the system material is simple in preparation process and excellent in biocompatibility, and can be endowed with multiple functions.

Description

technical field [0001] The invention belongs to the field of tissue filling and tissue engineering microcarrier materials, in particular to the preparation of a polysaccharide-active protein / polypeptide-based active hydrogel microsphere with high cell affinity, the obtained product and application. Background technique [0002] With the development of social economy and the improvement of people's anti-aging awareness, people's demand for cosmetic fillers is gradually increasing. In recent years, dermal fillers obtained by dispersing a certain proportion of microspheres in a gel matrix have received extensive attention. The microspheres are gradually encapsulated by fibroblasts in vivo, and at the same time, fibroblasts are stimulated to secrete collagen, and then the generated collagen Proteins fill in the voids created by the slow degradation of the microspheres. Because of the collagen-stimulating effect, the filling effect of the microsphere dispersion lasts for a long ...

Claims

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Application Information

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IPC IPC(8): A61L27/52A61L27/54A61L27/26A61L27/50
CPCA61L27/52A61L27/54A61L27/26A61L27/50A61L2300/412A61L2300/45A61L2300/236A61L2300/254C08L89/00C08L5/08
Inventor 魏长征谈浩琪郭家宏余青青蒋丽霞
Owner SHANGHAI QISHENG BIOLOGICAL PREPARATION CO LTD
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