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Preparation method of cartilage acellular matrix composite scaffold and application thereof

An acellular matrix and composite scaffold technology, which is applied in the field of preparation of cartilage acellular matrix composite scaffolds, can solve the problem that the composition and structure of extracellular matrix cannot be completely simulated, and achieves good biocompatibility and biodegradability, and can Strong controllability and evenly distributed effect

Active Publication Date: 2020-10-30
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most complex synthetic materials and preparation processes cannot completely simulate the composition and structure of the extracellular matrix of natural tissues, and the acellular matrix, as the acellular component of the tissue, stores substances such as cell adhesion proteins and growth factors, which can provide The proliferation, migration and differentiation of cells provide an ideal microenvironment, which is unmatched by any material

Method used

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  • Preparation method of cartilage acellular matrix composite scaffold and application thereof
  • Preparation method of cartilage acellular matrix composite scaffold and application thereof
  • Preparation method of cartilage acellular matrix composite scaffold and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] This embodiment discloses a method for preparing extracellular matrix particles, and the specific steps are as follows:

[0053] 1) Scrape hyaline cartilage tissue from pig articular cartilage (within 24 hours after pig slaughter), cut it into small pieces, put it in PBS solution for 5 minutes of ultrasonication, vacuum freeze-dry, and put it in a -20°C refrigerator for later use.

[0054] 2) Place the freeze-dried extracellular matrix obtained in step 1) in an EP tube, add grinding beads, and grind in a cryogenic grinder for 5-10 minutes. Wherein, the grinding temperature is -20° C., the frequency is 60 Hz, and the machine speed is 3000 r / min. The appearance and diameter of the particles after grinding are as follows: figure 1 As shown, its diameter is about 10-50 μm.

Embodiment 2

[0056] This embodiment discloses a gentle decellularization method, the specific steps are as follows:

[0057] 1) The granules prepared in Example 1 were placed in the hypotonic solution and the hypertonic solution, washed and shaken for 3 minutes, and repeated 3 times. Then the particles were frozen in liquid nitrogen, and then dissolved in a hot water bath at 37°C, and repeated 5 times.

[0058] 2) Move the particles into 1% CHAPS solution to wash 3 times, shake at 800r / min for 3h; then add to 100U / mL DNAse1 solution, shake for 9 at 37°C at 100r / min After 3 hours, change the fresh DNAse1 solution every 3 hours, and finally wash with 0.5% CHAPS solution for 15 minutes.

[0059] 3) Rinse the substrate well with distilled water to remove all detergent. The pelleted decellularized matrix (pdECM) was collected by centrifugation, lyophilized and stored at -20°C.

[0060] After decellularization treatment, no obvious cell debris remained, the cell removal rate reached 95% and a...

Embodiment 3

[0062] In this experiment, on the basis of Example 2, the effects of three different chemical reagents on the decellularization effect were compared (all the other conditions were in Example 2), which were polyethylene glycol octylphenyl ether (Triton X-100), ten Sodium dialkyl sulfate (SDS), 3-[(3-cholesteryl aminopropyl) dimethylamino]-1-propanesulfonic acid (CHAPS), the comparison method is as follows:

[0063] Table 1

[0064]

[0065] Among them, the concentrations of the three solutions are all 1%, and they are shaken under normal temperature conditions.

[0066] Quantitative analysis of DNA and GAG in six different ways, the results are as follows: figure 2 As shown, in general, under different time conditions, the decellularization effect of the CHAPS group is better than that of the other two groups, which is specifically manifested in the high DNA removal rate and the retention of more GAGs.

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Abstract

The invention belongs to the field of biomedical materials, and particularly relates to a preparation method of a cartilage acellular matrix composite scaffold and an application thereof. The structure of the cartilage acellular matrix composite scaffold is shown as a formula I, and the cartilage acellular matrix composite scaffold is a natural polymer or a derivative thereof and is a digested cartilage acellular matrix; wherein the mass percentage of the natural polymer or the derivative thereof is 25-75%, preferably 25-50%; wherein the mass percentage of the digested cartilage acellular matrix is 25-75%, preferably 50-75%. The invention discloses an acellular matrix / natural polymer composite material, after being implanted into a body, natural macromolecules or derivatives thereof are gradually degraded and absorbed in vivo, adhesion protein, growth factors and the like in acellular matrixes promote mesenchymal stem cells in bone marrow blood to differentiate into cartilage cells andmaintain the phenotype of the cartilage cells, the cartilage healing effect is effectively improved, a good operation curative effect is achieved, and the clinical application prospect is good.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, in particular to a preparation method and application of a cartilage decellularized matrix composite scaffold. Background technique [0002] There is no vascular tissue, lymphatic drainage and innervation in the articular cartilage, and the renewal rate of the matrix is ​​slow. Once the cartilage is damaged, it is difficult to heal itself. With the aging of the population, the proportion of patients with osteoarthritis gradually increases every year. According to the World Health Organization, about 10% of men over the age of 60 have symptoms of osteoarthritis. Due to differences in hormone secretion, the proportion of female patients is more serious. According to official statistics, the number of osteoarthritis patients in China is at least 100 million, and the number of patients who need surgery every year is as high as 20 million. On the other hand, the cartilage of the knee j...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/24A61L27/50A61L27/54A61L27/58
CPCA61L27/3633A61L27/3612A61L27/3654A61L27/3687A61L27/3691A61L27/24A61L27/50A61L27/54A61L27/58A61L2430/06A61L2430/40A61L2400/06A61L2300/414A61L2300/412
Inventor 王启光樊渝江卢艳梁洁
Owner SICHUAN UNIV
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