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Achilles tendon repairing patch and preparation method thereof

A patch and sheet-like technology, applied in the field of biomaterials, can solve the problems of poor repair effect of Achilles tendon injury and poor biocompatibility, and achieve good Achilles tendon repair effect, good biocompatibility, and good cell microenvironment Effect

Pending Publication Date: 2022-04-15
THE AFFILIATED HOSPITAL OF MEDICAL SCHOOL NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, polylactic acid and polyglycolic acid are the most widely used synthetic materials in tissue engineering tendon, which have poor biocompatibility and are acidic after degradation. Poor Achilles tendon injury repair

Method used

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  • Achilles tendon repairing patch and preparation method thereof
  • Achilles tendon repairing patch and preparation method thereof
  • Achilles tendon repairing patch and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1 Preparation of Achilles tendon repair patch

[0049] The present embodiment is the preparation method of Achilles tendon repair patch, comprising the following steps:

[0050] S1. Preparation of SIS matrix

[0051] S11. Collect pig small intestine, wash and cut into small sections of about 5cm-10cm, perform mechanical treatment, tear off the serosa and muscular layer of the small intestine, scrape off the mucous membrane and muscularis mucosa, and obtain the SIS of the small intestinal submucosa, soak the SIS in In PBS buffer, shake and wash 3 times on a shaker, 5-30min each time, such as figure 1 As shown in A.

[0052] S12. Soak the SIS obtained in step S11 in a mixture of methanol and chloroform, the volume ratio of methanol and chloroform in the mixture is 1:1-1:3, and shake on a shaker for 4-12 hours to extract bound esters, Take out the SIS and soak it in PBS buffer solution, shake and wash 3 times on a shaker, 5-30min each time. Repeat the above ...

Embodiment 2

[0068] Example 2 Comparison of small intestinal submucosa before and after decellularization

[0069] 1. Frozen section production

[0070] Take the SIS before and after decellularization obtained in Example 1 for comparison, cut the SIS before and after decellularization to a size of about 1cm×1cm, put it into an embedding box, add OCT embedding agent to completely cover the tissue, and put it in the ice cutter When it is completely solidified, take out the embedding block, place it on a small freezing platform, put it into the sample head to clamp, fine-tune the sample head, trim off excess tissue on the surface and start slicing at the target position, set the thickness to 5-10 μm, turn the handle to cut out the tissue, Coverslips are attached to the tissue.

[0071] 2. H&E staining

[0072] Take SIS frozen slides before and after decellularization, stain with hematoxylin for 10 minutes, rinse with tap water for 10 seconds, dry the slides, add differentiation solution for...

Embodiment 3

[0077] Example 3 Identification of Tendon Stem Cells

[0078] The third-generation tendon stem cells obtained in Example 1 were taken for identification. The tendon stem cells were washed with PBS, digested with trypsin (0.25% Trypsin-EDTA Gibco), centrifuged at 400 g for 5-30 min, and resuspended in 500 μl of ice-cold PBS (containing 10% Fetal bovine serum), take 1ug FITC, PE, AF647, PE-cy 7 labeled anti-rat antibody and isotype control IgG, and incubate at 4°C for 1 hour.

[0079] Using flow cytometry (BECKMAN CytoFlex) analysis, each sample counted about 5 × 10 4 , data processing using FlowJo_V10.8.0. The antibodies used in this example are Anti-CD29 (BD, 562153), Anti-CD44 (BD, 550974), Anti-CD90 (BD, 551401), Anti-CD34 (SA NTA CRUZ BIOTECHOLOGY sc-7324), Anti-CD31 ( Proteintech, FITC-65058). The result is as Figure 5 As shown, more than 90% of the tendon-derived cells were positive for mesenchymal stem cell markers CD44, CD29, and fibroblast marker CD90, and negativ...

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Abstract

The invention provides an achilles tendon repair patch and a preparation method thereof.The achilles tendon repair patch comprises a flaky SIS matrix, tendon stem cells are planted on the SIS matrix, the achilles tendon repair patch takes the flaky SIS matrix as a biological scaffold and the tendon stem cells as seed cells, has good biocompatibility and strong tendon differentiation capacity, and can be used for repairing achilles tendons. The sheet-shaped SIS matrix can wrap the achilles tendon broken end during tendon repair, so that adhesion between surrounding soft tissues and the achilles tendon broken end is isolated, meanwhile, the tendon stem cells carried by the sheet-shaped SIS matrix are accurately attached to the broken end, a good cell microenvironment is provided for proliferation and migration of the tendon stem cells, and a good achilles tendon repair effect is achieved.

Description

technical field [0001] The invention relates to the technical field of biomaterials, in particular to an Achilles tendon repair patch and a preparation method thereof. Background technique [0002] Achilles tendon injury is a relatively common sports injury clinically, ranking third among human tendon rupture injuries. According to statistics, the incidence rate of Achilles tendon injury is 18 / 10000, and the misdiagnosis rate of acute Achilles tendon rupture can be as high as 27% due to doctor's misdiagnosis and patient's negligence. If the missed diagnosis cannot be timely and properly treated for more than 4 weeks, it will develop into old Achilles tendon rupture. Occurs more frequently in athletes and people engaged in greater work intensity, which has seriously affected people's work and life. Common Achilles tendon injuries include acute Achilles tendon rupture, Achilles tendon defect, and chronic Achilles tendinopathy. For acute Achilles tendon rupture and Achilles ...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36A61L27/50
Inventor 毛海蛟赵基源毛旭锋姚立炜张西前
Owner THE AFFILIATED HOSPITAL OF MEDICAL SCHOOL NINGBO UNIV
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