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Tissue-specific meniscus extracellular matrix material as well as preparation method and application thereof

A tissue-specific, meniscus technology, applied in the field of biomedicine, can solve the problems of limited application, poor biocompatibility, and ignoring the protection of the overall function of the joint

Inactive Publication Date: 2021-01-22
钟刚
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the mechanical stress during meniscus buffering movement requires very strong mechanical strength, traditional natural materials are difficult to meet this condition. Therefore, the scaffold materials currently used for meniscus tissue engineering repair mainly use artificial synthetic materials, and artificial synthetic materials Poor biocompatibility, which greatly limits their use in meniscus repair
More importantly, current scaffold materials and meniscal repair strategies neglect the preservation of the overall function of the joint, especially the limitation of articular cartilage degeneration, which is extremely important for tissue regeneration and the quality of life of patients in later life

Method used

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  • Tissue-specific meniscus extracellular matrix material as well as preparation method and application thereof
  • Tissue-specific meniscus extracellular matrix material as well as preparation method and application thereof
  • Tissue-specific meniscus extracellular matrix material as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Preparation of mECM.

[0036] 1. Prepare mECM by decellularizing the inner 1 / 3 part of the fresh porcine meniscus, the specific method is as follows:

[0037] (1) Obtain fresh menisci of pigs aged 6-7 months at the slaughterhouse (acquired within 12 hours after slaughter), wash and take the inner 1 / 3 part, and carefully divide it into 5-10 mm in size 3 small pieces of tissue;

[0038] (2) Place the small tissue pieces in an acetic acid solution with a concentration of 0.02mol / L, and soak for 48 hours at 4°C;

[0039] (3) Freeze-dry the tissue pieces at -80°C for 24 hours, then rewarm at room temperature for 4 hours to obtain freeze-dried tissues, and then repeat the above steps to prepare three freeze-dried tissues;

[0040] (4) Grind the freeze-dried tissue into powder, place it in a mixed system of 2% SDS and 10mmol / l tris, stir at 25°C for 24 hours, discard the supernatant, and perform three cycles to obtain a precipitate;

[0041] (5) Treat the precipi...

Embodiment 2

[0057] Example 2. The experiment of mECM promoting the ability of BMSCs to differentiate into fibrocartilage.

[0058] In vitro experiments, this example mixed BMSCs with mECM and conventional cowhide-derived collagen hydrogel (the most commonly used natural scaffold material, Cowhide collagen) for three-dimensional culture, and checked the proliferation of BMSCs in mECM and Cowhidecollagen. Tissue staining, qPCR and Western blotting were used to evaluate the ability of mECM to induce BMSCs to differentiate into fibrocartilage.

[0059] 1. Experimental method:

[0060] (1) Isolation and culture of BMSCs: Three-day-old SD rats were euthanized, soaked in alcohol for two minutes, and the hind limbs of suckling mice were obtained and placed in penicillin and streptomycin solution for two minutes. backbone. Insert a one-mL syringe needle (22-gauge needle) filled with medium into the femoral shaft, flush and collect BMSCs in the femur. BMSCs were cultured in complete medium. The...

Embodiment 3

[0072] Example 3. The experiment of rat back mECM promoting the ability of BMSCs to differentiate into cartilage.

[0073] In this example, mECM was combined with BMSCs, cultured three-dimensionally in vitro for one week, transplanted to the back of the rat by microsurgery, and the tissue on the back of the rat was taken out one month later, and the tissue of the tissue was detected by HE, safranin and toluidine blue staining Morphology and chondrogenic differentiation.

[0074] 1. Experimental method:

[0075] (1) Transplantation of the three-dimensional culture complex to the back of rats: refer to the experimental method in Example 2 (2) Perform three-dimensional culture of BMSCs in mECM, and take out the three-dimensional culture complex for use after one week; the back of seven-week-old male rats Skin preparation and disinfection were carried out near the midline, and a longitudinal incision of about 1 cm in length was made along the midline of the back, and the mECM+BMS...

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Abstract

The invention relates to the technical field of biological medicine, in particular to a tissue-specific extracellular matrix material, a preparation method thereof and application thereof in protecting joint injury and degeneration after meniscus injury. The mECM is prepared by decellularizing the 1 / 3 part of the inner side of a porcine meniscus, the preparation method can thoroughly remove cellsand nucleic acid in the cells, and main components of an extracellular matrix are retained at the same time, so that the maintenance of the steady state of the cells is facilitated; an mECM material prepared by the method has excellent mechanical properties and biocompatibility, not only can repair meniscus injury, but also can effectively reduce fibrosis of an injured part; and the mECM can be used as a scaffold material to be compounded with bone marrow mesenchymal stem cells to play a role in protecting joint injury and degeneration after meniscus injury, and the method is of great significance to tissue regeneration and later life quality of patients.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a tissue-specific meniscus extracellular matrix material, a preparation method thereof, and an application in protecting joint damage and degeneration after meniscus injury. Background technique [0002] The meniscus is a C-shaped tissue composed of elastic cartilage located between the femoral condyle and the tibial plateau in the knee joint. Its main function is to buffer the mechanical pressure between the tibia and femur during movement and avoid direct mechanical impact on the articular cartilage. damage. With the increase of outdoor activities, the incidence of meniscal injury is increasing. After the meniscus is injured, its protective function on the joint is weakened, so it is often accompanied by soft tissue injuries of the knee, such as cruciate ligament injury, joint capsule injury and cartilage surface injury, etc., showing severe pain in the knee joint, unable ...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/36A61L27/38A61K35/32A61P19/02A61K35/28
CPCA61L27/3834A61L27/3633A61L27/3687A61L27/3691A61L27/40A61K35/32A61P19/02A61K35/28A61L2430/24A61K2300/00
Inventor 钟刚
Owner 钟刚
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