A kind of plga cell scaffold for articular cartilage repair and its preparation method and application

A cell scaffold and chondrocyte technology, applied in the field of medicine, can solve the problems of cell dedifferentiation, poor bionic effect, and excessive absorption of induced matrix.

Active Publication Date: 2021-07-09
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Chondrocyte transplantation technology to repair cartilage damage has made great progress in recent years. Unfortunately, due to the small size of the initial cartilage tissue sample, slow-proliferating chondrocytes must undergo long-term expansion to reach a certain number, and the treatment cycle is long. And unfavorable factors such as excessive absorption of induced matrix, poor cell recognition signal of matrix material, poor bionic effect, and dedifferentiation of cells lead to unsatisfactory treatment results

Method used

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  • A kind of plga cell scaffold for articular cartilage repair and its preparation method and application
  • A kind of plga cell scaffold for articular cartilage repair and its preparation method and application
  • A kind of plga cell scaffold for articular cartilage repair and its preparation method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1: Preparation of PLGA porous scaffold

[0078] 1. Preparation of PLGA porous scaffold. The scaffold part of the articular cartilage repair material uses polylactic acid-glycolic acid (PLGA) as the matrix material, which is composed of organic polymer polylactic acid (PLA) and polyglycolic acid (PGA), according to a certain gradient ratio (polylactic acid (PLA): Polyglycolic acid (PGA) = 100:0, 85:15, 75:25, 50:50, 25:75, 15:85, 0:100) polymerization product ( figure 1 ).

[0079] Polylactic acid (PLA), polyglycolic acid (PGA), polylactic acid-glycolic acid (PLGA) monomer chemical structure formula is as follows:

[0080] Polylactic acid [poly(lactic acid), PLA]:

[0081] Polyglycolic acid [poly(glycolic acid), PGA]:

[0082] Polylactic-co-glycolic acid [poly(lactic-co-glycolic acid), PLGA]:

[0083] Prepare the PLGA porous scaffold by freeze-drying technique, the steps are as follows:

[0084] a) PLA / The PGA polymerization product—polylactic ac...

Embodiment 2

[0110] Example 2: Primary culture of adult cells

[0111] 1. Chondrocytes. Adult chondrocytes are derived from articular hyaline cartilage in nonweight-bearing areas.

[0112] a) Cartilage sampling preparation. Items to be prepared include trypsin, type II collagenase, DMEM high-glucose medium, PBS buffer, pipettes, culture dishes or flasks of different capacities, centrifuge tubes, filters, gauze, surgical equipment such as tweezers, scissors, etc. for disinfection after supplies.

[0113] b) Cartilage sampling. After anesthesia, disinfect with povidone iodine and deiodine with 75% alcohol, take about 50-100 mg of articular hyaline cartilage from non-weight-bearing parts, and put it into a petri dish filled with PBS buffer.

[0114] c) Repeated pipetting and washing with cold PBS buffer containing antibiotics 3 times. After the third wash, let stand for 5 minutes, discard the upper liquid and floating tissue, and move the cartilage tissue into a clean sterile vessel.

...

Embodiment 3

[0134] Example 3: Preparation of platelet-rich plasma (PRP) and its release of growth-promoting factors

[0135] 1. Using a 20mL sterile syringe and a vacuum blood collection tube containing ACD-A anticoagulant, draw about 18mL of blood at room temperature (22°C), and put the blood into four 5mL centrifuge tubes, 4mL in each tube. Platelet-rich plasma preparation should be completed within 4 to 6 hours.

[0136] 2. Preparation of platelet-rich plasma. Platelet-rich plasma with effective concentration was prepared by secondary centrifugation, and the activity levels of cytokines PDGF and TGF-β1 in platelet plasma prepared by this method increased significantly (P<0.01).

[0137] a) Place 4mL of each tube symmetrically in a centrifuge, centrifuge for the first time at 200-400g×10min, take 3mm of plasma above and below the buffy coat after centrifugation, and put it in another centrifuge tube,

[0138] b) Perform a second centrifugation at 200-400 g×10 min, remove the supernata...

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Abstract

The invention discloses a PLGA cell scaffold for articular cartilage repair, a preparation method and application thereof. The PLGA cell scaffold includes a PLGA porous scaffold and chondrocytes, bone marrow mesenchymal stem cells and adipose stem cells planted in the PLGA porous scaffold; wherein, the PLGA porous scaffold is composed of polylactic acid (PLA) and A multi-layered composite scaffold composed of polyglycolic acid (PGA) polymerization products; the PLGA porous scaffold is divided into three regions: the outer region, the middle region, and the inner region. Inoculate chondrocytes and adipose stem cells, and inoculate chondrocytes and bone marrow mesenchymal stem cells into the pores in the inner area of ​​the PLGA scaffold. The scaffold material of the present invention uses a gradient ratio of artificially synthesized polymer materials combined with natural biological proteins to present a multi-level composite structure. Experiments have proved that the PLGA cell scaffold of the present invention has a good repair effect on cartilage defects.

Description

technical field [0001] The present invention relates to an articular cartilage repair material and its preparation method and application, in particular to a PLGA cell scaffold made by combining a composite scaffold with mechanical strength, tissue compatibility and biodegradability and adult cells , can be used for clinical treatment of cartilage damage caused by various reasons. The invention belongs to the technical field of medicine. Background technique [0002] Cartilage damage is a common and frequently-occurring disease in clinical practice, especially joint damage is very common in people over 50 years old. Diseased joints occur in different parts of the body, and can be single or multiple, with varying degrees of dysfunction. Different degrees of inflammatory reactions and cartilage defects can be found under arthroscopy. Under normal circumstances, doctors will treat symptoms, such as pain relief, anti-inflammation, physical therapy, etc.; if the lesion continu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/18A61L27/22A61L27/24A61L27/56
CPCA61L27/18A61L27/222A61L27/24A61L27/3817A61L27/3834A61L27/56A61L2430/06C08L67/04C08L77/04C08L89/00
Inventor 王克维王文波孙殿军
Owner HARBIN MEDICAL UNIVERSITY
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