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Mechanical stimulation method for improving in-vitro preservation effect of articular cartilage

A technology of articular cartilage and mechanics, applied in the field of biomedicine, can solve the problems of decreased activity of chondrocytes, achieve the effects of reducing chondrocyte apoptosis, improving utilization rate and cartilage repair technology level, and improving preservation effect

Active Publication Date: 2018-05-18
TAISHAN MEDICAL UNIV
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problem that the activity of chondrocytes significantly decreases after 2 weeks of preservation in the existing technology of preserving active articular cartilage in vitro, the present invention provides a mechanical stimulation method to improve the preservation effect of articular cartilage in vitro. Rolling pressure stimulation is applied in the cartilage tissue preservation environment to achieve the goal of improving the activity and mechanical properties of chondrocytes preserved in vitro tissue culture, so as to establish a new method of rolling pressure stimulation tissue bank for cartilage preservation in vitro tissue culture environment

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preparation example Construction

[0039] The tissue culture medium used in the present invention can also be TSMU culture medium, and the TSMU culture medium is a published Chinese patent application "a preservation solution for osteochondral grafts and its preparation method" (application number: CN201510093497.0) Osteochondral graft preservation solution described in , each 1000ml of osteochondral graft preservation solution includes: glucose 80-125mmol, amino acid 0.1-2.0mmol, antioxidant 0.2-2.0mmol, basic fibroblast growth factor 5-60nmol , Inorganic salt 1-120mmol, vitamin 1-9mmol, sodium pyruvate 1-2mmol, penicillin 50-70U, the rest is deionized water. The inorganic salt is selected from one or a combination of sodium chloride, potassium chloride and magnesium chloride. The specific preparation steps of the osteochondral graft preservation solution used in the present invention are:

[0040] Take glucose 14.4g, amino acid 0.21g, antioxidant 0.23g, basic fibroblast growth factor 0.0016g, sodium chloride...

Embodiment 1

[0048] (1) In vitro aseptically obtain fresh osteochondral tissue in the weight-bearing area of ​​the knee joint within 3 hours of animal death, with a length of 15 mm and a diameter of about 6.0 mm.

[0049] (2) Mechanical stimulation scheme at the initial stage of the experiment: place the articular cartilage tissue obtained in vitro in a rolling mechanical stimulation device containing DMEM tissue culture medium, and apply rolling mechanical stimulation to the cartilage tissue block; according to the stimulation frequency of 2HZ, the stimulation time The combined parameters of 30min and pressure of 1.5Mpa give cartilage mechanical stimulation, and then replace with fresh tissue culture medium and store in vitro; the environment required for the mechanical stimulation method is: under sterile conditions, the temperature is controlled at 0°C-37 ℃, 1 standard atmospheric pressure, humidity 40%-60%.

[0050] (3) On the 3rd day, 7th day, 14th day, and 21st day after mechanical s...

Embodiment 2

[0052] (1) Fix the knee joint sample on the operating table in accordance with the aseptic principle, disinfect the sample with povidone iodine, and spread a sterile towel. Make a midline incision of the knee joint, open the joint capsule, dissect the patellar ligament and the anterior and posterior cruciate ligaments, select the load-bearing area of ​​the medial and medial condyle of the femoral articular surface, and use a special osteochondral harvesting instrument to collect cylindrical osteochondral pieces (about 10 mm in length and 8 mm in diameter) , given continuous PBS washing during the sampling process), and then the bottom surface of the osteochondral block was trimmed and leveled, put into a sterile vessel and rinsed with PBS.

[0053] (2) Using the developed rolling pressure loading device, the cartilage tissue is loaded according to the loading frequency, time, and pressure as the loading conditions; the loading box is sterilized with ethylene oxide; the conditio...

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Abstract

The invention discloses a mechanical stimulation method for improving the in-vitro preservation effect of articular cartilage. The method comprises the steps as follows: articular cartilage tissue ofanimals or human is acquired sterilely in vitro; the articular cartilage tissue is preserved in a tissue culture solution under certain pressure, humidity and air temperature, cartilage transplant blocks are subjected to periodical mechanical stimulation every 3 days with a special rolling mechanical stimulation method according to the parameter combination (0.1-3 HZ, 1-120 min and 0.1-3 Mpa) of stimulation frequency, time and pressure intensity, and then, the culture solution is replaced for preservation; the cell survival rate and Young modulus of cartilage tissue are detected in different periods. By means of the method, the cell survival rate and mechanical property of the cartilage tissue are notably improved, the duration of the action is significantly prolonged, and the in-vitro preservation effect of the cartilage tissue is improved. The method can be applied to cartilage tissue culture preservation tissue banks, and the utilization rate of cartilage tissue banks and the repairquality of articular cartilage are significantly improved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a mechanical stimulation method for improving the preservation effect of articular cartilage in vitro. Background technique [0002] In recent years, with the comprehensive popularization of sports in our country, there are more and more patients with articular cartilage injury. However, articular cartilage injuries, especially severe cartilage defects involving subchondral bone, often lack ideal treatment methods. Therefore, the treatment of articular cartilage injuries is an urgent medical problem to be solved internationally. Currently, clinical methods for the treatment of articular cartilage injuries include: microfracture, autologous chondrocyte transplantation, tissue engineered cartilage transplantation, autologous and allogeneic articular cartilage transplantation, etc. It is observed clinically that the first three methods are difficult to form high-qua...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0226A01N1/0242
Inventor 亓建洪周路曲鹏玮刘延菊谢地宋洪强吴雅迪
Owner TAISHAN MEDICAL UNIV
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