Changes In The Content Of Type ¢ñ And ¢ò Collagen In The Intervertebral Disc Tissue Of Animal Models With Different Injury Types
Keywords
Injury, Cartilage Endplate, Type I And Type Ii Collagen, Reverse Transcription Polymerase Chain Reaction
Abstract
Objective To clarify the cellular function changes of intervertebral disc chondrocytes during the fibrosis repair process after intervertebral disc injury by quantitatively measuring the levels of collagen type I and II collagen in the cartilage endplate cells of the injured animal model. Methods A rabbit intervertebral disc injury model was established. 40 healthy New Zealand white rabbits were selected and randomly divided into an annulus fibrosus puncture group, a single upper cartilage endplate puncture group, a multiple upper and lower cartilage endplate puncture group, and a sham operation control group, respectively. On the second day after the operation and at 2, 8, 12, and 24 weeks after the operation, 2 experimental animals were randomly selected from each group to obtain intervertebral disc specimens, and reverse transcription polymerase chain reaction was used to determine the mRNA levels of type I and type II collagen in the cartilage endplate cells of the intervertebral disc. . First, RNA was extracted from the intervertebral disc cartilage endplate tissue, and the RNA concentration and the ratio of RNA concentration to protein concentration were measured using a nucleic acid analyzer. Revertaid M-Mulv reverse transcriptase was used for reverse transcription. Type I and II collagen primers were designed to establish a polymerase chain reaction. System, use 0.5¡ÁTBE electrophoresis buffer to prepare 2% agarose gel electrophoresis (120 V, 40 min), observe the amplification results under UV light, and measure the gray value of different electrophoresis bands. Results The mRNA levels of type ¢ñ collagen in each surgical group showed a progressively increasing trend 8 weeks after surgery, and the extent of the increase was related to the degree of injury. The mRNA level of type II collagen increased transiently within the first 2 weeks in the annulus fibrosus puncture group and the superior endplate single puncture group, and then began to decrease progressively; while in the superior and inferior cartilage endplate multiple puncture groups, the There has been a downward trend since the surgery. Conclusion The synthesis amount of type I collagen by chondrocytes in the nucleus pulposus tissue of injured intervertebral disc continues to increase over time, while the synthesis amount of type II collagen begins to decrease progressively after a slight increase. The formation and change patterns of type ¢ñ and ¢ò collagen in injured intervertebral disc cartilage cells are different from natural degeneration.
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Original research was done by Liu Hao, Liu Xi, Deng Yuxiao, Huang Fuguo
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