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脱钙骨移植诱导关节软骨再生活性相关特异性蛋白的研究

Study on specific proteins involved in articular cartilage regeneration activity induced by decalcified bone transplantation.

作者信息

Wang Min, Dai Tianming, Feng Zhencheng, Meng Qingqi, Wang Wen, Li Siming

机构信息

Department of Orthopaedics, Guangzhou Red Cross Hospital, Jinan University, Guangzhou, China.

出版信息

Ann Transl Med. 2021 Jan;9(2):160. doi: 10.21037/atm-20-8195.

Abstract

BACKGROUND

Through a comprehensive analysis of the joint synovial fluid produced in the process of rabbit articular cartilage regeneration, the role and characteristics of knee synovial fluid in the process of decalcified bone transplantation-induced articular cartilage regeneration were explored.

METHODS

Twenty New Zealand white rabbits (approximately 2.5 kg in weight) were selected, and bilateral distal femoral bones from two randomly selected rabbits were extracted. After decalcification, the bones were cut into 2 mm × 4 cm long decalcified bone strips. Meanwhile, the other 18 rabbits were randomly divided into three groups: the test group (8 rabbits), the positive control group (6 rabbits), and the blank group (4 rabbits). In the test group, the decalcified bone joint was transplanted into the rabbits at the articular cartilage defect; in the positive control group, the articular cartilage defect of the rabbits were treated and put aside; in the blank group, no rabbits were treated. On the day of transplantation, and on the 4th, 8th, 12th, and 16th weeks after transplantation, the joint synovial fluid of each group was taken for two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) analysis, and related database verification and identification, and compared with the positive control group and the blank group.

RESULTS

Using 2D-PAGE to separate various proteins in the synovial fluid of the rabbit knee joints, it was found that there were differential protein spots in the test group compared with the blank group and the positive control group. After conducting a comparative search and query in the UniProt database, through comprehensive analysis, it was finally found that three proteins with molecular weights of 23,429.4, 57,431.4, and 26,071.1 that may be related to the regeneration of articular cartilage appeared in the test group.

CONCLUSIONS

In the process of inducing the regeneration of articular cartilage using decalcified bone transplantation, knee joint synovial fluid produced specific proteins, which may play an important role in the regeneration of articular cartilage. These findings may offer novel ideas in laying a foundation for the in-depth study of articular cartilage regeneration.

摘要

背景

通过对兔关节软骨再生过程中产生的关节滑液进行综合分析,探讨膝关节滑液在脱钙骨移植诱导关节软骨再生过程中的作用及特点。

方法

选取20只新西兰白兔(体重约2.5 kg),随机选取2只兔子双侧股骨远端,脱钙后将骨切成2 mm×4 cm长的脱钙骨条。同时,将另外18只兔子随机分为三组:试验组(8只)、阳性对照组(6只)、空白组(4只)。试验组将脱钙骨关节移植于兔关节软骨缺损处;阳性对照组对兔关节软骨缺损处进行处理后搁置;空白组不做处理。于移植当天及移植后第4、8、12、16周,采集各组关节滑液进行二维聚丙烯酰胺凝胶电泳(2D-PAGE)、基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)分析及相关数据库验证鉴定,并与阳性对照组和空白组比较。

结果

用2D-PAGE分离兔膝关节滑液中的各种蛋白质,发现试验组与空白组及阳性对照组相比有差异蛋白点。在UniProt数据库中进行比较检索查询,经综合分析,最终在试验组中发现三种分子量分别为23429.4、57431.4和26071.1的可能与关节软骨再生相关的蛋白质。

结论

在利用脱钙骨移植诱导关节软骨再生过程中,膝关节滑液产生了特异性蛋白质,可能在关节软骨再生中起重要作用。这些发现可能为深入研究关节软骨再生奠定基础提供新思路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c372/7867953/6719517ac48d/atm-09-02-160-f1.jpg

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