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不同灌洗液冲洗对创伤性骨关节炎兔模型中瞬时受体电位香草酸 5 及关节内炎症的影响。

Effect of infusion irrigation with different irrigating solutions on transient receptor potential vanilloid 5 and intra-articular inflammation in a post-traumatic osteoarthritis rabbit model.

机构信息

Department of Anatomy, Hubei University of Medicine, Shiyan, 442000, Hubei, China.

Department of Traumatic Orthopedics, RenminHospital, Hubei University of Medicine, No.39 Chaoyang Road, Maojian District, Shiyan, 442000, Hubei, China.

出版信息

Eur J Med Res. 2021 Mar 12;26(1):24. doi: 10.1186/s40001-021-00491-0.

Abstract

BACKGROUND

The incidence of post-traumatic osteoarthritis (PTOA) after anterior cruciate ligament reconstruction (ACLR) is high, but there is still a lack of intra-operative preventive measures. This study aimed to evaluate the effect of different irrigating solutions continuous irrigation on intra-articular inflammation and cartilage degeneration.

METHODS

66 New Zealand rabbits were randomly divided into normal (N) group, no treatment (NT) group, sodium chloride (NaCl) group, magnesium sulfate (MgSO) group, and calcium chloride (CaCl) group. The right knee joint of the experimental group was utilized to construct the model of PTOA, and the left side was utilized as the normal control group. At different time points postoperatively, the blood concentration of hemoglobin and Mg2 + , the synovial fluid concentration of IL-1 β, TNF-α, tartrate-resistant acid phosphatase-5b (TRAP-5b), and Type II Collagen, the gene expression of IL-1 β and MMP-3, and the protein expression of TRPV5 and CaM were detected. Pearson's linear correlation was employed to identify the possible relationship between the expression of TRAP-5b and the expression of IL-1β, IL-6, TNF-α, and Type II collagen. The hematoxylin and eosin staining (HE), Masson's trichrome staining, and Alcian blue staining were performed at postoperative 35 days. Osteoarthritis Scoring (OA score) comprised categories including Alcian blue staining, cartilage histology, the cellular density of cartilage, degree of cell disintegration, and formation of chondrocyte cluster were blindly scored by trained researchers at postoperative 35 days.

RESULTS

There was no statistical difference (P > 0.05) in the hemoglobin concentration between different groups. The concentration of serum Mg in the MgSO group was higher than that of the other three groups (P < 0.05) on the same day of operation, then gradually decreased. The expression of IL-1 β, IL-6, and TRAP-5b in synovial fluid increased 5 days after the operation, decreased at 15 days, and then increased again with time in the NT group, NaCl group, and NT group and NaCl group. At 35 days after the operation, the expression of IL-1 β, IL-6, TRAP-5b, and type II collagen in the MgSO group were lower than that in the other three groups (except group N) (P < 0.05).The correlation analysis results showed that the TRAP-5b levels correlated positively with IL-1 β, IL-6, TNF-α, and type II collagen concentrations. The histological examination revealed that the surface smoothness of cartilage, the morphology of chondrocytes, the arrangement of collagen fibers, and the density of proteoglycan in the MgSO group were better than those in other experimental groups. At 35 days postoperatively, the gene expression of IL-1 β and MMP-3 and the protein expression of CaM and TRPV5 in synovium in the MgSO group was lower than that in the NaCl group and CaCl group.

CONCLUSION

Intra-operative irrigation with magnesium sulfate solution can inhibit the inflammatory factors and the expression of TRPV5, which can also reduce collagen loss and delay cartilage degeneration. Therefore, the use of magnesium sulfate in intra-operative irrigation may be an ideal choice to prevent PTOA.

摘要

背景

前交叉韧带重建(ACLR)后创伤后骨关节炎(PTOA)的发病率很高,但仍缺乏术中预防措施。本研究旨在评估不同冲洗液持续冲洗对关节内炎症和软骨退变的影响。

方法

66 只新西兰兔随机分为正常(N)组、无处理(NT)组、氯化钠(NaCl)组、硫酸镁(MgSO)组和氯化钙(CaCl)组。实验组右膝关节构建 PTOA 模型,左侧为正常对照组。术后不同时间点检测血红蛋白和 Mg2+浓度、关节滑液中 IL-1β、TNF-α、抗酒石酸酸性磷酸酶-5b(TRAP-5b)和 II 型胶原浓度、IL-1β和 MMP-3 基因表达以及 TRPV5 和 CaM 蛋白表达。采用 Pearson 线性相关分析鉴定 TRAP-5b 与 IL-1β、IL-6、TNF-α和 II 型胶原表达的可能关系。术后 35 天进行苏木精-伊红(HE)染色、Masson 三色染色和阿尔辛蓝染色。术后 35 天,由经过培训的研究人员对 Alcian 蓝染色、软骨组织学、软骨细胞密度、细胞解体程度和软骨细胞簇形成等进行盲法评分。

结果

不同组间血红蛋白浓度无统计学差异(P>0.05)。MgSO 组术后当天血清 Mg 浓度高于其他三组(P<0.05),然后逐渐下降。术后 5 天,NT 组、NaCl 组和 NT+NaCl 组关节滑液中 IL-1β、IL-6 和 TRAP-5b 的表达增加,15 天后降低,随后随时间再次增加。术后 35 天,MgSO 组 IL-1β、IL-6、TRAP-5b 和 II 型胶原的表达均低于其他三组(N 组除外)(P<0.05)。相关性分析结果表明,TRAP-5b 水平与 IL-1β、IL-6、TNF-α和 II 型胶原浓度呈正相关。组织学检查显示,MgSO 组软骨表面光滑度、软骨细胞形态、胶原纤维排列和软骨蛋白聚糖密度均优于其他实验组。术后 35 天,MgSO 组滑膜中 IL-1β和 MMP-3 的基因表达以及 CaM 和 TRPV5 的蛋白表达均低于 NaCl 组和 CaCl 组。

结论

术中硫酸镁溶液冲洗可抑制炎症因子和 TRPV5 的表达,减少胶原丢失,延缓软骨退变。因此,术中使用硫酸镁冲洗可能是预防 PTOA 的理想选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dceb/7953622/c167eb130c80/40001_2021_491_Fig1_HTML.jpg

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