Comparison of cellular activity and gene expression in repaired versus reconstructed anterior cruciate ligaments: A case–control study

IF 2.7 Q2 ORTHOPEDICS

Journal of Experimental Orthopaedics Pub Date : 2025-07-21 DOI:10.1002/jeo2.70364

Edoardo Monaco, Alessandro Annibaldi, Adnan Saithna, Fabio Marzilli, Alessandro Carrozzo, Vincenzo Visco, Andrea Ferretti, Danilo Ranieri

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Abstract

Purpose

The aim of this study was to compare gene expression and cell biology in biopsy specimens from repaired and reconstructed anterior cruciate ligaments

Methods

A biopsy of the repaired and reconstructed anterior cruciate ligaments was performed. Gene expression analysis of alpha smooth muscle alpha actin via immunofluorescence and gene expression analysis of Collagen I, Collagen III, Collagen I–III ratio, and p16 via reverse transcription polymerase chain reaction was performed. In addition, cell motility and migration were quantitatively assessed. Data analysis was performed using analysis of variance (ANOVA) and Tukey's multiple comparison tests.

Results

Twenty-five biopsies, 15 from repaired anterior cruciate ligaments and 10 from reconstructed anterior cruciate ligaments were obtained. Gene expression analysis showed a higher expression of Collagen I in the reconstruction group (p = 0.0027) while the expression of alpha smooth muscle actine, was significantly greater for the repair group (p = 0.001). Quantitative analysis demonstrated significantly higher cell motility in the cells from repaired ligament compared to cells cultured from grafts (43 vs. 54% residual open area from scratch, p = 0.0001 at 24 h; 10 vs. 17% residual open area from scratch, p = 0.001 at 48 h).

Conclusions

Molecular analysis of repaired and reconstructed anterior cruciate ligaments demonstrates significant differences with respect to collagen quality, α-SMA, and p16. dPCs cultured from biopsies tissue showed a greater cellular activation and migration capacity in the repaired ligament. The clinical importance of these findings is a confirmation that repaired anterior cruciate ligament tissue exhibits a cellular behaviour consistent with excellent biological healing potential.

Level of Evidence

Level III, case–control study.

Abstract Image

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修复与重建前交叉韧带细胞活性和基因表达的比较：一项病例对照研究

目的比较修复和重建的前交叉韧带活检标本的基因表达和细胞生物学特性。方法对修复和重建的前交叉韧带进行活检。免疫荧光法分析α -平滑肌α -肌动蛋白基因表达，逆转录聚合酶链反应法分析I型胶原、III型胶原、I - III型胶原比例、p16基因表达。此外，定量评估细胞运动和迁移。数据分析采用方差分析（ANOVA）和Tukey多重比较检验。结果25例活检，修复前交叉韧带15例，重建前交叉韧带10例。基因表达分析显示，修复组α -平滑肌肌动蛋白表达显著高于重建组（p = 0.0027），修复组α -平滑肌肌动蛋白表达显著高于修复组（p = 0.001）。定量分析表明，与移植物培养的细胞相比，修复韧带细胞的细胞运动性显著提高(43比54%的从头开始的剩余开放面积，p = 0.0001；10 vs. 17%从头开始的剩余开放面积，48小时p = 0.001)。结论修复和重建前交叉韧带的分子分析显示胶原质、α-SMA和p16有显著差异。活检组织培养的dPCs在修复韧带中显示出更大的细胞活化和迁移能力。这些发现的临床重要性是证实修复的前交叉韧带组织表现出与优异的生物愈合潜力一致的细胞行为。证据等级III级，病例对照研究。

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