BackgroundMechanical loading is purported to restore ligament biomechanics post-injury. But this is difficult to corroborate in clinical research when key ligament tissue properties (e.g. strength, stiffness), cannot be accurately measured. We reviewed experimental animal models, to evaluate if post-injury loading restores tissue biomechanics more favourably than immobilisation or unloading. Our second objective was to explore if outcomes are moderated by loading parameters (e.g. nature, magnitude, duration, frequency of loading).MethodsElectronic and supplemental searches were performed in April 2021 and updated in May 2023. We included controlled trials using injured animal ligament models, where at least one group was subjected to a mechanical loading intervention postinjury. There were no restrictions on the dose, time of initiation, intensity, or nature of the load. Animals with concomitant fractures or tendon injuries were excluded. Prespecified primary and secondary outcomes were force/stress at ligament failure, stiffness, laxity/deformation. The Systematic Review Center for Laboratory animal Experimentation tool was used to assess the risk of bias.ResultsThere were seven eligible studies; all had a high risk of bias. All studies used surgically induced injury to the medial collateral ligament of the rat or rabbit knee. Three studies recorded large effects in favour of ad libitum loading postinjury (vs. unloading), for force at failure and stiffness at 12-week follow up. However, loaded ligaments had greater laxity at initial recruitment (vs. unloaded) at 6 and 12 weeks postinjury. There were trends from two studies that adding structured exercise intervention (short bouts of daily swimming) to ad libitum activity further enhances ligament behaviour under high loads (force at failure, stiffness). Only one study compared different loading parameters (e.g. type, frequency); reporting that an increase in loading duration (from 5 to 15 min/day) had minimal effect on biomechanical outcomes.ConclusionThere is preliminary evidence that post-injury loading results in stronger, stiffer ligament tissue, but has a negative effect on low load extensibility. Findings are preliminary due to high risk of bias in animal models, and the optimal loading dose for healing ligaments remains unclear.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12891-023-06653-x.