Purpose: There exist promising opportunities to improve pharmaceutical efficiency of osteoarthritic (OA) drugs by advancing intra-articular drug delivery systems. However, the study of intra-articular fate of therapeutics in OA joints is limited by readily available in vivo models. Common models include chemical ablation of cellular activity in the joint, which poorly resembles the human condition, and surgical destabilization, which disrupts the integrity of the joint capsule, thereby impacting joint clearance and biodistribution of therapeutics. Here we present the development of a Non-Invasive Knee Injury (NIKI) device that delivers an external force in skeletally mature rats to rupture the anterior cruciate ligament (ACL), an injury that commonly leads to OA in humans. Non-invasive ACL ruptures have been successful in mice, but few studies have involved rats. In this study, device parameters needed for successful ACL rupture were assessed in rat cadavers, immediately post mortem. Future work will evaluate the biological, behavioral, and structural consequences of the injury. Ultimately, we aim to develop a biologically and clinically relevant model of PTOA with an undisrupted synovial capsule. Methods: A linear pneumatic thruster was mounted to a t-beam and the system was controlled by manually varying the pressure regulator voltage. Euthanized Lewis rats (322.9 g ± 22.3 g) were placed supine with the knee and foot secured in 3D-printed mounts such that a compressive vertical load translated the femur relative to the tibia. To evaluate if leg orientation affected ACL rupture rate, mount angle was laterally rotated such that the feet were 36°, 46°, 56°, or 66° relative to the body midline, and a compressive force of 70.8 N was delivered to the knee at 2.24 N/s. To evaluate the effect of loading rate, knees were subjected to 77.9 N at 1.93 N/s or 2.77 N/s. For all studies, ACL condition was evaluated by gross dissection and rated based on visual assessment of tear severity. The presence of tibial draw was also manually evaluated relative to uninjured knees. Results: Calibration by springs indicated that the NIKI could deliver a maximum compressive load of 120N, which exceeds forces needed to rupture the ACL in Sprague Dawley rats. Fig. 1 shows the NIKI device and representative images of ACL tear severity. While higher angles had a numerically greater mean tear severity, no significant difference in tear severity was observed between foot angles. At 66°, one unintended bone injury was observed. Loading rate analysis, which was conducted at a higher maximum force than foot angle studies, had 100% of animals with at least a medium-grade tear. There was no significant difference between the two loading rates studied, and two bone injuries were observed in both loading conditions. In knees exhibiting tibial draw, 97.2% of the cases had at least a medium-grade tear, while 88.9% of the cases with no observed draw did not have a tear. Conclusions: To obtain the greatest incidence of ACL tear without bone injury, there appears to be a critical balance between anatomical orientation and maximum force applied – higher forces yielded greater rate of ACL rupture, but also were associated with increased bone fracture. Future studies will assess more anatomical positions in combination with forces between 70N and 78N to determine the conditions that yield consistent tears without bone injuries. While tibial draw is a subjective assessment, the presence of tibial draw may be a strong indicator of ACL rupture. While the NIKI device is in early stages of development, the clinically relevant joint conditions could allow this model to be used for numerous investigations including studying intra-articular fate.View Large Image Figure ViewerDownload Hi-res image Download (PPT)