David D. McErlain

Forced mobilization accelerates pathogenesis: characterization of a preclinical surgical model of osteoarthritis

Preclinical osteoarthritis (OA) models are often employed in studies investigating disease-modifying OA drugs (DMOADs). In this study we present a comprehensive, longitudinal evaluation of OA pathogenesis in a rat model of OA, including histologic and biochemical analyses of articular cartilage degradation and assessment of subchondral bone sclerosis. Male Sprague-Dawley rats underwent joint destabilization surgery by anterior cruciate ligament transection and partial medial meniscectomy. The contralateral joint was evaluated as a secondary treatment, and sham surgery was performed in a separate group of animals (controls). Furthermore, the effects of walking on a rotating cylinder (to force mobilization of the joint) on OA pathogenesis were assessed. Destabilization-induced OA was investigated at several time points up to 20 weeks after surgery using Osteoarthritis Research Society International histopathology scores, in vivo micro-computed tomography (CT) volumetric bone mineral density analysis, and biochemical analysis of type II collagen breakdown using the CTX II biomarker. Expression of hypertrophic chondrocyte markers was also assessed in articular cartilage. Cartilage degradation, subchondral changes, and subchondral bone loss were observed as early as 2 weeks after surgery, with considerable correlation to that seen in human OA. We found excellent correlation between histologic changes and micro-CT analysis of underlying bone, which reflected properties of human OA, and identified additional molecular changes that enhance our understanding of OA pathogenesis. Interestingly, forced mobilization exercise accelerated OA progression. Minor OA activity was also observed in the contralateral joint, including proteoglycan loss. Finally, we observed increased chondrocyte hypertrophy during pathogenesis. We conclude that forced mobilization accelerates OA damage in the destabilized joint. This surgical model of OA with forced mobilization is suitable for longitudinal preclinical studies, and it is well adapted for investigation of both early and late stages of OA. The time course of OA progression can be modulated through the use of forced mobilization.

A bony connection signals laryngeal echolocation in bats

Echolocation is an active form of orientation in which animals emit sounds and then listen to reflected echoes of those sounds to form images of their surroundings in their brains. Although echolocation is usually associated with bats, it is not characteristic of all bats. Most echolocating bats produce signals in the larynx, but within one family of mainly non-echolocating species (Pteropodidae), a few species use echolocation sounds produced by tongue clicks. Here we demonstrate, using data obtained from micro-computed tomography scans of 26 species (n = 35 fluid-preserved bats), that proximal articulation of the stylohyal bone (part of the mammalian hyoid apparatus) with the tympanic bone always distinguishes laryngeally echolocating bats from all other bats (that is, non-echolocating pteropodids and those that echolocate with tongue clicks). In laryngeally echolocating bats, the proximal end of the stylohyal bone directly articulates with the tympanic bone and is often fused with it. Previous research on the morphology of the stylohyal bone in the oldest known fossil bat (Onychonycteris finneyi) suggested that it did not echolocate, but our findings suggest that O. finneyi may have used laryngeal echolocation because its stylohyal bones may have articulated with its tympanic bones. The present findings reopen basic questions about the timing and the origin of flight and echolocation in the early evolution of bats. Our data also provide an independent anatomical character by which to distinguish laryngeally echolocating bats from other bats.

Molecular and Histological Analysis of a New Rat Model of Experimental Knee Osteoarthritis

Abstract:  Articular cartilage degeneration is the most consistently observed feature of osteoarthritis (OA). Animal and human studies have shown that various forms of exercise influence the course of the disease in different ways. In addition, early changes in articular cartilage that influence the progression of OA, such as the expression of cytokines, require further investigation. We have used a surgically induced experimental model of knee OA to address these questions. Here, we discuss our recent studies investigating the effects of an exercise paradigm in surgically induced OA, which determined that the destabilized knee joint is susceptible to enhanced degeneration when subjected to low‐intensity, low‐impact exercise. Further, we investigated early global changes in gene expression in articular chondrocytes from degenerating cartilage. Identified candidate genes including genes involved in chemokine, endothelin, and transforming growth factor‐α signaling are discussed in the context of articular cartilage degeneration in early OA.

Subchondral cysts create increased intra-osseous stress in early knee OA: A finite element analysis using simulated lesions.

AIM OF STUDY To investigate the role of intra-osseous lesions in advancing the pathogenesis of Osteoarthritis (OA) of the knee, using Finite Element Modeling (FEM) in conjunction with high-resolution imaging techniques. METHODS Twenty early stage OA patients (≤ Grade 2 radiographic score) were scanned with a prototype, cone-beam CT system. Scans encompassed the mid-shaft of the femur to the diaphysis of the proximal tibia. Individual bones were segmented to create 3D geometric models that were transferred to FE software for loading experiments. Patient-specific, inhomogeneous material properties were derived from the CT images and mapped directly to the FE models. Duplicate models were also created, with a 3D sphere (range 3-12 mm) introduced into a weight-bearing region of the joint, mimicking the size, location, and composition of a subchondral bone cyst (SBC). A spherical shell extending 1mm radially around the SBC served as the sample volume for measurements of von Mises equivalent stress. Both models were vertically loaded with 750 N, or approximately 1 body weight during a single-leg stance. RESULTS All FE models exhibited a physiologically realistic weight-bearing distribution of stress, which initiated at the joint surface and extended to the cortical bone. Models that contained the SBC experienced a nearly two-fold increase in stress (0.934 ± 0.073 and 1.69 ± 0.159 MPa, for the non-SBC and SBC models, respectively) within the bone adjacent to the SBC. In addition, there was a positive correlation found between the diameter of the SBC and the resultant intra-osseous stress under load (p = 0.004). CONCLUSIONS Our results provide insights into the mechanism by which SBC may accelerate OA, leading to greater pain and disability. Based on these findings, we feel that patient-derived FE models of the OA knee - utilizing in vivo imaging data - present a tremendous potential for monitoring joint mechanics under physiological loads.

Integration and evaluation of a needle-positioning robot with volumetric microcomputed tomography image guidance for small animal stereotactic interventions

PURPOSE Preclinical research protocols often require insertion of needles to specific targets within small animal brains. To target biologically relevant locations in rodent brains more effectively, a robotic device has been developed that is capable of positioning a needle along oblique trajectories through a single burr hole in the skull under volumetric microcomputed tomography (micro-CT) guidance. METHODS An x-ray compatible stereotactic frame secures the head throughout the procedure using a bite bar, nose clamp, and ear bars. CT-to-robot registration enables structures identified in the image to be mapped to physical coordinates in the brain. Registration is accomplished by injecting a barium sulfate contrast agent as the robot withdraws the needle from predefined points in a phantom. Registration accuracy is affected by the robot-positioning error and is assessed by measuring the surface registration error for the fiducial and target needle tracks (FRE and TRE). This system was demonstrated in situ by injecting 200 microm tungsten beads into rat brains along oblique trajectories through a single burr hole on the top of the skull under micro-CT image guidance. Postintervention micro-CT images of each skull were registered with preintervention high-field magnetic resonance images of the brain to infer the anatomical locations of the beads. RESULTS Registration using four fiducial needle tracks and one target track produced a FRE and a TRE of 96 and 210 microm, respectively. Evaluation with tissue-mimicking gelatin phantoms showed that locations could be targeted with a mean error of 154 +/- 113 microm. CONCLUSIONS The integration of a robotic needle-positioning device with volumetric micro-CT image guidance should increase the accuracy and reduce the invasiveness of stereotactic needle interventions in small animals.

Veselka et al. reply

Replying to: N. B. Simmons, K. L. Seymour, J. Habersetzer & G. F. Gunnell 466, 10.1038/nature09219 (2010).We appreciate the comments of Simmons et al. and welcome the new information they have provided about the oldest fossil bat, Onychonycteris finneyi, as well as their confirmation of contact between the stylohyal and tympanic bones in Myzopoda aurita, an extant laryngeal echolocator. Two skeletal features—relatively large cochleae and contact between the stylohyal and tympanic bones—identify extant bats with the capacity for laryngeal echolocation. Although the size of the cochlea can be measured in O. finneyi, the stylohyals may or may not have contacted the tympanics. Simmons et al. disagree with our interpretation of the possible contact between the stylohyal and the tympanic bone in O. finneyi, which indicated that this Eocene bat may have had the capacity for laryngeal echolocation, and have a different interpretation of our results.