Source: UNIV OF MINNESOTA submitted to NRP
MAGNETIC RESONANCE IMAGING OF SUBCLINICAL OSTEOCHONDROSIS IN HORSES
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
1011096
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2016
Project End Date
Sep 30, 2019
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
UNIV OF MINNESOTA
(N/A)
ST PAUL,MN 55108
Performing Department
Veterinary Population Medicine
Non Technical Summary
Osteochondrosis (OC) and its clinically apparent form, osteochondrosis dissecans (OCD), are some of the most common developmental orthopaedic diseases affecting horses and have enormous impact on the equine industry. Osteochondrosis is a highly dynamic disease, with lesions developing and healing over the first year of life. Although early OC lesions may heal with conservative treatment, lesions that persist beyond joint-specific time points or that include clefts that extend through the articular cartilage are unlikely to resolve without surgical intervention.The primary objective of the proposed study is to use MRI to identify lesions of subclinical OC in foals predisposed to OCD and/or suspected to have OC based on radiographic screening. We hypothesize that quantitative MRI techniques (T2 cartilage maps) are able to identify early subclinical lesions of OC and can be used to monitor lesion healing or progression. We also hypothesize that MRI will allow identification of markers predictive of the healing potential of individual OC lesions.During the first stage of the study, stifle and hock joint specimens harvested from foals predisposed to OCD or suspected of suffering from subclinical OC will be evaluated. At predilection sites of OC, T2 cartilage maps will be obtained using a clinically available 3T MR scanner with the aim of identifying areas of epiphyseal cartilage necrosis characterized by increased T2 relaxation times. MRI findings will be validated by histological examination of the imaged sites. During the second stage of the study, MRI techniques evaluated during the cadaveric specimen studies will be tested forapplication in live foals. Young foals predisposed to OC and/or diagnosed with subclinical OC will undergoMRI to obtain T2 cartilage maps of their hock and stifle joints. Cartilage maps will be evaluated for changes consistent with the presence of epiphyseal cartilage necrosis. Progression or resolution of subclinical OC lesions will be monitored using consecutive MRI exams at 2 month intervals until 12 month of age, when lesions are considered permanent or until complete healing of the lesion, whichever occurs first. Lesion characteristics (size, location, T2 relaxation times) will be evaluated for their ability to predict lesion healing or progression. Twelve-month-old foals with MRI diagnosis of OCD will undergo arthroscopic surgery to confirm the diagnosis of OCD.
Animal Health Component
50%
Research Effort Categories
Basic
(N/A)
Applied
50%
Developmental
50%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
31138101050100%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3810 - Horses, ponies, and mules;

Field Of Science
1050 - Developmental biology;
Goals / Objectives
Specific Aim 1: Evaluate novel MRI sequences with immediate clinical availability for their ability to identify early lesions of OC.Hypothesis: T2 maps obtained at clinically relevant field strength (1.5 or 3 Tesla) are able to identify areas of epiphyseal cartilage necrosis, the hallmark of subclinical OC, in ex vivo equine joint specimens.Specific Aim 2: Identify and monitor progression vs. healing of subclinical OC lesions in young foals using bimonthly MRI exams with the aim of identifying specific markers of healing vs. progression.Hypothesis: T2 maps obtained in vivo are able to identify lesions of subclinical OC in foals; alterations in T2 relaxation time of OC lesions along with the size and location of the subclinical lesion can be used as predictors of lesion healing vs. progression
Project Methods
Specific aim 1: Evaluate novel MRI sequences with immediate clinical availability for their ability to identify early lesions of osteochondrosis (OC).In this aim, T2 cartilage maps will be generated in a clinically available MR scanner with the aim of identifying areas of ischemic cartilage necrosis, OC latens, ex vivo using equine stifle and hock joint explants.Stifle and hock joints will be harvested from cadavers of foals (n=5) aged 3 - 6 months predisposed to OC based on familial history of the disease. Specimens will be collected from foal cadavers presented to the University of Minnesota Veterinary Diagnostic Laboratory for diagnostic necropsy, as well as from foals euthanized on the field by veterinarians. Distal femora, tibiae and tali will be freed from their soft tissue attachments and will be stored wrapped in saline soaked sponges at -20 C prior to MRI studies.Collected specimens will undergo MRI at the College of Veterinary Medicine using the 3T GE MR scanner available on site. T2 cartilage maps will be obtained from known predilection sites of OC in each joint, including the proximal and middle third of the lateral and medial trochlear ridges of the femur, the distal intermediate ridge and medial malleolus of the distal tibia, and the distal aspects of the lateral and medial trochlear ridges of the talus. MRI findings will be evaluated for the presence of discrete areas of increased relaxation time in the T2 maps confined to the epiphyseal cartilage, which is consistent with epiphyseal cartilage necrosis (OC latens).At the conclusion of the MRI evaluations, specimens will be fixed in formalin for 48h then decalcified using EDTA solution. Decalcified specimens will be serially sectioned into 3 mm thick blocks matching the planes of the MRI and processed for routine histology. Five μm thick sections cut from the surface of the blocks, matching the location of the MRI slices, will be stained with hematoxylin and eosin and Safranin O. To validate the MRI findings, histological sections will be evaluated for the presence of necrotic epiphyseal cartilage, characterized by decreased Safranin O staining of the extracellular matrix and karyopyknosis on the H&E sections, in areas corresponding with regions of increased T2 relaxation time in the matching MRI slicesSpecific Aim 2: Identify and monitor progression vs. healing of subclinical OC lesions in young foals using bimonthly MRI exams with the aim of identifying specific markers of healing vs. progression.In these in vivo experiments foals predisposed to OC will undergo MRI of their hock and stifle joint using T2 maps to demonstrate the presence of epiphyseal cartilage necrosis and monitor its progression vs. healing.Foals (n=5) aged 4 to 6 months old with unilateral OC of the stifle and/or hock joints or with breeding history (OCD in dam and/or sire) consistent with high likelihood of OC will be enrolled in the study. Diagnosis of unilateral OC will be established based on the presence of unexplained joint effusion and associated mild lameness without marked radiographic changes in the affected joint (Dabareiner et al., 1993). At the time of enrollment foals will undergo in vivo MRI evaluation to obtain T2 cartilage maps both the affected and the contralateral stifle and hock joints. Foals confirmed with subclinical OC during the first MRI session will undergo repeated MRI at 2 month intervals until 1 year of age, at which time OC(D) lesions are expected to be either permanent or healed (Dik et al., 1999). Foals with persisting lesions of OCD will undergo treatment using arthroscopy, which will also serve to confirm the diagnosis.At the conclusion of the study, lesions identified using the T2 maps will be divided into two categories (healed OC lesion vs. clinical OCD) and the MRI findings will be evaluated retrospectively to identify markers predictive of lesion healing or progression. Parameters evaluated will include the T2 relaxation time of the lesion at the time of presentation, temporal changes in its T2 relaxation time (increasing vs. decreasing over time), as well as location and size of the OC lesion.

Progress 10/01/16 to 09/30/19

Outputs
Target Audience:During the second year off the study we have completed data acquisition which is currently being processed and evaluated. Until data processing and evaluation is complete no results can be reported. Thus, we have reached the target audience of this research project yet. Changes/Problems:Analysis of data obtained from studies aimed to accomplish specific aims 1 and 2 identified major problems which cannot be corrected retrospectively but gives us critical information for future study designs. A major problem encountered during completion of specific aim 1 was our inability to obtain data how long a given horse has been dead on farm before it was transported to University of Kentucky diagnostic lab. Consequently, advanced degree of autolysis made histological confirmation of the MRI findings from these cadaveric joint specimens difficult or impossible. In specific aim 2, time elapsed between radiographic identification of subclinical OC lesions in a horse and itsenrollment in the study and subsequent MRI was often too long - universallydue to difficulties obtaining owner consent for the MRI- therefore, by the time the firstMRI session was completed the lesion had healed and was no longer identifiable.? What opportunities for training and professional development has the project provided?Although the research project so far has not yielded the expected results, it provided us with the opportunity to develop close collaboration with theDepartment of Veterinary Science at theUniversity of Kentucky and with the Hagyard Medical Institute in Lexington, KY. Furthermore, the current study also gives us the necessary experience and preliminary data to facilitate submission of a new grant proposal addressing the shortcomings of the current design.? How have the results been disseminated to communities of interest?Data analysis for specific aims 1 and 2 are still both ongoing, thus no results of the study have been disseminated yet. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Specific aim 1: Equine cadaveric stifle and tarsocrural joint specimens were harvested from 11 foals aged 3 weeks to 6 months by our collaborator in Kentucky and shipped to us for ex vivo MRI. Ex vivo MRI identified lesions suggestiveof subclinical osteochondrosis (OC) in 3 specimens. Unfortunately, histological evaluation of the suspected lesions yielded equivocal results due to high degree of autolysis of the specimens. We are currently trying to identify alternative methods that allow us confirmation of the identity of the suspected OC lesions identified in MRI. Specific aim 2: Two consecutive In vivo MRI of the stifle and/or tarsocrural joints of three thoroughbred foals, suspected to be affected with subclinical OC based on results of radiographic screening, have been completed. The MRI data is currently undergoing analysis to identify the presence of subclinical OC lesions; however, preliminary data analysis suggests that none of the imaged foals had lesions consistent with subclinical OC at the time of the imaging.

Publications


    Progress 10/01/16 to 09/30/17

    Outputs
    Target Audience: Nothing Reported Changes/Problems:To accomplish sepcific aim 2 we need to have adequate number of young horses presenting with subtle, early lesions of osteochondrosis dissecnas. Unfortunately,these cases are not available at the University of Minnesota in a large enough numbers to allow completion of the study. Thus, we had to reach out to the eqine industry and eventually forge a collaboration with one of the premier equine hospitals in Lexington KY (Hagyard Medical Institute) to accomplish proposed enrollment goals. Enrollment of horses into the in vivo portion of the study (aim 2) is currently ongoing in KY. What opportunities for training and professional development has the project provided?The project leader (Ferenc Toth) has increasedf his knowledge about nover MRI sequences, suitable for evaluation of musculoskeletal tissues. New collaboration with the equine industry located in Lexington, KY (Gluck Center adn HAgyard Medical Institute)was also developed. A pre-vet, undergraduate student has gained exposure to veterinary research. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?Complete data analysis for specific aim 1 and publish results in a refereed journal. Complete enrollment of horses, and accomplish their sequential MRI as outlined for specific aim 2. Data analysis and reporting of results will follow.

    Impacts
    What was accomplished under these goals? We have completed data gathering for specific aim 1. We have collected joints from 8 juvenile horses and imaged themusing T2 cartilage maps and susceptibility weighted imaging in a 3T MR scanner. MR data is currently being processed, i.e., segmented by a student worker, for further analysis to be completed during year 2. For specific aim 2, after failing to enroll cases here at the University of Minnesota,we have developed collaboration with the Hagyard Medical Institute in Lexington KY, and are starting to enroll horses into the in vivo portion of the study.

    Publications