Progress 03/26/14 to 03/25/19
Outputs
Target Audience:Undergraduate, graduate and veterinary students, veterinarians, orthopedic specialists, general public, biotechnology companies. Changes/Problems:Because we have good results from our initial CCLD model and needed to pursue an ovariectomized (OVX) model as well, we decided not to pursue the risky Botulinum toxin approach during this project. We were able to incorporate gene therapy using a therapeutic transgene into both the CCLD (traumatic osteoarthritis) and OVX (menopausal, non-traumatic osteoarthritis) models in our sheep. The treadmill running has continued to produce clinical OA progression in a significantly reduced timeframe. What opportunities for training and professional development has the project provided?Graduate students Crystal Collier and Tracy Habermehl, veterinary students Peter Welsh and Adam Ustach and undergraduate students Kyleigh Taylor and Anisa Samhouri have been involved in planning and executing surgeries and subsequent management of the experimental animals and analysis. This project is the basis for an M.S. project for graduate student Crystal Collier and will contribute to Michael Vakula's PhD. work. Crystal has had previous exposure to this project as an undergraduate student and took over the management of this project until completion. Crystal has also been part of the pursuit of intellectual property protection for her work including open dialogue with our technology transfer department and interested biotech companies. Undergraduate student Anisa Samhouri worked closely with Crystal and is now a veterinary student at Colorado State University and is working with Dr. Laurie R. Goodrich, Professor of Equine Surgery and Lameness on gene therapy efforts in horses. This project also provided the opportunity for veterinary students Peter Welsh (first/second year), Kolby Talbot and Hannah Rodriguez (both fourth year students at WSU) to learn clinical veterinary skills in radiography that veterinary students do not learn while here at USU or after transfer to WSU. How have the results been disseminated to communities of interest?Dissemination of this work includes: Rachel Hill's invited poster entitled 'Development of a Translational Large Animal Osteoarthritis Model' at the 25th Annual Merial-NIH Veterinary Scholars Symposium, "One Health", Cornell University, Ithaca, NY a poster presentation entitled 'Efficacy of micro-computed tomography for analysis of osteoarthritis progression in sheep' at the Student Research Symposium, Office of Research and Graduate Studies. Utah State University, Logan, UT an interview regarding Rachel's work posted on the CAAS website. a peer-reviewed publication of the early work (Rachel J. Hill, Holly M. Mason, Gavin Yeip, Samer S. Merchant, Aaron L. Olsen, Rusty D. Stott, Arnaud J. Van Wettere, Eadric Bressel, and Jeffrey B. Mason. (2017). The Influence of Oblique Angle Forced Exercise in Surgically Destabilized Stifle Joints Is Synergistic with Bone, but Antagonistic with Cartilage in an Ovine Model of Osteoarthritis. Arthritis, vol. 2017, Article ID 7481619, doi:10.1155/2017/7481619). Crystal Collier Crystal Collier and Jeffrey Mason. (2018). Using rAAV-TIMP3 vector for the Improvement of Osteoarthritis in an Ovine Model. USU Student Research Symposium, Utah State University, Logan, UT. Crystal G. Collier, Peter Welsh, Kate C. Parkinson, Anisa Samhouri, McKenna R. Walters, Jeffrey B. Mason. (2018). Using a recombinant adeno-associated viral vector to deliver a Timp-3 Transgene for the Improvement of Osteoarthritis in an Ovine Model. ADVS Student Research Symposium, Utah State University, Logan, UT. Peter Welsh Peter Welsh, Crystal Collier, Holly Mason, Michael Clayton, Arnaud Van Wettere, Aaron Olsen and Jeffery B. Mason. (2018). Peroneus Tertius Lesions: Case Report Associated with Cranial Cruciate Ligament Desmotomies in Sheep. ADVS Student Research Symposium, Utah State University, Logan, UT. Peter Welsh Peter Welsh, Crystal Collier, Holly Mason, Michael Clayton, Arnaud Van Wettere, Aaron Olsen and Jeffery B. Mason. (2019). Peroneus Tertius Lesions: Case Report Associated with Cranial Cruciate Ligament Desmotomies in Sheep. Submitted as a case report to the Journal of the American Association for Laboratory Animal Science. Additional data will be disseminated by Crystal Collier in her M.S. thesis and as two anticipated publications covering the effect of gene therapy on 1) bony changes including osteophytosis and microCT and 2) biomarkers and soft tissue analysis. Gait analysis will be included in Michael Vakula's Ph.D. dissertation and potentially included in his/Crystal's publications. We were contacted by and had a conference call with Medtronic (Dwight Nelson and Dan Schuster) regarding our experimental model. What do you plan to do during the next reporting period to accomplish the goals?Future plans include completing analysis of samples and data collected from the experiments completed in 2018/2019, including the microCT being completed at the University of Michigan. If results from analysis at the University of Michigan and final results from our lab confirm our initial preliminary finding, we will pursue additional funding for planning and execution of a clinical trial using this model.
Impacts
What was accomplished under these goals?
The objectives of this proposal were to 1) develop an improved, non-surgical large animal osteoarthritis model by induction of a transient orthopedic insult by using a Botulinum toxin (Botox) model and 2) to investigate gene therapy and stem cell therapies for the treatment of joint disease in an induced large animal model of OA. The most immediate impact of our work will be on 1) biotech companies interested in pursuing this work for use in humans and 2) in dogs and horses, particularly performance animals who suffer from high incidences of OA. Undergraduate, graduate and veterinary students, orthopedic specialists and the general public will also benefit from our work. Development of our models included the development of a novel oblique angle treadmill forced exercise procedure to exacerbate the progression of clinically-detectable OA. Initial experiments included a classic CCLD model of OA in female sheep. Results from these initial studies using our novel forced exercise regime and the CCLD procedure produced clinically-detectable OA, which normally takes a year or more to develop, in 3-4 months. Because of this success and knowledge from other work involving the influence of ovarian function of development of OA (Mason et al., 2015) and the high costs associated with the proposed botox approach, we decided to proceed with our therapeutic gene therapy model in CCLD and in OVX sheep. Both CCLD and OVX sheep were injected with saline or a therapeutic Timp-3 rAAV transgene virus and subjected to forced exercise. After 16 weeks of forced exercise, both models displayed evidence of OA progression and positive effects of the Timp-3 gene therapy were seen in both models as well. Timp-3 gene therapy reduced the incidence of osteophytosis, the most common marker of OA. Differences in serum and urinary glycosaminoglycan were also detected between gene therapy-treated groups and controls. Our gene therapy-treated CCLD sheep showed that our therapeutic Timp-3 transgene could positively influence traumatic OA progression and our OVX model showed that our therapeutic Timp-3 transgene could positively influence non-traumatic, menopausal OA. These results were made possible by the application of our novel oblique angle forced exercise procedures. Substitution of an OVX model for the botox model has still provided a non-traumatic OA model with the additional benefit of being a menopausal model. This will allow the future use of reproductive manipulation to dissect the causative factors in menopausal OA.
Publications
|
Progress 10/01/17 to 09/30/18
Outputs
Target Audience:Undergraduate, graduate and veterinary students, veterinarians, orthopedic specialists, general public, biotechnology companies Changes/Problems:As this is the last stage of this project, no further experiments are planned. However, if results from analysis at the University of Michigan and final results from our lab confirm our initial preliminary finding, we will pursue additional funding for planning and execution of a clinical trial using this model. What opportunities for training and professional development has the project provided?Graduate students Crystal Collier and Tracy Habermehl, veterinary students Peter Welsh and Adam Ustach and undergraduate students Kyleigh Taylor and Anisa Samhouri have been involved in planning and executing surgeries and subsequent management of the experimental animals and analysis. This project will be the basis for an M.S. project for graduate student Crystal Collier and will contribute to Michael Vakula's PhD. work. Crystal has had previous exposure to this project as an undergraduate student and has taken on the management of this project and will continue with this for the next year. Undergraduate student Anisa Samhouri will continue to work closely with Crystal during the next year of this project. This phase of the project also provided the opportunity for veterinary students Peter Welsh (first year), Kolby Talbot and Hannah Rodriguez (both fourth year students at WSU) to learn clinical veterinary skills in radiography that veterinary students do not learn while here at USU or after transfer to WSU. How have the results been disseminated to communities of interest?Dissemination of this work includes presentations by: Crystal Collier Crystal Collier and Jeffrey Mason. (2018). Using rAAV-TIMP3 vector for the Improvement of Osteoarthritis in an Ovine Model. USU Student Research Symposium, Utah State University, Logan, UT. Crystal G. Collier, Peter Welsh, Kate C. Parkinson, Anisa Samhouri, McKenna R. Walters, Jeffrey B. Mason. (2018). Using a recombinant adeno-associated viral vector to deliver a Timp-3 Transgene for the Improvement of Osteoarthritis in an Ovine Model. ADVS Student Research Symposium, Utah State University, Logan, UT. Peter Welsh Peter Welsh, Crystal Collier, Holly Mason, Michael Clayton, Arnaud Van Wettere, Aaron Olsen and Jeffery B. Mason. (2018). Peroneus Tertius Lesions: Case Report Associated with Cranial Cruciate Ligament Desmotomies in Sheep. ADVS Student Research Symposium, Utah State University, Logan, UT. (manuscript currently in preparation). What do you plan to do during the next reporting period to accomplish the goals?Our plan of work includes completing analysis of samples and data collected from the experiments completed in 2018. Crystal is currently analyzing urine and serum samples collected weekly throughout the project. Her preliminary analysis suggests differences in aggrecan degredation between treated animals and controls in both types of treatments (mechanical destabilization and non-mechanical ovariectomy). Preliminary readiograph analysis have revealed a significant down-regulation of osteophytosis in treated animals, again in both types of treatment. Analysis of the biomarker samples is continuing as is the anaylsis of gait analysis by Ph.D. student Michael Vakula in the laboratory of Dr. Eadric Bressel. The stifle joint samples have been sent to collaborators (Dr. Kurt Hankenson, University of Michigan) for more in-depth orthopedic analysis at the University of Michigan's Orthopaedic Research Laboratories.
Impacts
What was accomplished under these goals?
The positive results of the influence of oblique treadmill running on the exacerbation of clinical osteoarthritis progression has prompted the acceleration of gene therapy testing in this model. We included a non-mechanical method to increase progression of osteoarthritis in ewes, the ovariectomy model, which is proven to produce clinical OA on a similar timeline as destabilization without mechanical disruption, with the additional advantage of providing a menopausal model of OA. Young ewes of uniform breeding, age and body size underwent a cranial cruciate ligament desmotomy, n=8 (as previously) or an ovariectomy procedure n=7. Surgeries took place the week of 12/18-12/22/17. Sheep were allowed to rest and adapt to the surgical manipulations for approximately 100 days prior to treadmill running. Seventy days postoperatively, three sheep in each group were intra-articularly injected into the left stifle joint with 6E+12 genome copies of a rAAV-coTimp-3 virus. The remaining sheep received a vehicle-only injection. One-hundred days postoperatively, sheep were run on a treadmill at an oblique angle (as previously) for 16 weeks, every other day for 32 min at 80 m/sec. Blood and urine were collected biweekly, digital radiographs and subjective lameness exams monthly and gait analysis at the beginning, middle and end of the 16 weeks. After 16 weeks of treadmill exercise, animals were collected and analysis includes circulating biomarkers, digital radiography, micro-computed tomography, soft tissue and gait analysis, virus biodistribution and transgene protein production and testing for the development of neutralizing antibodies to the injected virus. Treadmill running and barn housing of the surgically-modified sheep has been completed, samples have been collected and analysis is currently underway. This work included initial gene therapy testing by Master's student Crystal Collier and veterinary student Peter Welsh. This phase of the project provided the opportunity for veterinary students Peter Welsh (first year), Kolby Talbot and Hannah Rodriguez (both fourth year students at WSU) to learn clinical veterinary skills in radiography that veterinary students do not learn while here at USU or after transfer to WSU. Preliminary results from radiograph analysis conducted throughout the duration of the project have demonstrated a significant reduction of osteophytosis (a key marker of osteoarthritis progression) due to the rAAV therapeutic gene therapy treatment. These samples have been sent to our collaborator (Dr. Kurt Hankenson, Professor of Orthopaedic Surgery, Orthopaedic Research Laboratories) at the University of Michigan for in depth analysis of osteoarthritis progression. I was also contacted and had a conference call with two representatives from Medtronic (a large surgical biotech company) regarding our sheep model and a potential future collaboration. This prompted development of an additional method for gait analysis in collaboration with Dr. Eadric Bressel and Ph.D. student Michael Vakula.
Publications
|
Progress 10/01/16 to 09/30/17
Outputs
Target Audience:Undergraduate, graduate and veterinary students, orthopedic specialists, general public, biotechnology companies Changes/Problems:We are proceeding with changes as outlined last year by including an ovariectomy model and by injection of virus into the successful CCLD model. Significant delays in this project occurred due to major changes at the PPRL facility, including regulatory changes imposed the USDA in response the "Clay Station incident' and to the retirement of the director of the facility. The presumptive graduate student for this project decided to take an additional year to finish her undergraduate work and an administrative oversight delayed the progress on the production of the virus. All of these issues have been resolved and the animals are scheduled to be collected at the end of July 2018. What opportunities for training and professional development has the project provided?Graduate students Crystal Collier and Tracy Habermehl, veterinary students Peter Welsh and Adam Ustach and undergraduate students Kyleigh Taylor and Anisa Samhouri have been involved in planning for and will participate in the upcoming week of surgeries and subsequent management of the experimental animals. This project will be the basis for an M.S. project for graduate student Crystal Collier. Crystal has had previous exposure to this project as an undergraduate student and will now take on a significant portion of the management of this project and will continue with this for the next two years. Undergraduate student Anisa Samhouri will work closely with Crystal during the next year of this project. How have the results been disseminated to communities of interest?Dissemination of this work includes Rachel Hill's invited poster entitled 'Development of a Translational Large Animal Osteoarthritis Model' at the 25th Annual Merial-NIH Veterinary Scholars Symposium, "One Health", Cornell University, Ithaca, NY, a poster presentation entitled 'Efficacy of micro-computed tomography for analysis of osteoarthritis progression in sheep' at the Student Research Symposium, Office of Research and Graduate Studies. Utah State University, Logan, UT, an interview regarding Rachel's work posted on the CAAS website and a peer-reviewed publication of the work to date (Rachel J. Hill, Holly M. Mason, Gavin Yeip, Samer S. Merchant, Aaron L. Olsen, Rusty D. Stott, Arnaud J. Van Wettere, Eadric Bressel, and Jeffrey B. Mason. (2017). The Influence of Oblique Angle Forced Exercise in Surgically Destabilized Stifle Joints Is Synergistic with Bone, but Antagonistic with Cartilage in an Ovine Model of Osteoarthritis. Arthritis, vol. 2017, Article ID 7481619, doi:10.1155/2017/7481619). We were recently contacted by and had a conference call with Medtronic (Dwight Nelson and Dan Schuster) regarding our experimental model. What do you plan to do during the next reporting period to accomplish the goals?The positive results of the influence of oblique treadmill running on the exacerbation of clinical osteoarthritis progression has prompted the acceleration of gene therapy testing in this model. We will also include a non-mechanical method to increase progression of osteoarthritis in ewes, the ovariectomy model, which is proven to produce clinical OA on a similar timeline as destabilization without mechanical disruption, with the additional advantage of providing a menopausal model of OA. Young ewes of uniform breeding, age and body size will undergo a cranial cruciate ligament desmotomy, n=6 (as previously) or an ovariectomy procedure n=6 (surgeries scheduled for the week of 12/18-12/22). Sheep will be allowed to rest and adapt to the surgical manipulations for approximately 100 days prior to treadmill running. Seventy days postoperatively, three sheep in each group will be intra-articularly injected into the left stifle joint with 6E+12 genome copies of a rAAV-coTimp-3 virus. The remaining sheep will receive a vehicle-only injection. One-hundred days postoperatively, sheep will be run on a treadmill at an oblique angle (as previously) for 16 weeks, every other day for 32 min at 80 m/sec. Blood and urine will be collected biweekly, digital radiographs and subjective lameness exams monthly and gait analysis at the beginning and end of the 16 weeks. After 16 weeks of treadmill exercise, animals will be collected and analysis will include circulating biomarkers, digital radiography, micro-computed tomography, soft tissue and gait analysis, virus biodistribution and transgene protein production and testing for the development of neutralizing antibodies to the injected virus.
Impacts
What was accomplished under these goals?
The modeling of a surgically destabilized sheep model of osteoarthritis has been Published (Rachel J. Hill, Holly M. Mason, Gavin Yeip, Samer S. Merchant, Aaron L. Olsen, Rusty D. Stott, Arnaud J. Van Wettere, Eadric Bressel, and Jeffrey B. Mason. (2017). The Influence of Oblique Angle Forced Exercise in Surgically Destabilized Stifle Joints Is Synergistic with Bone, but Antagonistic with Cartilage in an Ovine Model of Osteoarthritis. Arthritis, vol. 2017, Article ID 7481619, doi:10.1155/2017/7481619). This publication includes a USU veterinary student and six USU faculty. Objective 1. Produce codon optimized Timp-3 plasmid and rAAV for transduction of sheep stifle joints. The codon optimized Timp-3 gene was transferred from the plasmid used for cell/tissue transfections to the plasmid used for production of the rAAV virus. The virus is currently in production using serotype 2/9 and a CB7 (chicken beta actin) promoter with a CMV enhancer region. The combination of this serotype and promoter/enhancer has proven efficacious in previous in vivo stifle transduction experiments. Sheep are unique in that they possess neutralizing antibodies to several rAAV capsids that most other large animals do not produce. Endogenous neutralizing antibodies to serotype 2/9 has not been detected in sheep to date. Objective 2. Acquire a young, skeletally mature and uniform cohort of sheep for the next group of experiments, perform surgeries and coordinate treadmill running and data collection tasks. Young ewes are the most desirable animals for production. Acquiring a cohort of young ewes took longer than expected. Surgeries are scheduled for the week of 12/18-12/22. Treadmill running and barn housing of the surgically-modified sheep has been arranged.
Publications
|
Progress 10/01/15 to 09/30/16
Outputs
Target Audience:Target Audience Undergraduate, graduate and veterinary students, orthopedic specialists, general public, biotechnology companies Changes/Problems:Changes/Problems In addition to considering using ovariectomized sheep with and without supplementation with specific estrogen receptor modulators (SERMs) to reduce variations in cyclic activity and input, we may proceed with gene therapy in surgically destabilized and/or Botox sheep. Gene therapy may be accomplished through direct rAAV virus injection or indirectly through injection of transgene-transduced mesenchymal stromal cells (MSCs) into the stifle joint. What opportunities for training and professional development has the project provided?Opportunities Veterinary student Rachel Hill and undergraduate students McKenna Walters and Crystal Collier were all involved in the processing and analysis of samples and the coordination between sample types. Rachel Hill additionally developed advanced skills in collecting and analysis of digital radiography and micro-computed tomography images and data. These skills are directly applicable in the veterinary medicine profession. Rachel and both undergraduate students additionally developed skills in sample processing, assay development and data analysis. Rachel also took a lead role in organizing the data for manuscript preparation and submission. This project also involved training on the application of micro-CT in a large animal model of disease for our USU veterinary and undergraduate students and graduate student Samer Merchant and undergraduate student Gavin Yeip at the University of Utah. How have the results been disseminated to communities of interest?Dissemination Dissemination of this work includes Rachel Hill's invited poster entitled 'Development of a Translational Large Animal Osteoarthritis Model' at the 25th Annual Merial-NIH Veterinary Scholars Symposium, "One Health", Cornell University, Ithaca, NY, a poster presentation entitled 'Efficacy of micro-computed tomography for analysis of osteoarthritis progression in sheep' at the Student Research Symposium, Office of Research and Graduate Studies. Utah State University, Logan, UT, and an interview regarding Rachel's work posted on the CAAS website. This work has recently been submitted for publication: Rachel J. Hill, Holly M. Mason, Gavin Yeip, Samer S. Merchant, Aaron L. Olsen, Rusty D. Stott, Arnaud J. Van Wettere, Eadric Bressel, and Jeffrey B. Mason. (2016). The influence of forced exercise in surgically destabilized stifle joints is synergistic with bone, but antagonistic with cartilage in an Ovine model of osteoarthritis. What do you plan to do during the next reporting period to accomplish the goals?Plan of Work The positive results of the influence of oblique treadmill running on the exacerbation of clinical osteoarthritis progression will provide an optimal platform for the testing and validation of our Botox model. We initially proposed that sheep would receive botulinum toxin A (Botox) injection in the gastrocnemius, tibialis anterior or the quadriceps femoris muscles. However, after observing the changes induced by cranial cruciate ligament desmotomy (CCLD) at the time of joint collection, we will now perform initial Botox injections into the gluteobiceps muscle. Paralysis of the gluteobiceps muscle will decrease tension in the lateral femoropatellar ligament, but will allow the sheep to maintain reasonable control of foot placement on the treadmill and combined with oblique-angle treadmill forced exercise, should increase progression of osteoarthritis similar to that observed with the CCLD surgery. In addition, because the gluteobiceps muscle lies on the lateral aspect of the thigh, this approach should simplify the injection process.
Impacts
What was accomplished under these goals?
Accomplishments The modeling of a surgically destabilized sheep model of osteoarthritis has been completed and submitted for publication. Objective 1. Stifle joints, serum and urine samples from control and all three treatment groups will be analyzed. Stifle joints were collected, including collection of synovial fluid samples when possible. Joints were fixed in NBF. To retain the integrity of the joint for micro-computed tomography scanning and analysis, we injected fixative inside the joint capsule to ensure complete fixation of the intact joint capsule. The joints were transferred to the University of Utah, Small Animal Imaging facility where, in collaboration with Dr. Ed Hsu, the Core Director, graduate student Samer Merchant and veterinary student Rachel Hill, methods to image sheep stifle joints were developed and all control and experimental stifle joints were imaged and analyzed. Upon completion of micro-CT analysis, the joints were returned to USU. The joints were subsequently disarticulated and macroscopically scored at the Utah Veterinary Diagnostic Laboratory in collaboration with Dr. Arnaud Van Wettere and Dr. Holly Mason and with the assistance of Undergraduate Student researchers McKenna Walters and Crystal Collier. We next developed a novel method to analyze the weekly sheep urine samples for glycosaminoglycan content and analyzed the weekly serum samples for aggrecan fragments. Combined, these measures allowed determination of both glycosaminoglycan synthesis and aggrecanase activity, both commonly accepted biomarkers of joint integrity. Dr. Holly Mason and myself analyzed and graded the biweekly digital radiographs of the stifle joints in all animals and Dr. Eadric Bressel (in the Department of Kinesiology & Health Science at USU) formalized his preliminary gait analysis of the sheep on the treadmill. Objective 2. Coordinate the finding of all collaborators and prepare a manuscript for publication. Radiographic data was coordinated with the micro-CT bone analysis and with the soft-tissue macroscopic scoring, while the urine glycosaminoglycan content and serum aggrecan fragments were coordinated on a timeline with the radiographs, gait analysis and treadmill running and time of surgery. The analysis has resulted in a manuscript submitted for publication.
Publications
|
Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Target Audience Undergraduate, graduate and veterinary students, orthopedic specialists, general public, biotechnology companies Changes/Problems:Changes/Problems Two of the ewes never started cycling at the end of the project when they should have exited anestrous. We will consider using ovariectomized sheep with and without supplementation with specific estrogen receptor modulators (SERMs) to reduce variations in cyclic activity and input. What opportunities for training and professional development has the project provided?Opportunities We may include introduction of a therapeutic transgene in destabilized joints. The gene has been isolated and is currently in plasmid form to be incorporated into a rAAV upon IRB approval. How have the results been disseminated to communities of interest?Dissemination Current results will be submitted for publication in a peer-reviewed journal. What do you plan to do during the next reporting period to accomplish the goals?Plan of Work Determine protocols for botox use to achieve appropriate joint destabilization, similar to that achieved with surgical destabilization. Potentially include marker and therapeutic rAAV transgene stifle injections.
Impacts
What was accomplished under these goals?
Accomplishments Objective 1. Osteoarthritis (OA) will be induced in control sheep using a common surgical approach. We performed multiple minimally-invasive cranial cruciate ligament desmotomy unilaterally in the sheep stifles to provide traditionally destabilized sheep for treadmill and control groups. Objective 2. Sheep with surgically destabilized stifle joints will be subjected to forced exercise on a treadmill. Sheep were on a treadmill, including switching between straight and oblique-angle running and were filmed while running to permit gait analysis. Preliminary gait analysis has demonstrated significant differences between the peak knee angle (maximal knee abduction or varus angle) for exercised-only sheep (86) compared with surgery + exercise sheep (77) over time. Peak knee range of motion appears to decrease over time (visits) for sheep 86 (about a 10% decline between visit 1 and 5), whereas peak range of motion for sheep 77 appears to be fairly constant until the final visit, where it decreased drastically. Serum samples are currently being analyzed for osteoarthritis biomarkers and micro-computed tomography is currently being conducted on all stifle joints in collaboration with the University of Utah. Digital radiographs are being scored collaboratively with the University of Pennsylvania.
Publications
|
Progress 03/26/14 to 09/30/14
Outputs
Target Audience:
Nothing Reported
Changes/Problems: Changes/Problems Objective 3. OA progression will be monitored with treadmill video, lameness examination, stifle joint ultrasound examination and changes in voluntary ambulatory/mobility parameters. Drs Holly Mason, Rusty Stott and myself and veterinary student Rachel Hill determined after using the current USU-available ultrasound units and additionally hosting testing demos for three other units, we have decided that digital radiographs will be more effective in recovering osteoarthritic changes in the sheep stifle joint. We have refined the methods required to collect digital radiographs. We have also, in collaboration with U of U, performed preliminary ex vivo micro-computed tomographic imaging of a collected sheep stifle joint. What opportunities for training and professional development has the project provided? Opportunities Veterinary student Rachel Hill and undergraduate student Porter Nielsen were both involved in the initial stages of project development. Rachel Hill additionally presented her work at the Merial Veterinary Scholars Symposium at Cornell University. Undergraduate student, Porter Nielsen is continuing to work in my lab. We have applied for funding to continue this project through the USTAR Veterinary Diagnostics and Infectious Diseases program (10/16/14). I have also spoken with Program Officer Dr. Bernadette Tyree at the NIAMS regarding this project and received positive feedback regarding submission of this work for NIH funding (10/24/14). We have also been contacted by Merial Limited regarding this arthritis model, which has, to date resulted in a confidentiality agreement and conference call with Merial and Genzyme. We have also met with the Utah State University Commercial Enterprises Office and have been urged by them to produce preliminary results from this model ASAP. As demonstrated by the level of interest from Merial, Genzyme and our Commercial Enterprises Office, this project is likely to produce IP with a high commercialization potential. How have the results been disseminated to communities of interest? Dissemination To date, dissemination has consisted of Rachel's poster at the Merial conference and a brief interview posted on the CAAS website. What do you plan to do during the next reporting period to accomplish the goals? Plan of Work Objective 1. Analyze traditionally destabilized sheep with and without treadmill exposure in vivo with digital radiographs and gait analysis. Gait analysis will be performed in collaboration with Dr. Eadric Bressel in the USU Biomechanics department. Radiographs will be performed by Dr. Holly Mason. Histological and pathological assessment will be conducted in collaboration with Dr. Arnaud Van Wettere at the UVDL. Objective 2. Botox-induced destabilization protocols will be developed in collaboration with Dr. Briedi Gillespie.
Impacts
What was accomplished under these goals?
Accomplishments Objective 1. Osteoarthritis (OA) will be induced in control sheep using a common surgical approach. Drs Aaron Olsen, Rusty Stott, Holly Mason and myself have developed the protocols for and optimized a minimally-invasive cranial cruciate ligament desmotomy unilaterally in the sheep stifle. We plan to perform multiple surgeries in January/February to provide traditionally destabilized sheep for treadmill and control groups. Objective 2. Sheep with surgically destabilized stifle joints will be subjected to forced exercise on a treadmill. Veterinary student Rachel Hill, undergraduate student Porter Nielsen and myself developed the equipment and protocols for running sheep on a treadmill, including switching between straight and oblique-angle running and for filming the sheep while running to permit gait analysis.
Publications
|
|