Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid for the...
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![Page 1: Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid for the Treatment of Lameness in Horses Sara Tracy Advisors: Dr.](https://reader036.fdocuments.us/reader036/viewer/2022062516/56649d455503460f94a21dcf/html5/thumbnails/1.jpg)
Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid
for the Treatment of Lameness in Horses
Sara TracyAdvisors: Dr. Skip Rochefort PhD and *Dr. Jill Parker VMD DACVS
Department of Chemical Engineering and *College of Veterinary MedicineOregon State University
Corvallis, Oregon
![Page 2: Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid for the Treatment of Lameness in Horses Sara Tracy Advisors: Dr.](https://reader036.fdocuments.us/reader036/viewer/2022062516/56649d455503460f94a21dcf/html5/thumbnails/2.jpg)
Objective
• Establish baselines for the properties of healthy joints• Correlate concentration and molecular weight of HA to
viscoelastic properties• Study the difference between healthy and diseased
synovial fluid
![Page 3: Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid for the Treatment of Lameness in Horses Sara Tracy Advisors: Dr.](https://reader036.fdocuments.us/reader036/viewer/2022062516/56649d455503460f94a21dcf/html5/thumbnails/3.jpg)
Motivation
• The equine industry looses up to one billion each year due to lameness
• Up to 14% of the horse population has problems with lameness
• Hyaluronic acid is used to treat lameness and joint pain in both horses and humans
Motivation
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Hyaluronic Acid (HA)
Sodium Hyaluronate, HyaluronanSodium Hyaluronate, Hyaluronan• Made up of repeating glucuronic acid and N-Made up of repeating glucuronic acid and N-
acetylglucosamine subunitsacetylglucosamine subunits• High molecular weight: 0.2 to 10 million DaltonHigh molecular weight: 0.2 to 10 million Dalton• Major component of synovial fluidMajor component of synovial fluid• Exhibits viscoelastic propertiesExhibits viscoelastic properties
![Page 5: Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid for the Treatment of Lameness in Horses Sara Tracy Advisors: Dr.](https://reader036.fdocuments.us/reader036/viewer/2022062516/56649d455503460f94a21dcf/html5/thumbnails/5.jpg)
Synovial FluidMultipurpose fluid surrounding all Multipurpose fluid surrounding all articular jointsarticular joints
Has both viscous and elastic Has both viscous and elastic properties properties
(viscoelastic)(viscoelastic)
ViscousViscous properties - lubrication properties - lubrication
ElasticElastic properties - shock absorption properties - shock absorption
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Skeletal View
![Page 7: Molecular and Rheological Characterization of Hyaluronic Acid and Equine Synovial Fluid for the Treatment of Lameness in Horses Sara Tracy Advisors: Dr.](https://reader036.fdocuments.us/reader036/viewer/2022062516/56649d455503460f94a21dcf/html5/thumbnails/7.jpg)
Common Equine Joint Problems
Degenerative Joint Disease – DJD• Damage to the articular cartilage • Commonly affects heavily worked and aged horses
Osteochondritis Dissecans – OCD• Failure of the bone underlying the smooth articular
cartilage to form properly• Commonly affects young horses
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Rheological Characterization
Steady shear rate test
•Plate rotates at increasing speeds (shear rate) while rheometer measures shear stress
•Viscosity = Shear rate
Shear stress___________
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35-189 Steady Shear Test Results For All Joints
0.01
0.10
1.00
10.00
100.00
0.10 1.00 10.00 100.00 1000.00
Shear Rate (1/s)
Viscosity (P)
RH
LH
LMC
RMC
LS
35-126 RS OCD
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Rheological Characterization
Dynamic oscillatory shear test
• Plate oscillates at increasing frequencies and specified strain (amount of rotation) while rheometer measures shear stress
G’ represents elastic (storage) modulus
G’’ represents viscous (loss) modulus
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35-189 Dynamic Test Results For All Joints
0.00
0.01
0.10
1.00
10.00
100.00
0.01 0.10 1.00 10.00 100.00
Frequency (Hz)
Modulus (dyn/cm^2)
LS G'
LS G''
LMC G'
LMC G''
RMC G'
RMC G''
LH G'
LH G''
RH G'
RH G''
35-126 RS OCD G'
35-126 RS OCD G''
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Molecular Characterization
• Molecules separated by size using porous gel column
• Molecular mass determined using light scattering
• Concentration determined using refractive index detector
SEC - MALLS
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-0.1
0.0
0.1
0.2
0.3
0.4
20 30 40 50 60 70
AUX, 90° Detector
T ime (m in)
Alignment - 35-189 LH #3
90°
AUX2
Volume Delay : 0.133 mL
Light Scattering
RI Detector
HA Peak
Protein Peak
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35-189 Steady Shear Test Results For All Joints
0.01
0.10
1.00
10.00
100.00
0.10 1.00 10.00 100.00 1000.00
Shear Rate (1/s)
Viscosity (P)
RH
LH
LMC
RMC
LS
35-126 RS OCD
Right Stifle (OCD)Mw=3.6E+06C=0.09 mg/ml
Right HockMw=2.5E+06C=0.57 mg/ml
Right Middle CarpalMw=1.7E+06C=1.00 mg/ml
Left StifleMw=2.3E+06C=0.66 mg/ml
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Preliminary Results
• Right and left joints from the same horse have similar rheological curves
• Stifle > Carpal > Hock
• OCD joints appear to have lower HA concentrations that healthy joints
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Conclusion
• Both molecular weight and concentration play an important role in the viscoelastic properties of joint fluid
– c*Mw = hydrodynamic interactions parameter
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Future Work
• Investigate the repeatability issue of SEC-MALLS with synovial fluid
• Explore the effects of HA supplements for the treatment of lameness on an equine test group
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Acknowledgements
• Howard Hughes Medical Institute
• Dr. Kevin Ahern
• Dr. Skip Rochefort, OSU Chemical Engineering Dept
• Dr. Jill Parker, OSU College of Veterinary Medicine
• Matt Wesley
• Katy and Anne Volmert