OBJECTIVES: (1) assess the molecular weight dependence of hyaluronan's (HA) cartilage boundary lubricating ability, alone and in combination with proteoglycan 4 (PRG4), at physiological concentrations; (2) determine if HA and PRG4 interact in solution via electrophoretic mobility shift assay (EMSA). METHODS: The cartilage boundary lubricating ability of a broad range of MW HA (20 kDa, 132 kDa, 780 kDa, 1.5 MDa, and 5 MDa) at 3.33 mg/ml, both alone and in combination with PRG4 at 450 μg/ml, was assessed using a previously described cartilage-on-cartilage friction test. Static, μ(static, Neq), and kinetic, <μ(kinetic, Neq)>, were calculated. An EMSA was conducted with PRG4 and monodisperse 150 kDa and 1,000 kDa HA. RESULTS: Friction coefficients were reduced by HA, in a MW-dependent manner. Values of <μ(kinetic, Neq)> in 20 kDa HA, 0.098 (0.089, 0.108), were significantly greater compared to both 780 kDa, 0.080 (0.072, 0.088), and 5 MDa, 0.079 (0.070, 0.089). Linear regression showed a significant correlation between both μ(static, Neq) and <μ(kinetic, Neq)>, and log HA MW. Friction coefficients were also reduced by PRG4, and with subsequent addition of HA; however the synergistic effect was not dependent on HA MW. Values of <μ(kinetic, Neq)> in PRG4, 0.080 (0.047, 0.113), were significantly greater than values of PRG4+various MW HA (similar in value, averaging 0.040 (0.033, 0.047)). EMSA indicated that migration of 150 kDa and 1,000 kDa HA was retarded when combined with PRG4 at high PRG4:HA ratios. CONCLUSIONS: These results suggest alterations in HA MW could significantly affect synovial fluid's cartilage boundary lubricating ability, yet this diminishment in function could be circumvented by physiological levels of PRG4 forming a complex, potentially in solution, with HA.
OBJECTIVES: (1) assess the molecular weight dependence of hyaluronan's (HA) cartilage boundary lubricating ability, alone and in combination with proteoglycan 4 (PRG4), at physiological concentrations; (2) determine if HA and PRG4 interact in solution via electrophoretic mobility shift assay (EMSA). METHODS: The cartilage boundary lubricating ability of a broad range of MW HA (20 kDa, 132 kDa, 780 kDa, 1.5 MDa, and 5 MDa) at 3.33 mg/ml, both alone and in combination with PRG4 at 450 μg/ml, was assessed using a previously described cartilage-on-cartilage friction test. Static, μ(static, Neq), and kinetic, <μ(kinetic, Neq)>, were calculated. An EMSA was conducted with PRG4 and monodisperse 150 kDa and 1,000 kDa HA. RESULTS: Friction coefficients were reduced by HA, in a MW-dependent manner. Values of <μ(kinetic, Neq)> in 20 kDa HA, 0.098 (0.089, 0.108), were significantly greater compared to both 780 kDa, 0.080 (0.072, 0.088), and 5 MDa, 0.079 (0.070, 0.089). Linear regression showed a significant correlation between both μ(static, Neq) and <μ(kinetic, Neq)>, and log HA MW. Friction coefficients were also reduced by PRG4, and with subsequent addition of HA; however the synergistic effect was not dependent on HA MW. Values of <μ(kinetic, Neq)> in PRG4, 0.080 (0.047, 0.113), were significantly greater than values of PRG4+various MW HA (similar in value, averaging 0.040 (0.033, 0.047)). EMSA indicated that migration of 150 kDa and 1,000 kDa HA was retarded when combined with PRG4 at high PRG4:HA ratios. CONCLUSIONS: These results suggest alterations in HA MW could significantly affect synovial fluid's cartilage boundary lubricating ability, yet this diminishment in function could be circumvented by physiological levels of PRG4 forming a complex, potentially in solution, with HA.
Authors: Saleem Abubacker; Dragana Ponjevic; Hyun O Ham; Phillip B Messersmith; John R Matyas; Tannin A Schmidt Journal: Connect Tissue Res Date: 2015-12-02 Impact factor: 3.417
Authors: Kimberly A Waller; Kaitlyn E Chin; Gregory D Jay; Ling X Zhang; Erin Teeple; Scott McAllister; Gary J Badger; Tannin A Schmidt; Braden C Fleming Journal: Am J Sports Med Date: 2017-01-27 Impact factor: 6.202
Authors: Y Asakawa-Tanne; S Su; R Kunimatsu; N Hirose; T Mitsuyoshi; Y Okamoto; E Tanaka; K Tanne; K Tanimoto Journal: J Dent Res Date: 2014-12-10 Impact factor: 6.116
Authors: Bridgette T Peal; Rachel Gagliardi; Jin Su; Lisa A Fortier; Michelle L Delco; Alan J Nixon; Heidi L Reesink Journal: J Orthop Res Date: 2020-02-09 Impact factor: 3.494