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Fig. 6 | Arthritis Research & Therapy

Fig. 6

From: Hyaluronic acid synthesis, degradation, and crosslinking in equine osteoarthritis: TNF-α-TSG-6-mediated HC-HA formation

Fig. 6

TSG6-mediated intermolecular crosslinking of HA. Size exclusion chromatography-multi-angle light scattering (SEC-MALS) was used to determine how the quaternary structure of monodisperse, 150 kDa HA was altered in the presence of TSG-6 and inter-alpha-inhibitor protein (IαIp), with one replicate (n = 1) for each treatment condition. A Control solution: HA and IαIp. Two significant peaks (HA and IαIp) were observed at their calculated MW of 141 kDa and 154 kDa, respectively. B, C HA, TSG-6, and IαIp. C was created by re-scaling the MW y-axis of B to show the high MW of HA aggregates. The light scattering (LS) shows 2 peaks with higher MW of ~ 220 kDa (equivalent to the MW of HC-HA complex, HC + HA = 75 kDa + 150 kDa) a weighted average of ~ 3288 kDa. The bold, dotted blue line indicates the calculated molecular weight across each peak, corresponding to MW (kDa) on the right y-axis. The dashed blue line represents the expected MW of 150 kDa for the monodisperse HA solution. D Viscosity increased in the TSG6-mediated HC-HA crosslinking reaction mixture (HA + IαI + TSG6) as compared to all other 3 controls. Data are displayed as scatter plots representing the mean ± SD (n = 5 technical replicates)

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