Surgically induced osteoarthritis in the rat results in the development of both osteoarthritis-like joint pain and secondary hyperalgesia S.E. Bove,

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Surgically induced osteoarthritis in the rat results in the development of both osteoarthritis-like joint pain and secondary hyperalgesia S.E. Bove, M.S., K.D. Laemont, M.S., R.M. Brooker, B.S., M.N. Osborn, B.S., B.M. Sanchez, B.S., R.E. Guzman, D.V.M, Ph.D., K.E. Hook, M.S., P.L. Juneau, M.S., J.R. Connor, M.S., K.S. Kilgore, Ph.D. Osteoarthritis and Cartilage Volume 14, Issue 10, Pages 1041-1048 (October 2006) DOI: 10.1016/j.joca.2006.05.001 Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 1 Effect of MMT surgery on change in HPWD. Change in HPWD was assessed by use of an incapacitance tester 1 day before and 3, 7, 14 and 21 days post MMT, sham or MCL sham surgery. Statistically significant differences were determined by RMANOVA (*P<0.0001) vs sham-operated rats. N=8 rats per group. Representative of three separate experiments. Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 2 Effect of MMT surgery on mechanical (punctate) allodynia. A. Mechanical allodynia was assessed using von Frey monofilaments and expressed as PWT (in grams). PWT was determined 1 day before and 3, 7, 14 and 21 days post MMT or sham surgery. Statistically significant differences were determined by RMANOVA (*P<0.001) vs sham-operated rats. B. Percent of MMT rats exhibiting allodynia was calculated as follows: % incidence of rats exhibiting allodynia=((# of rats with PWT <16g)/(# of rats with PWT ≥16g))×100. N=16 rats per group. Combination of two separate experiments. Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 3 Effect of MMT surgery on thermal hyperalgesia. Thermal hyperalgesia was assessed by use of a Hargreaves apparatus and expressed as PWL (in seconds). PWL was determined 1 day before and 3, 7, 14 and 21 days post MMT or sham surgery. N=8 rats per group. Representative of two separate experiments. Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 4 Effect of MMT surgery on mechanical (blunt pressure) hyperalgesia. Mechanical hyperalgesia was evaluated by the Randall–Selitto test using a paw pressure analgesia instrument and expressed as the amount of force (g) necessary to elicit a stereotyped flinch response. Mechanical hyperalgesia was determined 1 day before and 3, 7, 14 and 21 days post MMT or sham surgery. N=8 rats per group. Representative of two separate experiments. Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 5 Histology of sham, MCL sham- and MMT-operated rats 21 days post surgery. Frontal sections of the medial aspect of rat knee joints were stained with Toluidine Blue. (A and B) Sham and MCL sham surgery. Knees have normal healthy cartilage, subchondral bone and synovium. (C) MMT surgery. Prominent chondrophyte/osteophyte present adjacent to severe cartilage erosion. Chondrocyte loss is complete to the deep zone and significant proteoglycan loss can be detected all the way to the tidemark. Extensive subchondral sclerosis and bone marrow fibrosis is present with loss of some bone marrow spaces. No synovial inflammation is present. Magnification ×5. Representative of two separate experiments (n=3–6 rats per group). Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 6 Effect of rofecoxib and gabapentin on change in HPWD. MMT surgery was performed on day 0. On day 21 the rats were assessed on an incapacitance tester for change in HPWD to establish pre-dose values. Rats were then administered 10mg/kg rofecoxib, 100mg/kg gabapentin, or vehicle (PO) and reassessed 2h later. Statistically significant differences were determined using one-way ANOVA followed by Dunnett's multiple comparison's procedure (*P<0.05 vs vehicle at the same time point). N=8 rats per group. Representative of three separate experiments. Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions

Fig. 7 Effect of rofecoxib and gabapentin on mechanical (punctate) allodynia. MMT or sham surgery was performed on day 0. On day 21, PWT was determined. The rats were then administered 10mg/kg rofecoxib, 100mg/kg gabapentin, or vehicle (PO) and reassessed 2h later. Statistically significant differences were determined using one-way ANOVA followed by Dunnett's multiple comparison's procedure (*P<0.05 vs vehicle). N=8 (vehicle and sham) or six (rofecoxib and gabapentin) rats per group. Representative of two separate experiments. Osteoarthritis and Cartilage 2006 14, 1041-1048DOI: (10.1016/j.joca.2006.05.001) Copyright © 2006 OsteoArthritis Research Society International Terms and Conditions