Targeting Bone Alleviates Osteoarthritis in Osteopenic Mice and Modulates Cartilage Catabolism

Subchondral bone modifications occur early in the development of osteoarthritis (OA). The level of bone resorption might impact cartilage remodeling. We therefore assessed the in vivo and in vitro effects of targeting bone resorption in OA and cartilage metabolism.OA was induced by meniscectomy (MNX) in ovariectomized osteopenic mice (OP) treated with estradiol (E2), pamidronate (PAM), or phosphate buffered saline (PBS) for 6 weeks. We assessed the subchondral bone and cartilage structure and the expression of cartilage matrix proteases. To assess the involvement of bone soluble factors in cartilage metabolism, supernatant of human bone explants pre-treated with E2 or PAM were transferred to cartilage explants to assess proteoglycan release and aggrecan cleavage. OPG/RANKL mRNA expression was assessed in bone explants by real-time quantitative PCR. The role of osteoprotegerin (OPG) in the bone-cartilage crosstalk was tested using an OPG neutralizing antibody.Bone mineral density of OP mice and osteoclast number were restored by E2 and PAM (p<0.05). In OP mice, E2 and PAM decreased ADAMTS-4 and -5 expression, while only PAM markedly reduced OA compared to PBS (2.0±0.63 vs 5.2±0.95; p<0.05). OPG/RANKL mRNA was increased in human bone explants treated with both drugs (2.2-3.7-fold). Moreover, supernatants from bone explants cultured with E2 or PAM reduced aggrecan cleavage and cartilage proteoglycan release (73±8.0% and 80±22% of control, respectively, p<0.05). This effect was reversed with osteoprotegerin blockade.The inhibition of bone resorption by pamidronate in osteopenic mice alleviates the histological OA score with a reduction in the expression of aggrecanases. Bone soluble factors, such as osteoprotegerin, impact the cartilage response to catabolic factors. This study further highlights the importance of subchondral bone in the regulation of joint cartilage damage in OA

OPG exerts protective effects on cartilage catabolism.

<p><b>A.</b> OPG/RANKL mRNA ratio in IL1-β stimulated bone explants cultured with estradiol at 0.01 µM (E-8) and 1 µM (E-6), and with pamidronate (PAM). The graph represents the mRNA ratio to controls in 3 different experiments. <b>B.</b> Effect of OPG blockade by a neutralizing antibody (OPG-ab) on ARGS³⁷⁴ sequences expression in supernatants of cartilage explants cultured with conditioned bone media. Image of a representative Western blot, and the graph shows the mean quantifications ± SEM of 4 different experiments. Quantification was done on the 50 kDa bands. *: p<0.05.</p

Effect of bone resorption inhibitors on bone indices in osteopenic mice with joint instability.

§<p>: compared to controls, p<0.05.</p>*<p>: compared to PBS, p<0.05.</p

Histological assessment of cartilage lesions.

<p>A. OA scored according to the 2010 OARSI recommendations. The score represents the sum of the grades at the tibia and femur in sagittal sections. B. Magnifications of slides stained with safranin-O (40×) in control mice (a: Sham operated knees; b: MNX knees; n = 8) and osteopenic (OP) mice treated with phosphate buffered saline (PBS) (c; n = 10), estradiol (E₂) (d; n = 6), or pamidronate (PAM) (e; n = 5). *: p<0.05.</p