Evidence that Contamination by Lipopolysaccharide Confounds in Vitro Studies of Adiponectin Activity in Bone

Adiponectin, a hormone produced and secreted from adipose tissue, circulates at levels that are inversely related to visceral fat mass and bone mineral density. Adiponectin receptors are expressed in bone cells, and several studies have shown that adiponectin affects bone phenotype and might play a role in the cross talk between fat and bone tissues. In the current study, we determined global changes in gene expression induced by adiponectin in mouse bone marrow cells, in order to identify the molecular mechanisms that mediate adiponectin's effect to inhibit osteoclast differentiation in these cultures. The gene signature that was produced by microarray analysis was very similar to a signature produced by activation of type I interferons (IFN), and we therefore tested the hypothesis that the adiponectin preparation, although marketed as lipopolysaccharide (LPS) free, was contaminated with LPS that induced an IFN response in the bone marrow cells. Heat inactivation of the adiponectin preparation and the use of small interfering RNA to knockdown the AdipoR1 receptor had not diminished the activity of the adiponectin preparation to induce the IFN target genes Ccl5 and Irf7. Thus, the changes in gene expression determined in the bone marrow cultures are likely to be the result of a combination of adiponectin and LPS effects. Our study suggests that the purity of commercially available proteins needs to be verified and that experimental results of adiponectin activity in vitro should be interpreted cautiously. (Endocrinology 153: 2076-2081, 2012)