Non-enzymatic Glycation of Bone Collagen Modifies Osteoclastic Activity and Differentiation

Abstract

Type I collagen, the major organic component of bone matrix, undergoes a series of post-translational modifications that occur with aging, such as the non-enzymatic glycation. This spontaneous reaction leads to the formation of advanced glycation end products (AGEs), which accumulate in bone tissue and affect its structural and mechanical properties. We have investigated the role of matrix AGEs on bone resorption mediated by mature osteoclasts and the effects of exogenous AGEs on osteoclastogenesis. Using in vitro resorption assays performed on control-and AGE-modified bone and ivory slices, we showed that the resorption process was markedly inhibited when mature osteoclasts were seeded on slices containing matrix pentosidine, a well characterized AGE. More specifically, the total area resorbed per slice, and the area degraded per resorption lacuna created by osteoclasts, were significantly decreased in AGE-containing slices. This inhibition of bone resorption was confirmed by a marked reduction of the release of type I collagen fragments generated by the collagenolytic enzymes secreted by osteoclasts in the culture medium of AGE-modified mineralized matrices. This effect is likely to result from decreased solubility of collagen molecules in the presence of AGEs, as documented by the reduction of pepsin-mediated digestion of AGE-containing collagen. We found that AGE-modified BSA totally inhibited osteoclastogenesis in vitro, most likely by impairing the commitment of osteoclast progenitors into pre-osteoclastic cells. Although the mechanisms remain unknown, AGEs might interfere with osteoclastic differentiation and activity through their interaction with specific cell-surface receptors, because we showed that both osteoclast progenitors and mature osteoclasts expressed different AGEs receptors, including receptor for AGEs (RAGEs). These results suggest that AGEs decreased osteoclast-induced bone resorption, by altering not only the structural integrity of bone matrix proteins but also the osteoclastic differentiation process. We suggest that AGEs may play a role in the alterations of bone remodeling associated with aging and diabetes.Non-enzymatic glycation is a common post-translational modification of proteins induced by the spontaneous condensation of reducing sugars (e.g. glucose) and metabolic intermediates (e.g. triose phosphates, glyoxal, and methylglyoxal) with free amino groups in lysine or arginine residues. The early step of the so-called Maillard reaction is the formation of a Schiff base adduct to protein. This early glycation product undergoes a reversible rearrangement to form an Amadori product adduct to protein (i.e. an intermediate glycation product). Schiff bases and Amadori products then undergo a complex series of rearrangements, oxidations, and/or dehydrations along different chemical pathways to produce a class of irreversible adducts to proteins, the advanced glycation end products (AGEs) 3 (reviewed in Refs. 1 and 2). A direct consequence of these highly diverse reaction pathways leadin...

High d(+)glucose concentration inhibits RANKL-induced osteoclastogenesis

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The contribution of collagen crosslinks to bone strength

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