Hyperglycemia exerts disruptive effects on the secretion of TGF-β1 and its matrix ligands, decorin and biglycan, by mesenchymal sub-populations and macrophages during bone repair

Introduction Bone has a high capacity for repair, but for patients with uncontrolled type 2 diabetes mellitus (T2DM), the associated hyperglycemia can significantly delay osteogenic processes. These patients respond poorly to fracture repair and bone grafts, leading to lengthy care plans due to arising complications. Mesenchymal stromal cells (MSCs) and M2 macrophages are both major sources of transforming growth factor-β1 (TGF-β1), a recognized mediator for osteogenesis and whose bioavailability and activities are further regulated by matrix small leucine-rich proteoglycans (SLRPs), decorin and biglycan. The aim of this study was to investigate how in vivo and in vitro hyperglycemic (HGly) environments can influence levels of TGF-β1, decorin, and biglycan during bone repair, with additional consideration for how long-term glucose exposure and cell aging can also influence this process. Results Following bone healing within a T2DM in vivo model, histological and immuno-labeling analyses of bone tissue sections confirmed delayed healing, which was associated with significantly elevated TGF-β1 levels within the bone matrices of young diabetic rats, compared with normoglycemic (Norm) and aged counterparts. Studies continued to assess in vitro effects of normal (5.5 mM) and high (25 mM) glucose exposure on the osteogenic differentiation of compact bone derived mesenchymal stromal cells (CB-MSCs) at population doubling (PD)15, characterized to contain populations of lineage committed osteoblasts, and at PD150, where transit-amplifying cells predominate. Short-term glucose exposure increased TGF-β1 and decorin secretion by committed osteoblasts but had a lesser effect on transit-amplifying cells. In contrast, the long-term exposure of CB-MSCs to high glucose was associated with decreased TGF-β1 and increased decorin secretion. Similar assessments on macrophage populations indicated high glucose inhibited TGF-β1 secretion, preventing M2 formation. Discussion Collectively, these findings highlight how hyperglycemia associated with T2DM can perturb TGF-β1 and decorin secretion by MSCs and macrophages, thereby potentially influencing TGF-β1 bioavailability and signaling during bone repair. GRAPHICAL ABSTRACT Bone healing follows an initial inflammatory phase followed by a reparative phase during which TGF-β1 within the healing extracellular environment promotes MSC proliferation and early commitment of MSCs to the osteoblast lineage. Conversely, TGF-β1 signaling inhibits later stages of osteoblast differentiation during the deposition of a bone matrix. Increased and prolonged levels of TGF-β1 and decorin and biglycan within the diabetic bone healing environment have the potential to delay the development of mature osteoblasts. Likewise, the decreased levels of TGF-β1 associated with the aged diabetic healing environment hinders the early stages of osteoblast development and hence bone formation. While they are not the only source of TGF-β1, hyperglycemic environments impede the development of the M2 phenotype that are known to play important roles in bone formation during the reparative stage.