Local Application of Isogenic Adipose-Derived Stem Cells Restores Bone Healing Capacity in a Type 2 Diabetes Model

Abstract

Bone regeneration is typically a reliable process without scar formation. The endocrine disease type 2 diabetes prolongs and impairs this healing process. In a previous work, we showed that angiogenesis and osteogenesis-essential steps of bone regeneration-are deteriorated, accompanied by reduced proliferation in type 2 diabetic bone regeneration. The aim of the study was to improve these mechanisms by local application of adipose-derived stem cells (ASCs) and facilitate bone regeneration in impaired diabetic bone regeneration. The availability of ASCs in great numbers and the relative ease of harvest offers unique advantages over other mesenchymal stem cell entities. A previously described unicortical tibial defect model was utilized in diabetic mice (Lepr db2/2 ). Isogenic mouse adiposederived stem cells (mASCs) db2/db2 were harvested, transfected with a green fluorescent protein vector, and isografted into tibial defects (150,000 living cells per defect). Alternatively, control groups were treated with Dulbecco's modified Eagle's medium or mASCs WT . In addition, wild-type mice were identically treated. By means of immunohistochemistry, proteins specific for angiogenesis, cell proliferation, cell differentiation, and bone formation were analyzed at early (3 days) and late (7 days) stages of bone regeneration. Additionally, histomorphometry was performed to examine bone formation rate and remodeling. Histomorphometry revealed significantly increased bone formation in mASC db2/db2 -treated diabetic mice as compared with the respective control groups. Furthermore, locally applied mASCs db2/db2 significantly enhanced neovascularization and osteogenic differentiation. Moreover, bone remodeling was upregulated in stem cell treatment groups. Local application of mACSs can restore impaired diabetic bone regeneration and may represent a therapeutic option for the future. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:836-844 SIGNIFICANCEThis study showed that stem cells obtained from fat pads of type 2 diabetic mice are capable of reconstituting impaired bone regeneration in type 2 diabetes. These multipotent stem cells promote both angiogenesis and osteogenesis in type 2 diabetic bony defects. These data might prove to have great clinical implications for bony defects in the ever-increasing type 2 diabetic patient population.

Inhibition of GDF8 (Myostatin) accelerates bone regeneration in diabetes mellitus type 2

“…mASC of Lepr db−/− Black 6 mice were differentiated in osteogenic medium as described before for 7 days to evaluate the pro-osteogenic effect of Follistatin application 35 . Differentiation medium combined with Follistatin or Myostatin neutralizing antibody (3 µg/ml) showed significantly enhanced osteogenic differentiation, while Activin neutralizing antibody (7.5 µg/ml) showed a less osteogenic differentiative effect.…”

“…Alizarin Red staining (28 days after osteogenic differentiation) was performed as previously described 28 , 35 .…”

Notch‑1 inhibition reduces proliferation and promotes osteogenic differentiation of bone marrow mesenchymal stem cells

“…Due to the unique properties of BMSCs, including easy isolation, in vitro amplification, differentiation and immune tolerance (3), this cell type has therapeutic potential in vascularization and tissue repair. Therefore, BMSCs are widely used in cell therapy clinical trials (4,5) and pre-clinical studies (6,7). In addition, the prospect of applications in alternative therapy is broad.…”

Coculture of endothelial progenitor cells and mesenchymal stem cells enhanced their proliferation and angiogenesis through PDGF and Notch signaling

“…Mesenchymal stem cells (MSCs) are multipotent stem cells capable of self‐renew, differentiating, and participating in angiogenesis. Both MSCs and EPCs take part in vascularization and tissue repair and have been widely employed for cell‐based therapy in both preclinical studies and clinical trials .…”