Abstract
Peripheral artery disease is an atherosclerotic disease that causes limb ischemia and has few effective treatments. Stem cell therapy is a promising treatment option, but concomitant diabetes may limit its effectiveness. PURPOSE: To evaluate the therapeutic potential of skeletal muscle pericytes to augment postischemic neovascularization following the induction of limb ischemia in wild type (WT) and type 2 diabetic (T2DM) mice. METHODS: Pericytes were isolated via fluorescence activated cell sorting for CD45-CD34-CD146+ cells, and pericyte phenotype was confirmed via surface marker expression, gene expression, and in vitro differentiation potential. WT C57BL/6 (n=10) and db/db T2DM (n=8) mice underwent unilateral femoral artery ligation to induce limb ischemia. 24 hrs post-ligation, pericytes or vehicle control were transplanted into the muscles of the ischemic hindlimbs. Postischemic neovascularization was assessed via foot blood flow at pre-surgery, post-surgery, and postoperative days 3, 7, 14, 21, and 28 using laser Doppler perfusion imaging. Differences in gene expression were determined using t-tests; differences in blood flow were determined using linear mixed models. RESULTS: CD45-CD34-CD146+ pericytes were positive for mesenchymal stem cell markers CD90 (74%) and CD105 (65%) and weakly positive for the pericyte marker PDGFRβ (42%) and the endothelial cell marker CD144 (36%). Pericytes transdifferentiated into skeletal myocytes, adipocytes, osteocytes, and endothelial cells. Pericytes had significantly (p<0.05) upregulated Sca1 (4.0-fold) gene expression, downregulated CD31 (0.2-fold) gene expression, and no difference in MyoD (1.0-fold), Pax3 (1.3-fold), or Pax7 (1.0-fold) expression. Blood flow recovery in WT mice was significantly higher after pericyte transplantation than after vehicle control (p=0.03; 81.1±6% vs. 64.7±9% at postoperative day 28). Blood flow recovery in T2DM mice after pericyte transplantation was not different than after vehicle control (p=0.44; 50.1±4% vs. 53.5±4% at postoperative day 28). CONCLUSION: T2DM negates the capacity of pericytes to augment neovascularization after limb ischemia. Further study is required to determine the mechanism by which T2DM impairs this pericyte function. Support: AAUW American Dissertation Fellowship
Published Version
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