Therapeutic angiogenesis in diabetic apolipoprotein E-deficient mice using bone marrow cells, functional hemangioblasts and metabolic intervention.
OBJECTIVE
Peripheral arterial disease (PAD) is a major health problem especially when associated to concomitant diabetes and hypercholesterolemia. Hyperglycemia with an overwhelming generation of oxygen radicals and formation of glycation end-products exacerbates oxidation-sensitive mechanisms activated by tissue ischemia. Administration of autologous bone marrow cells (BMC) is an increasing notable intervention to induce therapeutic angiogenesis, ameliorated by metabolic intervention (MT). Recently, hemangioblasts (HS) with functional properties were isolated.


METHODS
The effects of integrate regimen with intravenous BMC, HS, and MT (1.0% vitamin E, 0.05% vitamin C, and 6% l-arginine) were examined in the ischemic hindlimb of ApoE(-/-) diabetic and non-diabetic. Blood flow ratio was monitored by use of a laser Doppler blood flowmeter. Capillary density was determined in sections of the adductor and semimembranous muscles with antibody against CD31.


RESULTS
BMC or HS alone, and BMC plus HS increased blood flow and capillary densities and decreased interstitial fibrosis. These effects were amplified by additional MT, at least in part, through the nitric oxide pathway, reduction of systemic oxidative stress and macrophage infiltration. Investigation of molecular mechanisms in bone marrow (BM)-derived progenitor cells from mice revealed that BMC therapy and, more consistently, in combination with MT ameliorated functional activity via decreased cellular senescence and increased telomerase and chemokine CXCR4 activities. Telomerase activity was also increased by HS alone or HS+MT and, more consistently, by BMC+HS alone or in combination with MT.


CONCLUSIONS
Intravenous autologous BMC and HS intervention together with MT increased therapeutic angiogenesis in the ApoE(-/-) diabetic mouse hindlimb.
Balestrieri
Maria Luisa
ML
Department of Biochemistry and Biophysics, 1st School of Medicine, II University of Naples, 80138 Naples, Italy. marialuisa.balestrieri@unina2.it
Lu
Shi-Jiang
SJ
de Nigris
Filomena
F
Giovane
Alfonso
A
Williams-Ignarro
Sharon
S
D'Armiento
Francesco Paolo
FP
Feng
Qiang
Q
Fiorito
Carmela
C
Testa
Gianluca
G
Pastore
Lucio
L
Cacciatore
Francesco
F
Mancini
Francesco Paolo
FP
Servillo
Luigi
L
De Rosa
Gaetano
G
Pagliarulo
Caterina
C
Rienzo
Monica
M
Minucci
Pellegrino Biagio
PB
Farzati
Bartolomeo
B
Salvatore
Francesco
F
Rengo
Franco
F
Ignarro
Louis Joseph
LJ
Giordano
Antonio
A
Baker
Andrew
A
Lanza
Robert
R
Napoli
Claudio
C
D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency; D001120:Arginine / Q000627:therapeutic use; D001205:Ascorbic Acid / Q000627:therapeutic use; D016026:Bone Marrow Transplantation; D003921:Diabetes Mellitus, Experimental / Q000628:therapy*; D055018:Hemangioblasts / Q000637:transplantation*; D006614:Hindlimb / Q000098:blood supply*; D007511:Ischemia / Q000628:therapy; D051379:Mice; D018919:Neovascularization, Physiologic / Q000502:physiology*; D016491:Peripheral Vascular Diseases / Q000628:therapy*; D012039:Regional Blood Flow; D014810:Vitamin E / Q000627:therapeutic use
D001057:Apolipoproteins E; D014810:Vitamin E; D001120:Arginine; D001205:Ascorbic Acid