Therapeutic angiogenesis by implantation of a capillary structure constituted of human adipose tissue microvascular endothelial cells.
OBJECTIVE
We previously reported a novel technology for the engineering of a capillary network using an optical lithographic technique. To apply this technology to the therapy of ischemic diseases, we tested human omental microvascular endothelial cells (HOMECs) as an autologous cell source and decellularized human amniotic membranes (DC-AMs) as a pathogen-free and low immunogenic transplantation scaffold.


RESULTS
Human umbilical vein endothelial cells were aligned on a patterned glass substrate and formed a capillary structure when transferred onto an amniotic membrane (AM). In contrast, HOMECs were scattered and did not form a capillary structure on AMs. Treatment of HOMECs with sphingosine 1-phosphate (S1P) inhibited HOMEC migration and enabled HOMEC formation of a capillary structure on AMs. Using quantitative RT-PCR and Western blot analyses, we demonstrated that the main S1P receptor in HOMECs is S1P(2), which is lacking in human umbilical vein endothelial cells, and that inhibition of cell migration by S1P is mediated through an S1P(2)-Rho-Rho-associated kinase signaling pathway. Implantation of capillaries engineered on DC-AMs into a hindlimb ischemic nude mouse model significantly increased blood perfusion compared with controls.


CONCLUSIONS
A capillary network consisting of HOMECs on DC-AMs can be engineered ex vivo using printing technology and S1P treatment. This method for regeneration of a capillary network may have therapeutic potential for ischemic diseases.
Yoshida
Tomoko
T
Department of Nanomedicine, Tokyo Medical and Dental University, Tokyo, Japan.
Komaki
Motohiro
M
Hattori
Hideshi
H
Negishi
Jun
J
Kishida
Akio
A
Morita
Ikuo
I
Abe
Mayumi
M
D000273:Adipose Tissue / Q000098:blood supply*; D000650:Amnion / Q000637:transplantation*; D000818:Animals; D015153:Blotting, Western; D002465:Cell Movement; D002478:Cells, Cultured; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism / Q000637:transplantation*; D006614:Hindlimb; D006801:Humans; D007511:Ischemia / Q000503:physiopathology / Q000601:surgery*; D008246:Lysophospholipids / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D055806:Microvessels / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000637:transplantation*; D018482:Muscle, Skeletal / Q000098:blood supply*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009852:Omentum / Q000098:blood supply*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D049349:Receptors, Lysosphingolipid / Q000235:genetics / Q000378:metabolism; D012039:Regional Blood Flow; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism; D000081025:Sphingosine-1-Phosphate Receptors; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014182:Transplantation, Autologous; D020741:rho GTP-Binding Proteins / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
D008246:Lysophospholipids; D047428:Protein Kinase Inhibitors; D049349:Receptors, Lysosphingolipid; C477522:S1PR2 protein, human; D000081025:Sphingosine-1-Phosphate Receptors; C060506:sphingosine 1-phosphate; D054460:rho-Associated Kinases; D020741:rho GTP-Binding Proteins; D013110:Sphingosine