VEGF analysis induced by endothelialized gas-plasma treated D,L-PLA scaffolds.
OBJECTIVE
Vascular endothelial growth factor (VEGF) isoforms play different roles in the temporal sprouting of endothelial-lined vessels in a nude mouse peritoneal model as cells respond to nontreated control and gas-plasma-treated bioresorbable poly-D,L-lactide acid 3D scaffolds with human aortic endothelial cells (HAEC).


METHODS
Nude mice peritoneums were incubated with HAEC (CW = control; TW = gas-plasma treated) or polymer scaffolds (Cp = control; Tp = treated) for 12, 24 and 72 days. Cytoplasmic and nuclear protein fractions were isolated using NER, electrophoresized using NuPAGE-MES and analyzed by WesternBreeze Chemiluminescent.


RESULTS
Prominent VEGF bands included 28, 45 and 62 kDa; 52-kDa VEGF observed in cytoplasmic TW fractions contributed about 18.6% at 12 days, 20.0% at 24 days and 13.1% at 72 days of the total VEGF signal. Yet, it was only noted in CW at 72 days where it accounted for 6.9%. A unique 32-kDa band appeared in both Cp (24.6%) and Tp (18.3%). Significant differences between band densities occurred for cytoplasmic nuclear CW24- TW24 (P = .022), CW72-TW72 (P = .011) and, also, cytoplasmic Cp24-Tp24 (P = .038).


CONCLUSIONS
The temporal and spatial organization of the TW isoforms results in more angiogenesis.
Polan
Jodie L
JL
Janey Briscoe Center for Cardiovascular Research, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
Morse
Brian
B
Wetherold
Suzanne
S
Villanueva-Vedia
Rosa E
RE
Phelix
Clyde
C
Barera-Roderiquiz
Edwin
E
Waggoner
Douglas
D
Goswami
Nilesh
N
Munoz
Oscar
O
Agrawal
C Mauli
CM
Bailey
Steve R
SR
D000818:Animals; D001011:Aorta / Q000187:drug effects*; D001672:Biocompatible Materials / Q000494:pharmacology*; D003470:Culture Media / Q000494:pharmacology; D016228:Endothelial Growth Factors / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008819:Mice, Nude; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D034521:Nuclear Matrix-Associated Proteins / Q000494:pharmacology*; D010537:Peritoneum / Q000187:drug effects*; D011091:Polyesters / Q000494:pharmacology*; D020033:Protein Isoforms / Q000494:pharmacology*; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
D001672:Biocompatible Materials; D003470:Culture Media; D016228:Endothelial Growth Factors; D034521:Nuclear Matrix-Associated Proteins; D011091:Polyesters; D020033:Protein Isoforms; D042461:Vascular Endothelial Growth Factor A; C033616:poly(lactide)