Angiogenic bFGF expression from gas-plasma treated scaffolds.
OBJECTIVE
In vivo experiments indicate that gas-plasma-treated D,L-polylactide polymers expressing basic fibroblast growth factor (bFGF) exhibit enhanced angiogenesis. bFGF is not a single entity, but it is instead a family of isoforms. Consequently, we sought to determine which bFGF isoforms and levels initiate angiogenesis in nude mice peritoneums.


METHODS
Cytoplasmic and nuclear bFGF were characterized for nude mice peritoneums incubated with nontreated scaffolds containing HAEC (CW), its respective polymer-only scaffolds (Cp) and gas-plasma treated scaffolds with HAEC (TW) and without cells (Tp). NuPAGE electrophoresis and WesternBreeze Chemiluminescent kits were used to analyze relative bFGF densities and molecular weights. VEGF was quantified using ImageJ.


RESULTS
bFGF bands were located at molecular weights of 24, 48, 58, 72 and 80 kDa, depending on whether they were from cytoplasms or nuclei. At 12, 24 and 72 days, 58-kDa bFGF bands were observed from nuclei of TW and Tp, 80-kDa bFGF bands were only observed in cytoplasmic fractions < or = 24 days. Total cytoplasmic and nuclear bFGF intensities increased from 12 to 24 days, then declined by 72 days.


CONCLUSIONS
(1) Gas-plasma treated scaffolds up-regulate bFGF isoforms. (2) bFGF was expressed in the nuclei; however, 80-kDa bFGF was seen only in cytoplasms.
Bailey
Steve R
SR
Department of Medicine, Janey Briscoe Center for Cardiovascular Research, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA. baileys@uthscsa.edu
Polan
Jodie L
JL
Morse
Brian
B
Wetherold
Suzanne
S
Villanueva-Vedia
Rosa E
RE
Waggoner
Douglas
D
Phelix
Clyde
C
Barera-Roderiquiz
Edwin
E
Goswami
Nilesh
N
Munoz
Oscar
O
Agrawal
C Mauli
CM
D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects*; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage / Q000235:genetics* / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects* / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000235:genetics*; D034521:Nuclear Matrix-Associated Proteins / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology; D010537:Peritoneum / Q000187:drug effects*; D011091:Polyesters / Q000494:pharmacology*; D020033:Protein Isoforms / Q000008:administration & dosage / Q000235:genetics* / Q000494:pharmacology*
D001672:Biocompatible Materials; D034521:Nuclear Matrix-Associated Proteins; D011091:Polyesters; D020033:Protein Isoforms; D016222:Fibroblast Growth Factor 2; C033616:poly(lactide)