Proliferation and beta-tubulin for human aortic endothelial cells within gas-plasma scaffolds.
OBJECTIVE
We determined if human aortic endothelial cells (HAEC) enhanced proliferative and angiogenic phenotypes within gas-plasma treated bioresorbable D,L-polylactic acid (D,L-PLA) three-dimensional scaffolds.


METHODS
6 x 10(3) HAEC (N=120) were incubated for 6, 12 or 18 days within either non-treated control or treated scaffolds. Before removing media, unstained wells were observed for apparent cell densities. Quantitative colorimetric WST-1 mitochondrial assays were determined for pooled conditioned media from both HAEC attached to wells and their respective HAEC-containing scaffolds. Fixed HAEC in scaffolds were examined using non-quantitative laser confocal microcopy with FITC-conjugated consensus, Types-I/II or Type-III beta-tubulin.


RESULTS
WST-1 indicated that significantly (p<0.05) less mitochondria were on cell culture plates than inside scaffolds but for different reasons. For example, a 12-18 days comparison between WST-1 and beta-tubulin indicated that wells decreased because of overgrowth apotosis; whereas, mitochondrial activity inside treated scaffolds decreased with increased tubulogenesis. Observed with consensus and Type-I/II beta-tubulin, HAEC-treated scaffolds exhibited increased cell-cell interconnections and angiogenic cords undergoing tubulogenesis to form vessels with central lumens as well as increased Type-III beta-tubulin, predominantly in cells of smaller surface areas. Moreover, beta-tubulin inside HAEC-treated scaffolds appeared in discrete cytoskeletal and podial regions; yet, beta-tubulin for HAEC-control scaffolds was located in more diffuse cytoplasmic regions especially at 18 days.


CONCLUSIONS
HAEC-treated scaffolds undergo increased migration, proliferation, beta-tubulin expression and quiescent cord formation. HAEC in scaffolds represent a potential model to study mechanisms for vascular cord progression into tubes. WST-1 does not represent accurate cell densities in three-dimensional scaffold matrices.
Bailey
Steven R
SR
UTHSCSA: Janey Briscoe Center for Cardiovascular Research, 78229, USA. baileys@uthscsa.edu
Polan
Jodie L
JL
Munoz
Oscar C
OC
Agrawal
Mauli C
MC
Goswami
Nilesh J
NJ
D020341:Absorbable Implants; D001011:Aorta / Q000166:cytology / Q000187:drug effects*; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects*; D005740:Gases; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D010641:Phenotype; D011091:Polyesters; D013194:Staining and Labeling; D013499:Surface Properties; D023822:Tissue Engineering; D014404:Tubulin / Q000378:metabolism*
D001672:Biocompatible Materials; D005740:Gases; D036341:Intercellular Signaling Peptides and Proteins; D011091:Polyesters; D014404:Tubulin; C033616:poly(lactide)