Evaluation of angiogenic activities of hyaluronan oligosaccharides of defined minimum size.
OBJECTIVE
Oligosaccharides of hyaluronan (o-HAs) were proved to have pro-angiogenic activities. The aim of this study was to obtain four hyaluronan oligosaccharides (o-HAs) of defined molecular weight including 4 saccharide residues (o-HA4), 6 saccharide residues (o-HA6), 8 saccharide residues (o-HA8), and 10 saccharide residues (o-HA10) under optimum conditions, and compare their angiogenic activities.


METHODS
The four o-HAs were prepared by digesting the native high molecular weight HA with hyaluronidase under optimum conditions. The effects of the four o-HAs on human umbilical vein endothelial cell (HUVEC) proliferation were measured using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenil tetrazolium bromide (MTT) assay. Angiogenic effects of the four o-HAs were evaluated using chicken chorioallantoic membrane (CAM) assay. The effects of the four o-HAs on the vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) messenger ribonucleic acid (mRNA) expressions were detected by semi-quantitative polymerase chain reaction (PCR).


RESULTS
O-HA6, o-HA8 and o-HA10 could effectively promote the proliferation of HUVEC, induce angiogenesis in the CAM assay and increase VEGF mRNA levels in HUVEC. No significant effects were found with o-HA4.


CONCLUSIONS
These results suggested that o-HA6, o-HA8, and o-HA10, but not o-HA4 were effective angiogenic factors.
Cui
Xiangzhen
X
Institute of Biochemical and Biotechnological Drugs, Shandong University, Jinan 250012, Shandong, China.
Xu
Huanli
H
Zhou
Shuai
S
Zhao
Ting
T
Liu
Aihua
A
Guo
Xueping
X
Tang
Wei
W
Wang
Fengshan
F
D043925:Angiogenesis Inducing Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D001681:Biological Assay; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D066298:In Vitro Techniques; D008970:Molecular Weight; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis
D043925:Angiogenesis Inducing Agents; D009844:Oligosaccharides; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; D006820:Hyaluronic Acid