This Article Full Text Full Text (PDF) Data Supplement All Versions of this Article: 26/9/1998    most recent 01.ATV.0000233359.74484.77v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Ishikawa, T. Articles by Asahara, T. Search for Related Content PubMed PubMed Citation Articles by Ishikawa, T. Articles by Asahara, T.

(Arteriosclerosis, Thrombosis, and Vascular Biology. 2006;26:1998.)
© 2006 American Heart Association, Inc.

Vascular Biology

Establishment of a Functionally Active Collagen-Binding Vascular Endothelial Growth Factor Fusion Protein In Situ

Tetsuya Ishikawa; Masamichi Eguchi; Mika Wada; Yo Iwami; Kayoko Tono; Hideki Iwaguro; Haruchika Masuda; Tetsuro Tamaki; Takayuki Asahara

From the Department of Regenerative Medicine (T.I., M.E., M.W., Y.I., K.T., H.I., H.M., T.T, T.A.), Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa, Japan and Stem Cell Translational Research, Institute of Biomedical Research; Innovation/RIKEN Center for Developmental Biology (T.A.), Chuo-ku, Kobe, Hyogo, Japan.

Correspondence to Takayuki Asahara or Tetsuya Ishikawa, Department of Regenerative Medicine, Tokai University School of Medicine, Bohseidai, Isehara, Kanagawa 259-1193 Japan. E-mail Asa777{at}aol.com or tecchan@is.icc.u-tokai.ac.jp

Objective— Tissue regeneration requires both growth factor and extracellular matrix such as collagen, serving as a scaffold for cell growth. We established FNCBD-VEGF121, consisting of the fibronectin collagen-binding domain (FNCBD) and vascular endothelial growth factor (VEGF) 121, and investigated its properties.

Methods and Results— FNCBD-VEGF121 specifically bound to gelatin and type I, II, III, IV, and V collagen. This collagen-bound FNCBD-VEGF121 captured soluble VEGF receptor 2 (VEGFR-2)/Fc chimeric protein. Cell growth-promoting activity of FNCBD-VEGF121 was almost identical to that of VEGF121. The VEGF fusion protein significantly enhanced the expression of VEGFR-2 (71.6±0.8%) on endothelial progenitor cells (EPCs) derived from umbilical cord blood. Expectably, the collagen-bound VEGF fusion protein not only promoted the growth of endothelial cells (ECs) but also induced the expression of VEGFR-2 (63.7±0.8%) on non-adherent cells expanded from bone marrow CD34⁺ cells. Moreover, the VEGF fusion protein enhanced sprout formation of ECs in a matrigel model. In vivo experiments revealed that FNCBD-VEGF121 had local effects but not systemic effect on EPC mobilization.

Conclusions— These results suggest that FNCBD-VEGF121 stably maintains an optimally high and local concentration of VEGF with collagen matrix and stimulates both ECs and EPCs in situ, supplying a vascular regeneration niche.

Tissue regeneration requires both growth factor and extracellular matrix. We established FNCBD-VEGF121, consisting of the fibronectin collagen-binding domain and vascular endothelial growth factor 121. FNCBD-VEGF121 stably maintains an optimally high and local concentration of VEGF with collagen matrix and stimulates cellular activity in situ, supplying a vascular niche regeneration.

Key Words: collagen • endothelium • growth substance • proteins