Premium
Improved angiogenic response in pig heart following ischaemic injury using human skeletal myoblast simultaneously expressing VEGF 165 and angiopoietin‐1
Author(s) -
Ye Lei,
Haider Husnain Kh.,
Jiang Shujia,
Tan Ru San,
Ge Ruowen,
Law Peter K.,
Sim Eugene K.W.
Publication year - 2007
Publication title -
european journal of heart failure
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.149
H-Index - 133
eISSN - 1879-0844
pISSN - 1388-9842
DOI - 10.1016/j.ejheart.2006.04.008
Subject(s) - immunostaining , myocyte , angiogenesis , ex vivo , medicine , in vivo , neovascularization , pathology , andrology , immunohistochemistry , biology , microbiology and biotechnology
Objective To achieve angiogenic interaction between VEGF 165 and angiopoietin‐1 (Ang‐1) using a novel adenoviral bicistronic vector (Ad‐Bic) encoding the two factors and delivered ex vivo using sex‐mismatched human skeletal myoblasts. Methods and results: A myocardial infarction model was developed in 29 female pigs; randomised into four groups: DMEM (group‐1, n =6); Adenovirus null (Ad‐null) vector‐myoblast (group‐2, n =5); Ad‐Ang‐1 myoblast (group 3, n =7) and Ad‐Bic‐myoblast (group‐4, n =11). Three weeks later, 5 ml DMEM without myoblasts or containing 3 × 10 8 myoblasts carrying lac‐z gene and transduced with Ad‐null, Ad‐Ang‐1 or Ad‐Bic were injected intra‐myocardially in and around the infarct. 2D‐echocardiography and fluorescent microsphere studies 6‐ and 12‐weeks post‐treatment revealed significantly improved cardiac performance and regional blood flow in groups 3 and 4. Histological studies and Y‐chromosome analysis revealed extensive survival of lac‐z positive myoblasts staining positive for human proteins in the pig heart. ELISA, immunostaining and RT‐PCR revealed that Ad‐Bic transduced myoblasts concomitantly but transiently expressed hVEGF 165 and Ang‐1 both in vitro and in vivo. Double fluorescent immunostaining of the tissue sections for vWFactor‐III and smooth muscle actin showed significantly higher vascular density of mature blood vessels per low power microscopic field in groups 3 and 4 at 6‐ and 12‐weeks. Conclusion: Our combined approach led to enhanced angiogenesis with a greater percentage of functionally mature blood vessels in a porcine heart.