Regulation of new blood vessel growth into ischemic skeletal muscle

Diagrammatic illustration of the surgical procedure performed on the rabbits. A muscle flap based on the left deep inferior epigastric artery is sutured onto muscle of the right thigh after ligating the right common iliac artery. (Courtesy of Pevec WC, Hendricks D, Rosenthal MS, Shestak KC, Steed DL, Webster MW: Revascularization of an ischemic limb by use of a muscle pedicle flap: A rabbit model. J Vasc Surg 1991;13:385–90.)

Microvessel density at the muscle flap-hind limb interface.

Histology of vessel density in the muscle-flap region of control and bFGF-treated animals. A, The flap-limb interface of a control animal, 50× magnification (hematoxylin and eosin stain). Microvessels can be seen, but are scarce and arranged in clusters. B, The flap-limb interface of a bFGF-treated animal, 50× magnification (hematoxylin-eosin stain). The distribution pattern of microvessels is diffuse, and vessel density is increased.

Immunohistochemical staining for bFGF at the muscle-flap interface in control and bFGF-treated animals. A, Control tissue, 40× magnification, showing intense pericellular staining with minimal staining in the muscle-flap interface. Brown stain indicates positive reaction for bFGF. B, bFGF-treated tissue, 40× magnification, has both pericellular staining and diffuse extracellular staining throughout the flap-limb interface.

Actin and bFGF mRNA expression analyzed by PCR. Lane 1—DNA marker; control animal, lanes 2–5: lane 2—positive control, lane 3—DNAse treated, lane 4—RNAse treated, lane 5—negative control; bFGF-treated animal, lanes 6–9: lane 6—positive control, lane 7—DNAse treated, lane 8—RNAse treated, lane 9—negative control; lane 10—bovine brain. The primer pair used is shown above each panel. Arrow indicates position of correct PCR fragment.