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. 2003 Nov 14;554(Pt 3):721–730. doi: 10.1113/jphysiol.2003.057059

Figure 4. Endothelium-dependent relaxation of smooth muscle-embedded, endothelium-overlaid collagen gels.

Figure 4

A, ACh (10 μm) induced Ca2+ transients in BAECs (a). A similar degree of [Ca2+]i elevation was obtained with a higher concentration of ACh in BBECs (100 μm, b). Representative traces of 30 (BAEC) or 28 (BBEC) cells are shown. Ba, in vitro model vessel consisting of bovine aortic smooth muscle cells (BASMC) embedded in a collagen gel lattice with overlaid endothelium. BASMC were embedded in type I collagen gel, and BAECs or BBECs were overlaid after 24 h. A gel contraction assay was performed after a further 24 h (right). Noradrenaline (NAd, 1 μm) induced a rapid contraction of the gels. Subsequent application of 10 μm ACh induced relaxation of precontracted gels when BAEC was overlaid (b, ○, n = 20). Note that l-NAME (0.1 mm) inhibited ACh-induced gel relaxation but did not affect initial gel contraction (b, •, n = 6). BBEC-overlaid gels did not show relaxation in response to 100 μm ACh both in control (○, n = 22) and l-NAME-treated gels (c, •, n = 6).