Detection of myocardial cell damage in isolated rat hearts with near-infrared spectroscopy.
One hallmark of cell death resulting from prolonged ischemia is cell membrane disruption. We apply optical spectroscopy to gauge membrane disruption in isolated rat hearts by monitoring (1) the washout of myoglobin (Mb) and (2) the accumulation of an exogenous contrast agent in permeabilized cells. The contrast agent, a neodymium (Nd) chelate, has several absorptions in the visible and near-IR, and when present in the perfusate, it cannot penetrate cellular membranes. When membrane integrity is disrupted, however, it is expected to accumulate within the intracellular space; moreover, cellular Mb is expected to wash out. To test this hypothesis, rat hearts (n=12) are perfused with Krebs-Henseleit buffer (KHB), followed by perfusion with KHB in which a 5 mM Nd-DTPA solution is present. Membrane damage is then induced by infusion of digitonin into the Nd-KHB perfusate to provide a digitonin concentration of 2.5, 5, or 10 microg/mL. After 30 min of infusion, Mb levels fall to 46+/-14% of baseline levels and Nd-DTPA rises to 161+/-19% of predigitonin levels. No apparent dependence of total membrane disruption on digitonin concentration over the concentration range studied is found, although higher concentrations do lead to more rapid membrane disruption.
Nighswander-Rempel
Stephen P
SP
Institute for Biodiagnostics, 435 Ellice Avenue, Winnipeg, Manitoba R3B 1Y6 Canada.
Shaw
R Anthony
RA
Kuzio
Bozena
B
Kupriyanov
Valery V
VV
D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D004072:Digitonin; D006321:Heart / Q000187:drug effects; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D066298:In Vitro Techniques; D008560:Membrane Fluidity / Q000187:drug effects; D008657:Metabolic Clearance Rate; D017682:Myocardial Stunning / Q000139:chemically induced / Q000175:diagnosis* / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism; D009211:Myoglobin / Q000032:analysis / Q000378:metabolism*; D009354:Neodymium; D051381:Rats; D017207:Rats, Sprague-Dawley; D013055:Spectrophotometry, Infrared / Q000379:methods*
D015415:Biomarkers; D009211:Myoglobin; D009354:Neodymium; D004072:Digitonin