sepharose and Myocardial-Infarction

The purpose was to assess effects of intrapericardially deposited adipose-derived stem cells (ADSC) as a source of angiogenic factors on cryoinjury infarction. To enhance this effect and reduce incorporation of ADSC into tissue, the cells were immobilized in agarose gel patches transplanted onto cryoinjured epicardium. In domestic pigs (15-20 kg) the left ventricular (LV) anterior wall of exposed hearts was cryoinjured using aluminum rod (Φ=25 mm) cooled in liquid nitrogen. Sterilized circular patches made of agarose gel were placed in a nylon bag and sutured to cryoinjured epicardium. In 4 pigs, the patches contained 650,000 human ADSCs; in control animals patches were cell-free (n=2) or no patches were implanted (n=2). Cine and T(1)-weighted MRI was performed in vivo weekly (4 weeks) after injury using a 3 T imager. Following baseline imaging, a double bolus of gadopentate dimeglumine was injected (GdDTPA, 0.05 and 0.15 mmol/kg) and serial short axis images were acquired. In the 4-week ADSC-treated group, 2 pigs were assessed using GdDTPA and 2 pigs were assessed using MnCl(2) (70 micromol/kg/14 min). In all pigs, 5 × 10(6) NIR fluorescent microspheres (15 µm FMS, 645/680 nm) were injected into the hearts, which were excised, sliced and examined with fluorescence imaging for FMS content. Triphenyltetrazolium chloride (TTC) staining was used to determine necrotic areas. Four-week infarction areas were hypokinetic and appeared hyperintensive on Gd-enhanced MR images, hypointensive on Mn-enhanced images, and were TTC-negative. First-pass Gd enhancement kinetics was faster in the infarct area of ADSC-treated hearts: 152 ± 89 vs 54 ± 5.3% of normal in control (p = 0.03). Accordingly, FMS fluorescence was much higher in the treated infarcts (144 ± 59% of remote, n = 4) relative to control hearts (58 ± 13%, n = 4), which correlated with 3 times higher microvascular density in treated hearts. LV wall thickening was partially restored by ADSC treatment. ADSC-containing patches attached to cryoinjured epicardium greatly improved perfusion and microvascular density of scar tissue.

Topics: Adipose Tissue; Animals; Disease Progression; Freezing; Gadolinium DTPA; Gels; Heart Ventricles; Humans; Kinetics; Magnetic Resonance Imaging; Myocardial Infarction; Myocardium; Pericardium; Postmortem Changes; Sepharose; Stem Cell Transplantation; Stem Cells; Sus scrofa; Time Factors

Topics: Amidines; Cross-Linking Reagents; Humans; Immunoprecipitation; Myocardial Infarction; Sepharose; Troponin T

Lipoprotein(a) (Lp[a]) is associated with an increased cardiovascular risk. It is similar to low-density lipoprotein with an additional molecule of apo A covalently linked to apo B-100 by one disulfide bridge. Apo A is highly homologous to plasminogen. The kringle 4 motive of plasminogen is repeated between 10 and 40 times in apo (a). Currently, there is no drug therapy available to lower Lp(a). Since October 1993, we have carried out over 160 immunoadsorption treatments on 3 patients with elevated Lp(a) as their only risk factor and a history of myocardial infarction. Lp(a) was removed from plasma by sepharose coupled anti-Lp(a) columns. Lp(a) levels were lowered from above 170 mg/dl to below 30 mg/dl immediately after Lp(a) apheresis. To achieve this, the patient's plasma volume had to be treated 2 to 3 times. Nonspecific protein loss during column changes remained negligible. There were no serious unwanted effects during or after treatment. Minor circulatory problems (tachycardia, flush) occurred in 11% of the treatments but only with plasma flow rates above 55 ml/min. In 1 patient, coronary angiography after 2 years and in another patient after 1 year showed no progression. The third patient has not yet had repeat coronary angiography. Like the others, he reported subjective improvement after 1 year of apheresis. It is concluded that Lp(a) apheresis may retard progression of atherosclerosis in patients with selective Lp(a) elevation. Further studies to support this hypothesis are needed.

Topics: Adult; Blood Component Removal; Blood Volume; Coronary Angiography; Coronary Disease; Electrophoresis; Humans; Immunosorbents; Lipoprotein(a); Longitudinal Studies; Male; Middle Aged; Myocardial Infarction; Risk Factors; Sepharose; Treatment Outcome

Administration of growth factors is emerging as a new therapeutic approach for the enhancement of collateral vessel formation in the ischemic heart. We have investigated the effects of intramyocardial delivery of FGF-2 in the presence and absence of heparin on angiogenesis in a porcine model of myocardial infarction. Yorkshire pigs were subjected to myocardial infarction by the placement of an embolization coil in the left anterior descending artery (n = 5). Four to five weeks after creation of an infarct, FGF-2 (10 micrograms) alone or in complex with heparin, heparan sulfate, or heparin agarose beads was injected either into the normal myocardium or along the infarct border area. Histologic evaluation of each injection site was performed 4 to 5 weeks post-injection. The effect of FGF-2 on angiogenesis was evaluated by determining the number of capillaries (diameter < 20 microns (and arterioles (> 20 microns with tunica media) in each area observed. The number of capillaries were not affected by the treatment of FGF-2 both in normal myocardium and infarct border area. However, in the normal myocardium, the number of arterioles were increased with the treatment of FGF-2 alone (85 +/- 59%, P < 0.04), FGF-2 plus heparin (281 +/- 193%, P < 0.004) and FGF-2-coated heparin beads (241 +/- 141%, P < 0.01), as compared to control. Delivery of FGF-2 into the infarct border area, also increased the number of arterioles when FGF-2 was given with heparin (736 +/- 154%, P < 0.001) or heparin beads (700 +/- 109%, P < 0.001), as compared to control. FGF-2 administered with heparin was the most effective method of enhancing angiogenesis as compared to FGF-2 alone, FGF-2 plus heparan sulfate, or FGF-2 coated heparin agarose beads.

Topics: Affinity Labels; Animals; Capillaries; Collateral Circulation; Coronary Vessels; Dose-Response Relationship, Drug; Fibroblast Growth Factor 2; Follow-Up Studies; Heart; Hemodynamics; Heparitin Sulfate; Injections, Intramuscular; Male; Myocardial Infarction; Neovascularization, Pathologic; Recombinant Proteins; Sepharose; Swine

Myocardial infarction (MI) progresses from the acute death of myocytes and the infiltration of inflammatory cells into granulation, followed by scars. During the healing process, the myocardial interstitial cell population in the infarcted tissues increases markedly and then decreases. We postulated that apoptosis is responsible for this process. Twenty-four male Japanese white rabbits underwent a 30-minute occlusion of the left coronary artery followed by reperfusion for 2 days, 2 weeks, or 4 weeks (n=8 each). The histological features consisted of dead cardiomyocytes and marked leukocyte infiltration at 2 days after MI and granulation consisting of numerous alpha-smooth muscle actin-positive myofibroblasts, macrophage antigen-positive macrophages, and neovascularization at 2 weeks. At 4 weeks, the cellularity decreased markedly, and scars were evident. Interstitial cells with positive nick end labeling were significantly more frequent at the light microscopic level in the 2-day MI samples (5.3+/-3.6% in the center and 6.9+/-3.3% in the periphery of the infarct region) than in the 2-week (2.5+/-1.0%) and 4-week (0.5+/-0.5%) samples. DNA electrophoresis showed a clear ladder in tissues from the ischemic areas at 2 days after MI but not at 2 and 4 weeks after MI. Ultrastructurally, typical apoptotic figures, including apoptotic bodies and condensed nuclei without ruptured plasma membranes, were detected in leukocytes from all hearts with 2-day MI and in myofibroblasts, endothelial cells, and macrophages from all hearts with 2-week MI. In the electron microscopic in situ nick end labeling, immunogold particles intensely labeled the condensed chromatin of the typical apoptotic nuclei. These particles were also accumulated on nuclei of the interstitial cells showing homogeneous density but not definite condensation as typical apoptotic nuclei, suggesting an early stage of apoptosis. Thus, apoptosis plays an important role in the disappearance of both the infiltrated leukocytes and the proliferated interstitial cells after MI. This finding may have therapeutic implications for postinfarct ventricular remodeling through apoptosis handling during the healing stage of MI.

Topics: Animals; Apoptosis; Biotin; Deoxyuracil Nucleotides; DNA; DNA Fragmentation; Electrophoresis, Agar Gel; Endothelium, Vascular; Leukocytes; Male; Microscopy, Electron; Muscle Fibers, Skeletal; Myocardial Infarction; Rabbits; Sepharose; Staining and Labeling; Wound Healing

The effect of chronic, periadventitial administration of basic (b) fibroblast growth factor (FGF) on endothelial dysfunction in the collateral-dependent and normally perfused coronary microcirculation was examined. Ameroid constrictors were placed on the proximal left circumflex coronary artery (LCX) in 23 pigs. In 11 pigs, bFGF was released from calcium alginate microcapsules into the perivascular space of the proximal left anterior descending coronary artery (LAD) and LCX. After 5-8 wk, coronary arterial microvessels (80-170 microns) were studied in a pressurized (40 mmHg) no-flow state with video microscopy. Receptor-mediated endothelium-dependent relaxations to ADP and serotonin were reduced while contraction to acetylcholine was enhanced in the collateral-dependent LCX microvessels of non-bFGF-treated control hearts. Relaxation of vessels to the non-receptor-mediated, endothelium-dependent agent A-23187; endothelium-independent relaxation to nitroprusside; and contraction to KCl were similar in all groups. Chronic treatment with bFGF normalized responses to ADP, serotonin, and acetylcholine in the collateral-dependent LCX region but had no effect on the responses of vessels in the normally perfused LAD region. Arteriolar density in the collateral-perfused LCX region of bFGF-treated hearts was markedly increased (4-fold compared with that in untreated hearts, suggesting a link between the angiogenic effect of bFGF and its action on endothelial preservation. Thus the periadventitial, sustained delivery of bFGF preserves receptor-mediated, endothelium-dependent responses in the collateral-dependent LCX region but has no effect on responses of microvessels in the normally perfused LAD region or on non-receptor-mediated endothelium-dependent relaxation.

Topics: 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; Acetylcholine; Adenosine Diphosphate; Alginates; Animals; Arginine; Calcimycin; Coronary Circulation; Coronary Vessels; Drug Carriers; Endothelium, Vascular; Female; Fibroblast Growth Factor 2; Glucuronic Acid; Heart; Hexuronic Acids; Indomethacin; Ketanserin; Male; Microcirculation; Muscle Relaxation; Myocardial Infarction; Myocardium; Nitroarginine; Nitroprusside; Potassium Chloride; Prostaglandin Endoperoxides, Synthetic; Sepharose; Serotonin; Swine; Thromboxane A2; Vasodilation

The subforms of MM isozyme of creatine kinase (ATP:creatine N-phosphotransferase, EC 2.7.3.2, CK) in sera obtained from healthy adults and patients were determined by agarose gel isoelectric focusing (IEF). The patients were classified into six groups according to serum CK-MM activities and IEF patterns. The IEF spectra offered useful information on cell hyperplasia, augmented cell membrane permeability, cell destruction and release time of CK-MM in the circulation from the cells for diagnosis, progress observation and prognosis, especially in the cases of chronic hepatic diseases, acute myocardial infarction and muscular dystrophy. Macro CKs were also determined by IEF. Macro CKs could be completely distinguished from each other, and CK isozymes consisting of macro CK type 1 could be presumed by isoelectric points.

Topics: Chromatography, Gel; Creatine Kinase; Health Status; Humans; Immunoenzyme Techniques; Isoelectric Focusing; Isoenzymes; Liver Diseases; Muscular Dystrophies; Myocardial Infarction; Neoplasms; Prognosis; Sepharose

Topics: Counterimmunoelectrophoresis; Dextrans; Humans; Immunoelectrophoresis; Myocardial Infarction; Myoglobin; Sepharose

Topics: Chromatography, Ion Exchange; Creatine Kinase; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Evaluation Studies as Topic; Humans; Isoenzymes; Myocardial Infarction; Sepharose