Preparation of low density lipoprotein from large apheresis cartridges for induction of cell death in Saos2 osteoblasts.
Atherosclerosis is epidemiologically associated with postmenopausal osteoporosis presumably by common etiologic factors, reflecting a state of comorbidity in aging. Osteoblasts make a significant facet of this comorbidity state. The present study shows that LDL (native and oxidized) separated by conventional density ultracentrifugation induces osteoblast cell growth arrest in culture. Since the density unltracentrifugation is a tedious procedure we examined, in the present study, the option of LDL purification by ionic strength elution from LDL-apheresis cartridges. We tested the ability of LDL and oxidized LDL (oxLDL) from apheresis columns to induce apoptosis in human Saos2 osteoblasts. Isotonic NaCl effluent washed from LDL-apheresis columns (before starting elution of LDL) induced cell proliferation. In some of the effluent fractions that stimulated Saos2 osteoblasts, up to 15% of the stimulation levels could be significantly inhibited with antilipoprotein A antibodies. After the isotonic washing (150 mM NaCl), upon elution with high ionic strength, 0.2-0.3 M NaCl, some front-runner LDL eluate fractions also induced cell growth and others did not inhibit Saos2 cell growth. This indicates that these fractions might have been contaminated with apolipoprotein A or with other mitogenic compounds. In contrast, the late-to-elute (last 1/3) LDL portion, with a mean density of 1.042 g/mL, killed the cells as expected. This suggests that only the very last one third of LDL eluted by high ionic strength (0.3-0.5 M) is free of osteoblast-mitogenic compounds or lipoprotein-A containing particles. This approach to LDL purification might serve as a convenient and economic method for studying the composition of individual LDL particles and their interaction with cells in culture.
Klein
Benjamin Y
BY
Laboratory of Experimental Surgery, Hadassah University Hospital Ein-Kerem, Jerusalem, Israel. byklein@hadassah.org.il
Kerem
Zohar
Z
Rojansky
Nathan
N
D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects*; D002852:Chromatography, Ion Exchange / Q000379:methods; D006382:Hemadsorption; D006801:Humans; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D010006:Osteoblasts / Q000187:drug effects*
D008077:Lipoproteins, LDL