ABSTRACT
Increased concentration of lipoprotein(a) is a risk factor of coronary heart disease. lipoprotein(a) consists of LDL-like and highly polymorphic apolipoprotein(a). Here we studied the effect of lipoprotein(a)-containing sera with different apolipoprotein(a) phenotypes on lipid accumulation by THP-1 monocyte-like cells. Cholesterol concentration in lysates of THP-1 cells was significantly higher after their incubation with lipoprotein(a)-containing serum samples with low-molecular-weight phenotype of apolipoprotein(a) in comparison with samples with a high-molecular-weight apolipoprotein(a) phenotype irrespective of initial cholesterol level as well as serum concentrations of apoB-100, oxidized LDL, and circulating immune complexes. The presence of the most atherogenic small dense LDL subfractions in examined sera in addition to a low-molecular-weight apolipoprotein(a) phenotype resulted in significant elevation of cholesterol accumulation by THP-1 cells. The data obtained explain greater atherogenicity of lipoprotein(a) with low-molecular-weight apolipoprotein(a) phenotype.
Subject(s)
Apolipoproteins A/metabolism , Cholesterol/metabolism , Apolipoproteins/metabolism , Culture Media, Serum-Free , Humans , Macrophages/metabolism , Monocytes/metabolism , THP-1 CellsABSTRACT
We determined conditions for effective transduction of multipotent mesenchymal stromal cells from human adipose tissue with adenoviral constructs carrying the gene of human bone morphogenetic protein BMP-2. The peak of transgene transcription and BMP-2 protein secretion in the transduced cultures was observed on day 6 after infection. The maximum transcription of BMP-2 gene and genes of osteogenic markers (bone sialoprotein, osteopontin, and osteocalcin) was observed in the medium containing sodium ß-glycerophosphate and ascorbic acid. Addition of D 3 vitamin did not enhance the expression of BMP-2 gene in transduced cells. The obtained cell cultures with high osteogenic potential can be used in bone tissue engineering.
Subject(s)
Adenoviridae/genetics , Bone Morphogenetic Protein 2/genetics , Mesenchymal Stem Cells/metabolism , Adipose Tissue, White/cytology , Antigens, CD/metabolism , Bone Morphogenetic Protein 2/biosynthesis , Bone Morphogenetic Protein 2/metabolism , Cell Differentiation , Cells, Cultured , Humans , Integrin-Binding Sialoprotein/metabolism , Mesenchymal Stem Cells/physiology , Osteocalcin/metabolism , Osteopontin/metabolism , Transcription, Genetic , Transduction, GeneticABSTRACT
We studied of osteogenic differentiation of multipotent mesenchymal stromal cells from human adipose tissue. Experiments showed that 1α,25-dihydroxycalciferol is a more effective inductor of osteogenesis than dexamethasone. Comparative analysis revealed activation of gene expression for the major osteogenic markers on day 7 of culturing in a medium containing 1α,25-dihydroxycalciferol. It was found that transcription of genes encoding type 1 collagen proteins, osteopontin, osteocalcin, and bone sialoprotein peaked on day 14 in culture, while the expression of alkaline phosphatase and bone morphogenetic protein-2 genes increased over 21 days. Intensive mineralization of the extracellular matrix was observed starting from day 14 in culture. On the basis of the analysis of these data, optimal terms for osteogenic induction (day 14) and an optimal inductor (1α,25-dihydroxycalciferol) were chosen and the protocol of effective osteogenic differentiation of multipotent mesenchymal stromal cells from human adipose tissue was developed for creation of tissue-engineered bone equivalents.
