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. 2009 Dec;5(4):380-9.

In Vitro Effects of PDGF Isoforms (AA, BB, AB and CC) on Migration and Proliferation of SaOS-2 Osteoblasts and on Migration of Human Osteoblasts

Alessandra Colciago  1 , Fabio Celotti, Lavinia Casati, Rinaldo Giancola, Stefano M Castano, Guido Antonini, Maria Cristina Sacchi, Paola Negri-Cesi
Affiliations

In Vitro Effects of PDGF Isoforms (AA, BB, AB and CC) on Migration and Proliferation of SaOS-2 Osteoblasts and on Migration of Human Osteoblasts

Alessandra Colciago et al. Int J Biomed Sci. 2009 Dec.

Abstract

PDGF is a major constituent of platelet rich plasma (PRP), responsible of chemotactic and possibly of mitogenic effects of PRP on osteoblasts. PDGF family includes 5 isoforms: PDGF-AA, PDGF-AB, PDGF-BB, PDGF-CC and PDGF-DD, all expressed in platelets except PDGF-DD. Aim of this study was to analyze the effect of recombinant hPDGF-A, -AB, -B and -C, on migration and proliferation of a human osteoblastic cell line, SaOS-2. Preliminary observations on cell migration were also done in primary cultures of human osteoblasts. In vitro microchemotaxis and (3)H-thymidine mitogenic assays were used. While PDGF-AB is active at concentrations present in PRP, PDGF-AA and BB are chemotactic only at much higher doses. PDGF-C is totally inactive alone or together with the active isoforms. PDGF-AA, PDGF-BB and PDGF-C stimulate SaOS-2 proliferation only at the highest dose tested, while PDGF-AB is ineffective. Primary osteoblasts are less sensitive than SaOS-2 and progressively lose responsiveness with increasing passages in culture, in line with loss of cell differentiation. The different PDGF isoforms act differentially on osteoblasts, the-AB isoform appearing the major responsible of the PRP chemiotaxis. PDGF, at the concentrations present in PRP, does not affect cell proliferation.

Keywords: PDGF isoforms;; PRP; SaOS-2-cells; migration; osteoblasts.

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Figures

Figure 1
Figure 1
Expression of PDGF receptor alpha and beta in SaOS-2 cells. Data are the Ct values of each standard point. The line represents the regression curve.
Figure 2
Figure 2
Effect of different PDGF isoforms on SaOS-2 cells migration. Values are means ± SD of the rate of migrated cells compared to DMEM. The number of samples analyzed is shown in the squares at the bottom of the columns. *p<0.05 vs. DMEM.
Figure 3
Figure 3
Effect of different PDGF-C doses on SaOS-2 cells migration. Values are means ± SD of the rate of migrated cells compared to DMEM. The number of samples analyzed is shown in the squares at the bottom of the columns.
Figure 4
Figure 4
Effect of different PDGF isoforms on SaOS-2 cells proliferation. Values are means ± SD of the rate of [3H]-thymidine incorporation compared to DMEM. The number of samples analyzed is shown in the squares at the bottom of the columns. *p<0.05 vs. DMEM.
Figure 5
Figure 5
Effect of different PDGF-C doses on SaOS-2 cells proliferation. Values are means ± SD of the rate of [3H]-thymidine incorporation compared to DMEM. The number of samples analyzed is shown in the squares at the bottom of the columns. *p<0.05 vs. DMEM.
Figure 6
Figure 6
Photomicrographies of primary cultures of human osteoblasts, labelled with osteopontin (OPN). Panels a and b: from 2nd to 4th passage in culture the presence of OPN, a phenotypic marker for osteoblasts, is indicative of a good differentiation of these cells as osteoblasts. Panels c and d: from the 5th passage in vitro, the OPN positive cells show a progressive loss of differentiation. Panels a and d: original magnification 40×; panels b and c: original magnification 20×.
Figure 7
Figure 7
Sensitivity of primary human osteoblasts to the chemotactic effect of PRP after different passages in culture. Values are means ± SD of the rate of migrated cells compared to DMEM.
Figure 8
Figure 8
Effect of different PDGF isoforms on primary human osteoblast migration. Values are means ± SD of the rate of migrated cells compared to DMEM. The number of samples analyzed is shown in the squares at the bottom of the columns. *p<0.05 vs. DMEM
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