Page 104 - Magnesium-based supports for stem cell therapy of vascular disease - Mónica Echeverry Rendón

CHAPTER 6
Fig.6. Effect of exposure of Mg in ASC on angiovasculogenesis in vitro. ASCs (Red: SM22α , blue: DAPI) pre- treated with Mg extracts from coated samples were able to form networks after the addition of HUVECS (Green:CD31, blue: DAPI). Oppositely, cells treated with c.p Mg suffered a high decrement in cell population and only an isolated HUVECs added prost treatment were observed.
The influence of leachables on already established vascular networks by ASC – HUVEC co-cultures was, however, vulnerable to exposure to leachables. The morphological integrity of control vascular networks remained intact over the entire period of the experiment (Fig. 7). The exposure of pre-established vascular networks to c.p Mg leachables completely obliterated their integrity; only cellular debris was visible at the end of the experiment. The exposure of pre-established vascular networks to leachables from surface-coated materials compromised the integrity of the endothelial tubes (Fig. 7, green) while the ASCs’ integrity (Fig. 7, red) appeared to be unaffected. The HUVEC that comprised the tubes had lost intercellular connections and had a rounded appearance suggestive of apoptosis. The ASC retained their typical mesenchymal, fibroblast appearance and showed no morphological signs of apoptosis.
Fig.7 Endothelial vascular networks are compromised by leachables from surface-coated materials, while ASC are refractory. Leachables from surface-coated materials and c.p Mg were added to pre-established vascular networks and the influence assessed by immunofluorescent staining; HUVEC in green (CD31) and ASC in red (SM22α). Nuclei in blue were counterstained with DAPI. Leachables from surface-coated materials disrupted the in- tegrity of the networks by induction of HUVEC, visible as single and rounded cells. Exposure to c.p Mg leachables caused massive cell death and thus loss of networks as well as support cells (ASC). Non-treated networks remained intact.
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