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Human mesenchymal stem cells labelled with dye-loaded amorphous silica nanoparticles: long-term biosafety, stemness preservation and traceability in the beating heart.

Gallina C, Capelôa T, Saviozzi S, Accomasso L, Catalano F, Tullio F, Martra G, Penna C, Pagliaro P, Turinetto V, Giachino C - J Nanobiotechnology (2015)

Bottom Line: However, there are still controversies on their effective biosafety on cellular systems.During prolonged culture hMSCs do not undergo cyto- or genotoxicity, preserve their proliferative potential and their stemness/differentiation properties.The demonstrated biosafety, combined with proper cell labelling and visualization in histological sections, make these SiO2-NPs optimal candidates for the purpose of stem cell tracking inside heart tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Biological Sciences, University of Turin, 10, Regione Gonzole, CAP 10043, Orbassano, TO, Italy. clara.gallina@gmail.com.

ABSTRACT

Background: Treatment of myocardial infarction with mesenchymal stem cells (MSCs) has proven beneficial effects in both animal and clinical studies. Engineered silica nanoparticles (SiO2-NPs) have been extensively used as contrast agents in regenerative medicine, due to their resistance to degradation and ease of functionalization. However, there are still controversies on their effective biosafety on cellular systems. In this perspective, the aims of the present study are: 1) to deeply investigate the impact of amorphous 50 nm SiO2-NPs on viability and function of human bone marrow-derived MSCs (hMSCs); 2) to optimize a protocol of harmless hMSCs labelling and test its feasibility in a beating heart model.

Results: Optimal cell labelling is obtained after 16 h exposure of hMSCs to fluorescent 50 nm SiO2-NPs (50 µg mL(-1)); interestingly, lysosomal activation consequent to NPs storage is not associated to oxidative stress. During prolonged culture hMSCs do not undergo cyto- or genotoxicity, preserve their proliferative potential and their stemness/differentiation properties. Finally, the bright fluorescence emitted by internalized SiO2-NPs allows both clear visualization of hMSCs in normal and infarcted rat hearts and ultrastructural analysis of cell engraftment inside myocardial tissue.

Conclusions: Overall, 50 nm SiO2-NPs display elevated compatibility with hMSCs in terms of lack of cyto- and genotoxicity and maintenance of important features of these cells. The demonstrated biosafety, combined with proper cell labelling and visualization in histological sections, make these SiO2-NPs optimal candidates for the purpose of stem cell tracking inside heart tissue.

No MeSH data available.


Related in: MedlinePlus

Assessment of cytotoxic and genotoxic stress in cultured hMSCs. a Representative flow cytometry histograms of UT- and SiO2-NPs -treated hMSCs at t0: forward scatter (FS) in function of propidium iodide fluorescence (PI) allows discriminating living cells (bottom population) from dead cells (top population). b Quantification of hMSCs viability during in vitro culture, represented as percentage variation respect to UT cells at the same time point. c, d Representative immunofluorescence experiments for γH2AX in UT- and SiO2-NPs -treated hMSCs at t0. Magnification 63×, scale bar 20 µm. e Quantification of percentage positive cells for nuclear γH2AX foci during culture
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Fig2: Assessment of cytotoxic and genotoxic stress in cultured hMSCs. a Representative flow cytometry histograms of UT- and SiO2-NPs -treated hMSCs at t0: forward scatter (FS) in function of propidium iodide fluorescence (PI) allows discriminating living cells (bottom population) from dead cells (top population). b Quantification of hMSCs viability during in vitro culture, represented as percentage variation respect to UT cells at the same time point. c, d Representative immunofluorescence experiments for γH2AX in UT- and SiO2-NPs -treated hMSCs at t0. Magnification 63×, scale bar 20 µm. e Quantification of percentage positive cells for nuclear γH2AX foci during culture

Mentions: Demonstrated the absence of a detectable SiO2-NPs -induced oxidative stress at t0, the possibility of long-term toxicity was then investigated by analysing cell survival during prolonged in vitro culture (7 days) via propidium iodide (PI) staining (Fig. 2a, b). UT- and SiO2-NPs -treated hMSCs displayed a similar small amount of dead cells, as can be seen in the representative flow cytometry histograms at t0 (Fig. 2a, top populations). Quantification of percentage cell survival then revealed the same trend also after 1 (R 1d), 3 (R 3d) and 7 (R 7d) days of recovery in standard medium (Fig. 2b), thereby indicating that SiO2-NPs did not lead to significant cytotoxicity in hMSCs both after their uptake and at later time points of in vitro culture.Fig. 2


Human mesenchymal stem cells labelled with dye-loaded amorphous silica nanoparticles: long-term biosafety, stemness preservation and traceability in the beating heart.

Gallina C, Capelôa T, Saviozzi S, Accomasso L, Catalano F, Tullio F, Martra G, Penna C, Pagliaro P, Turinetto V, Giachino C - J Nanobiotechnology (2015)

Assessment of cytotoxic and genotoxic stress in cultured hMSCs. a Representative flow cytometry histograms of UT- and SiO2-NPs -treated hMSCs at t0: forward scatter (FS) in function of propidium iodide fluorescence (PI) allows discriminating living cells (bottom population) from dead cells (top population). b Quantification of hMSCs viability during in vitro culture, represented as percentage variation respect to UT cells at the same time point. c, d Representative immunofluorescence experiments for γH2AX in UT- and SiO2-NPs -treated hMSCs at t0. Magnification 63×, scale bar 20 µm. e Quantification of percentage positive cells for nuclear γH2AX foci during culture
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4625930&req=5

Fig2: Assessment of cytotoxic and genotoxic stress in cultured hMSCs. a Representative flow cytometry histograms of UT- and SiO2-NPs -treated hMSCs at t0: forward scatter (FS) in function of propidium iodide fluorescence (PI) allows discriminating living cells (bottom population) from dead cells (top population). b Quantification of hMSCs viability during in vitro culture, represented as percentage variation respect to UT cells at the same time point. c, d Representative immunofluorescence experiments for γH2AX in UT- and SiO2-NPs -treated hMSCs at t0. Magnification 63×, scale bar 20 µm. e Quantification of percentage positive cells for nuclear γH2AX foci during culture
Mentions: Demonstrated the absence of a detectable SiO2-NPs -induced oxidative stress at t0, the possibility of long-term toxicity was then investigated by analysing cell survival during prolonged in vitro culture (7 days) via propidium iodide (PI) staining (Fig. 2a, b). UT- and SiO2-NPs -treated hMSCs displayed a similar small amount of dead cells, as can be seen in the representative flow cytometry histograms at t0 (Fig. 2a, top populations). Quantification of percentage cell survival then revealed the same trend also after 1 (R 1d), 3 (R 3d) and 7 (R 7d) days of recovery in standard medium (Fig. 2b), thereby indicating that SiO2-NPs did not lead to significant cytotoxicity in hMSCs both after their uptake and at later time points of in vitro culture.Fig. 2

Bottom Line: However, there are still controversies on their effective biosafety on cellular systems.During prolonged culture hMSCs do not undergo cyto- or genotoxicity, preserve their proliferative potential and their stemness/differentiation properties.The demonstrated biosafety, combined with proper cell labelling and visualization in histological sections, make these SiO2-NPs optimal candidates for the purpose of stem cell tracking inside heart tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Biological Sciences, University of Turin, 10, Regione Gonzole, CAP 10043, Orbassano, TO, Italy. clara.gallina@gmail.com.

ABSTRACT

Background: Treatment of myocardial infarction with mesenchymal stem cells (MSCs) has proven beneficial effects in both animal and clinical studies. Engineered silica nanoparticles (SiO2-NPs) have been extensively used as contrast agents in regenerative medicine, due to their resistance to degradation and ease of functionalization. However, there are still controversies on their effective biosafety on cellular systems. In this perspective, the aims of the present study are: 1) to deeply investigate the impact of amorphous 50 nm SiO2-NPs on viability and function of human bone marrow-derived MSCs (hMSCs); 2) to optimize a protocol of harmless hMSCs labelling and test its feasibility in a beating heart model.

Results: Optimal cell labelling is obtained after 16 h exposure of hMSCs to fluorescent 50 nm SiO2-NPs (50 µg mL(-1)); interestingly, lysosomal activation consequent to NPs storage is not associated to oxidative stress. During prolonged culture hMSCs do not undergo cyto- or genotoxicity, preserve their proliferative potential and their stemness/differentiation properties. Finally, the bright fluorescence emitted by internalized SiO2-NPs allows both clear visualization of hMSCs in normal and infarcted rat hearts and ultrastructural analysis of cell engraftment inside myocardial tissue.

Conclusions: Overall, 50 nm SiO2-NPs display elevated compatibility with hMSCs in terms of lack of cyto- and genotoxicity and maintenance of important features of these cells. The demonstrated biosafety, combined with proper cell labelling and visualization in histological sections, make these SiO2-NPs optimal candidates for the purpose of stem cell tracking inside heart tissue.

No MeSH data available.


Related in: MedlinePlus