Cardiovascular Drugs and Therapy

, Volume 26, Issue 1, pp 9–21 | Cite as

Stem Cell Therapy for Arterial Restenosis: Potential Parameters Contributing to the Success of Bone Marrow-Derived Mesenchymal Stromal Cells

  • Amalia Forte
  • Barbara Rinaldi
  • Loredana Sodano
  • Liberato Berrino
  • Francesco Rossi
  • Mauro Finicelli
  • Mario Grossi
  • Gilda Cobellis
  • Chiara Botti
  • Marisa De Feo
  • Pasquale Santè
  • Umberto Galderisi
  • Marilena Cipollaro
Article

Abstract

Purpose

Restenosis is a complex and heterogeneous pathophysiological phenomenon occurring in patients submitted to revascularization procedures. Previous studies proved the antirestenotic properties of injected allogenic mesenchymal stromal cells (MSCs) in an experimental model of rat carotid (re)stenosis induced through arteriotomy. In this study we describe some of the effects subsequent to MSC treatment of rats submitted to carotid arteriotomy and possibly responsible for their antirestenotic effect.

Methods

Rat MSCs were isolated from bone marrow, expanded in vitro and characterized. Subsequently, we evaluated the effects of MSC administration via tail vein at 3 and 7 days after carotid arteriotomy both in rat serum and in injured carotids, focusing on DNA oxidative damage (8-oxo-dG detection), cell proliferation index (BrdU incorporation assay), apoptotic index (TUNEL assay), the expression of inflammation- and proliferation-related genes (RT-PCR), the release of growth factors and of inflammation-related cytokines (antibody arrays and ELISA).

Results

MSC administration induced a greater cell proliferation in carotids after arteriotomy, together with an increased level of VEGF in the serum and with the higher expression of VEGF mRNA in injured carotids. Serum analysis also revealed a decreased level of the pro-inflammatory cytokines CXCL1, CXCL5, L-Selectin, ICAM-1 and LIX, and of TIMP1 and SDF-1alpha in MSC-treated rats. The MSC immunomodulatory activity was confirmed by the decreased expression of TLR2 and TLR4 in injured carotids.

Conclusions

MSCs play an immunomodulatory paracrine role when injected in rats submitted to carotid arteriotomy, accompanied by the release of VEGF, possibly contributing to the accelerated repair of the injured vascular wall.

Key words

Mesenchymal stromal cells Restenosis Inflammation Vascular endothelial growth factor 

Notes

Acknowledgements

We are grateful to Dr. Karen English for support with ELISA serum analysis, to Dr. Monica Mattia for excellent care of animal welfare and to Ms. M.R. Cipollaro for administrative assistance.

Grants

This work has been partially funded by a SHRO grant to U.G.

Supplementary material

10557_2011_6359_Fig6_ESM.jpg (12 kb)
Supplemental Figure 1

Representative cross-sections of carotids from DiI-labeled MSC-treated rats harvested at 3 and 7 days after arteriotomy. A, Uninjured carotid; B, rat carotid harvested at 7 days after arteriotomy and DiI-labeled MSCs-treatement. Representative DiI-labeled MSCs emitting red fluorescence are indicated by white arrows. Nuclei were counterstained with Hoechst 33258, emitting blue fluorescence. 40x magnification. (JPEG 11 kb)

10557_2011_6359_MOESM1_ESM.tif (12 mb)
High resolution (TIFF 12267 kb)
10557_2011_6359_MOESM2_ESM.doc (38 kb)
Supplemental Table 1Summary of RT-PCR primer sequences, position, annealing temperature and PCR product length of the target genes analysed in rat carotids. (DOC 38 kb)
10557_2011_6359_MOESM3_ESM.doc (36 kb)
Supplemental Table 2Summary of data concerning the characterization of bone marrow-derived MSCs expanded in vitro. TOP: RT-PCR analysis of PPAR-g and osteopontin on mRNA samples extracted from undifferentiated MSCs and from MSCs induced to differentiate to adipocytes and osteocytes with different media. Data are expressed as arbitrary densitometric units, mean ± SEM, n = 3 for each group. BOTTOM: FACS analysis of MSCs cultured for 15 days from passage zero. (DOC 36 kb)

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Amalia Forte
    • 1
    • 5
  • Barbara Rinaldi
    • 1
    • 5
  • Loredana Sodano
    • 1
    • 5
  • Liberato Berrino
    • 1
    • 5
  • Francesco Rossi
    • 1
    • 5
  • Mauro Finicelli
    • 4
  • Mario Grossi
    • 1
    • 5
  • Gilda Cobellis
    • 3
    • 5
  • Chiara Botti
    • 3
    • 5
  • Marisa De Feo
    • 2
    • 5
  • Pasquale Santè
    • 2
    • 5
  • Umberto Galderisi
    • 1
    • 5
  • Marilena Cipollaro
    • 1
    • 5
  1. 1.Department of Experimental MedicineSecond University of NaplesNaplesItaly
  2. 2.Department of Cardiothoracic and Respiratory SciencesSecond University of NaplesNaplesItaly
  3. 3.Department of General PathologySecond University of NaplesNaplesItaly
  4. 4.Human Health FoundationTerniItaly
  5. 5.Excellence Research Center for Cardiovascular DiseasesSecond University of NaplesNaplesItaly

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