Angiogenically stimulated alternative monocytes maintain their pro-angiogenic and non-inflammatory phenotype in long-term co-cultures with HUVEC

Issue title: Selected Presentations held at the 33th Annual Conference of the German Society for Clinical Microcirculation and Hemorheology, Villingen-Schwenningen, Germany, 14-15 November, 2014

Article type: Research Article

Authors: Krüger, Anne | Mayer, Anke | Roch, Toralf^; | Schulz, Christian | Lendlein, Andreas^; | Jung, Friedrich^;

Affiliations: Institute of Biomaterial Science and Berlin-Brandenburg Center for Regenerative Therapies, Helmholtz-Zentrum Geesthacht, Teltow, Germany | Helmholtz Virtual Institute – Multifunctional Biomaterials for Medicine, Teltow and Berlin, Germany

Note: [] Present address: German Center for Health Services Research in Dermatology, Institute for Health Services Research in Dermatology and Nursing, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

Note: [] Corresponding authors: Friedrich Jung and Andreas Lendlein, Institute of Biomaterial Science and Berlin-Brandenburg Center for Regenerative Therapies, Helmholtz-Zentrum Geesthacht, Kantstraße 55, 14513 Teltow, Germany. Tel.: +49 3328 352 450; Fax: +49 3328 352 452; E-mails: [email protected] (Friedrich Jung); [email protected] (Andreas Lendlein).

Note: [] Corresponding authors: Friedrich Jung and Andreas Lendlein, Institute of Biomaterial Science and Berlin-Brandenburg Center for Regenerative Therapies, Helmholtz-Zentrum Geesthacht, Kantstraße 55, 14513 Teltow, Germany. Tel.: +49 3328 352 450; Fax: +49 3328 352 452; E-mails: [email protected] (Friedrich Jung); [email protected] (Andreas Lendlein).

Abstract: Angiogenically stimulated alternative monocytes (aMO2) could be established as cellular release system accelerating the endothelialization of polymers rendering their surfaces hemocompatibility in a short-term study. However, for their clinical application it is essential that aMO2 do not switch back to the MO1 state sustaining their capability as cellular release system over an extended period of time. We explored whether aMO2 can maintain their differentiation state over 21 days in a mono- and in a co-culture with HUVEC. In comparison, the influence of recombinant VEGF-A165 on the endothelialization of biomaterials was assessed including endothelial cell (HUVEC) density, organisation of the endothelial cytoskeleton, cytokine secretion profile and release of prostacyclin, thromboxane A2 and matrix metalloproteinases. In mono-culture aMO2 secreted high amounts of VEGF and other growth factors/cytokines. Co-cultured with HUVEC, aMO2 accelerated the formation of a confluent HUVEC monolayer. Furthermore, no pro-inflammatory cytokines were found, neither in aMO2-mono, nor in co-cultures with HUVEC indicating that the majority of the aMO2 remained stable in their aMO2 state during the 21 days of cultivation. In contrast, the addition of recombinant VEGF-A165 instead of the co-culture with aMO2 resulted in the formation of stress fibres, dissociated marginal filament bands, and a detachment of HUVEC. In addition, the profile of bioactive agents of HUVEC (e.g. prostacyclin, thromboxane A2, matrix metalloproteinases, IFN-γ and TNF-α) was influenced by the VEGF-A165 treatment inducing the detachment of HUVEC. In conclusion, in co-culture with HUVEC aMO2 remained stable in their type 2 state over 21 days confirming the suitability of aMO2 as biological release system for the endothelialization of biomaterial surfaces with constant release of angiogenic factors but without secretion of pro-inflammatory cytokines over three weeks. Therefore, this endothelialization approach seems to be appropriate to improve the hemocompatibility of cardiovascular implant materials in vitro, and proved to be superior to the use of recombinant VEGF-A165.

Keywords: Angiogenically stimulated alternative monocytes, endothelialization, biomaterials, hemocompatible surface

DOI: 10.3233/CH-141875

Journal: Clinical Hemorheology and Microcirculation, vol. 58, no. 1, pp. 229-240, 2014