title: Pharmacologically active microcarriers associated with thermosensitive hydrogel as a growth factor releasing biomimetic 3D scaffold for cardiac tissue-engineering.
abstract: The challenge of tissue engineering of the infarcted heart is how to improve stem cell engraftment, survival, homing, and differentiation for myocardial repair. We here propose to integrate human adipose-derived stem cells (ADSCs) and pharmacologically active microcarriers (PAMs), a three-dimensional (3D) carrier of cells and growth factors, into an injectable hydrogel (HG), to obtain a system that stimulates the survival and/or differentiation of the grafted cells toward a cardiac phenotype. PAMs are biodegradable and non-cytotoxic poly(lactic-co-glycolic acid) (PLGA) microspheres conveying cells on their 3D surface that deliver continuously and in a controlled manner a growth factor (GF) acting on the transported cells and on the microenvironment to improve engraftment. The choice of the appropriate GF and its protection during the formulation process and delivery are essential. In this study two GFs, hepatocyte growth factor (HGF) and insulin-like growth factor (IGF-1), have been encapsulated under a solid state in order to limit their interaction with the polymer and conserve their integrity. GF precipitation conditions and release profile from PAMs have been first investigated before combining them to ADSCs. The released IGF-1 and HGF induced the protein synthesis of cardiac differentiation markers GATA4, Nkx2.5, cTnI and CX43 after 1week in vitro. Moreover, the GFs accelerated cell cycle progression, as suggested by the increased expression of Cyclin D1 mRNA and the widespread distribution of Ki67 protein. Integrating PAMs within the thermosensitive P407 hydrogel increased their elastic properties but decreased the transcription of most cardiac markers. In contrast, CX43 expression increased in ADSC-PAM-GF complexes embedded within the hydrogel compared to the ADSCs cultured alone in the absence of P407. These results suggest that particulate scaffolds releasing HGF and IGF-1 may be beneficial for applications in tissue-engineering strategies for myocardial repair and the association with a P407 hydrogel can increase substrate elasticity and junction connections in ADSCs.
authors.affiliation: LUNAM Université, UMR S-1066, F-49933 Angers, France; INSERM U1066, MINT "Micro et Nanomédecines Biomimétiques", F-49933 Angers, France; National Institute for Cardiovascular Research (INRC), I-40126 Bologna, Italy; Department of Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, 40126 Bologna, Italy.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Karam
authors.affiliation: National Institute for Cardiovascular Research (INRC), I-40126 Bologna, Italy; Department of Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, 40126 Bologna, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Muscari
authors.affiliation: LUNAM Université, UMR S-1066, F-49933 Angers, France; INSERM U1066, MINT "Micro et Nanomédecines Biomimétiques", F-49933 Angers, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Sindji
authors.affiliation: LUNAM Université, UMR S-1066, F-49933 Angers, France; INSERM U1066, MINT "Micro et Nanomédecines Biomimétiques", F-49933 Angers, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Bastiat
authors.affiliation: National Institute for Cardiovascular Research (INRC), I-40126 Bologna, Italy; Department of Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, 40126 Bologna, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Bonafè
authors.affiliation: LUNAM Université, UMR S-1066, F-49933 Angers, France; INSERM U1066, MINT "Micro et Nanomédecines Biomimétiques", F-49933 Angers, France.
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Venier-Julienne
authors.affiliation: LUNAM Université, UMR S-1066, F-49933 Angers, France; INSERM U1066, MINT "Micro et Nanomédecines Biomimétiques", F-49933 Angers, France. Electronic address: claudia.montero-menei@univ-angers.fr.
authors.forename: N Claudia
authors.initials: NC
authors.lastname: Montero-Menei
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D032701:Biomimetics; D002454:Cell Differentiation; D002478:Cells, Cultured; D017228:Hepatocyte Growth Factor / Q000008:administration & dosage*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D007334:Insulin-Like Growth Factor I / Q000008:administration & dosage*; D019344:Lactic Acid / Q000737:chemistry; D051379:Mice; D008958:Models, Molecular; D009206:Myocardium / Q000166:cytology*; D041681:NIH 3T3 Cells; D011100:Polyglycolic Acid / Q000737:chemistry; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013696:Temperature; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011100:Polyglycolic Acid; D019344:Lactic Acid; D017228:Hepatocyte Growth Factor; D007334:Insulin-Like Growth Factor I
keywords: Adipose-derived stem cells; Glycofurol (PubMed CID: 110717); Growth factors; Hydrogel; Laminin (PubMed CID: 44342165); Myocardial infarction; Pharmacologically active microcarriers; Poloxamer 188 (PubMed CID: 24751); Poly(dl-lactic-co-glycolic acid) (PubChem CID: 23111554); Poly-d-lysine hydrobromide (PubMed CID: 16219815)