title: Efficient generation of smooth muscle cells from adipose-derived stromal cells by 3D mechanical stimulation can substitute the use of growth factors in vascular tissue engineering.
abstract: Occluding artery disease causes a high demand for bioartificial replacement vessels. We investigated the combined use of biodegradable and creep-free poly (1,3-trimethylene carbonate) (PTMC) with smooth muscle cells (SMC) derived by biochemical or mechanical stimulation of adipose tissue-derived stromal cells (ASC) to engineer bioartificial arteries. Biochemical induction of cultured ASC to SMC was done with TGF-β1 for 7d. Phenotype and function were assessed by qRT-PCR, immunodetection and collagen contraction assays. The influence of mechanical stimulation on non-differentiated and pre-differentiated ASC, loaded in porous tubular PTMC scaffolds, was assessed after culturing under pulsatile flow for 14d. Assays included qRT-PCR, production of extracellular matrix and scanning electron microscopy. ASC adhesion and TGF-β1-driven differentiation to contractile SMC on PTMC did not differ from tissue culture polystyrene controls. Mesenchymal and SMC markers were increased compared to controls. Interestingly, pre-differentiated ASC had only marginal higher contractility than controls. Moreover, in 3D PTMC scaffolds, mechanical stimulation yielded well-aligned ASC-derived SMC which deposited ECM. Under the same conditions, pre-differentiated ASC-derived SMC maintained their SMC phenotype. Our results show that mechanical stimulation can replace TGF-β1 pre-stimulation to generate SMC from ASC and that pre-differentiated ASC keep their SMC phenotype with increased expression of SMC markers.
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, the Netherlands.
authors.forename: Mojtaba
authors.initials: M
authors.lastname: Parvizi
authors.affiliation: Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.
authors.forename: Lydia A M
authors.initials: LA
authors.lastname: Bolhuis-Versteeg
authors.affiliation: Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands.
authors.forename: André A
authors.initials: AA
authors.lastname: Poot
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, the Netherlands. m.c.harmsen@umcg.nl.
authors.forename: Martin C
authors.initials: MC
authors.lastname: Harmsen
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D001696:Biomechanical Phenomena; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000166:cytology*; D010641:Phenotype; D017154:Stromal Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
chemical_list: D053773:Transforming Growth Factor beta1
keywords: Adipose tissue-derived stromal cells (ASC); Poly(1,3-trimethylene carbonate) (PTMC); Smooth muscle cells (SMC); Transforming Growth Factor beta 1 (TGF-β1); Vascular tissue engineering