Spatial patterning of BMP-2 and BMP-7 on biopolymeric films and the guidance of muscle cell fate.
In the cellular microenvironment, growth factor gradients are crucial in dictating cell fate. Towards developing materials that capture the native microenvironment we engineered biomimetic films that present gradients of matrix-bound bone morphogenetic proteins (BMP-2 and BMP-7). To this end layer-by-layer films composed of poly(L-lysine) and hyaluronan were combined in a simple microfluidic device enabling spatially controlled growth factor diffusion along the film. Linear long-range gradients of both BMPs induced the trans-differentiation of C2C12 myoblasts towards the osteogenic lineage in a dose dependent manner with a different signature for each BMP. The osteogenic marker alkaline phosphatase (ALP) increased in a linear manner for BMP-7 and non-linearly for BMP-2. Moreover, an increased expression of the myogenic marker troponin T was observed with decreasing matrix-bound BMP concentration, providing a substrate that it is both osteo- and myo-inductive. Lastly, dual parallel matrix-bound gradients of BMP-2 and -7 revealed a complete saturation of the ALP signal. This suggested an additive or synergistic effect of the two BMPs. This simple technology allows for determining quickly and efficiently the optimal concentration of matrix-bound growth factors, as well as for investigating the presentation of multiple growth factors in their solid-phase and in a spatially controlled manner.
CNRS UMR 5628 (LMGP), MINATEC, 3 parvis Louis Néel, 38016 Grenoble, France; Université de Grenoble Alpes, Grenoble Institute of Technology, 3 parvis Louis Néel, 38016 Grenoble, France.
Jorge
J
Almodóvar
CNRS UMR 5628 (LMGP), MINATEC, 3 parvis Louis Néel, 38016 Grenoble, France; Université de Grenoble Alpes, Grenoble Institute of Technology, 3 parvis Louis Néel, 38016 Grenoble, France.
Raphaël
R
Guillot
CNRS UMR 5628 (LMGP), MINATEC, 3 parvis Louis Néel, 38016 Grenoble, France; Université de Grenoble Alpes, Grenoble Institute of Technology, 3 parvis Louis Néel, 38016 Grenoble, France.
Claire
C
Monge
Institute Albert Bonniot, Grenoble, France.
Julien
J
Vollaire
Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA 02139, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Seila
S
Selimović
Institute Albert Bonniot, Grenoble, France.
Jean-Luc
JL
Coll
Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA 02139, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
Ali
A
Khademhosseini
CNRS UMR 5628 (LMGP), MINATEC, 3 parvis Louis Néel, 38016 Grenoble, France; Université de Grenoble Alpes, Grenoble Institute of Technology, 3 parvis Louis Néel, 38016 Grenoble, France. Electronic address: catherine.picart@grenoble-inp.fr.
Catherine
C
Picart
D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism*; D055419:Bone Morphogenetic Protein 7 / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D006820:Hyaluronic Acid / Q000737:chemistry; D008567:Membranes, Artificial; D051379:Mice; D046210:Microfluidic Analytical Techniques; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011107:Polylysine / Q000737:chemistry; D011108:Polymers / Q000737:chemistry* / Q000494:pharmacology
D055396:Bone Morphogenetic Protein 2; D055419:Bone Morphogenetic Protein 7; D008567:Membranes, Artificial; D011108:Polymers; D011107:Polylysine; D006820:Hyaluronic Acid
Bone morphogenetic proteins; Gradient; Layer-by-layer; Microfluidic; Myogenesis; Osteoinduction