title: Effect of hydrogel porosity on marrow stromal cell phenotypic expression.
abstract: This study describes investigation of porous photocrosslinked oligo[(polyethylene glycol) fumarate] (OPF) hydrogels as potential matrix for osteoblastic differentiation of marrow stromal cells (MSCs). The porosity and interconnectivity of porous hydrogels were assessed using magnetic resonance microscopy (MRM) as a noninvasive investigative tool that could image the water construct inside the hydrogels at a high-spatial resolution. MSCs were cultured onto the porous hydrogels and cell number was assessed using PicoGreen DNA assay. Our results showed 10% of cells initially attached to the surface of scaffolds. However, cells did not show significant proliferation over a time period of 14 days. MSCs cultured on porous hydrogels had increased alkaline phosphatase activity as well as deposition of calcium, suggesting successful differentiation and maturation to the osteoblastic phenotype. Moreover, continued expression of type I collagen and osteonectin over 14 days confirmed osteoblastic differentiation of MSCs. MRM was also applied to monitor osteogenesis of MSCs on porous hydrogels. MRM images showed porous scaffolds became consolidated with osteogenic progression of cell differentiation. These findings indicate that porous OPF scaffolds enhanced MSC differentiation leading to development of bone-like mineralized tissue.
authors.affiliation: Department of Orthopedic Surgery, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
authors.forename: Mahrokh
authors.initials: M
authors.lastname: Dadsetan
authors.forename: Theresa E
authors.initials: TE
authors.lastname: Hefferan
authors.forename: Jan P
authors.initials: JP
authors.lastname: Szatkowski
authors.forename: Prasanna K
authors.initials: PK
authors.lastname: Mishra
authors.forename: Slobodan I
authors.initials: SI
authors.lastname: Macura
authors.forename: Lichun
authors.initials: L
authors.lastname: Lu
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Yaszemski
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001696:Biomechanical Phenomena; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002113:Calcification, Physiologic / Q000502:physiology; D002454:Cell Differentiation; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry / Q000494:pharmacology; D024042:Collagen Type I / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D005650:Fumarates / Q000737:chemistry; D020100:Hydrogels / Q000737:chemistry*; D008297:Male; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology; D015676:Osteonectin / Q000378:metabolism; D010641:Phenotype; D011092:Polyethylene Glycols / Q000737:chemistry; D016062:Porosity; D051381:Rats; D017207:Rats, Sprague-Dawley; D017550:Spectroscopy, Fourier Transform Infrared; D017154:Stromal Cells / Q000502:physiology* / Q000648:ultrastructure; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D020099:Coated Materials, Biocompatible; D024042:Collagen Type I; D003432:Cross-Linking Reagents; D005650:Fumarates; D020100:Hydrogels; D015676:Osteonectin; D011092:Polyethylene Glycols; C032005:fumaric acid; D000469:Alkaline Phosphatase