title: Novel mineral contrast agent for magnetic resonance studies of bone implants grown on a chick chorioallantoic membrane.
abstract: Magnetic resonance imaging (MRI) studies of tissue engineered constructs prior to implantation clearly demonstrate the utility of the MRI technique for studying the bone formation process. To test the utility of our MRI protocols for explant studies, we present a novel test platform in which osteoblast-seeded scaffolds were implanted on the chorioallantoic membrane of a chick embryo. Scaffolds from the following experimental groups were examined by high-resolution MRI: (a) cell-seeded implanted scaffolds (CIM), (b) unseeded implanted scaffolds (UCIM), (c) cell-seeded scaffolds in static culture (CIV) and (d) unseeded scaffolds in static culture (UCIV). The reduction in water proton transverse relaxation times and the concomitant increase in water proton magnetization transfer ratios for CIM and CIV scaffolds, compared to UCIV scaffolds, were consistent with the formation of a bone-like tissue within the polymer scaffold, which was confirmed by immunohistochemistry and fluorescence microscopy. However, the presence of angiogenic vessels and fibrotic adhesions around UCIM scaffolds can confound MRI findings of bone deposition. Consequently, to improve the specificity of the MRI technique for detecting mineralized deposits within explanted tissue engineered bone constructs, we introduce a novel contrast agent that uses alendronate to target a Food and Drug Administration-approved MRI contrast agent (Gd-DOTA) to bone mineral. Our contrast agent termed GdALN was used to uniquely identify mineralized deposits in representative samples from our four experimental groups. After GdALN treatment, both CIM and CIV scaffolds, containing mineralized deposits, showed marked signal enhancement on longitudinal relaxation time-weighted (T1W) images compared to UCIV scaffolds. Relative to UCIV scaffolds, some enhancement was observed in T1W images of GdALN-treated UCIM scaffolds, subjacent to the dark adhesions at the scaffold surface, possibly from dystrophic mineral formed in the fibrotic adhesions. Notably, residual dark areas on T1W images of CIM and UCIM scaffolds were attributable to blood inside infiltrating vessels. In summary, we present the efficacy of GdALN for sensitizing the MRI technique to the deposition of mineralized deposits in explanted polymeric scaffolds.
authors.affiliation: Department of Biophysics, Armed Forces Institute of Pathology Annex, Rockville, MD 20850, USA.
authors.forename: Ingrid E
authors.initials: IE
authors.lastname: Chesnick
authors.forename: Carol B
authors.initials: CB
authors.lastname: Fowler
authors.forename: Jeffrey T
authors.initials: JT
authors.lastname: Mason
authors.forename: Kimberlee
authors.initials: K
authors.lastname: Potter
mesh_terms: D019386:Alendronate / Q000494:pharmacology; D000818:Animals; D018786:Bone Substitutes / Q000737:chemistry*; D001842:Bone and Bones / Q000473:pathology; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000378:metabolism*; D003287:Contrast Media / Q000494:pharmacology*; D005355:Fibrosis / Q000473:pathology; D006571:Heterocyclic Compounds / Q000494:pharmacology; D007150:Immunohistochemistry / Q000379:methods; D008279:Magnetic Resonance Imaging / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D009942:Organometallic Compounds / Q000494:pharmacology; D010006:Osteoblasts / Q000166:cytology; D010063:Ovum / Q000473:pathology; D011108:Polymers / Q000737:chemistry; D019736:Prostheses and Implants; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds
chemical_list: D018786:Bone Substitutes; D003287:Contrast Media; D006571:Heterocyclic Compounds; D009942:Organometallic Compounds; D011108:Polymers; C050823:gadolinium 1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetate; D019386:Alendronate