In vivo noninvasive monitoring of a tissue engineered construct using 1H NMR spectroscopy.
Direct, noninvasive monitoring of tissue engineered substitutes containing live, functional cells would provide valuable information on dynamic changes that occur postimplantation. Such changes include remodeling both within the construct and at the interface of the implant with the surrounding host tissue, and may result in changes in the number of viable cells in the construct. This study investigated the use of 1H NMR spectroscopy in noninvasively monitoring the viable cell number within a tissue engineered construct in vivo. The construct consisted of mouse betaTC3 insulinomas in a disk-shaped agarose gel, surrounded by a cell-free agarose gel layer. Localized 1H NMR spectra were acquired from within implanted constructs, and the total choline resonance was measured. Critical issues that had to be addressed in accurately quantifying total choline from the implanted cells included avoiding signal from host tissue and correcting for interfering signal from diffusing solutes. In vivo NMR measurements were correlated with MTT assays and NMR measurements performed in vitro on explanted constructs. Total choline measurements accurately and noninvasively quantified viable betaTC3 cell numbers in vivo, in the range of 1 x 10(6) to more than 14 x 10(6) cells, and monitored changes in viable cell number that occurred in the same construct over time. This is the first study using NMR techniques to monitor viable cell numbers in an implanted tissue substitute. It established architectural characteristics that a construct should have to be amenable to NMR monitoring, and it set the foundation for future in vivo investigations with other tissue engineered implants.
Stabler
C L
CL
Georgia Institute of Technology/Emory University Center for the Engineering of Living Tissues, 315 Ferst Drive, Atlanta, GA 30332-0363, USA.
Long
R C
RC
Constantinidis
I
I
Sambanis
A
A
D000818:Animals; D008279:Magnetic Resonance Imaging / Q000379:methods; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D051379:Mice; D008991:Monitoring, Physiologic / Q000379:methods; D019397:Pancreas, Artificial; D023822:Tissue Engineering