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1.
Biotechnol Prog ; 27(4): 1115-25, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21608139

ABSTRACT

Noninvasive in vivo monitoring of tissue implants provides important correlations between construct function and the observed physiologic effects. As oxygen is a key parameter affecting cell and tissue function, we established a monitoring method that utilizes (19) F nuclear magnetic resonance (NMR) spectroscopy, with perfluorocarbons (PFCs) as oxygen concentration markers, to noninvasively monitor dissolved oxygen concentration (DO) in tissue engineered implants. Specifically, we developed a dual PFC method capable of simultaneously measuring DO within a tissue construct and its surrounding environment, as the latter varies among animals and with physiologic conditions. In vitro studies using an NMR-compatible bioreactor demonstrated the feasibility of this method to monitor the DO within alginate beads containing metabolically active murine insulinoma ßTC-tet cells, relative to the DO in the culture medium, under perfusion and static conditions. The DO profiles obtained under static conditions were supported by mathematical simulations of the system. In vivo, the dual PFC method was successful in tracking the oxygenation state of entrapped ßTC-tet cells and the surrounding peritoneal DO over 16 days in normal mice. DO measurements correlated well with the extent of cell growth and host cell attachment examined postexplantation. The peritoneal oxygen environment was found to be variable and hypoxic, and significantly lower in the presence of metabolically active cells. The significance of the dual PFC system in providing critical DO measurements for entrapped cells and other tissue constructs, in vitro and in vivo, is discussed.


Subject(s)
Fluorocarbons , Oxygen/analysis , Oxygen/metabolism , Tissue Engineering , Animals , Cell Line, Tumor , Magnetic Resonance Spectroscopy , Mice , Models, Theoretical
2.
Tissue Eng Part C Methods ; 17(9): 887-94, 2011 Sep.
Article in English | MEDLINE | ID: mdl-21486202

ABSTRACT

The function of an implanted tissue-engineered pancreatic construct is influenced by many in vivo factors; however, assessing its function is based primarily on end physiologic effects. As oxygen significantly affects cell function, we established a dual perfluorocarbon method that utilizes (19)F nuclear magnetic resonance spectroscopy, with perfluorocarbons as oxygen concentration markers, to noninvasively monitor dissolved oxygen concentration (DO) in ßTC-tet cell-containing alginate beads and at the implantation milieu. Beads were implanted in the peritoneal cavity of normal and streptozotocin-induced diabetic mice. Using this method, the feasibility of acquiring real-time in vivo DO measurements was demonstrated. Results showed that the mouse peritoneal environment is hypoxic and the DO is further reduced when ßTC-tet cell constructs were implanted. The DO within cell-containing beads decreased considerably over time and could be correlated with the relative changes in the number of viable encapsulated cells. The reduction of construct DO due to the metabolic activity of the ßTC-tet cells was also compatible with the implant therapeutic function, as observed in the reversal of hyperglycemia in diabetic mice. The importance of these findings in assessing implant functionality and host animal physiology is discussed.


Subject(s)
Implants, Experimental , Monitoring, Physiologic/methods , Oxygen/analysis , Pancreas, Artificial , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Animals , Blood Glucose/metabolism , Cell Survival , Diabetes Mellitus, Experimental/blood , Diabetes Mellitus, Experimental/therapy , Fluorocarbons/chemistry , Magnetic Resonance Spectroscopy , Mice , Mice, Inbred BALB C , Peritoneal Cavity/pathology , Solubility
3.
J Biotechnol ; 150(2): 232-9, 2010 Oct 15.
Article in English | MEDLINE | ID: mdl-20804794

ABSTRACT

Due to the high solubility of oxygen in perfluorocarbons (PFCs), these compounds have been explored for improved cell and tissue oxygenation. The goal of this study is to investigate the effects of a PFC emulsion on cellular growth and function in a tissue engineered construct. A perfluorotributylamine (PFTBA) emulsion was co-encapsulated at 10 vol% with mouse ßTC-tet insulinoma cells in calcium alginate beads and cultured under normoxic and severely hypoxic conditions. The number of metabolically active cells and the induced insulin secretion rate were measured over time for up to 16 days. Results showed no significant effect of PFTBA relative to the PFTBA-free control. The alginate-PFC-cell system was also modeled mathematically, and simulations tracked the number of viable cells over time under the same conditions used experimentally. Simulations revealed only a small, likely experimentally undetectable difference in cell density between the PFC-containing and PFC-free control beads. It is concluded that PFTBA up to 10 vol% has no significant effect on the growth and function of encapsulated ßTC-tet cells under normoxic and hypoxic conditions.


Subject(s)
Cell Proliferation/drug effects , Cell Survival/drug effects , Fluorocarbons/pharmacology , Models, Biological , Oxygen/metabolism , Animals , Cell Hypoxia , Cell Line, Tumor , Computer Simulation , Insulinoma , Mice
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