Metabolite profiling and pharmacokinetic evaluation of hydrocortisone in a perfused three-dimensional human liver bioreactor.
Endotoxin lipopolysaccharide (LPS) is known to cause liver injury primarily involving inflammatory cells such as Kupffer cells, but few in vitro culture models are applicable for investigation of inflammatory effects on drug metabolism. We have developed a three-dimensional human microphysiological hepatocyte-Kupffer cell coculture system and evaluated the anti-inflammatory effect of glucocorticoids on liver cultures. LPS was introduced to the cultures to elicit an inflammatory response and was assessed by the release of proinflammatory cytokines, interleukin 6 and tumor necrosis factor α. A sensitive and specific reversed-phase-ultra high-performance liquid chromatography-quadrupole time of flight-mass spectrometry method was used to evaluate hydrocortisone disappearance and metabolism at near physiologic levels. For this, the systems were dosed with 100 nM hydrocortisone and circulated for 2 days; hydrocortisone was depleted to approximately 30 nM, with first-order kinetics. Phase I metabolites, including tetrahydrocortisone and dihydrocortisol, accounted for 8-10% of the loss, and 45-52% consisted of phase II metabolites, including glucuronides of tetrahydrocortisol and tetrahydrocortisone. Pharmacokinetic parameters, i.e., half-life, rate of elimination, clearance, and area under the curve, were 23.03 hours, 0.03 hour(-1), 6.6 × 10(-5) l⋅hour(-1), and 1.03 (mg/l)*h, respectively. The ability of the bioreactor to predict the in vivo clearance of hydrocortisone was characterized, and the obtained intrinsic clearance values correlated with human data. This system offers a physiologically relevant tool for investigating hepatic function in an inflamed liver.
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Ujjal
U
Sarkar
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Dinelia
D
Rivera-Burgos
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Emma M
EM
Large
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
David J
DJ
Hughes
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Kodihalli C
KC
Ravindra
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Rachel L
RL
Dyer
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Mohammad R
MR
Ebrahimkhani
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.) srt@mit.edu wishnok@mit.edu.
John S
JS
Wishnok
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.).
Linda G
LG
Griffith
Department of Biological Engineering (U.S., D.R.-B., K.C.R., R.L.D., M.R.E., J.S.W., L.G.G., S.R.T.), Department of Chemistry (S.R.T.), and Department of Mechanical Engineering (L.G.G.), Massachusetts Institute of Technology, Cambridge, Massachusetts; and CN Bio Innovations, Oxford University Begbroke Science Park, Begbroke, Oxfordshire, United Kingdom (E.M.L., D.J.H.) srt@mit.edu wishnok@mit.edu.
Steven R
SR
Tannenbaum
D019149:Bioreactors; D018920:Coculture Techniques; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D020719:Glucuronides / Q000378:metabolism; D006207:Half-Life; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism* / Q000493:pharmacokinetics*; D007249:Inflammation / Q000139:chemically induced / Q000378:metabolism; D015848:Interleukin-5 / Q000096:biosynthesis; D007728:Kupffer Cells / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D016207:Cytokines; D020719:Glucuronides; D015848:Interleukin-5; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D051544:Cytochrome P-450 CYP3A; D006854:Hydrocortisone