title: Complex permittivity measurement as a new noninvasive tool for monitoring in vitro tissue engineering and cell signature through the detection of cell proliferation, differentiation, and pretissue formation.
abstract: In in vitro tissue engineering, microporous scaffolds are commonly used to promote cell proliferation and differentiation in three-dimensional structures. Classic measurement methods are particularly time consuming, difficult to handle, and destructive. In this study, a new nondestructive method based on complex permittivity measurement (CPM) is proposed to monitor and track the osteoblast and macrophage differentiation through their morphological variation upon cell attachment and proliferation inside the microporous scaffolds. CPM is performed using a vector network analyzer and a dielectric probe under sterile conditions in a laminar-flow hood. A suitable effective medium approximation (EMA) is applied to fit the data in order to extract the parameters of the different constituents. Our data show that the EMA depolarization factor can be monitored to assess the variation of cell morphology characterizing cell attachment. Discrimination between two batches of scaffolds seeded, respectively, with 2 million and 1 million osteoblast cells is possible; the ratio of their CPM-derived cell volume fractions is in agreement with the ratio of their cell seeding numbers. In addition, cell proliferation inside scaffolds seeded with osteoblasts cultured in alpha minimum essential medium and inside scaffolds seeded with osteoblasts cultured in alpha minimum essential medium supplemented to induce the formation of extracellular matrix is monitored via CPM over several days. CPM-determined cell volume fraction is compared to DNA assay cell counts. Extracellular matrix formation and cell presence was confirmed by scanning electron microscopy. A set of three signature parameters (epsilon'mem, epsilon'cyt, kappa'cyt) characteristic of cell line is extracted from CPM. Distinct signatures are recorded for osteoblasts and macrophages, thus confirming the ability of CPM to discriminate between different cell types. This study demonstrates the potential of CPM as a diagnostic tool to monitor quickly and noninvasively cell growth and differentiation inside microporous scaffolds. Our findings suggest that the use of CPM could be extended to many biomedical applications, such as drug detection and automation of tissue and bacterial cultures in bioreactors.
authors.affiliation: Department of Biomedical Engineering, McGill University, Montreal, QC H3A 2B4, Canada. bagnaninchi_po@yahoo.fr
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Bagnaninchi
authors.forename: Maria
authors.initials: M
authors.lastname: Dikeakos
authors.forename: Teodor
authors.initials: T
authors.lastname: Veres
authors.forename: Maryam
authors.initials: M
authors.lastname: Tabrizian
mesh_terms: D016475:3T3 Cells; D000465:Algorithms; D000818:Animals; D018786:Bone Substitutes / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D048429:Cell Size; D048271:Chitosan / Q000737:chemistry; D017097:Electric Impedance; D008422:Materials Testing / Q000379:methods*; D051379:Mice; D008954:Models, Biological; D010006:Osteoblasts / Q000145:classification* / Q000502:physiology* / Q000648:ultrastructure; D010012:Osteogenesis / Q000502:physiology; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D018786:Bone Substitutes; D048271:Chitosan