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

被引:23
作者
Bagnaninchi, PO
Dikeakos, M
Veres, T
Tabrizian, M [1 ]
机构
[1] McGill Univ, Dept Biomed Engn, Ctr Biorecognit & Biosensors, Montreal, PQ H3A 2B4, Canada
[2] McGill Univ, Dept Biomed Engn, Montreal, PQ H3A 2B4, Canada
[3] Keele Univ, Inst Sci & Technol Med, Stoke On Trent ST4 7QB, Staffs, England
[4] Natl Res Council Canada, Inst Mat Inst, Boucherville, PQ JAB 6Y4, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
cell differentiation; cell growth; cell proliferation; complex permittivity; effective medium approximation; microporous scaffold; tissue engineering;
D O I
10.1109/TNB.2004.837901
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:243 / 250
页数:8
相关论文
共 26 条
[1]   Characterization of biological cells by dielectric spectroscopy [J].
Asami, K .
JOURNAL OF NON-CRYSTALLINE SOLIDS, 2002, 305 (1-3) :268-277
[2]   Characterization of heterogeneous systems by dielectric spectroscopy [J].
Asami, K .
PROGRESS IN POLYMER SCIENCE, 2002, 27 (08) :1617-1659
[3]   Towards on-line monitoring of cell growth in microporous scaffolds: Utilization and interpretation of complex permittivity measurements [J].
Bagnaninchi, PO ;
Dikeakos, M ;
Veres, T ;
Tabrizian, M .
BIOTECHNOLOGY AND BIOENGINEERING, 2003, 84 (03) :343-350
[4]  
BAGNANINCHI PO, 2003, P S MAT RES SOC, V773
[5]   Dielectric spectroscopy of erythrocyte cell suspensions. A comparison between Looyenga and Maxwell-Wagner-Hanai effective medium theory formulations [J].
Bordi, F ;
Cametti, C ;
Gili, T .
JOURNAL OF NON-CRYSTALLINE SOLIDS, 2002, 305 (1-3) :278-284
[6]   Dielectric spectroscopy of blood [J].
Chelidze, T .
JOURNAL OF NON-CRYSTALLINE SOLIDS, 2002, 305 (1-3) :285-294
[7]   On-line determination of animal cell concentration in two industrial high-density culture processes by dielectric spectroscopy [J].
Ducommun, P ;
Kadouri, A ;
von Stockar, U ;
Marison, IW .
BIOTECHNOLOGY AND BIOENGINEERING, 2002, 77 (03) :316-323
[8]  
FITZGERALD JJ, 1997, DIELECTRIC SPECTROSC
[9]  
FOSTER KR, 1989, CRIT REV BIOMED ENG, V17, P25
[10]  
Freshney IR, 2000, CULTURE ANIMAL CELLS