Journal
BIOMATERIALS
Volume 30, Issue 17, Pages 3068-3074Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2009.02.021
Keywords
Tissue engineering; Hypoxia; Optical sensors; Microspheres; Poly(dimethylsiloxane)
Funding
- NINDS NIH HHS [R01 NS065205-01A1, R01 NS065205] Funding Source: Medline
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We present the development and characterization of fluorescent oxygen-sensing microparticles designed for measuring oxygen concentration in microenvironments existing within standard cell culture and transparent three-dimensional (3D) cell scaffolds. The microparticle synthesis employs poly(dimethylsiloxane) to encapsulate silica gel particles bound with an oxygen-sensitive luminophore as well as a reference or normalization fluorophore that is insensitive to oxygen. We developed a rapid, automated and non-invasive sensor analysis method based on fluorescence microscopy to measure oxygen concentration in a hydrogel scaffold. We demonstrate that the microparticles are non-cytotoxic and that their response is comparable to that of a traditional dissolved oxygen meter. Microparticle size (5-40 mu m) was selected for microscale-mapping of oxygen concentration to allow measurements local to individual cells. Two methods of calibration were evaluated and revealed that the sensor system enables characterization of a range of hypoxic to hyperoxic conditions relevant to cell and tissue biology (i.e., pO(2) 10-160 mm Hg). The calibration analysis also revealed that the microparticles have a high fraction of quenched luminophore (0.90 +/- 0.02), indicating that the reported approach provides significant advantages for sensor performance. This study thus reports a versatile oxygen-sensing technology that enables future correlations of local oxygen concentration with individual cell response in cultured engineered tissues. (C) 2009 Elsevier Ltd. All rights reserved.
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