There are many drivers behind cell-based research including tissue engineering, in vitro toxicology, drug delivery, but also antibacterial coatings.
Surface plasmon resonance has been originally commercialized for protein interaction measurements. It is only recently that researchers started developing the SPR for measurements on living cells. We are proud to be the first ones to enable measurements of trans- and paracellular uptake.
“MP-SPR allows us to work with living cell monolayers grown directly on the sensor surface or with the aid of e.g. fibronectin and other growth promoting proteins. With MP-SPR, we are able to observe and quantify the differences in cell uptake kinetics of nanoparticles in dependence with the surface characteristics of the nanoparticle and their targeting." - Asst. Prof. Tapani Viitala, University of Helsinki, Finland
Assessment of T-cell specificity to tumor associated antigens is crucial for the development of personalized immunotherapies against cancer. Multi-Parametric Surface Plasmon Resonance (MP-SPR) was applied for characterization of interaction of T-cell receptors (TCR) from tumor-specific CD8+ T-cells. Intact live cells were captured onto biomimetic surfaces composed of artificial cell membranes functionalized with peptide-major histocompatibility complexes (pMHC). Real-time affinity analysis of immunological synapse formation betweenTCR and pMHC was completed from 4 different T-cell populations.
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Figure 3. Uptake of mesoporous silica nanoparticles in human epithelial cervical cancer cells (HeLa). Positively charged nanoparticles (C-SiNPs) showed a more efficient uptake and caused a larger response in MP-SPR than negatively charged NPs (P-SiNPs). There was a rapid initial response when SiNPs reached the flow-cell, due to some remaining stock solvent dimethyl sulfoxide (DMSO) after dilutions.
Nanoparticles (NPs) are extensively studied as drug delivery systems. NPs enter the cells usually by active transport, i.e. endocytosis. Multi-Parametric Surface Plasmon Resonance (MP-SPR) is previously used for protein-protein interactions is now used in pioneering NP – living cell studies. Confluent monolayers of human epithelial cervical cancer cells (HeLa) were grown on sensor slides. Uptake of mesoporous silica nanoparticles (SiNPs), branched polyethyleneimine–DNA polyplexes (bPEI–DNA PPs), and extracellular vesicles (EVs) were studied using MP-SPR. Uptake was measured at different temperatures and the activation energy of the cell uptake was calculated using Arrhenius plots.
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Figure 1. (A) Binding of stem cells (AD-MSC) on ceramic hydroxyapatite surface was determined for orthopedic prosthesis development. (B) Binding of breast cancer cells (MCF7) to target peptide was measured to develop biosensor for cancer detection from blood sample.
Implants are covered with proteins and cells when introduced into the body. To understand interactions on these interfaces, in vitro tools are utilized. Real-time label-free platforms allowing dynamic and static flow-conditions are exploited to understand cell adhesion and in this way improve compatibility of implants. The same features are advantageous also for biosensor development based on cell detection. Cells can also be used as an analyte in a clinical biosensor (for cancer) study.
Attachment of human mesenchymal stem cells (AD-MSC) and lysozyme protein on a few tens of micrometers thick hydroxyapatite (HA), coated by plasma spraying, was measured by Multi-Parametric Surface Plasmon Resonance (MP-SPR). Hydroxyapatite is a component present in tooth and bone and in its synthetic form it is widely used in orthopedic prosthesis to enhance implant osseointegration (connection to a living bone). The results of MP-SPR measurements showed that cells favor binding on HA coating as opposed to gold.
In a separate experiment, a biosensor was developed to detect tumor cells. Binding of breast cancer cells (MCF7) and non-cancerous cells (MCF-10A) to a surface bound targeting peptide (18-4) and a reference peptide were measured. The biosensor surface was able to distinguish cancer cells from normal cells.
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Figure 1. MDCKII cell monolayer was deposited on a gold sensor slide. Drug interaction with the cell monolayer was measured in constant flow conditions.
Cell monolayer was deposited on a SPR sensor slide. MultiParametric Surface Plasmon Resonance (MP-SPR) was used to measure drugs (propranolol and D-mannitol) interaction with the cell monolayer in real time and in controlled flow conditions. It was possible to distinguish between paracellular and transcellular drug absorption routes.
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