Matthew Dixon Oct 13, ’16 ~ 11 min


How do nanoparticles interact with cells?

How nanomaterials interact with the environment after they have been disposed of has many implications for potential toxicity and health concerns. Whether nanomaterials are being incorporated into commercial goods for their anti-microbial properties such as in work-out clothes or used for targeted drug therapies their overall prevalence is increasing. Therefore, the likelihood of someone coming into contact with these materials is also increasing.Figure 1:  How does a nanoparticle interact with a model cell membrane?

QCM-D (Quartz crystal microbalance with dissipation monitoring) can be used as a step in better understanding the interactions of nanoparticles with cells by approximating the cell surface with a supported lipid bilayer (SLB) and then flowing nanoparticles across the model cell membrane to probe the nanoparticle-cell membrane interaction. Using this cell membrane mimic allows the researcher to simplify the cell, and to investigate specific aspects of the membrane by building up the complexity. A multitude of variables from either the standpoint of the model cell membrane or the nanoparticle itself can be investigated. These include:

  • membrane composition
  • presence of membrane proteins or other constituents such as cholesterol
  • nanoparticle type
  • nanoparticle size
  • nanoparticle coating
  • pH
  • ionic strength
  • temperature
  • flow rate
  • additives in nanoparticle solution

In this post I will briefly give an introduction to this research area and highlight some key references and exciting recent work.

Supported lipid bilayer formation via QCM-D - Key references

One of the great things about this kind of research is that the assay design details are already outlined, and there are published recipes on how to form a supported lipid bilayer onto a QCM-D sensor.

The first paper illustrating how QCM-D can uniquely probe lipid vesicle deposition and the mechanism by which the vesicles can rupture to form a SLB onto a solid substrate was written by “Surface specific kinetics of lipid vesicle adsorption measured with a quartz crystal microbalance” Keller C. A.; Kasemo B. Biophys J. 1998, 75(3), 1397-1402. In this reference, three different surfaces were investigated:

  1. hydrophobic surface
  2. silicon dioxide surface and
  3. gold surface.

On the (1) hydrophobic surface, a monolayer of lipid was found to deposit. On the (3) gold surface, the vesicles adsorbed intact. Yet on the (2) silicon dioxide surface, the vesicles initially adsorbed intact until a significant surface concentration was reached and then the vesicles ruptured to form a lipid bilayer.

Several years later a description of the step by step protocols for how to form SLBs onto different surfaces including silicon dioxide, titanium dioxide, and gold were detailed in “Quartz crystal microbalance with dissipation monitoring of supported lipid bilayers on various substrates”Cho, NJ; Frank, C. W.; Kasemo, B.; Höök, F. Nature Protocols 2010, 5, 1096–1106. This reference is extremely useful for those new to lipid bilayers that are looking for detailed step by step instructions for how to prepare these kinds of surfaces.

The mechanism of supported lipid bilayer formation characterized by QCM-D

Figure 2: Example QCM-D data showing vesicle surface adsorption (B) and rupture to form a supported lipid bilayer (C).  The resonant frequency (blue line, left Y axis) of the Q-Sensor decreases as mass is bound to the surface and increases as the vesicle ruptures releasing associated water.  The dissipation (red line, right Y axis) relates to the viscoelasticity (softness or rigidity) and initially increases as the vesicles adsorb intact until a critical coverage is reached whereby the vesicles rupture and form a rigid bilayer.  The cartoon shows an intact vesicle coming into contact with the surface and then rupturing to form a SLB.  The sample is introduced at arrow 1 and washed with buffer at arrow 2.  The structure of DOPC is shown in the inset.Example QCM-D data, demonstrating lipid bilayer formation, along with a corresponding illustration, is shown in the figure to the right.

  • The lipid in this example is 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)
  • Solution of 0.25 mg/ml DOPC in PBS at 7.4 pH was prepared and flown across an SiO2 QSensor at 25°C with a flow rate of 0.1 ml/min.  
  • The 7th harmonic is shown for clarity with the arrows indicating vesicles being introduced into the system (step 1) and when the system was washed with buffer (step 2).  

"The vesicle rupture to lipid bilayer “fingerprint” is unique to the QCM-D technology"

The key feature of QCM-D data when making a SLB is the characteristic rupture step shown at (B) in the figure.  After a critical coverage of surface adsorbed vesicles is reached, or when an external stimulus is applied such as a reactive peptide or a change in osmotic pressure, the frequency increases as the mass of associated solvent inside the vesicle is lost as the vesicle ruptures and a lipid bilayer forms. This is also why the dissipation decreases, because the adsorbed layer is going from a soft viscoelastic vesicle film with quite a bit of associated solvent to a rigid lipid bilayer with little to no associated solvent. This vesicle rupture to lipid bilayer “fingerprint” is unique to the QCM-D technology and allows probing the mechanism for bilayer formation.  Surface support composition, ionic strength, pH, vesicle complexity, and constituents such as cholesterol or membrane proteins can be altered and their effects probed on how the resulting lipid bilayer forms.

Investigating the effects of nanoparticles on the SLB model cell membrane - five examples of publications

After the lipid bilayer (or intact vesicle layer) is prepared then usually a particle solution is flown across the top and the resulting signal changes correspond to how the particle interacts with the membrane surface. The effects of particle type, size, concentration, outer coating chemistry, solution ionic strength, pH, and temperature can all be measured. A number of different groups are working in this area. Below I list five different studies:

  1. Terri Camesano’s group at Worcester Polytechnic Institute recently published how both the size of a nanoparticle and the presence of natural organic matter (NOM) affects how the particle interacts with a lipid membrane. They found that the lack of NOM caused very little nanoparticle - surface interactions but that the presence of this NOM caused significant interactions leading in some cases to significant membrane removal with the largest nanoparticles tested.
    “Size dependence of gold nanoparticle interactions with a supported lipid bilayer: A QCM-D study” Christina M. Bailey, Elaheh Kamaloo, Kellie L. Waterman, Kathleen F. Wang, Ramanathan Nagarajan, Terri A. Camesano Biophysical Chemistry 2015, 203–204, 51–61. 
  2. Joel Pedersen’s group from the University of Wisconsin recently published work detailing how ordered membrane domains (or lipid rafts with phase segregated domains) affect charged nanoparticle interactions. They found that the presence of liquid ordered domains increased the attachment between positively charged particles and the underlying membrane.  They even have their own blog post about this work that can be found here:

    nanomaterial-blog-post.gifOpen access “Formation of supported lipid bilayers containing phase-segregated domains and their interaction with gold nanoparticles” E. S. Melby, et al., Environ. Sci.: Nano2016, 3, 45-55.  Published by The Royal Society of Chemistry..
  3. Kai Loon Chen’s group from Johns Hopkins University recently illustrated how the outer coating of a nanoparticle can affect the way it interacts with a SLB.  Specifically, they were interested in how an outer protein layer that could encapsulate a silver nanoparticle in the human body would affect the particle’s interaction with cells.  They found the protein layer caused disruption of the electrostatic interactions between the particle and the bilayer and therefore caused less overall interaction. 

    nanoparticles-outer-coating-affects-interaction.gifPublished with permission from “Influence of Solution Chemistry and Soft Protein Coronas on the Interactions of Silver Nanoparticles with Model Biological Membranes” Q. Wang, et al., Environ. Sci. Technol. 2016, 50 (5), 2301–2309.  Copyright 2016 American Chemical Society.”  
  4. Sofia Svedhem’s group from Chalmers Institute of Technology recently published an example of how certain nanoparticles (in this case TiO2) can actually tear holes in SLBs in the presence of calcium ions. A mechanism of the TiO2 particle – calcium ion - lipid bilayer interaction was proposed with the particles interacting with the membrane near where calcium had loosened the ionic interaction between the lipid and the surface.  The particle could then remove a portion of the bilayer due to this weakened attraction between the bilayer and surface.

    “TiO2 nanoparticle interactions with supported lipid membranes – an example of removal of membrane patches” F. Zhao, et. al., RSC Adv., 2016, 6, 91102-91110. 

  5. Nathalie Tufenkji’s group from McGill University recently published work showing how decreasing the interaction of a SLB with a substrate by tailoring the solution chemistry affected how nanoparticles interacted with said bilayer. The idea being that these free floating bilayers could potentially be more representative of an actual cell membrane and thus this method may be a better way to make and study the effects of nanoparticles with cell membranes overall. 

    Published with permission from “Toward More Free-Floating Model Cell Membranes: Method Development and Application to Their Interaction with Nanoparticles” N. Yousefi, et al., ACS Appl. Mater. Interfaces, 2016, 8 (23), pp 14339–14348.  Copyright 2016 American Chemical Society.”


Read more about the assessment and characterization of the interaction of nanomaterials with biological systems in our application note:

Application note  Screening nanoparticle - protein interactions  Download

Read review articles on nanoparticles


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