scholarly journals Applicability and Limitations in the Characterization of Poly-Dispersed Engineered Nanomaterials in Cell Media by Dynamic Light Scattering (DLS)

Materials ◽  
2019 ◽  
Vol 12 (23) ◽  
pp. 3833 ◽  
Author(s):  
Arianna Marucco ◽  
Elisabetta Aldieri ◽  
Riccardo Leinardi ◽  
Enrico Bergamaschi ◽  
Chiara Riganti ◽  
...  
Keyword(s):  
Light Scattering ◽  
Dynamic Light Scattering ◽  
Culture Media ◽  
Raw 264.7 ◽  
Pre Treatment ◽  
The Stability ◽  
Definition Of ◽  
Cellular Tests

The dispersion protocol used to administer nanomaterials (NMs) in in vitro cellular tests might affect their toxicity. For this reason, several dispersion procedures have been proposed to harmonize the toxicological methods, allowing for the comparison of the data that were obtained by different laboratories. At the same time, several techniques and methods are available to monitor the identity of the NMs in the cell media. However, while the characterization of suspensions of engineered NMs having narrow size distribution may be easily performed, the description of aggregated NMs forming polydispersions is still challenging. In the present study, sub-micrometric/nanometric TiO2, SiO2, and CeO2 were dispersed in cell media by using two different dispersion protocols, with and without albumin (0.5%) and with different sonication procedures. Dynamic Light Scattering (DLS) was used to characterize NMs in stock solutions and culture media. Pitfalls that affect DLS measurements were identified and, guidance on a critical analysis of the results provided. The NMs were then tested for their cytotoxicity (LDH leakage) toward murine macrophages (RAW 264.7) and PMA-activated human monocytes (THP-1). As markers of pro-inflammatory response, nitric oxide (NO) and cytokine IL-1β production were measured on RAW 264.7 and THP-1 cells, respectively. The pre-treatment with albumin added to a strong sonication treatment increases the stability and homogeneity of the suspensions of nanometric samples, but not of the submicrometric-samples. Nevertheless, while TiO2 and CeO2 were non-cytotoxic in any conditions, differences in cytotoxicity, NO, and IL-1β releases were found for the SiO2, depending upon the protocol. Overall, the results suggest that there is no one-fits-all method valid for all NMs, since each class of NMs respond differently. The definition of validated procedures and parameters for the selection of the most appropriate method of dispersion for each class of NM appears to be a more efficacious strategy for the harmonization of the dispersion protocols.

scholarly journals Characterization of Nanomaterial Dispersion in Solution Prior to In Vitro Exposure Using Dynamic Light Scattering Technique

2007 ◽  
Vol 101 (2) ◽  
pp. 239-253 ◽  
Author(s):  
Richard C. Murdock ◽  
Laura Braydich-Stolle ◽  
Amanda M. Schrand ◽  
John J. Schlager ◽  
Saber M. Hussain
Keyword(s):  
Light Scattering ◽  
Dynamic Light Scattering ◽  
In Vitro Exposure ◽  
Scattering Technique ◽  
Dynamic Light Scattering Technique ◽  
Light Scattering Technique

scholarly journals Characterization of gelification of chitosan solutions by dynamic light scattering

2012 ◽  
Vol 87 (4) ◽  
pp. 2376-2380 ◽  
Author(s):  
W.A. de Morais ◽  
M.R. Pereira ◽  
J.L.C. Fonseca
Keyword(s):  
Light Scattering ◽  
Dynamic Light Scattering

scholarly journals Characterization of Recombinant Adeno-Associated Viruses (rAAVs) for Gene Therapy Using Orthogonal Techniques

Pharmaceutics ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 586
Author(s):  
Liam Cole ◽  
Diogo Fernandes ◽  
Maryam T. Hussain ◽  
Michael Kaszuba ◽  
John Stenson ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Light Scattering ◽  
Dynamic Light Scattering ◽  
Viral Vector ◽  
Structural Characteristics ◽  
Genetic Material ◽  
Label Free ◽  
Gene Therapies ◽  
Multiple Challenges

Viruses are increasingly used as vectors for delivery of genetic material for gene therapy and vaccine applications. Recombinant adeno-associated viruses (rAAVs) are a class of viral vector that is being investigated intensively in the development of gene therapies. To develop efficient rAAV therapies produced through controlled and economical manufacturing processes, multiple challenges need to be addressed starting from viral capsid design through identification of optimal process and formulation conditions to comprehensive quality control. Addressing these challenges requires fit-for-purpose analytics for extensive characterization of rAAV samples including measurements of capsid or particle titer, percentage of full rAAV particles, particle size, aggregate formation, thermal stability, genome release, and capsid charge, all of which may impact critical quality attributes of the final product. Importantly, there is a need for rapid analytical solutions not relying on the use of dedicated reagents and costly reference standards. In this study, we evaluate the capabilities of dynamic light scattering, multiangle dynamic light scattering, and SEC–MALS for analyses of rAAV5 samples in a broad range of viral concentrations (titers) at different levels of genome loading, sample heterogeneity, and sample conditions. The study shows that DLS and MADLS® can be used to determine the size of full and empty rAAV5 (27 ± 0.3 and 33 ± 0.4 nm, respectively). A linear range for rAAV5 size and titer determination with MADLS was established to be 4.4 × 1011–8.7 × 1013 cp/mL for the nominally full rAAV5 samples and 3.4 × 1011–7 × 1013 cp/mL for the nominally empty rAAV5 samples with 3–8% and 10–37% CV for the full and empty rAAV5 samples, respectively. The structural stability and viral load release were also inferred from a combination of DLS, SEC–MALS, and DSC. The structural characteristics of the rAAV5 start to change from 40 °C onward, with increasing aggregation observed. With this study, we explored and demonstrated the applicability and value of orthogonal and complementary label-free technologies for enhanced serotype-independent characterization of key properties and stability profiles of rAAV5 samples.

Noninvasive characterization of metabolites secreted in culture media by bovine embryos during in vitro production

Metabolomics ◽  
2016 ◽  
Vol 12 (5) ◽  
Author(s):  
Érika Cristina dos Santos ◽  
Camila Bruna de Lima ◽  
Kelly Annes ◽  
Marcella Pecora Milazzotto
Keyword(s):  
Culture Media ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Noninvasive Characterization ◽  
Vitro Production

scholarly journals Detection Limits of DLS and UV-Vis Spectroscopy in Characterization of Polydisperse Nanoparticles Colloids

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Emilia Tomaszewska ◽  
Katarzyna Soliwoda ◽  
Kinga Kadziola ◽  
Beata Tkacz-Szczesna ◽  
Grzegorz Celichowski ◽  
...  
Keyword(s):  
Light Scattering ◽  
Dynamic Light Scattering ◽  
Point Of View ◽  
Spectroscopy Analysis ◽  
Force Microscopy ◽  
Atomic Force ◽  
Transmission Electron ◽  
Vis Spectroscopy ◽  
The One

Dynamic light scattering is a method that depends on the interaction of light with particles. This method can be used for measurements of narrow particle size distributions especially in the range of 2–500 nm. Sample polydispersity can distort the results, and we could not see the real populations of particles because big particles presented in the sample can screen smaller ones. Although the theory and mathematical basics of DLS technique are already well known, little has been done to determine its limits experimentally. The size and size distribution of artificially prepared polydisperse silver nanoparticles (NPs) colloids were studied using dynamic light scattering (DLS) and ultraviolet-visible (UV-Vis) spectroscopy. Polydisperse colloids were prepared based on the mixture of chemically synthesized monodisperse colloids well characterized by atomic force microscopy (AFM), transmission electron microscopy (TEM), DLS, and UV-Vis spectroscopy. Analysis of the DLS results obtained for polydisperse colloids reveals that several percent of the volume content of bigger NPs could screen completely the presence of smaller ones. The presented results could be extremely important from nanoparticles metrology point of view and should help to understand experimental data especially for the one who works with DLS and/or UV-Vis only.

Characterization of amphiphilic poly-N-vinylpyrrolidone derivatives by dynamic light scattering

2006 ◽  
Vol 352 (42-49) ◽  
pp. 5055-5059 ◽  
Author(s):  
Apostolos K. Rizos ◽  
Ioannis Tsikalas ◽  
Aristidis M. Tsatsakis ◽  
Michail I. Shtilman
Keyword(s):  
Light Scattering ◽  
Dynamic Light Scattering
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