Response Surface Methodology to Efficiently Optimize Intracellular Delivery by Photoporation

Photoporation is an up-and-coming technology for the gentle and efficient transfection of cells. Inherent to the application of photoporation is the optimization of several process parameters, such as laser fluence and sensitizing particle concentration, which is typically done one factor at a time...

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Bibliographic Details
Published in:International journal of molecular sciences 2023-02, Vol.24 (4), p.3147
Main Authors: Goemaere, Ilia, Punj, Deep, Harizaj, Aranit, Woolston, Jessica, Thys, Sofie, Sterck, Karen, De Smedt, Stefaan C, De Vos, Winnok H, Braeckmans, Kevin
Format: Article
Language:English
Subjects:
Analysis
Animals
Case studies
cell therapy
Design of experiments
Dextran
Dextrans
Fluence
Graphene
intracellular delivery
Lasers
Light
Macrophages
Methods
Mice
Nanoparticles
Optimization
photoporation
polydopamine nanoparticles
Process parameters
Quantum dots
Response surface methodology
Scanning electron microscopy
Surface analysis (chemical)
Technology application
Transfection
Variance analysis
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Summary:Photoporation is an up-and-coming technology for the gentle and efficient transfection of cells. Inherent to the application of photoporation is the optimization of several process parameters, such as laser fluence and sensitizing particle concentration, which is typically done one factor at a time (OFAT). However, this approach is tedious and runs the risk of missing a global optimum. Therefore, in this study, we explored whether response surface methodology (RSM) would allow for more efficient optimization of the photoporation procedure. As a case study, FITC-dextran molecules of 500 kDa were delivered to RAW264.7 mouse macrophage-like cells, making use of polydopamine nanoparticles (PDNPs) as photoporation sensitizers. Parameters that were varied to obtain an optimal delivery yield were PDNP size, PDNP concentration and laser fluence. Two established RSM designs were compared: the central composite design and the Box-Behnken design. Model fitting was followed by statistical assessment, validation, and response surface analysis. Both designs successfully identified a delivery yield optimum five- to eight-fold more efficiently than when using OFAT methodology while revealing a strong dependence on PDNP size within the design space. In conclusion, RSM proves to be a valuable approach to efficiently optimize photoporation conditions for a particular cell type.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms24043147