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1.
Cell Mol Life Sci ; 80(4): 93, 2023 Mar 16.
Article in English | MEDLINE | ID: mdl-36929461

ABSTRACT

Multicellular tumor spheroids are rapidly emerging as an improved in vitro model with respect to more traditional 2D culturing. Microwell culturing is a simple and accessible method for generating a large number of uniformly sized spheroids, but commercially available systems often do not enable researchers to perform complete culturing and analysis pipelines and the mechanical properties of their culture environment are not commonly matching those of the target tissue. We herein report a simple method to obtain custom-designed self-built microwell arrays made of polydimethylsiloxane or agarose for uniform 3D cell structure generation. Such materials can provide an environment of tunable mechanical flexibility. We developed protocols to culture a variety of cancer and non-cancer cell lines in such devices and to perform molecular and imaging characterizations of the spheroid growth, viability, and response to pharmacological treatments. Hundreds of tumor spheroids grow (in scaffolded or scaffold-free conditions) at homogeneous rates and can be harvested at will. Microscopy imaging can be performed in situ during or at the end of the culture. Fluorescence (confocal) microscopy can be performed after in situ staining while retaining the geographic arrangement of spheroids in the plate wells. This platform can enable statistically robust investigations on cancer biology and screening of drug treatments.


Subject(s)
Neoplasms , Spheroids, Cellular , Humans , Cell Line , Cell Line, Tumor
2.
Int J Mol Sci ; 23(21)2022 Nov 05.
Article in English | MEDLINE | ID: mdl-36362358

ABSTRACT

In this work, the effects of femtosecond laser irradiation and doping with plasmonic gold nanorods on the degree of conversion (DC) of a urethane dimethacrylate (UDMA)-triethylene glycol dimethacrylate (TEGDMA) nanocomposite were investigated. The UDMA-TEGDMA photopolymer was prepared in a 3:1 weight ratio and doped with dodecanethiol- (DDT) capped gold nanorods of 25 × 75 or 25 × 85 nm nominal diameter and length. It was found that the presence of the gold nanorods alone (without direct plasmonic excitation) can increase the DC of the photopolymer by 6-15%. This increase was found to be similar to what could be achieved with a control heat treatment of 30 min at 180 °C. It was also shown that femtosecond laser impulses (795 nm, 5 mJ pulse energy, 50 fs pulse length, 2.83 Jcm-2 fluence), applied after the photopolymerization under a standard dental curing lamp, can cause a 2-7% increase in the DC of undoped samples, even after thermal pre-treatment. The best DC values (12-15% increase) were obtained with combined nanorod doping and subsequent laser irradiation close to the plasmon resonance peak of the nanorods (760-800 nm), which proves that the excited plasmon field can directly facilitate double bond breakage (without thermoplasmonic effects due to the short pulse length) and increase the crosslink density independently from the initial photopolymerization process.


Subject(s)
Nanocomposites , Nanotubes , Gold , Lasers
3.
Biosensors (Basel) ; 11(8)2021 Aug 23.
Article in English | MEDLINE | ID: mdl-34436094

ABSTRACT

This review summarizes and compares the available surface treatment and bonding techniques (e.g., corona triggered surface activation, oxygen plasma surface activation, chemical gluing, and mixed techniques) and quality/bond-strength testing methods (e.g., pulling test, shear test, peel test, leakage test) for bonding PDMS (polydimethylsiloxane) with other materials, such as PDMS, glass, silicon, PET (polyethylene terephthalate), PI (polyimide), PMMA (poly(methyl methacrylate)), PVC (polyvinyl chloride), PC (polycarbonate), COC (cyclic olefin copolymer), PS (polystyrene) and PEN (polyethylene naphthalate). The optimized process parameters for the best achievable bond strengths are collected for each substrate, and the advantages and disadvantages of each method are discussed in detail.


Subject(s)
Microfluidic Analytical Techniques , Cycloparaffins , Dimethylpolysiloxanes , Microfluidics , Polycarboxylate Cement , Polymers , Polymethyl Methacrylate , Silicon , Surface Properties , Temperature
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