Abstract
In this study, a paper-integrated analytical device that combined a paper-based colorimetric assay with a paper-based cell culture platform was developed for the in situ detection of hydrogen sulfide (H2S) in three-dimensional (3D)-cultured, live prostate cancer cells. Two kinds of paper substrates were fabricated using a simple wax-printing methodology to form the cell culture and detection zones, respectively. LNCaP cells were seeded directly on the paper substrate and grown in the paper-integrated analytical device. The cell viability and H2S production of LNCaP cells were assessed using a simple water-soluble tetrazolium salt colorimetric assay and H2S-sensing paper, respectively. The H2S-sensing paper showed good sensitivity (sensitivity: 6.12 blue channel intensity/μM H2S, R2 = 0.994) and a limit of quantification of 1.08 μM. As a result, we successfully measured changes in endogenous H2S production in 3D-cultured, live LNCaP cells within the paper-integrated analytical device while varying the duration of incubation and substrate concentration (L-cysteine). This paper-integrated analytical device can provide a simple and effective method to investigate H2S signaling pathways and drug screening in a 3D culture model.
Highlights
Prostate cancer is the second most frequently occurring cancer in men and the fifth leading cause of death worldwide [1]
We introduce a paper-integrated analytical device that combines a paperbased colorimetric assay with a paper-based cell culture platform for the in situ detection of Hydrogen sulfide (H2 S) in 3D-cultured prostate cancer LNCaP cells
Given its porous internal microstructure and diverse surface morphology, paper is an interesting alternative to traditional 3D cell culture with hydrogels or porous scaffolds
Summary
Prostate cancer is the second most frequently occurring cancer in men and the fifth leading cause of death worldwide [1]. Prostate cancer is a hormonally regulated malignancy, and androgen receptor signaling is essential for its evolution from the beginning androgendependent state to the late aggressive androgen-resistant state [2]. Androgen deprivation therapy (ADT) is the main treatment option for early-stage prostate cancer. ADT is palliative, and prostate cancer eventually progresses to an androgen-independent state that is more aggressive and fatal, for which hormone blockade therapy fails [3]. As no effective therapy for prostate cancer is available, new diagnostic methods and preventive interventions must be developed urgently to lower morbidity, mortality, and medical costs related with this tumor. Hydrogen sulfide (H2 S) acts as a gaseous transmitter in organisms, and abnormal
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