An In-House-Built and Light-Emitting-Diode-Based Photodynamic Therapy Device for Enhancing Verteporfin Cytotoxicity in a 2D Cell Culture Model.

This paper describes a novel, simple, and low-cost device to perform in vitro photodynamic therapy (PDT) assays, named the PhotoACT. The device was built using a set of conventional programmable light-emitting diodes (LEDs), a liquid crystal display (LCD) module, and a light sensor connected to a commercial microcontroller board. The box-based structure of the prototype was made with medium-density fiberboards (MDFs). The internal compartment can simultaneously allocate four cell culture multiwell microplates. As a proof of concept, we studied the cytotoxic effect of the photosensitizer (PS) verteporfin against the HeLa cell line in two-dimensional (2D) culture. HeLa cells were treated with increasing concentrations of verteporfin for 24 h. The drug-containing supernatant medium was discarded, the adherent cells were washed with phosphate-buffered saline (PBS), and drug-free medium was added. In this study, the effect of verteporfin on cells was examined either without light exposure or after exposure for 1 h to light using red-green-blue (RGB) values of 255, 255, and 255 (average fluence of 49.1 ± 0.6 J/cm2). After 24 h, the cell viability was assessed by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay. Experimental results showed that exposure of cells treated with verteporfin to the light from the device enhances the drug's cytotoxic effect via a mechanism mediated by reactive oxygen species (ROS). In addition, the use of the prototype described in this work was validated by comparing the results with a commercial PDT device. Thus, this LED-based photodynamic therapy prototype represents a good alternative for in vitro studies of PDT.

A method for generating 3D thermal models with decoupled acquisition.

BACKGROUND AND OBJECTIVE: Both thermal imaging and 3D scanning offer convenient advantages for medical applications, namely, being contactless, non-invasive and fast. Consequently, many approaches have been proposed to combine both sensing modalities in order to acquire 3D thermal models. The predominant approach is to affix a 3D scanner and a thermal camera in the same support and calibrate them together. While this approach allows straightforward projection of thermal images over the 3D mesh, it requires their simultaneous acquisition. In this work, a method for generation of 3D thermal models that allows combination of separately acquired 3D mesh and thermal images is presented. Among the advantages of this decoupled acquisition are increased modularity of acquisition procedures and reuse of legacy equipment and data. METHODS: The proposed method is based on the projection of thermal images over a 3D mesh. Unlike previous methods, it is considered that the 3D mesh and the thermal images are acquired separately, so camera pose estimation is required to determine the correct spatial positioning from which to project the images. This is done using Structure from Motion, which requires a series of interest points correspondences between the images, for which the SIFT method was used. As thermal images of human skin are predominantly homogeneous, an intensity transformation is proposed to increase the efficacy of interest point detection and make the approach feasible. Before projection, the adequate alignment of the 3D mesh in space is determined using Particle Swarm Optimization. For validation of the method, the design and implementation of a test object is presented. It can be used to validate other methods and can be reproduced with common printed circuit board manufacturing processes. RESULTS: The proposed approach is accurate, with an average displacement error of 1.41  mm (s = 0.74  mm) with the validation test object and 4.58 mm (s = 2.12  mm) with human subjects. CONCLUSIONS: The proposed method is able to combine separately a acquired 3D mesh and thermal images into an accurate 3D thermal model. The results with human subjects suggest that the method can be successfully employed in medical applications.