Byproducts of normal mitochondrial metabolism and homeostasis include the buildup of potentially damaging levels of reactive oxygen species (ROS), Ca2+, etc., which must be normalized. Evidence suggests that brief mitochondrial permeability transition pore (mPTP) openings play an important physiological role maintaining healthy mitochondria homeostasis. Adaptive and maladaptive responses to redox stress may involve mitochondrial channels such as mPTP and inner membrane anion channel (IMAC). Their activation causes intra- and intermitochondrial redox-environment changes leading to ROS release. This regenerative cycle of mitochondrial ROS formation and release was named ROS-induced ROS release (RIRR). Brief, reversible mPTP opening-associated ROS release apparently constitutes an adaptive housekeeping function by the timely release from mitochondria of accumulated potentially toxic levels of ROS (and Ca2+). At higher ROS levels, longer mPTP openings may release a ROS burst leading to destruction of mitochondria, and if propagated from mitochondrion to mitochondrion, of the cell itself. The destructive function of RIRR may serve a physiological role by removal of unwanted cells or damaged mitochondria, or cause the pathological elimination of vital and essential mitochondria and cells. The adaptive release of sufficient ROS into the vicinity of mitochondria may also activate local pools of redox-sensitive enzymes involved in protective signaling pathways that limit ischemic damage to mitochondria and cells in that area. Maladaptive mPTP- or IMAC-related RIRR may also be playing a role in aging. Because the mechanism of mitochondrial RIRR highlights the central role of mitochondria-formed ROS, we discuss all of the known ROS-producing sites (shown in vitro) and their relevance to the mitochondrial ROS production in vivo.

Mitochondrial Reactive Oxygen Species (ROS) and ROS-Induced ROS Release

Photoacoustic imaging (also called optoacoustic or thermoacoustic imaging) has the potential to image animal or human organs, such as the breast and the brain, with simultaneous high contrast and high spatial resolution. This article provides an overview of the rapidly expanding field of photoacoustic imaging for biomedical applications. Imaging techniques, including depth profiling in layered media, scanning tomography with focused ultrasonic transducers, image forming with an acoustic lens, and computed tomography with unfocused transducers, are introduced. Special emphasis is placed on computed tomography, including reconstruction algorithms, spatial resolution, and related recent experiments. Promising biomedical applications are discussed throughout the text, including (1) tomographic imaging of the skin and other superficial organs by laser-induced photoacoustic microscopy, which offers the critical advantages, over current high-resolution optical imaging modalities, of deeper imaging depth and higher absorptioncontrasts, (2) breast cancerdetection by near-infrared light or radio-frequency–wave-induced photoacoustic imaging, which has important potential for early detection, and (3) small animal imaging by laser-induced photoacoustic imaging, which measures unique optical absorptioncontrasts related to important biochemical information and provides better resolution in deep tissues than optical imaging.

/pdf/photoacoustic-imaging-in-biomedicine-5c6h0us7a9.pdf

Photoacoustic imaging in biomedicine

1.1 Photodynamic Therapy and Imaging
The purpose of this review is to present the current state of the role of imaging in photodynamic therapy (PDT). In order for the reader to fully appreciate the context of the discussions embodied in this article we begin with an overview of the PDT process, starting with a brief historical perspective followed by detailed discussions of specific applications of imaging in PDT. Each section starts with an overview of the specific topic and, where appropriate, ends with summary and future directions. The review closes with the authors’ perspective of the areas of future emphasis and promise. The basic premise of this review is that a combination of imaging and PDT will provide improved research and therapeutic strategies.

PDT is a photochemistry-based approach that uses a light-activatable chemical, termed a photosensitizer (PS), and light of an appropriate wavelength, to impart cytotoxicity via the generation of reactive molecular species (Figure 1a). In clinical settings, the PS is typically administered intravenously or topically, followed by illumination using a light delivery system suitable for the anatomical site being treated (Figure 1b). The time delay, often referred to as drug-light interval, between PS administration and the start of illumination with currently used PSs varies from 5 minutes to 24 hours or more depending on the specific PS and the target disease. Strictly speaking, this should be referred to as the PS-light interval, as at the concentrations typically used the PS is not a drug, but the drug-light interval terminology seems to be used fairly frequently. Typically, the useful range of wavelengths for therapeutic activation of the PS is 600 to 800 nm, to avoid interference by endogenous chromophores within the body, and yet maintain the energetics necessary for the generation of cytotoxic species (as discussed below) such as singlet oxygen (1O2). However, it is important to note that photosensitizers can also serve as fluorescence imaging agents for which activation with light in the 400nm range is often used and has been extremely useful in diagnostic imaging applications as described extensively in Section 2 of this review. The obvious limitation of short wavelength excitation is the lack of tissue penetration so that the volumes that are probed under these conditions are relatively shallow.






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Figure 1


(A) A schematic representation of PDT where PS is a photoactivatable multifunctional agent, which, upon light activation can serve as both an imaging agent and a therapeutic agent. (B) A schematic representation of the sequence of administration, localization and light activation of the PS for PDT or fluorescence imaging. Typically the PS is delivered systemically and allowed to circulate for an appropriate time interval (the “drug-light interval”), during which the PS accumulates preferentially in the target lesion(s) prior to light activation. In the idealized depiction here the PS is accumulation is shown to be entirely in the target tissue, however, even if this is not the case, light delivery confers a second layer of selectivity so that the cytotoxic effect will be generated only in regions where both drug and light are present. Upon localization of the PS, light activation will result in fluorescence emission which can be implemented for imaging applications, as well as generation cytotoxic species for therapy. In the former case light activation is achieved with a low fluence rate to generate fluorescence emission with little or no cytotoxic effect, while in the latter case a high fluence rate is used to generate a sufficient concentration of cytotoxic species to achieve biological effects.

/pdf/imaging-and-photodynamic-therapy-mechanisms-monitoring-and-76rc7acuq5.pdf

Imaging and photodynamic therapy: mechanisms, monitoring, and optimization.

There have been three basic approaches to optical tomography since the early 1980s: diffraction tomography, diffuse optical tomography and optical coherence tomography (OCT). Optical techniques are of particular importance in the medical field, because these techniques promise to be safe and cheap and, in addition, offer a therapeutic potential. Advances in OCT technology have made it possible to apply OCT in a wide variety of applications but medical applications are still dominating. Specific advantages of OCT are its high depth and transversal resolution, the fact, that its depth resolution is decoupled from transverse resolution, high probing depth in scattering media, contact-free and non-invasive operation, and the possibility to create various function dependent image contrasting methods. This report presents the principles of OCT and the state of important OCT applications. OCT synthesises cross-sectional images from a series of laterally adjacent depth-scans. At present OCT is used in three different fields of optical imaging, in macroscopic imaging of structures which can be seen by the naked eye or using weak magnifications, in microscopic imaging using magnifications up to the classical limit of microscopic resolution and in endoscopic imaging, using low and medium magnification. First, OCT techniques, like the reflectometry technique and the dual beam technique were based on time-domain low coherence interferometry depth-scans. Later, Fourier-domain techniques have been developed and led to new imaging schemes. Recently developed parallel OCT schemes eliminate the need for lateral scanning and, therefore, dramatically increase the imaging rate. These schemes use CCD cameras and CMOS detector arrays as photodetectors. Video-rate three-dimensional OCT pictures have been obtained. Modifying interference microscopy techniques has led to high-resolution optical coherence microscopy that achieved sub-micrometre resolution. This report is concluded with a short presentation of important OCT applications. Ophthalmology is, due to the transparent ocular structures, still the main field of OCT application. The first commercial instrument too has been introduced for ophthalmic diagnostics (Carl Zeiss Meditec AG). Advances in using near-infrared light, however, opened the path for OCT imaging in strongly scattering tissues. Today, optical in vivo biopsy is one of the most challenging fields of OCT application. High resolution, high penetration depth, and its potential for functional imaging attribute to OCT an optical biopsy quality, which can be used to assess tissue and cell function and morphology in situ. OCT can already clarify the relevant architectural tissue morphology. For many diseases, however, including cancer in its early stages, higher resolution is necessary. New broad-bandwidth light sources, like photonic crystal fibres and superfluorescent fibre sources, and new contrasting techniques, give access to new sample properties and unmatched sensitivity and resolution.

/pdf/optical-coherence-tomography-principles-and-applications-17y32cabbm.pdf

Optical coherence tomography - principles and applications

Author(s): Vogel, Alfred; Venugopalan, Vasan | Abstract: The mechanisms of pulsed laser ablation of biological tissues were studied. The transiently empty space created between the fiber tip and the tissue surface improved the optical transmission to the target and thus increased the ablation efficiency. It was found that the structure and morphology also affect the energy transport among tissue constituents.

/pdf/mechanisms-of-pulsed-laser-ablation-of-biological-tissues-58cltfa63z.pdf

Mechanisms of pulsed laser ablation of biological tissues.

The laser irradiation of cartilage results in a plastic deformation of the tissue allowing for the creation of new stable shapes. During photothermal stimulation, mechanically deformed cartilage undergoes a temperature dependent phase transition, which results in accelerated stress relaxation of the tissue matrix. Cartilage specimens thus reshaped can be used to recreate the underlying framework of structures in the head and neck. Optimization of this process has required an understanding of the biophysical processes accompanying reshaping and also determination of the laser dosimetry parameters, which maintain graft viability. Extensive in vitro, ex-vivo, and in vivo animal investigations, as well as human trials, have been conducted. This technology is now in use to correct septal deviations in an office-based setting. While the emphasis of clinical investigation has focused on septoplasty procedures, laser mediated cartilage reshaping may have application in surgical procedures involving the trachea, laryngeal framework, external ear, and nasal tip. Future directions for research and device design are discussed.

/pdf/cartilage-reshaping-an-overview-of-the-state-of-the-art-pot2x4o642.pdf

Cartilage reshaping: An overview of the state of the art

A phase-resolved polarization sensitive optical coherence tomography (PS-OCT) system was used to determine the burn depth of in vivo tissue non-invasively. The phase retardation information was obtained by processing the analytical interference fringe signals from two perpendicular polarization-detection channels. From the birefringence images for the four reference polarization states, the birefringence reduction in the rat skin due to thermal damage can be measured. The determined burn depth showed good agreement with histological result.

Determination of burn depth by phase-resolved polarization-sensitive optical coherence tomography

Cryogen spray cooling (CSC) is used to pre-cool the epidermis during laser dermatological procedures such as treatment of port wine stain (PWS) birthmarks. It is known that PWS patients with medium to high epidermal melanin concentrations are at a high risk of epidermal thermal damage after laser irradiation. To avoid this complication, it is necessary to maximize CSC efficiency and, thus, essential to understand the mechanical and thermal interactions of cryogen droplets with the sprayed surface. It has been observed that cryogen sprays exhibit droplet rebound as droplets impinge on the skin surface. Studies of water droplet impact on hard surfaces have shown that droplet rebound may be suppressed by dissolving small amounts (a few percent) of diverse polymer or surfactant solutions prior to atomization. To investigate the possibility of suppressing the rebound of cryogen droplets in a similar way, we have constructed a device that allows observation of the impact, spreading, and rebound of individual water and cryogen droplets with and without these solutions, and their influence on cryogen/surface dynamics and heat transfer. Our preliminary studies show that dissolving a 4% non-ionic surfactant in water reduces droplet rebound and thickness of the residual liquid layer. The maximum spread of water droplets after impact can be described within 20% accuracy by a previously developed theoretical model. The same model provides an even more accurate prediction of the maximum spread of cryogen droplets. This study will aid the analysis of future results and design conditions of new studies, which will recreate conditions to determine if added surfactant solutions suppress droplet rebound and lead to improved CSC efficiency.

Design and Construction of Experimental Device to Study Cryogen Droplet Deposition and Heat Transfer

Precise knowledge of the acoustic field in biological tissue after pulsed laser irradiation is necessary in order to perform medical imaging. Often, the photoacoustic pulse is delivered as a circular, stress confined laser spot incident upon a planar surface, such as skin. The resulting photoacoustic wave propagates in the tissue and may be detected by a detector or array of detectors. The detected signals can then be used to reconstruct the optical properties and, perhaps, the structure of the tissue. In this paper, we study the beam profile of a circular laser spot incident upon tissue
phantoms and propose a paradigm for further investigations, comparing the photoacoustic source as a baffled acoustic piston. Two phantom geometries were studied, a hemisperical acrylamide gel and a dyed water solution. An acoustic transducer with an active area of 200 μm was scanned about the circumference of the hemisphere. The laser spot was incident on the face. For the dye solution, the transducer was submerged and rotated about the laser spot on the water
surface. The beam profile of the gel showed some diffractive lobes while the profile from the dyed solution did not, though the main lobe was more robust.

Photoacoustic diffraction studies of planar tissue phantoms

Background: The chick chorioallantoic membrane (CAM) model allows direct observation of vascularization acutely in explanted or cultured tissues in an immunologically isolated environment. In vivo, angioinvasion of the tissue matrix does not occur in viable cartilage tissue, whereas denatured or nonviable grafts are readily vascularized and/or resorbed.

/pdf/angioresistance-of-thermally-modified-cartilage-grafts-in-2rk14a6hvl.pdf

J. Stuart Nelson

Papers

Cartilage reshaping: An overview of the state of the art

Determination of burn depth by phase-resolved polarization-sensitive optical coherence tomography

Design and Construction of Experimental Device to Study Cryogen Droplet Deposition and Heat Transfer

Photoacoustic diffraction studies of planar tissue phantoms

Angioresistance of thermally modified cartilage grafts in the chick chorioallantoic membrane model.