Projection mapping system for laser speckle contrast image: feasibility study for clinical application.

Significance: Laser speckle contrast images (LSCIs) have been utilized to monitor blood flow perfusion. However, they have conventionally been observed on monitor screens, resulting in potential spatial mismatching between the imaging region of interest (IROI) and monitor screen. Aim: This study proposes a projection mapping (PM) system for LSCIs (PMS_LSCI) that projects LSCIs to directly observe the blood flow perfusion in the IROI. Approach: The PMS_LSCI consists of a camera, imaging optics, a laser projector, and graphic user interface software. The spatial matching in the regions of interest was performed by adjusting the software screen of the LSCI in the IROI and evaluated by conducting in-vitro and in-vivo studies. An additional in-vivo study was performed to investigate the feasibility of real-time PM of the LSCI. Results: The spatial mismatching in the regions of interest was ranged from 2.74% to 6.47% depending on the surface curvature. The PMS_LSCI could enable real-time PM of LSCI at four different blood flow states depending on blood pressure. Conclusions: The PMS_LSCI projects the LSCI in the IROI by interacting with a projector instead of the monitor screen. The PMS_LSCI presented clinical feasibility in the in-vitro and in-vivo studies.

Handheld microfocused ultrasound device for facial lifting: A preliminary study of ULTIGHT.

OBJECTIVE: Although focused ultrasound modalities have achieved positive clinical results in noninvasive skin rejuvenation, they presented various side effects and particularly severe pain during treatment. This study introduces a microfocused ultrasound (MFU) device, ULTIGHT, to overcome the severe pain issue, providing quasi-facial lifting. MATERIALS AND METHODS: Transducer surface was imaged with a scanning electron microscope. The energies of four treatment cartridges were measured using an ultrasound power meter. In vitro experiments were performed to quantitatively evaluate the MFU thermal zones (MFUTZs) and treatment line (TL) of 10 MFUTZs. Ex vivo experiments were performed to evaluate the MFUTZs and temperature rise in tissue. Clinical trials using eight volunteers were performed to qualitatively evaluate facial lifting. RESULTS: The MFU transducer clearly showed a smooth and no air gap surface. ULTIGHT produced 10 discrete MFUTZs in a TL of length 10 mm. In ex vivo tissue, discrete linear MFUTZs were clearly observed at lower number of TLs; however, they started to aggregate at higher number of TLs. The temperature rise was linearly increased as a function of the number of treatments. A single MFUTZ resulted in a temperature rise of 3°C-10°C that could cause hyperthermia for body temperature. In the clinical trials, the volunteers showed quasi-facial lifting right after treatment on the lower facial region. CONCLUSIONS: ULTIGHT provides relatively low energy, which may be advantageous or disadvantageous depending on clinical applications. Additionally, it has the advantage of being pain-free even without anesthetic during treatment, providing quasi-facial lifting right after treatment.

Development of a Customized Endoscopic Dual-Diffusing Optical Fiber Probe for Pancreatic Cancer Therapy: Toward Clinical Use.

Objective: We developed a dual-diffusing optical fiber probe (DDOFP), capable of uniformly illuminating the anatomical structure of pancreatic duct for photodynamic therapy (PDT) of pancreatic cancer in clinical settings. Background: Optical fiber presents a unique route for pancreatic PDT by enabling access to the pancreatic duct. For effective pancreatic PDT, the optical fiber should produce a uniform illumination covering of the pancreatic duct, while maintaining its transmission property under thermomechanical stresses in surgical environments. Methods: The transmission profiles of DDOFP were measured using a charge-coupled device (CCD) camera at two directions: front-spherical and side-cylindrical areas of the optical fiber. We simulated the change in transmission property by curved tube structures using optically transparent phantom. DDOFP was integrated with 19-gauge needle catheter that is commercially used as an optical guide to treat pancreatic cancer. The temperature of DDOFP was measured at the end face using a thermistor probe in the bovine tissue, while delivering laser energy of over 200 and 500 J. Results: DDOFP was customized to secure the inner diameter of the 19-gauge needle catheter of 686 µm to be integrated as a clinical device. The round ball lens fiber tip minimized the back-burn effect caused by blood carbonization during surgery and induced front-spherical diffusion. DDOFP produced uniform light illumination with intensity difference of <10%. When DDOFP was bent with a small curvature <15 mm, the transmission intensity was consistent. Under high-power laser transmission, DDOFP was found to be robust to cracking or deformation. Conclusions: DDOFP was customized for pancreatic PDT with superior thermomechanical property and uniform light illumination at both the front-spherical and side-cylindrical areas. This is the smallest clinically available optical fiber per our knowledge and officially approved by the Korea Food and Drug Administration (item approval number: 17-516). DDOFP can contribute immensely toward the efficient delivery of pancreatic PDT and photothermal therapy.

Non-invasive postoperative monitoring of pedicled rat skin flap using laser speckle contrast imaging.

PURPOSE: This study used non-invasive laser speckle contrast imaging (LSCI) modality to evaluate the blood flow changes in murine flap model and to investigate the clinical feasibility of the LSCI for postoperative monitoring. METHODS: Ten of 6-8 weeks old Spraque-Dawley rats with superficial inferior epigastric vessel based pedicled skin flaps were used in this experiment. The color changes of skin flap were evaluated by naked eyes and the LSCI modality 6, 24, and 48 h after surgery. RESULTS: In vessel ligated region of skin flap, skin color began to change to a bluish color immediately postoperatively. At 24 h postoperatively, skin necrosis was detectable with the naked eye and total necrosis occurred at 48 h postoperatively. Changes in laser speckle signal were consistent with changes observed with the naked eye, and blood flow index also presented significant differences between the ligated and non-ligated region. CONCLUSION: These correlated laser speckle signal patterns suggest that non-invasive monitoring of perfusion by LSCI is a useful technology that may be used to identify the ischemic skin flap.

Monitoring blood-flow in the mouse cochlea using an endoscopic laser speckle contrast imaging system.

Laser speckle contrast imaging (LSCI) enables continuous high-resolution assessment of microcirculation in real-time. We applied an endoscope to LSCI to measure cochlear blood-flow in an ischemia-reperfusion mouse model. We also explored whether using xenon light in combination with LSCI facilitates visualization of anatomical position. Based on a previous preliminary study, the appropriate wavelength for penetrating the thin bony cochlea was 830 nm. A 2.7-mm-diameter endoscope was used, as appropriate for the size of the mouse cochlea. Our endoscopic LSCI system was used to illuminate the right cochlea after dissection of the mouse. We observed changes in the speckle signals when we applied the endoscopic LSCI system to the ischemia-reperfusion mouse model. The anatomical structure of the mouse cochlea and surrounding structures were clearly visible using the xenon light. The speckle signal of the cochlea was scattered, with an intensity that varied between that of the stapes (with the lowest signal), the negative control, and the stapedial artery (with the highest signal), the positive control. In the cochlear ischemia-reperfusion mouse model, the speckle signal of the cochlea decreased during the ischemic phase, and increased during the reperfusion phase, clearly reflecting cochlear blood-flow. The endoscopic LSCI system generates high-resolution images in real-time, allowing visualization of blood-flow and its changes in the mouse cochlea. Anatomical structures were clearly matched using LSCI along with visible light.

Development of single-channel optical video stereomicroscopy.

This study introduces a single-channel optical video stereomicroscope based on a transparent rotating deflector (TRD) for high-resolution and high-magnification stereomicroscopy. The existing stereomicroscopes have some limitations, such as limited resolution and magnification, fixed optical channel, and the necessity to use higher quality and higher cost optical channel components compared with the conventional optical microscopes. The goal of this study was to develop a method for improved stereo imaging and stereovision for optical microscopy. Here, we demonstrate the generation of stereo video images of left and right pairs by the refraction of light passing through a motorized TRD. We estimated the corresponding rotation angles for human stereovision and the required torque. In addition, we evaluated the image quality stability under the TRD rotation.

Fabrication of various optical tissue phantoms by the spin-coating method.

Although numerous studies have been performed to fabricate various optical tissue phantom (OTP) models, the fabrication of OTPs that simulate skin layers is laborious and time-consuming owing to the intricate characteristics of skin tissue. This study presents various OTP models that optically and structurally simulate the epidermis­dermis skin layer. The spin-coating method was employed to reproduce a uniform thin layer that mimics the epidermis layer, and the fabrication parameters were optimized for epoxy and silicone reference materials. Various OTP models simulating blood vessels and hyperpigmentation lesions were fabricated using the two reference materials to determine their feasibility. The suitability of each of the two reference materials for OTP fabrication was qualitatively evaluated by comparing the quality of the OTP models.

Sonophoresis Using Ultrasound Contrast Agents: Dependence on Concentration.

Sonophoresis can increase skin permeability to various drugs in transdermal drug delivery. Cavitation is recognized as the predominant mechanism of sonophoresis. Recently, a new logical approach to enhance the efficiency of transdermal drug delivery was tried. It is to utilize the engineered microbubble and its resonant frequency for increase of cavitation activity. Actively-induced cavitation with low-intensity ultrasound (less than ~1 MPa) causes disordering of the lipid bilayers and the formation of aqueous channels by stable cavitation which indicates a continuous oscillation of bubbles. Furthermore, the mutual interactions of microbubble determined by concentration of added bubble are also thought to be an important factor for activity of stable cavitation, even in different characteristics of drug. In the present study, we addressed the dependence of ultrasound contrast agent concentration using two types of drug on the efficiency of transdermal drug delivery. Two types of experiment were designed to quantitatively evaluate the efficiency of transdermal drug delivery according to ultrasound contrast agent concentration. First, an experiment of optical clearing using a tissue optical clearing agent was designed to assess the efficiency of sonophoresis with ultrasound contrast agents. Second, a Franz diffusion cell with ferulic acid was used to quantitatively determine the amount of drug delivered to the skin sample by sonophoresis with ultrasound contrast agents. The maximum enhancement ratio of sonophoresis with a concentration of 1:1,000 was approximately 3.1 times greater than that in the ultrasound group without ultrasound contrast agent and approximately 7.5 times greater than that in the control group. These results support our hypothesis that sonophoresis becomes more effective in transdermal drug delivery due to the presence of engineered bubbles, and that the efficiency of transdermal drug delivery using sonophoresis with microbubbles depends on the concentration of microbubbles in case stable cavitation is predominant.

Observation of vasculature alternation by intense pulsed light combined with physicochemical methods.

Intense pulsed light (IPL) with low energy insufficient to completely destroy a vasculature was applied to rabbit ears to investigate vasculature alteration. Glycerol was combined with IPL to enhance the transfer efficacy of IPL energy. Both trans-illumination and laser speckle contrast images were obtained and analyzed after treatment. The application of IPL and glycerol combination induced vasodilation and improvement in blood flow. Moreover, such phenomenon was maintained over time. IPL may be applied to treat blood circulatory diseases by inducing vasodilation and to improve blood flow.

Decreased Bone Volume and Bone Mineral Density in the Tibial Trabecular Bone Is Associated with Per2 Gene by 405 nm Laser Stimulation.

Low-level laser therapy/treatment (LLLT) using a minimally invasive laser needle system (MILNS) might enhance bone formation and suppress bone resorption. In this study, the use of 405 nm LLLT led to decreases in bone volume and bone mineral density (BMD) of tibial trabecular bone in wild-type (WT) and Per2 knockout (KO) mice. Bone volume and bone mineral density of tibial trabecular bone was decreased by 405 nm LLLT in Per2 KO compared to WT mice at two and four weeks. To determine the reduction in tibial bone, mRNA expressions of alkaline phosphatase (ALP) and Per2 were investigated at four weeks after 405 nm laser stimulation using MILNS. ALP gene expression was significantly reduced in the LLLT-stimulated right tibial bone of WT and Per2 KO mice compared to the non-irradiated left tibia (p < 0.001). Per2 mRNA expression in WT mice was significantly reduced in the LLLT-stimulated right tibial bone compared to the non-irradiated left tibia (p < 0.001). To identify the decrease in tibial bone mediated by the Per2 gene, levels of runt-related transcription factor 2 (Runx2) and ALP mRNAs were determined in non-irradiated WT and Per2 KO mice. These results demonstrated significant downregulation of Runx2 and ALP mRNA levels in Per2 KO mice (p < 0.001). Therefore, the reduction in tibial trabecular bone resulting from 405 nm LLLT using MILNS might be associated with Per2 gene expression.

Single-channel stereoscopic video imaging modality based on transparent rotating deflector.

In this study, we developed a single-channel stereoscopic video imaging modality based on a transparent rotating deflector (TRD). Sequential two-dimensional (2D) left and right images were obtained through the TRD synchronized with a camera, and the components of the imaging modality were controlled by a microcontroller unit. The imaging modality was characterized by evaluating the stereoscopic video image generation, rotation of the TRD, heat generation by the stepping motor, and image quality and its stability in terms of the structural similarity index. The degree of depth perception was estimated and subjective analysis was performed to evaluate the depth perception improvement. The results show that the single-channel stereoscopic video imaging modality may: 1) overcome some limitations of conventional stereoscopic video imaging modalities; 2) be a potential economical compact stereoscopic imaging modality if the system components can be miniaturized; 3) be easily integrated into current 2D optical imaging modalities to produce a stereoscopic image; and 4) be applied to various medical and industrial fields.

Continuous monitoring of arthritis in animal models using optical imaging modalities.

Given the several difficulties associated with histology, including difficulty in continuous monitoring, this study aimed to investigate the feasibility of optical imaging modalities­cross-polarization color (CPC) imaging, erythema index (EI) imaging, and laser speckle contrast (LSC) imaging­for continuous evaluation and monitoring of arthritis in animal models. C57BL/6 mice, used for the evaluation of arthritis, were divided into three groups: arthritic mice group (AMG), positive control mice group (PCMG), and negative control mice group (NCMG). Complete Freund's adjuvant, mineral oil, and saline were injected into the footpad for AMG, PCMG, and NCMG, respectively. LSC and CPC images were acquired from 0 through 144 h after injection for all groups. EI images were calculated from CPC images. Variations in feet area, EI, and speckle index for each mice group over time were calculated for quantitative evaluation of arthritis. Histological examinations were performed, and the results were found to be consistent with those from optical imaging analysis. Thus, optical imaging modalities may be successfully applied for continuous evaluation and monitoring of arthritis in animal models.

The effects of a minimally invasive laser needle system on complete Freund's adjuvant-induced arthritis.

The present study aimed to investigate the effects of a minimally invasive laser needle system (MILNS) on the acute progression of arthritis. Previous studies showed controversial clinical results regarding the effects of low-level laser therapy on arthritis, with the outcomes depending upon stimulation parameters such as laser wavelength and dosage. Based on the positive effects of MILNS on osteoporotic mice, we hypothesized that MILNS could potentially suppress the progression of arthritis owing to its biostimulation effects. Eight C57BL/6 mice with complete Freund's adjuvant (CFA)-induced arthritis were used as acute progression arthritis models and divided into the laser and control groups (n = 4 each). In the laser group, after minimally invasive laser stimulation, laser speckle contrast images (LSCIs) were obtained every 6 h for a total of 108 h. The LSCIs in the control group were obtained without laser stimulation. The effects of MILNS on the acute progression of arthritis were indirectly evaluated by calculating the paw area and the average laser speckle index (LSI) at the arthritis-induced area. Moreover, the macrophage population was estimated in the arthritis-induced area. Compared to the control group, the laser group showed (1) lower relative variations of the paw area, (2) lower average LSI in the arthritis-induced area, and (3) lower macrophage population in the arthritis-induced area. These results indicate that MILNS may suppress the acute progression of CFA-induced arthritis in mice and may thus be used as a potential treatment modality of arthritis in clinics.

Three-dimensional stereoscopic imaging modality based on a single optical channel and detector.

A three-dimensional stereoscopic imaging modality (3D-SIM) based on a single optical channel and detector was developed to overcome some of the limitations of conventional 3D-SIM. It produces 3-D stereoscopic images by adjusting the angle of a transparent rotating deflector (TRD) to generate disparity between left and right images. The angular effect of the TRD was demonstrated to investigate the feasibility of the proposed method in 3-D stereoscopic image generation. Results indicate that image disparity increased as a function of the rotation angles of the TRD, while maintaining adequate 3-D perception. These results are expected to facilitate the practical use of a 3D-SIM in medicine.

Fabrication of a thin-layer solid optical tissue phantom by a spin-coating method: pilot study.

Solid optical tissue phantoms (OTPs) have been widely used for many purposes. This study introduces a spin-coating method (SCM) to fabricate a thin-layer solid OTP (TSOTP) with epidermal thickness. TSOTPs are fabricated by controlling the spin speed (250 to 2500 rpm), absorber concentration (0.2% to 1.0%), and the number of layers. The results show that the thicknesses of the TSOTPs are homogeneous in the region of interest. The one-layer TSOTP achieves maximum and minimum thicknesses of 65±0.28 µm (250 rpm) and 5.1±0.17 µm (2500 rpm), respectively, decreasing exponentially as a function of the spin speed. The thicknesses of the multilayer TSOTPs increases as a function of the number of layers and are correlated strongly with the spin speed (R2≥0.95). The concentration of the OTP mixture does not directly affect the thickness of the TSOTP; however, the absorption coefficients exponentially increase as a function of absorber concentration (R2≥0.98). These results suggest that the SCM can be used to fabricate homogeneous TSOTPs with various thicknesses by controlling the spin speed and number of layers. Finally, a double-layer OTP that combines epidermal TSOTP and dermal OTP is manufactured as a preliminary study to investigate the practical feasibility of TSOTPs.

The effects of minimally invasive laser needle system on suppression of trabecular bone loss induced by skeletal unloading.

This study was aimed to evaluate the effects of low-level laser therapy (LLLT) in the treatment of trabecular bone loss induced by skeletal unloading. Twelve mice have taken denervation operation. At 2 weeks after denervation, LLLT (wavelength, 660 nm; energy, 3 J) was applied to the right tibiae of 6 mice (LASER) for 5 days/week over 2 weeks by using a minimally invasive laser needle system (MILNS) which consists of a 100 µm optical fiber in a fine needle (diameter, 130 µm) [corrected]. Structural parameters and histograms of bone mineralization density distribution (BMDD) were obtained before LLLT and at 2 weeks after LLLT. In addition, osteocyte, osteoblast, and osteoclast populations were counted. Two weeks after LLLT, bone volume fraction, trabeculae number, and trabeculae thickness were significantly increased and trabecular separations, trabecular bone pattern factor, and structure model index were significantly decreased in LASER than SHAM (p < 0.05). BMDD in LASER was maintained while that in SHAM was shifted to lower mineralization. Osteocyte and osteoblast populations were significantly increased but osteoclast population was significantly decreased in LASER when compared with those in SHAM (p < 0.05). The results indicate that LLLT with the MILNS may enhance bone quality and bone homeostasis associated with enhancement of bone formation and suppression of bone resorption.

Low-level laser therapy using the minimally invasive laser needle system on osteoporotic bone in ovariectomized mice.

This study tested the effectiveness of low-level laser therapy (LLLT) in preventing and/or treating osteoporotic trabecular bone. Mice were ovariectomized (OVX) to induce osteoporotic bone loss. The tibiae of eight OVX mice were treated for 5 days each week for 2 weeks by LLLT (660 nm, 3 J) using a minimally invasive laser needle system (MILNS) which is designed to minimize loss of laser energy before reaching bone (LASER group). Another eight mice received a sham treatment (SHAM group). Structural parameters of trabecular bone were measured with in vivo micro-computed tomography images before and after laser treatment. After LLLT for 2 weeks, the percentage reduction (%R) was significantly lower in BV/TV (bone volume fraction) and Tb.N (trabecular number, p<0.05 and p<0.05) and significant higher in Tb.Sp (trabecular separation) and SMI (structure model index, p<0.05 and p<0.05) than in the SHAM group. The %R in BV/TV at sites directly treated by LLLT was significantly lower in the LASER group than the SHAM group (p<0.05, p<0.05). These results indicated that LLLT using MILNS may be effective for preventing and/or treating trabecular bone loss and the effect may be site-dependent in the same bone.

Cross-evaluation of facial hyperpigmented lesions based on fluorescence color image and cross-polarized color image.

BACKGROUND/PURPOSE: Hyperpigmentation is a common skin problem that looks darker than normal skin regions. Accurate evaluation of a hyperpigmented lesion (HPL) is of clinical importance because proper choice of treatment can be dependent on it. This study aimed to differentiate between epidermal and dermal HPLs. METHODS: Cross-polarized color images (CPCIs) and fluorescence color images (FCIs) were acquired from the same facial regions. Contrast-limited adaptive histogram equalization (CLAHE) was employed to enhance the image contrast and a fuzzy c-means algorithm was implemented to extract the HPLs. The HPLs were superimposed to investigate the difference between CPCI and FCI. RESULTS: The HPL was successfully extracted by applying both CLAHE and fuzzy c-means algorithms. CPCI and FCI resulted in a slightly different HPL, even from the same facial region, indicating a greater percentage area of HPL in FCI than CPCI. CONCLUSION: CPCI and FCI may be utilized to differentiate HPLs that exist in different skin layers. Thus, this approach may contribute to the effective treatment of HPLs.

Real-time measurement of skin erythema variation by negative compression: pilot study.

Skin erythema has been widely used as a diagnostic parameter in dermatology. This study describes a methodology for real-time measurement of skin erythema variation induced by negative compression. This study developed an optical measurement probe, which includes a RGB color sensor that translates in the vertical direction, with the magnitude of vertical translation dependening on the amount of skin deformation. Real-time measurement of erythema variation as a function of both negative compression and time was performed in vivo on 10 measurement sites located on the back of each of 12 volunteers who participated in this study. Negative compression was sequentially applied from -30 to -80 kPa and continuously at a constant magnitude (-80 kPa) condition. The results showed that skin erythema was uniformly induced at the measurement sites and linearly increased as a function of both negative compression and time. A wide range of individual variation was noted for skin erythema, which may be due to variations in anisotropic skin properties between volunteers. This study demonstrated the clinical feasibility of a novel optical device for skin erythema measurement. Future studies are needed to investigate the clinical applications of this device.

Development of polarization dental imaging modality and evaluation of its clinical feasibility.

OBJECTIVES: In the evaluation of tooth color, the specular reflection caused by roughness or saliva on the tooth surface may cause artefacts in image analysis. In this study, a polarization dental imaging modality (PDIM) was developed to obtain cross-polarized images and, therefore, to address the problem of specular reflection. Its clinical validity was evaluated by performing 3 studies of shade tab selection for implant, plaque distribution detection, and evaluation of tooth whitening. METHODS: In vivo human tooth and shade guide color images were obtained, and the minimum color difference between them was calculated for the best color matching shade tab selection. A dental plaque disclosing agent was used to differentiate plaque regions on teeth, and plaque distribution was detected by applying the K-means algorithm. In vivo human teeth were treated with a commercial tooth whitening gel, and tooth whitening was quantitatively evaluated using the PDIM images. RESULTS: The PDIM produced repeatable glare-free tooth color images by effectively removing the specular reflection from the tooth surface. The cross-polarized tooth images were successfully utilized for shade guide selection, plaque detection, and tooth whitening by minimizing artefacts in the quantitative image analysis. The PDIM could simultaneously provide both qualitative and quantitative assessment of the tooth condition in clinical diagnosis. CONCLUSIONS: The clinical feasibility of the PDIM was successfully verified in 3 clinical studies by showing its clinical efficacy as a new imaging modality.