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"Sato, Shunichi"
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A novel strategy to engineer pre-vascularized 3-dimensional skin substitutes to achieve efficient, functional engraftment
2019
Autologous split-thickness skin grafts are the preferred treatment for excised burn wounds, but donor sites for autografting are often limited in patients with extensive burns. A number of alternative treatments are already in use to treat large burns and ulcers. Despite intense efforts to develop tissue-engineered skin, delayed or absent vascularization is one of the major reasons for tissue-engineered skin engraftment failure. To overcome these problems, we developed a scaffold-free 3-dimensional (3D) skin substitute containing vascular networks that combine dermal fibroblasts, endothelial cells, and epidermal keratinocytes based on our layer-by-layer cell coating technique. We transplanted the pre-vascularized 3D skin substitutes onto full-thickness skin defects on severe combined immunodeficiency mice to assess their integration with the host tissue and effects on wound healing. We used non-vascularized 3D skin substitutes as a control. Vessels containing red blood cells were evident in the non-vascularized control by day 14. However, blood perfusion of the human-derived vasculature could be detected within 7 days of grafting. Moreover, the pre-vascularized 3D skin substitutes had high graft survival and their epidermal layers were progressively replaced by mouse epidermis. We propose that a novel dermo-epidermal 3D skin substitute containing blood vessels can promote efficient reconstruction of full-thickness skin defects.
Journal Article
Multi-beam ultrafast laser processing of free-standing nanofilms
by
Miwa, Taito
,
Kozawa, Yuichi
,
Kadoguchi, Naohiro
in
Ablation
,
Aperture
,
Characterization and Evaluation of Materials
2023
In this study, femtosecond laser-based multi-beam interference laser processing on nanofilms with nanometer thicknesses was demonstrated. The resulting multi-hole, two-dimensional lattice pattern reflected a laser interference fringe formed on the surface of the nanofilm, with no breaks or cracks. In anticipation of the actual nanostructure fabrication, additional laser processing was performed to drill additional holes in the spaces between the existing holes, resulting in high-density multi-point hole drilling beyond the interference fringe pitch. Notably, processing materials with thicknesses close to 100 nm or less is difficult even with a state-of-the-art focused-ion-beam system. The presented method, in contrast, allows instantaneous, submicrometer-scale multi-point hole drilling of nanofilms over a large area, opening up a new frontier of nanoengineering. Future applications will include the fabrication of electron phase plates, membrane-based optomechanical devices, microelectromechanical systems, and engineering of atomic layer materials.
Journal Article
Effect of shock wave power spectrum on the inner ear pathophysiology in blast-induced hearing loss
2021
Blast exposure can induce various types of hearing impairment, including permanent hearing loss, tinnitus, and hyperacusis. Herein, we conducted a detailed investigation of the cochlear pathophysiology in blast-induced hearing loss in mice using two blasts with different characteristics: a low-frequency dominant blast generated by a shock tube and a high-frequency dominant shock wave generated by laser irradiation (laser-induced shock wave). The pattern of sensorineural hearing loss (SNHL) was low-frequency- and high-frequency-dominant in response to the low- and high-frequency blasts, respectively. Pathological examination revealed that cochlear synaptopathy was the most frequent cochlear pathology after blast exposure, which involved synapse loss in the inner hair cells without hair cell loss, depending on the power spectrum of the blast. This pathological change completely reflected the physiological analysis of wave I amplitude using auditory brainstem responses. Stereociliary bundle disruption in the outer hair cells was also dependent on the blast’s power spectrum. Therefore, we demonstrated that the dominant frequency of the blast power spectrum was the principal factor determining the region of cochlear damage. We believe that the presenting models would be valuable both in blast research and the investigation of various types of hearing loss whose pathogenesis involves cochlear synaptopathy.
Journal Article
Synthesis of Noble Metals and Their Alloy Nanoparticles by Laser-Induced Nucleation in a Highly Intense Laser Field
by
Sato, Shunichi
,
Yamazaki, Yuki
,
Nakamura, Takahiro
in
Ablation
,
Aqueous solutions
,
Catalytic activity
2022
The synthesis of noble metals and their alloy nanoparticles by laser-induced nucleation is described. Femtosecond laser pulses with an energy on the order of mJ were tightly focused to create an intensity of 1014 W/cm2 or more in an aqueous solution of noble metal ions. The intense laser field generated solvated electrons and hydrogen radicals that have a highly reducing ability, resulting in nucleation through the reduction of the noble metal ions and particle growth through ripening. This laser-induced nucleation method can be performed without any reducing agents. Excess irradiation of chloroauric acid solution led to the formation of a stable colloidal solution of gold nanoparticles without any surfactants. Additionally, the irradiation of a mixed solution of different noble metal ions formed solid–solution alloy nanoparticles, even though these metals were immiscible in the bulk. Moreover, the laser-induced nucleation made it possible to form quinary solid-solution alloy nanoparticles of noble metals. The mechanism of superior catalytic activity found for alloy nanoparticles by using Rh–Pd–Pt solid–solution nanoparticles is discussed in terms of elemental distributions inside the particles.
Journal Article
In vivo visualization of burn depth in skin tissue of rats using hemoglobin parameters estimated by diffuse reflectance spectral imaging
by
Saitoh, Daizoh
,
Nishidate, Izumi
,
Parvez, Md. Anowar
in
Animals
,
Burns - diagnostic imaging
,
Diagnostic Imaging
2024
Burn injuries represent a global public health problem that kills an estimated 180,000 people annually. Non-fatal burns result in prolonged hospitalization, disfigurement, and disability. The most common, convenient, and widely used method for assessing burn depth is physical or visual examination, but the accuracy of this method is reportedly poor (60% to 75%). Rapid, correct assessment of burn depth is very important for the optimal management and treatment of burn patients. New methods of burn depth assessment that are inexpensive, simple, rapid, non-contact, and non-invasive are therefore needed.
The aim of this study was to propose an approach to visualize the spatial distribution of burn depth using hemoglobin parameters estimated from spectral diffuse reflectance imaging and to demonstrate the feasibility of the proposed approach for differentiating burn depth in a rat model of scald burn injury.
The new approach to creating a spatial map of burn depth was based on canonical discriminant analysis (CDA) of total hemoglobin concentration, tissue oxygen saturation, and methemoglobin saturation as estimated from spectral diffuse reflectance images. Burns of three different degrees of severity were created in rat dorsal skin by 10-s exposure to water maintained at 70°C, 78°C, and 98°C, respectively. Spectral images for dorsal regions were acquired under anesthesia immediately after burn injury and at 24 h, 48 h, and 72 h after injury.
Most areas of images in the group with skin exposed to 70°C water and 98°C water were classified as 70°C burn and 98°C burn, respectively. In contrast, no significant difference between areas classified as 78°C burn and 98°C burn from 24 h to 72 h was evident in the group with skin exposed to 78°C water, suggesting that burn depth was heterogeneous.
The proposed approach combining diffuse reflectance spectral imaging and CDA appears promising for differentiating 70°C burns from 78°C burns and 98°C burns, and 98°C burns from 70°C burns and 78°C burns at 24 to 72 h after burn injury in a rat model of scald burn injury.
Journal Article
Chemical surface modification of graphene oxide by femtosecond laser pulse irradiation in aqueous suspensions
by
Muttaqin
,
Nishina, Yuta
,
Sato, Shunichi
in
absorption
,
Carbon
,
Characterization and Evaluation of Materials
2017
Reduction of graphene oxide (GO) by femtosecond laser pulse irradiation of an aqueous suspension was studied. Different laser parameters such as laser fluence and irradiation time were scanned to obtain the optimum reduced graphene oxide (rGO) with fewer defect sites and lower electrical resistivity. The fabricated rGO samples were characterized using several techniques such as X-ray diffraction, UV–Visible absorption spectrometry, Raman spectroscopy, X-ray photoelectron spectroscopy, and others. The XRD profiles of rGO revealed that the interplanar spacing between carbon layers significantly decreased to 3.51 Å, which is close to that of pristine graphite. Furthermore, the intensity ratio of
D
and
G
bands of rGO measured by Raman spectroscopy was more than 20 % smaller than that of GO, indicating the enhancement of sp
2
domains. It is noted that the defect sites and the disorder carbon double bond networks on the basal graphene plane were relatively decreased after reduction. In addition, the electrical resistivity of rGO significantly decreased to 3.3 Ω·cm under the optimum condition. From these results, femtosecond laser can be used as a suitable tool for GO reduction because it is a simple, controllable, and flexible method for getting highly reduced graphene oxide.
Journal Article
Multispectral imaging of cortical vascular and hemodynamic responses to a shock wave: observation of spreading depolarization and oxygen supply-demand mismatch
by
Kawauchi, Satoko
,
Nishidate, Izumi
,
Okuda, Wataru
in
Animals
,
Blast Injuries - diagnostic imaging
,
Blast Injuries - physiopathology
2019
Blast-induced traumatic brain injury has been a recent major concern in neurotraumatology. However, its pathophysiology and mechanism are not understood partly due to insufficient information on the brain pathophysiology during/immediately after shock wave exposure. We transcranially applied a laser-induced shock wave (LISW, ∼19 Pa · s) to the left frontal region in a rat and performed multispectral imaging of the ipsilateral cortex through a cranial window (n = 4). For the spectral data obtained, we conducted multiple regression analysis aided by Monte Carlo simulation to evaluate vascular diameters, regional hemoglobin concentration (rCHb), tissue oxygen saturation (StO2), oxygen extraction fraction, and light-scattering signals as a signature of cortical spreading depolarization (CSD). Immediately after LISW exposure, rCHb and StO2 were significantly decreased with distinct venular constriction. CSD was then generated and was accompanied by distinct hyperemia/hyperoxemia. This was followed by oligemia with arteriolar constriction, but it soon recovered (within ∼20 min). However, severe hypoxemia was persistently observed during the post-CSD period (∼1 h). These observations indicate that inadequate oxygen supply and/or excessive oxygen consumption continued even after blood supply was restored in the cortex. Such a hypoxemic state and/or a hypermetabolic state might be associated with brain damage caused by a shock wave.
Journal Article
The cause of acute lethality of mice exposed to a laser-induced shock wave to the brainstem
2022
Air embolism is generally considered the most common cause of death within 1 h of a blast injury. Shock lung, respiratory arrest, and circulatory failure caused by vagal reflexes contribute to fatal injuries that lead to immediate death; however, informative mechanistic data are insufficient. Here we used a laser-induced shock wave (LISW) to determine the mechanism of acute fatalities associated with blast injuries. We applied the LISW to the forehead, upper neck, and thoracic dorsum of mice and examined their vital signs. Moreover, the LISW method is well suited for creating site-specific damage. Here we show that only mice with upper neck exposure, without damage elsewhere, died more frequently compared with the other injured groups. The peripheral oxygen saturation (SpO
2
) of the former mice significantly decreased for < 1 min [
p
< 0.05] but improved within 3 min. The LISW exposure to the upper neck region was the most lethal factor, affecting the respiratory function. Protecting the upper neck region may reduce fatalities that are related to blast injuries.
Journal Article
Comparison of Therapeutic Effects between Pulsed and Continuous Wave 810-nm Wavelength Laser Irradiation for Traumatic Brain Injury in Mice
by
Hamblin, Michael R.
,
Xuan, Weijun
,
Whalen, Michael J.
in
Adenosine Triphosphate - metabolism
,
Analysis
,
Animals
2011
Transcranial low-level laser therapy (LLLT) using near-infrared light can efficiently penetrate through the scalp and skull and could allow non-invasive treatment for traumatic brain injury (TBI). In the present study, we compared the therapeutic effect using 810-nm wavelength laser light in continuous and pulsed wave modes in a mouse model of TBI.
TBI was induced by a controlled cortical-impact device and 4-hours post-TBI 1-group received a sham treatment and 3-groups received a single exposure to transcranial LLLT, either continuous wave or pulsed at 10-Hz or 100-Hz with a 50% duty cycle. An 810-nm Ga-Al-As diode laser delivered a spot with diameter of 1-cm onto the injured head with a power density of 50-mW/cm(2) for 12-minutes giving a fluence of 36-J/cm(2). Neurological severity score (NSS) and body weight were measured up to 4 weeks. Mice were sacrificed at 2, 15 and 28 days post-TBI and the lesion size was histologically analyzed. The quantity of ATP production in the brain tissue was determined immediately after laser irradiation. We examined the role of LLLT on the psychological state of the mice at 1 day and 4 weeks after TBI using tail suspension test and forced swim test.
The 810-nm laser pulsed at 10-Hz was the most effective judged by improvement in NSS and body weight although the other laser regimens were also effective. The brain lesion volume of mice treated with 10-Hz pulsed-laser irradiation was significantly lower than control group at 15-days and 4-weeks post-TBI. Moreover, we found an antidepressant effect of LLLT at 4-weeks as shown by forced swim and tail suspension tests.
The therapeutic effect of LLLT for TBI with an 810-nm laser was more effective at 10-Hz pulse frequency than at CW and 100-Hz. This finding may provide a new insight into biological mechanisms of LLLT.
Journal Article
Single-scan volumetric imaging throughout thick tissue specimens by one-touch installable light-needle creating device
by
Yamasaki, Miwako
,
Sato, Shunichi
,
Nemoto, Tomomi
in
631/1647/245/2225
,
631/1647/328/2057
,
631/1647/328/2235
2022
Biological tissues and their networks frequently change dynamically across large volumes. Understanding network operations requires monitoring their activities in three dimensions (3D) with single-cell resolution. Several researchers have proposed various volumetric imaging technologies. However, most technologies require large-scale and complicated optical setups, as well as deep expertise for microscopic technologies, resulting in a high threshold for biologists. In this study, we propose an easy-to-use light-needle creating device for conventional two-photon microscopy systems. By only installing the device in one position for a filter cube that conventional fluorescent microscopes have, single scanning of the excitation laser light beam excited fluorophores throughout over 200 μm thickness specimens simultaneously. Furthermore, the developed microscopy system successfully demonstrated single-scan visualization of the 3D structure of transparent YFP-expressing brain slices. Finally, in acute mouse cortical slices with a thickness of approximately 250 μm, we detected calcium activities with 7.5 Hz temporal resolution in the neuronal population.
Journal Article