RESUMO
Significance: 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. Aim: 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. Approach: 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. Results: 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. Conclusions: 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.
Assuntos
Queimaduras , Pele , Humanos , Ratos , Animais , Pele/química , Hemoglobinas/análise , Diagnóstico por Imagem , Água , Queimaduras/diagnóstico por imagemRESUMO
INTRODUCTION: To demonstrate the feasibility of our previously proposed Diffuse reflectance spectral imaging (DRSI) method for in vivo monitoring of oxygenated hemoglobin, deoxygenated hemoglobin, methemoglobin, tissue oxygen saturation, and methemoglobin saturation in a rat scald burn wound model and assess whether the method could be used for differentiating the burn depth groups in rats based on the hemoglobin parameters. METHODOLOGY: Superficial dermal burns (SDBs), deep dermal burns (DDBs), and deep burns (DBs) were induced in rat dorsal skin using a Walker-Mason method. An approach based on multiple regression analysis for spectral diffuse reflectance images aided by Monte Carlo simulations for light transport was used to quantify the hemoglobin parameters. Canonical discriminant analysis (CDA) was performed to discriminate SDB, DDB, and DB. RESULTS: CDA using the total hemoglobin concentration, tissue oxygen saturation, and methemoglobin saturation as the independent variables showed good performance for discriminating the SDB, DDB, and DB groups immediately after burn injury and the SDB group from the DDB and DB groups 24-72 h after burn injury. CONCLUSIONS: The DRSI method with multiple regression analysis for quantification of oxygenated hemoglobin, deoxygenated hemoglobin, and methemoglobin proved to be reliable for monitoring these hemoglobin derivatives in the rat experimental burn injury model. The parameters of tissue oxygen saturation, methemoglobin saturation, and total hemoglobin concentration are promising for the differentiating the degree of burn injury using CDA.
Assuntos
Queimaduras , Metemoglobina , Ratos , Animais , Oxigênio , Queimaduras/diagnóstico por imagem , Hemoglobinas/análiseRESUMO
Prostate cancer (PCa) is known as one of the most prevalent and fatal cancer types. This report describes an MRI-compatible photoacoustic/ultrasound (PA/US) imaging platform to improve the diagnosis of PCa. In the proposed solution, PA imaging, which offers real-time, non-ionizing imaging with high sensitivity and specificity, is combined with MRI, aiming to overcome PA's limited field of view (FOV) and make PA scalable for translation to clinical settings. Central to the design of the system is a reflector-based transrectal probing mechanism composed of MRI-compatible materials. The linear transducer with a center hole for optical fiber delivery can be mechanically actuated to form a multi-angled scan, allowing PA/US imaging from varied cross-sectional views. Performance assessment was carried out in phantom and ex-vivo settings. We confirmed the MRI compatibility of the system and demonstrated the feasibility of its tri-modal imaging capability by visualizing a tubing phantom containing contrast agents. The ex-vivo evaluation of targeted tumor imaging capability was performed with a mouse liver sample expressing PSMA-positive tumors, affirming the system's compatibility in spectroscopic PA (sPA) imaging with biological tissue. These results support the feasibility of the in-bore MRI-compatible transrectal PA and US and the potential clinical adaptability.
RESUMO
BACKGROUND: We previously developed a site-specific transvascular drug delivery system (DDS) based on photomechanical waves (PMWs) or laser-induced stress/shock waves (LISWs). In this study, we investigated the validity of this method to deliver a clinical photosensitizer, talaporfin sodium (TS), to subcutaneous tumors in mice and to enhance the efficacy of photodynamic therapy (PDT). METHODS: TS solution (2.5 mg/kg) was intravenously injected into mice. Immediately thereafter, PMWs were applied to the tumor by irradiating a laser target with a Q-switched ruby laser pulse (0.8 J/cm2). Five hours after TS administration, some tumors were excised to evaluate the depth distribution of the delivered TS under a fluorescence microscope. Other tumors were subjected to PDT by irradiating the tissues with a 665 nm continuous-wave laser diode (75 mW/cm2, 667 s) at this timepoint. The effects of PDT were evaluated on the basis of the two primary therapeutic mechanisms of TS-mediated PDT: i) damage to tumor cells and ii) damage to endothelial cells of tumor vessels, i.e., the vascular shutdown effect on tumors. RESULTS: PMW application significantly increased the accumulation of TS in the tumor parenchyma but not in the tumor vessel walls; the endothelial cell junctions of tumor vessels should be the route of TS delivery enhanced by PMWs. Thus, as a result of PMW application followed by PDT, while the vascular shutdown effect on the tumors was not enhanced, direct damage to the tumor cells was increased, resulting in significant tumor growth retardation without body weight loss for 7 days after treatment.
Assuntos
Fotoquimioterapia , Porfirinas , Camundongos , Animais , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Fotoquimioterapia/métodos , Células Endoteliais , LasersRESUMO
Purpose: We previously developed a site-selective transvascular drug delivery system based on nanosecond pulsed laser-induced photomechanical waves (PMWs). In this study, we applied this method to the delivery of cisplatin (cis-diamminedichloroplatinum, CDDP) to a subcutaneous tumor in a mouse and examined its antitumor effects. Methods: A mouse tumor model with subcutaneous inoculation of human head and neck cancer cells (FaDu cells) was used. The mice were divided into four groups: control without any treatment (control), CDDP application only (CDDP only), PMW application only (PMW only) and combined application of PMWs and CDDP (PMW+CDDP). A PMW was generated by irradiating a laser target, which was placed on the skin over the tumor, with a ruby laser pulse (fluence, 1.6 J/cm2). A CDDP solution was intraperitoneally injected into the mice (2.5 mg/kg). Results: Until 7 days posttreatment, the tumor volume in the control group monotonically increased, while the tumor volumes in the CDDP-only group and PMW-only group did not change greatly and that in the PMW+CDDP group slightly decreased. Afterward, the tumors started to regrow in all treatment groups, but the tumor growth rate was considerably low in the PMW+CDDP group. There was a significant difference in the time courses of tumor volume between the PMW+CDDP group and the control group for up to 14 days posttreatment. The ratio of the Ki-67-positive (proliferative) areas to the whole tumor regions in the PMW+CDDP group was significantly smaller than that in the control group at 7 days posttreatment. These results are attributable to the synergistic effects of enhanced extravasation of CDDP and mechanical tumoricidal effect by PMWs. Conclusion: The combined application of CDDP and PMWs significantly improved the antitumor effects on mouse subcutaneous tumors.
RESUMO
For the treatment of irreversible, extensive skin damage, artificial skins or cultured skins are useful when allogeneic skins are unavailable. However, most of them lack vasculature, causing delayed perfusion and hence delay or failure in engraftment of the tissues. We previously developed a prevascularized three-dimensional (3D) cultured skin based on the layer-by-layer cell coating technique (LbL-3D skin), in which cells are seeded and laminated on a porous polymer membrane for medium supply to the thick cultured tissue. Recent animal studies have demonstrated that LbL-3D skin can achieve rapid perfusion and high graft survival after transplantation. However, there were practical issues with separating LbL-3D skins from the membranes before transplantation and the handling separated LbL-3D skins for transplantation. To address these problems, in this study, we examined the use of biodegradable porous polymer membranes that enabled the transplantation of LbL-3D skins together with the membranes, which could be decomposed after transplantation. Thin films made from poly (lactic-co-glycolic acid) (PLGA) were irradiated with femtosecond laser pulses to create micro through-holes, producing porous membranes. We designed and fabricated culture inserts with the PLGA membranes and cultivated LbL-3D skins with 2 × 106 neonatal normal human dermal fibroblasts and 1 × 104 human umbilical vein endothelial cells in the dermis of 20 cell layers and 1 × 105 neonatal human epidermal keratinocytes in the epidermis. Histological analyses revealed that the skins cultured on the PLGA membranes had thickness of about 400 µm and that there were no defects in the quality of the skins cultured on the PLGA membranes when compared with those cultured on the conventional (nonbiodegradable) commercial membranes. The cultured LbL-3D skins were then transplanted together with the PLGA membranes onto full-thickness excisional wounds in mice. At 7 days posttransplantation onto a mouse, the tissues above and below the membrane were connected through the holes with collagen-positive fibers that appeared to migrate from both the host and donor sides, and favorable reepithelization was observed throughout the transplanted skin region. However, insufficient engraftment was observed in some cases. Thus, further optimization of the membrane conditions would be needed to improve the transplantation outcome.
Assuntos
Pele Artificial , Pele , Humanos , Camundongos , Animais , Queratinócitos , Células Endoteliais da Veia Umbilical Humana , PolímerosRESUMO
Accurate burn depth assessment is crucial to determine treatment plans for burn patients. We have previously proposed a method for performing burn depth assessments based on photoacoustic (PA) imaging, and we have demonstrated the validity of this method, which allows the successful detection of PA signals originating from the blood under the bloodless burned tissue, using rat burn models. Based on these findings, we started a clinical study in which we faced two technical issues: (1) When the burn depth was shallow, PA signals due to skin contamination and/or melanin in the epidermis (surface signals) could not be distinguished from PA signals originating from the blood in the dermis; (2) the size of the system was too large. To solve these issues, we propose a burn depth diagnosis based on dual-wavelength light emitting diodes (LEDs)-excited PA imaging. The use of LEDs rendered the system compact compared to the previous one that used a conventional solid-state laser. We replicated human burned skin by applying a titrated synthetic melanin solution onto the wound surface in albino rat burn models and measured their burn depths by PA excitation at 690 and 850 nm, where melanin and haemoglobin show greatly different absorption coefficients. As a result, the surface signals were eliminated by subtracting the PA signals at 690 nm from those at 850 nm. The resultant estimated burn depths were strongly correlated with the histological assessment results. The validity of the proposed method was also examined using a burn model of rats with real melanin.
Assuntos
Queimaduras , Técnicas Fotoacústicas , Ratos , Humanos , Animais , Cicatrização , Técnicas Fotoacústicas/métodos , Melaninas , Pele/diagnóstico por imagem , Pele/patologia , Queimaduras/patologiaRESUMO
Three-dimensional (3D) cultured skin containing vascular networks is a useful skin substitute that enables rapid reperfusion after transplantation. During its cultivation, however, insufficient nutrient delivery to the thick cultured tissue from the surrounding culture medium decreases the tissue viability. To solve this problem, in this study, we applied photobiomodulation (PBM), which can optically activate the electron transport chain of mitochondria, to human 3D skin cultures constructed using the layer-by-layer cell coating technique. PBM was applied once 5 days after the start of epidermal differentiation using a light-emitting diode array with a center wavelength of 440, 523, 658 or 823 nm at a constant light intensity of 15 mW cm-2 for 50 or 600 s. Two days after PBM, we assessed the viability of the tissues by a water-soluble tetrazolium-8 assay, adenosine triphosphate measurements and live/dead cell imaging, and the results showed that the PBM at 823 nm for 50 s (0.75 J cm-2 ) significantly improved the viability of the 3D-cultured skin.
Assuntos
Terapia com Luz de Baixa Intensidade , Pele Artificial , Humanos , Diferenciação Celular , PeleRESUMO
SIGNIFICANCE: Pseudomonas(P.) aeruginosa, a common cause of infection in burns, acquires antibiotic resistance easily and forms biofilms efficiently. Thus, it is difficult to control P. aeruginosa infection in burn wounds, which causes lethal septicemia. Antimicrobial photodynamic therapy (aPDT) is attractive as a new strategy to treat burn wound infections with drug-resistant bacteria. AIM: We examined the efficacy of methylene blue (MB)-mediated aPDT with various additives in a tissue depth-resolved manner to find conditions that minimize the bacterial invasion. APPROACH: We applied MB-mediated aPDT with LED array illumination to an extensive, full-thickness burn infected with P. aeruginosa in rats for three consecutive days (days 0, 1, and 2). On day 2, the depth distributions of bacteria were assessed based on the histological analysis using Gram staining. We examined how the addition of ethylenediaminetetraacetic acid (EDTA), ethanol, and dimethyl sulfoxide (DMSO) affected the efficacy of aPDT. RESULTS: Pure MB-mediated aPDT significantly reduced the numbers of bacteria with biofilms on the wound surface and in the epidermis compared with those for the control tissue (saline only). However, there were many bacteria in the deeper region of the tissue. In contrast, MB/EDTA/ethanol/DMSO-mediated aPDT minimized the numbers of bacteria in the broad depth region of the tissue. Still, a limited number of bacteria was observed in the subcutaneous tissue. CONCLUSIONS: The depthwise analysis of bacteria demonstrated the efficacy of the MB-mediated aPDT with the addition of EDTA, ethanol, and DMSO in controlling burn wound infections. However, further improvement of the therapy is needed to suppress bacterial migration into the deep tissue completely.
Assuntos
Queimaduras , Fotoquimioterapia , Infecção dos Ferimentos , Animais , Biofilmes , Queimaduras/tratamento farmacológico , Azul de Metileno/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Pseudomonas aeruginosa , Ratos , Infecção dos Ferimentos/tratamento farmacológicoRESUMO
The state-of-the-art configurations for acoustic-resolution photoacoustic (PA) microscope (AR-PAM) are large in size and expensive, hindering their democratization. While previous research on AR-PAMs introduced a low-cost light source to reduce the cost, few studies have investigated the possibility of optimizing the sensor actuation, particularly for the AR-PAM. Additionally, there is an unmet need to evaluate the image quality deterioration associated with the actuation inaccuracy. A low-cost actuation device is introduced to reduce the system size and cost of the AR-PAM while maintaining the image quality by implementing the advanced beamformers. This work proposes an AR-RAM incorporating the delta configuration actuator adaptable from a low-cost off-the-shelf 3D printer as the sensor actuation device. The image degradation due to the data acquisition positioning inaccuracy is evaluated in the simulation. We further assess the mitigation of potential actuation precision uncertainty through advanced 3D synthetic aperture focusing algorithms represented by the Delay-and-Sum (DAS) with Coherence Factor (DAS+CF) and Delay-Multiply-and-Sum (DMAS) algorithms. The simulation study demonstrated the tolerance of image quality on actuation inaccuracy and the effect of compensating the actuator motion precision error through advanced reconstruction algorithms. With those algorithms, the image quality degradation was suppressed to within 25% with the presence of 0.2 mm motion inaccuracy. The experimental evaluation using phantoms and an ex-vivo sample presented the applicability of low-cost delta configuration actuators for AR-PAMs. The measured full width at half maximum of the 0.2 mm diameter pencil-lead phantom were 0.45 ± 0.06 mm, 0.31 ± 0.04 mm, and 0.35 ± 0.07 mm, by applying the DAS, DAS+CF, and DMAS algorithms, respectively. AR-PAMs with a compact and low-cost delta configuration provide high-quality PA imaging with better accessibility for biomedical applications. The research evaluated the image degradation contributed by the actuation inaccuracy and suggested that the advanced beamformers are capable of suppressing the actuation inaccuracy.
RESUMO
BACKGROUND AND OBJECTIVES: Control of burn wound infection is difficult due to the increase in drug-resistant bacteria and deteriorated immune responses. In this study, we examined the usefulness of methylene blue (MB)-mediated antimicrobial photodynamic therapy (aPDT) with illumination by a light-emitting diode (LED) array for controlling invasive infections from the wound to inside the body for rats with an extended deep burn infected with Pseudomonas aeruginosa. STUDY DESIGN/MATERIALS AND METHODS: An MB solution with the addition of ethanol, ethylene-diamine-tetra-acetic acid disodium salt, and dimethyl sulfoxide was used as a photosensitizer (PS). An extended deep burn was made on the dorsal skin in rats and the wounds were infected with P. aeruginosa. The rats were divided into three groups: control (no treatment; n = 14), PS mixture application alone (PS alone group; n = 10), and aPDT group (n = 14). For aPDT, after the PS mixture was applied onto the surface of infected wounds, the wounds were illuminated with a 665-nm LED array at an intensity of 45 mW/cm2 three times per treatment, with an illumination duration of 20 minutes and an interval of 10 minutes. The treatment was repeated each day for 7 consecutive days (day 0-day 6). Bacterial numbers on the wound surface and the weights and survival rates of the animals were evaluated daily. At the endpoints, bacterial numbers in the liver and blood were counted. Since the PS mixture showed high dark toxicity against P. aeruginosa in vitro, the influence of the PS mixture application onto healthy skin was also examined in vivo. RESULTS: Even in the aPDT group, rapid bacterial regrowth was observed on the wound surface after each day's treatment, but the geometric mean values of the bacterial numbers before and after each aPDT were considerably lower than those in the control group. Application of the PS mixture alone showed a clear bactericidal effect only at day 0, which is attributable to the formation of biofilms after day 1. Rats in the aPDT group showed a smaller weight loss, a higher ratio of no bacterial migration at the endpoints, and significantly higher survival rates than those in the other two groups. Effects of repeated application of the PS mixture onto healthy skin were not evident. CONCLUSIONS: Application of MB-mediated aPDT with illumination by a high-intensity LED array daily for seven consecutive days was effective for suppressing invasive infection from the wound to inside the body in rats with an extensive deep burn infected with P. aeruginosa, resulting in significant improvement of their survival. Lasers Surg. Med. © 2021 Wiley Periodicals LLC.
Assuntos
Queimaduras , Fotoquimioterapia , Infecção dos Ferimentos , Animais , Queimaduras/complicações , Queimaduras/tratamento farmacológico , Iluminação , Azul de Metileno/farmacologia , Azul de Metileno/uso terapêutico , Fármacos Fotossensibilizantes/uso terapêutico , Ratos , Infecção dos Ferimentos/tratamento farmacológicoRESUMO
Antimicrobial photodynamic treatment (aPDT) for infection with drug-resistant bacteria has received much attention. For P. aeruginosa, however, efficient formation of biofilms and the nature of Gram-negative bacteria often limit the efficacy of aPDT. In this study, we investigated the effects of ethanol and ethylenediaminetetraacetic acid (EDTA) as additives on bacterial viability, biofilm biomass, and structures of bacteria and biofilms in methylene blue (MB)-mediated aPDT in vitro. Matured P. aeruginosa biofilms were incubated with 32-µm MB solutions with different concentrations of additives and then illuminated with 665-nm light from an LED array. The combined addition of 10% ethanol and 10 mm EDTA to MB resulted in significantly greater bactericidal effects than those of MB alone and of MB with 10% ethanol or 10 mm EDTA. Crystal violet assays showed significant reductions in biofilm biomass by aPDT with addition of both ethanol and EDTA compared to that in the case of aPDT with MB alone. Scanning electron microscopy showed broken bacterial cells and reduction in the cell density and amount of biofilm under those conditions. Ethanol addition alone did not improve aPDT efficacy. Reduced amount of biofilm by EDTA addition would have improved the transportation of MB and ethanol to bacteria.
Assuntos
Pseudomonas aeruginosa , Antibacterianos/farmacologia , Biofilmes , Ácido Edético/farmacologia , Etanol/farmacologia , Azul de Metileno/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologiaRESUMO
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.
Assuntos
Fibroblastos/citologia , Queratinócitos/citologia , Pele/irrigação sanguínea , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Linhagem Celular , Células Epidérmicas/citologia , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Camundongos SCID , Neovascularização Fisiológica , Pele/citologia , Pele/ultraestrutura , Transplante de PeleRESUMO
Indocyanine green lactosome (ICG-lactosome) is an attractive new-generation agent for photodynamic therapy (PDT) that is characterized by a near-infrared excitation wavelength and high stability in the bloodstream. Fluorescence imaging has been used to examine its pharmacokinetics in vivo, but no depth-resolved information can be obtained with this method. In this study, we applied photoacoustic (PA) imaging to visualize the depth distribution of ICG-lactosome in a mouse subcutaneous tumor model. With this method, the depth distribution of blood vessels can also be visualized, enabling detection of vascular shutdown effects due to PDT. We performed PA imaging of both the distributions of ICG-lactosome and blood vessels in a tumor before and after PDT, and we found that PA signals originating from ICG-lactosome were greatly increased at 18 h after drug injection but rapidly decreased after PDT. These results indicate efficient accumulation of ICG-lactosome and rapid photobleaching due to the PDT reaction in the tumor, respectively. After PDT, PA amplitudes of hemoglobin were significantly decreased, being attributable to vascular shutdown effects. These results show the usefulness of PA imaging for monitoring not only photosensitizer accumulation and bleaching but also vascular responses in PDT with ICG-lactosome. This method can be applied to the diagnosis of many types of PDT processes.
Assuntos
Neoplasias de Cabeça e Pescoço/diagnóstico por imagem , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Verde de Indocianina/farmacologia , Técnicas Fotoacústicas , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Modelos Animais de Doenças , Feminino , Verde de Indocianina/farmacocinética , Camundongos , Camundongos Nus , Fármacos Fotossensibilizantes/farmacocinéticaRESUMO
Information on the state of edema is important for treating severe burn injuries, but a method for noninvasive real-time quantitative diagnosis of edema is not available. Thus, in vivo spatiotemporal characteristics of serum albumin, which would behave differently from water in burned tissue, are not fully understood. In this study, we used a photoacoustic (PA) imaging method to visualize depth distribution of albumin in a rat deep burn model, for which Evans blue was used as a nontoxic molecular probe. Water content in the tissue and urine volume were also measured for reference. We performed PA imaging of albumin in three regions in the rats, burn and nonburn regions and their boundary, and the imaging showed that albumin started to leak out of the vessels in the boundary and diffused within the burned tissue. Diffusion of albumin into the nonburn region, where water content was increased, was limited. In the burn and boundary regions, albumin-originating PA signal increased in two phases: immediately after making burns and from 24 to 72 h after burn. The second increase is attributable to the selective return of water to the vessels, resulting in increased concentration of albumin in extravascular tissue.
Assuntos
Queimaduras/patologia , Edema/sangue , Técnicas Fotoacústicas/métodos , Albumina Sérica/análise , Animais , Biomarcadores/sangue , Modelos Animais de Doenças , Edema/etiologia , Ratos , Pele/metabolismo , Água/análiseRESUMO
Pulsed laser interaction with small metallic and dielectric particles has been receiving attention as a method of drug delivery to many cells. However, most of the particles are attended by many risks, which are mainly dependent upon particle size. Unlike other widely used particles, biodegradable particles have advantages of being broken down and eliminated by innate metabolic processes. In this paper, the perforation of cell membrane by a focused spot with transparent biodegradable microspheres excited by a single 800 nm, 80 fs laser pulse is demonstrated. A polylactic acid (PLA) sphere, a biodegradable polymer, was used. Fluorescein isothiocyanate (FITC)-dextran and short interfering RNA were delivered into many human epithelial carcinoma cells (A431 cells) by applying a single 80 fs laser pulse in the presence of antibody-conjugated PLA microspheres. The focused intensity was also simulated by the three-dimensional finite-difference time-domain method. Perforation by biodegradable spheres compared with other particles has the potential to be a much safer phototherapy and drug delivery method for patients. The present method can open a new avenue, which is considered an efficient adherent for the selective perforation of cells which express the specific antigen on the cell membrane.