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In vivo multiphoton microscopy detects longitudinal metabolic changes associated with delayed skin wound healing
摘要： Chronic wounds are difficult to diagnose and characterize due to a lack of quantitative biomarkers. Label-free multiphoton microscopy has emerged as a useful imaging modality capable of quantifying changes in cellular metabolism using an optical redox ratio of FAD/(NADH+FAD) autofluorescence. However, the utility of an optical redox ratio for long-term in vivo monitoring of tissue metabolism has not been robustly evaluated. In this study, we demonstrate how multiphoton microscopy can be used to monitor changes in the metabolism of individual full-thickness skin wounds in vivo. 3D optical redox ratio maps and NADH fluorescence lifetime images identify differences between diabetic and control mice during the re-epithelialization of wounds. These metabolic changes are associated with a transient increase in keratinocyte proliferation at the wound edge. Our study demonstrates that high-resolution, non-invasive autofluorescence imaging can be performed in vivo and that optical redox ratios can serve as quantitative optical biomarkers of impaired wound healing.
关键词： metabolism,optical redox ratio,autofluorescence,multiphoton microscopy,in vivo imaging,diabetes,FAD,NADH,wound healing
更新于2025-09-23 15:23:52
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Live imaging of angiogenesis during cutaneous wound healing in adult zebrafish
摘要： Angiogenesis, the growth of new blood vessels from pre-existing vessels, is critical for cutaneous wound healing. However, it remains elusive how endothelial cells (ECs) and pericytes (PCs) establish new blood vessels during cutaneous angiogenesis. We set up a live-imaging system to analyze cutaneous angiogenesis in adult zebrafish. First, we characterized basic structures of cutaneous vasculature. In normal skin tissues, ECs and PCs remained dormant to maintain quiescent blood vessels, whereas cutaneous injury immediately induced angiogenesis through the vascular endothelial growth factor signaling pathway. Tortuous and disorganized vessel networks formed within a few weeks after the injury and subsequently normalized through vessel regression in a few months. Analyses of the repair process of injured single blood vessels revealed that severed vessels elongated upon injury and anastomosed with each other. Thereafter, repaired vessels and adjacent uninjured vessels became tortuous by increasing the number of ECs. In parallel, PCs divided and migrated to cover the tortuous blood vessels. ECs sprouted from the PC-covered tortuous vessels, suggesting that EC sprouting does not require PC detachment from the vessel wall. Thus, live imaging of cutaneous angiogenesis in adult zebrafish enables us to clarify how ECs and PCs develop new blood vessels during cutaneous angiogenesis.
关键词： Pericytes,Cutaneous wound healing,Angiogenesis,Endothelial cells,Zebrafish
更新于2025-09-23 15:23:52
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Evaluation of Tear Film Lipid Layer Thickness Measurements Obtained Using an Ocular Surface Interferometer in Nasolacrimal Duct Obstruction Patients
摘要： Pre-vascularised cell sheets have been used to promote early angiogenesis and graft survival. However, the use of pre-vascularised mucosal cell sheets for burn wounds has been rarely evaluated. Therefore, we examined the applicability of an oral pre-vascularised mucosal cell sheet that we had previously developed for the treatment of cutaneous burn wounds. Methods: Mucosal keratinocytes, fibroblasts, and endothelial progenitor cells were isolated from the oral mucosa and peripheral blood and were expanded in vitro. Mucosal cell sheets were generated by seeding cultured keratinocytes onto a mixture of fibroblasts, endothelial cells, and fibrin. Third-degree burn wounds were created on the backs of rats and were covered with the cell sheets, skin grafts, or silastic sheets as a control. Gross and microscopic findings and gene expression profiles of wounds were compared among the groups. Results: CD31-positive microvessels were observed in the fibrin-matrix layer of the cell sheet. In the cutaneous burn wound model, the cell sheets promoted wound healing, with accelerated wound closure and less scarring than with silastic sheets and skin grafts. The cell sheets had more microvessels and proliferating cells and less neutrophil infiltration and fibrotic features than the controls or skin grafts. The cell sheet induced higher mRNA expression of KRT14, VEGFA, IL10, and AQP3 and lower mRNA expression of TGFB1, IL6, ICAM1, ACTA2, and FN1 than did the controls or skin grafts. Conclusions: The pre-vascularised mucosal cell sheet promotes cutaneous burn wound healing.
关键词： Skin,wound healing,cell sheet,oral mucosa,pre-vascularisation
更新于2025-09-23 15:22:29
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Volumetric measurement of dentoalveolar defects by means of intraoral 3D scanner and gravimetric model
摘要： The aim of this study was to evaluate the accuracy in volumetric measurements obtained on an experimental model using an intraoral scanner and a gravimetric method. Three identical partial dentate maxillary acrylic models with three fabricated alveolar defects, in anterior and posterior regions, were scanned using an intraoral scanner (20 scans/defects). The defects differed in terms of size and distance of neighbouring teeth. As references, replicas of each defect were created using a dimensional stable silicone impression material. After measuring the mass of each replica, the volume was calculated by dividing the mass of each replica by the density of the impression material. The defects had a volume, according to the gravimetric method, ranging from 40.5 to 143.7 mm3. The scans were imported to metrology software for analyses. Accuracy was determined in terms of trueness and precision. The mean trueness for all defect types was 0.168 mm3 (SD 0.691, range 2.82). There was no statistical significant difference between the mean trueness for all defects measured (p = 0.910). The mean precision for all defect types was 0.147 mm3 (SD 0.524, range 2.86). There were no statistical significant differences between the dental models in regard to mean precision (p = 0.401), however, there were statistical significant differences between defects in position 1 and 2 (p = 0.002) and 1 and 3 (p = 0.001). Based on the findings of this study, the intraoral scanner utilized in the current study presented an acceptable level of accuracy when measuring volume of defects.
关键词： Volumetric measurement,Intraoral scanning,Surgical techniques,Wound healing,Tissue alterations
更新于2025-09-23 15:22:29
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Effects of Carbon Arc Lamp Irradiation on Wound Healing in a Rat Cutaneous Full-Thickness Wound Model
摘要： Objective: The objective of the present study was to investigate the application of a carbon arc lamp on wound healing in a rat cutaneous full-thickness wound model. Background data: In clinical practice, wound healing has been promoted by irradiation with a carbon arc lamp. However, the corresponding mechanism has not been clearly defined. Methods: A cutaneous full-thickness wound on the back of rats was irradiated using a carbon arc lamp at a wavelength peak range of 620–740 nm with 54 J/cm2. Injured sham-irradiated control rats were used as the control. The rats were euthanized after 7, 14, and 21 days, while wound reepithelialization and healing quality were examined by histological analyses with comparison between groups. Cell proliferation was observed by 5-bromo-2￠-deoxyuridine (BrdU) immunohistochemical staining. Results: Irradiation by the carbon arc lamp significantly accelerated wound healing. The wound-healing rate in the treated group at day 21 was 98.42% – 0.56%, compared with 93.58% – 1.26% in the control group ( p < 0.05). Significant increases in the length of epithelial edges, collagen content, and microvessel density were observed in the wound sites in the treated group at days 7, 14, and 21 ( p < 0.05). Moreover, the number of BrdU-labeled cells increased in the wound edge at days 7 and 14 due to irradiation ( p < 0.05). Conclusions: The results demonstrated that the carbon arc lamp can promote wound healing together with improvement in its quality by stimulating cell proliferation.
关键词： wound healing,cell proliferation,carbon arc lamp
更新于2025-09-23 15:22:29
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Wound healing of laser injured skin with glycerol monooleicate cubic liquid crystal
摘要： Laser has found increasingly wider applications in the medical filed, but laser is likely to cause damage to patients’ skin. In this experiment, we were surprised to find that glyceryl monooleate (GMO)-based cubic liquid crystal had excellent healing effect on the skin of guinea pigs damaged by laser. Transepidermal water loss (TEWL), H.E. pathology, Masson trichrome dyeing, interleukin-6 (IL-6) levels and the percutaneous depth of fluorescein isothiocyanate (FITC) dyeing were used to evaluate the therapeutic effect of GMO-based cubic liquid crystals against laser damage of different degrees among guinea pigs. GMO-based cubic liquid crystals had an obvious effect in the treatment of slight and moderate laser damage. This finding may provide a effective medical treatment protocols for laser skin damage.
关键词： Cubic liquid crystal,Wound healing,Glyceryl monooleate (GMO),Laser
更新于2025-09-23 15:21:01
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Lack of relationship between epidermal denervation by capsaicin and incisional pain behaviours: A laser scanning confocal microscopy study in rats
摘要： BACKGROUND: Spontaneous pain after surgical incision is a significant problem for most post-operative patients. Pain management that relies on opioids is hindered by numerous side effects, fueling interest in non-opioid alternatives and multimodal approaches. Subcutaneous capsaicin infiltration has shown potential for reducing post-operative pain, but there are unanswered questions about safety and possible side effects. In adult rats, we characterized the analgesic effects of pre-operative capsaicin infiltration into the skin prior to plantar incision and assessed wound healing and epidermal innervation. METHODS: The surgical site on the plantar surface of the rat hind paw was infiltrated with 1% capsaicin or vehicle 30 minutes or 1 week prior to surgical incision. Spontaneous and evoked pain behaviors were assessed. Digital images of incised hind paws were used to quantify the surface area of the wound after suture removal. Epidermal nerve fiber quantification was performed on peri-incisional tissue biopsies. RESULTS: Intraplantar administration of capsaicin 30 minutes before surgical incision attenuated spontaneous pain behaviors, heat hyperalgesia, epidermal innervation, but it did not alter the rate of wound healing. Incisional pain hypersensitivity returned to baseline 2 weeks post-incision, at a time when no recovery of epidermal innervation is observed. CONCLUSIONS: Subcutaneous infiltration of capsaicin prior to surgical incision attenuated incision-induced pain behaviors and reduced epidermal innervation around the incision site. The long-lasting epidermal denervation by capsaicin had no impact in the rate of wound healing and recovery from pain behaviors.
关键词： epidermal innervation,capsaicin,rats,wound healing,post-operative pain
更新于2025-09-23 15:21:01
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Flexible organic lighta??emittinga??diodea??based photonic skin for attachable phototherapeutics
摘要： Phototherapeutics is both safely noninvasive and can be employed to treat a light variety of sites and diseases. Current rigid and bulky conventional light sources, such as LED or laser-based phototherapy devices, are difficult to transport and use for regular irradiation treatments. To solve this problem, flexible organic light-emitting diode (OLED) light sources are the best candidates, and if applied very thinly as a skin-like platform, the ultimate attachable phototherapeutics can be realized. We demonstrated a very thin flexible OLED-based photonic skin with a total thickness of 6 μm for application in attachable phototherapeutics. It was optimized by controlling the peak wavelengths (600–700 nm) and irradiation interval of the flexible OLED thus improving the regeneration effect of the artificial skin by up to 70%. In addition, when the flexible OLED-based photonic skin was attached to a dressing film before being applied to the skin, it delivered the same electro-optical properties, while protecting against external contamination. The OLED skin on the dressing film had an operating lifetime of more than 100 h. These results confirmed the applicability of flexible OLED-based photonic skin to various light treatment areas, such as surgical wounds that require periodic irradiation.
关键词： wound-healing,regeneration,photonic Skin,dressing,flexible OLED,phototherapeutics
更新于2025-09-23 15:19:57
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Laser tissue bondinga??a new venture into primary periodontal wound closure techniques
摘要： Purpose To evaluate the efficacy of Laser tissue bonding (LTB) by comparing with the routinely used sutures for closure of intraoral surgical wounds after periodontal surgery. Methods Patients enrolled in the study were divided into 2 groups. Seventeen patients were included for a split mouth study where modified periodontal flap operation was performed, and in test side, LTB was carried out whereas in control, conventional suturing. Laser welding of the gingiva was performed with an 810 diode laser with a power of 1.5 W in contact mode. Results A highly statistically significant difference was observed in LTB (test) subgroup when compared with conventional suturing, depicting superiority in terms of Power Doppler analysis of vascularisation, wound healing scores, time consumption and microbial colonization. Conclusion Within the limitations of the study, it was concluded that LTB was effective in achieving wound closure with no morbid complications associated. More long-term randomized controlled trials are required to further prove the efficacy of this novel technique.
关键词： Suturing,Laser tissue bonding,Wound healing,Periodontal flap
更新于2025-09-23 15:19:57
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Optical coherence tomography angiography monitors human cutaneous wound healing over time
摘要： Background: In vivo imaging of the complex cascade of events known to be pivotal elements in the healing of cutaneous wounds is a difficult but essential task. Current techniques are highly invasive, or lack the level of vascular and structural detail required for accurate evaluation, monitoring and treatment. We aimed to use an advanced optical coherence tomography (OCT)-based angiography (OCTA) technique for the non-invasive, high resolution imaging of cutaneous wound healing. Methods: We used a clinical prototype OCTA to image, identify and track key vascular and structural adaptations known to occur throughout the healing process. Specific vascular parameters, such as diameter and density, were measured to aid our interpretations under a spatiotemporal framework. Results: We identified multiple distinct, yet overlapping stages, hemostasis, inflammation, proliferation, and remodeling, and demonstrated the detailed vascularization and anatomical attributes underlying the multifactorial processes of dermatologic wound healing. Conclusions: OCTA provides an opportunity to both qualitatively and quantitatively assess the vascular response to acute cutaneous damage and in the future, may help to ascertain wound severity and possible healing outcomes; thus, enabling more effective treatment options.
关键词： microvasculature,wound healing,skin,Optical coherence tomography angiography (OCTA)
更新于2025-09-19 17:15:36