Multiphoton FLIM imaging of NADH and FAD to analyze cellular metabolic activity of reconstructed human skin in response to UVA light

Pena, Ana‐Maria; Boulade, Marine; Brizion, Sébastien; Tissot, Nicolas; Bornschlögl, Thomas; Galey, Jean‐Baptiste; Bernerd, Françoise; Planel, Emilie

doi:10.1117/12.2508858

Cited by 10 publications

(11 citation statements)

References 16 publications

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“…Fluorescence spectroscopy (27), fluorescence lifetime imaging (28) and two-photon fluorescence imaging (29) are the most common in vivo fluorescence imaging technologies used. We have previously shown that our custom-designed in vivo fluorescence imager can successfully be used as an optical indicator of the complex wound healing process in diabetes (30).…”

Section: Introductionmentioning

confidence: 99%

670 nm photobiomodulation improves the mitochondrial redox state of diabetic wounds

Mehrvar¹,

Mostaghimi²,

Foomani³

et al. 2021

Quant Imaging Med Surg

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Background: Photobiomodulation (PBM) by far-red (FR) to near-infrared (NIR) light has been demonstrated to accelerate diabetic wound healing in preclinical and clinical studies. Mitochondrial dysfunction and oxidative stress play key roles in impaired diabetic wound healing, and the effect of PBM on the metabolic state of diabetic wounds remains to be elucidated.Methods: In this study, a custom-designed in vivo fluorescence imaging technique was used to quantitatively assess the effect of FR-PBM on the mitochondrial bioenergetics of diabetic wounds. The intrinsic fluorescence of two mitochondrial co-enzymes, nicotinamide adenine dinucleotide (NADH) and oxidized flavin adenine dinucleotide (FAD), was monitored to quantify the redox ratio (RR) (NADH/FAD) of wounds over time.Results: Using an excisional model of wound healing, we demonstrated that 670 nm (FR) PBM improved mitochondrial bioenergetics and stimulated the rate of wound healing in diabetic db/db mice. Wound closure and the RR of diabetic wounds in response to 670 nm PBM (4.5 J/cm 2 , 60 mW/cm 2 for 90 s per day, 5 days/week) were compared to the sham-treated group. At day 9 of post-wounding, we observed a 43% decrease in the wound area and a 75% increase in RR in FR-treated diabetic mice compared to sham-treated diabetic mice. Conclusions:We conclude that the increase in mitochondrial RR and the related decrease in oxidative stress may be an important factor in FR-PBM mediated acceleration of wound healing in diabetic mice.

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Section: Introductionmentioning

confidence: 99%

670 nm photobiomodulation improves the mitochondrial redox state of diabetic wounds

Mehrvar¹,

Mostaghimi²,

Foomani³

et al. 2021

Quant Imaging Med Surg

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“…VivaScope images were studied primarily to visualize melanin, 30 while MPTflex images were used to visualize keratinocytes 31 . Similar to the previous report, 32 FLIM images and photon counts were acquired at two different wavelengths (760 and 890 nm) to accurately discriminate and quantitate the NADH from other autofluorophores in the skin like melanin and FAD. FLIM images were processed offline using SPCImage software from Becker and Hickl GmbH, Germany, as reported previously 33 …”

Section: Methodsmentioning

confidence: 99%

A novel professional‐use synergistic peel technology to reduce visible hyperpigmentation on face: Clinical evidence and mechanistic understanding by computational biology and optical biopsy

Bhardwaj,

Handler,

Mao

et al. 2024

Experimental Dermatology

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Topicals and chemical peels are the standard of care for management of facial hyperpigmentation. However, traditional therapies have come under recent scrutiny, such as topical hydroquinone (HQ) has some regulatory restrictions, and high concentration trichloroacetic acid (TCA) peel pose a risk in patients with skin of colour. The objective of our research was to identify, investigate and elucidate the mechanism of action of a novel TCA‐ and HQ‐free professional‐use chemical peel to manage common types of facial hyperpigmentation. Using computational modelling and in vitro assays on tyrosinase, we identified proprietary multi‐acid synergistic technology (MAST). After a single application on human skin explants, MAST peel was found to be more effective than a commercial HQ peel in inhibiting melanin (histochemical imaging and gene expression). All participants completed the case study (N = 9) without any adverse events. After administration of the MAST peel by a dermatologist, the scoring and VISIA photography reported improvements in hyperpigmentation, texture and erythema, which could be linked to underlying pathophysiological changes in skin after peeling, visualized by non‐invasive optical biopsy of face. Using reflectance confocal microscopy (VivaScope®) and multiphoton tomography (MPTflex™), we observed reduction in melanin, increase in metabolic activity of keratinocytes, and no signs of inflammatory cells after peeling. Subsequent swabbing of the cheek skin found no microbiota dysbiosis resulting from the chemical peel. The strong efficacy with minimum downtime and no adverse events could be linked to the synergistic action of the ingredients in the novel HQ‐ and TCA‐free professional peel technology.

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“…Indeed, 2PEF has been conjugated with liquid‐chromatography mass‐spectrometry (LC/MS) to validate redox states in differentiated stem cells [35] or in conjugation with spatial frequency domain imaging for the detection of NAD(P)H in keratinocytes in the Human skin [50]. Since then, time‐domain FLIM had been used, for example, to detect the metabolic activity of cells in engineered skin after UVA exposure [46].…”

Section: Multiphoton Fluorescence Imaging and Harmonic Generationmentioning

confidence: 99%

“…In general, given detectable fluorescence intensities, fluorescence lifetimes serve as indicators of fluorophores' environment such as local pH [43], or changes in calcium local concentration [44]. Combined with 2PEF, FLIM has been used in metabolic studies to study the effect of glucose concentration on cellular metabolic states, detecting molecular sub‐cellular populations of NAD(P)H [45], examining metabolic states of reconstituted human skin models after exposure to UVA radiation [46] to detect metabolic cell death pathways in artificial skin models [26] and to conduct optical redox ratio measurements through fluorescence lifetime of NAD(P)H and FAD to classify T‐cell activation [47, 48].…”

Section: Multiphoton Fluorescence Imaging and Harmonic Generationmentioning

confidence: 99%

Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

Hilzenrat

Gill

McArthur

2022

Journal of Biophotonics

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The past decade has seen an increasing demand for more complex, reproducible and physiologically relevant tissue cultures that can mimic the structural and biological features of living tissues. Monitoring the viability, development and responses of such tissues in real‐time are challenging due to the complexities of cell culture physical characteristics and the environments in which these cultures need to be maintained in. Significant developments in optics, such as optical manipulation, improved detection and data analysis, have made optical imaging a preferred choice for many three‐dimensional (3D) cell culture monitoring applications. The aim of this review is to discuss the challenges associated with imaging and monitoring 3D tissues and cell culture, and highlight topical label‐free imaging tools that enable bioengineers and biophysicists to non‐invasively characterise engineered living tissues.

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Multiphoton FLIM imaging of NADH and FAD to analyze cellular metabolic activity of reconstructed human skin in response to UVA light

Cited by 10 publications

References 16 publications

670 nm photobiomodulation improves the mitochondrial redox state of diabetic wounds

670 nm photobiomodulation improves the mitochondrial redox state of diabetic wounds

A novel professional‐use synergistic peel technology to reduce visible hyperpigmentation on face: Clinical evidence and mechanistic understanding by computational biology and optical biopsy

Imaging approaches for monitoring three‐dimensional cell and tissue culture systems

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