Modeling Aberrant Wound Healing Using Tissue-Engineered Skin Constructs and Multiphoton Microscopy

Torkian, Behrooz A.; Yeh, Alvin T.; Engel, Rodney A.; Sun, Carlos; Tromberg, Bruce J.; Wong, Brian J. F.

doi:10.1001/archfaci.6.3.180

Cited by 51 publications

(40 citation statements)

References 22 publications

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“…As a versatile, fast and affordable imaging tool, white light microscopy is frequently used in TE. Besides basic bright-field illumination [36,60], enhanced contrast methods such as dark-field illumination, phase-contrast and differential interference contrast techniques have been applied [61][62][63][64]. White light microscopy is primarily used for routine analysis (monitoring cell number and morphology after seeding of scaffolds) and for imaging stained tissue slices (histology [65]; various stains such as H&E, Von Kossa, trichrome, alcian blue and others are available [39,50,[66][67][68][69][70][71]).…”

Section: Transillumination and Fluorescence Microscopymentioning

confidence: 99%

“…Alexa Fluor, Cy, MFP or Qdot series) are used in many TE studies [85][86][87][88]. CFM is used to stain both fixed and live cells with nuclear, cytoplasmic, membrane-bound dyes or-for protein-specific detectionantibody-coupled dyes [59,64,65,68,69,76] (immunohistochemistry: [39,89]). In principle, any cellular structure can nowadays be specifically visualized and combinations of dyes are possible (multicolour imaging, multiplexing) when spectral overlap is minimal between dyes.…”

Section: Transillumination and Fluorescence Microscopymentioning

confidence: 99%

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Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine

Vielreicher

Schürmann

Detsch

et al. 2013

J. R. Soc. Interface.

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This review focuses on modern nonlinear optical microscopy (NLOM) methods that are increasingly being used in the field of tissue engineering (TE) to image tissue non-invasively and without labelling in depths unreached by conventional microscopy techniques. With NLOM techniques, biomaterial matrices, cultured cells and their produced extracellular matrix may be visualized with high resolution. After introducing classical imaging methodologies such as µCT, MRI, optical coherence tomography, electron microscopy and conventional microscopy two-photon fluorescence (2-PF) and second harmonic generation (SHG) imaging are described in detail (principle, power, limitations) together with their most widely used TE applications. Besides our own cell encapsulation, cell printing and collagen scaffolding systems and their NLOM imaging the most current research articles will be reviewed. These cover imaging of autofluorescence and fluorescence-labelled tissue and biomaterial structures, SHG-based quantitative morphometry of collagen I and other proteins, imaging of vascularization and online monitoring techniques in TE. Finally, some insight is given into state-of-the-art three-photon-based imaging methods (e.g. coherent anti-Stokes Raman scattering, third harmonic generation). This review provides an overview of the powerful and constantly evolving field of multiphoton microscopy, which is a powerful and indispensable tool for the development of artificial tissues in regenerative medicine and which is likely to gain importance also as a means for general diagnostic medical imaging.

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Section: Transillumination and Fluorescence Microscopymentioning

confidence: 99%

Section: Transillumination and Fluorescence Microscopymentioning

confidence: 99%

Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine

Vielreicher

Schürmann

Detsch

et al. 2013

J. R. Soc. Interface.

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“…As a new approach to identify the transdermal fluorescent model drug spatial distributions, multiphoton excitation imaging shows that oleic acid can induce increases in the hydrophilic intracorneocyte diffusion and in the localization of the hydrophobic model drug diffusion to the intercellular region (Yu et al, 2003;Sun et al, 2004). In the wounded skin, the intravital imaging technique was used to study the fibroblast activity and collagen production in the intact tissue-engineered skin (Torkian et al, 2004). More recently, a first clinical use of multiphoton endoscopes was reported for deep-tissue imaging as well as for intracorporal imaging.…”

Section: In Vivo Drug Screening and Administration In Skin And Kidneymentioning

confidence: 99%

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Wang

König

2010

Journal of Microscopy

180

131

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SummaryMultiphoton excitation laser scanning microscopy, relying on the simultaneous absorption of two or more photons by a molecule, is one of the most exciting recent developments in biomedical imaging. Thanks to its superior imaging capability of deeper tissue penetration and efficient light detection, this system becomes more and more an inspiring tool for intravital bulk tissue imaging. Two-photon excitation microscopy including 2-photon fluorescence and second harmonic generated signal microscopy is the most common multiphoton microscopic application. In the present review we take diverse ocular tissues as intravital samples to demonstrate the advantages of this approach. Experiments with registration of intracellular 2-photon fluorescence and extracellular collagen second harmonic generated signal microscopy in native ocular tissues are focused. Data show that the in-tandem combination of 2-photon fluorescence and second harmonic generated signal microscopy as two-modality microscopy allows for in situ co-localization imaging of various microstructural components in the whole-mount deep intravital tissues. New applications and recent developments of this high technology in clinical studies such as 2-photon-controlled drug release, in vivo drug screening and administration in skin and kidney, as well as its uses in tumourous tissues such as melanoma and glioma, in diseased lung, brain and heart are additionally reviewed. Intrinsic emission two-modal 2-photon microscopy/tomography, acting as an efficient and sensitive non-injurious imaging approach featured by high contrast and subcellular spatial resolution, has been proved to be a promising tool for intravital deep tissue imaging and clinical studies. Given the level of its performance, we believe Correspondence to B.-G. Wang. Tel: +49 3641 938496; fax: +49 3641 938552; e-mail: Baogui.Wang@mti.uni-jena.de that the non-linear optical imaging technique has tremendous potentials to find more applications in biomedical fundamental and clinical research in the near future.

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“…Advances in microscopy have provided some solutions, but these are not yet very common. A wound-model has been analyzed using two-photon microscopy with some success (Torkian et al, 2004). e use of spectromicroscopy may also provide a solution; Raman microscopy has been used to look at epidermal permeability , and may prove useful for several types of constructs.…”

Section: Skin In the Clinicmentioning

confidence: 99%

Fibroblast Differentiation and Models of Human Skin

Rakar¹

2016

Linköping University Medical Dissertations

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Denna bok avhandlar en riktning av biomedicinsk forskning som i det långa loppet sy ar till att nå bättre och banbrytande sätt att läka våra kroppar. Forskningen vilar på en tredelad grund -viktiga kunskaper som leder mot en generell förståelse för hur celler blir till vävnader, hur vävnad-er och organ kan manipuleras på ett kliniskt användbart sätt, och till sist hur denna kunskap kan appliceras på människans hudorgan.Den första delen handlar om cellers identitet, och den stora potentialen som våra egna stamceller har för framtidens läkekonst. Genom att förstå hur celler egentligen fungerar på arvsmassenivå kan vi utöka arsenalen av verktyg vi har för att få celler att bete sig på de sätt som behövs för att skapa nya organ. Vi har jobbat med att karakterisera och manipulera broblaster, en av de vanligaste celltyperna i människokroppen, och bland annat tagit fram olika typer av fettceller.Den andra delen handlar om hur man använder celler, biokemiska faktorer och biokompatibla material för att konstruera vävnader och organ. Innan forskningen når klinisk användbarhet har vi möjlighet att bygga begränsade modellsystem för att studera människans biologi. Cellsystem på labbet är mycket viktiga verktyg för att föra forskningen på vävnadsbyg-gande framåt.Den tredje delen behandlar hudforskning speci kt, och här samlas kunskaper från tidigare delarna som är relevanta för mänsklig hud. Här beskrivs användningen av två olika hudmodeller för att undersöka proceser viktiga vid sårläkning. Vidare beskrivs planerna kring ett större påbörjat projekt som sy ar till att använda alla tidigare lärdomar och tekniker för att åstadkomma en ny terapi för sårläkning.Abstract is thesis combines three publications and one manuscript, covering two principal topics: functional di erentiation of human broblasts and, laboratory models of human skin. e two topics favourably unite in the realm of tissue engineering. is thesis is therefore split into three main parts: 1. a discussion of phenotypic plasticity as it pertains to broblasts and the stem cell continuum; 2. a short review of engineered tissue, with particular focus on soluble factors and materials; and, 3. a motivated review of the biology, diversity and culture of skin, including skin construction. e intended goal of our research endeavor was to achieve the formulation of a bioactive therapy for skin regeneration. e main hypothesis was that broblast-to-keratinocyte di erentiation would facilitate wound healing, and that the protocol for such a method could be adapted to clinical translation. e foundation for the hypothesis lay in the di erentiation capabilities of primary dermal broblasts (Paper I). However, the goal has not yet been achieved. Instead, intermediate work on the construction of skin for the purpose of creating a model test-bed has resulted in two other publications. e use of excised human skin, a formidable reference sample for tissue engineered skin, has been used to investigate a gelatin-based material in re-epithelialization (Paper II). A rst attempt at standardizi...

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Modeling Aberrant Wound Healing Using Tissue-Engineered Skin Constructs and Multiphoton Microscopy

Cited by 51 publications

References 22 publications

Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine

Taking a deep look: modern microscopy technologies to optimize the design and functionality of biocompatible scaffolds for tissue engineering in regenerative medicine

Two‐photon microscopy of deep intravital tissues and its merits in clinical research

Fibroblast Differentiation and Models of Human Skin

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