Reepithelialization in focus: Non-invasive monitoring of epidermal wound healing in vitro

Kiesewetter, L.; Littau, Laura; Walles, Heike; Boccaccını, Aldo R.; Groeber‐Becker, Florian

doi:10.1016/j.bios.2019.111555

Cited by 23 publications

(16 citation statements)

References 30 publications

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“…The newly formed epidermis did not only show the corresponding cellular morphology, confirmed by H&E staining, but also the presence of the basal and supra-basal keratin network (K 10 and K14). In addition, the progression and speed of reepithelialization was similar to previously reported burn—and punch wounds in in vitro skin equivalents [ 23 , 33 ].…”

Section: Discussionsupporting

confidence: 84%

“…The media was collected at days 0, 1, 3, 7, 10, and 14, with fresh media as a control. Glucose consumption was calculated, as previously described [ 23 ].…”

Section: Methodsmentioning

confidence: 99%

“…These include skin irritation [ 20 ], skin sensitization [ 21 ] and skin corrosion [ 22 ]. Additionally, studies of cutaneous wound healing have been performed using in vitro skin models [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

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A 3D In Vitro Model for Burn Wounds: Monitoring of Regeneration on the Epidermal Level

et al. 2021

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Burns affect millions every year and a model to mimic the pathophysiology of such injuries in detail is required to better understand regeneration. The current gold standard for studying burn wounds are animal models, which are under criticism due to ethical considerations and a limited predictiveness. Here, we present a three-dimensional burn model, based on an open-source model, to monitor wound healing on the epidermal level. Skin equivalents were burned, using a preheated metal cylinder. The healing process was monitored regarding histomorphology, metabolic changes, inflammatory response and reepithelialization for 14 days. During this time, the wound size decreased from 25% to 5% of the model area and the inflammatory response (IL-1β, IL-6 and IL-8) showed a comparable course to wounding and healing in vivo. Additionally, the topical application of 5% dexpanthenol enhanced tissue morphology and the number of proliferative keratinocytes in the newly formed epidermis, but did not influence the overall reepithelialization rate. In summary, the model showed a comparable healing process to in vivo, and thus, offers the opportunity to better understand the physiology of thermal burn wound healing on the keratinocyte level.

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Section: Discussionsupporting

confidence: 84%

“…The media was collected at days 0, 1, 3, 7, 10, and 14, with fresh media as a control. Glucose consumption was calculated, as previously described [ 23 ].…”

Section: Methodsmentioning

confidence: 99%

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A 3D In Vitro Model for Burn Wounds: Monitoring of Regeneration on the Epidermal Level

et al. 2021

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“…Очень активно эквиваленты кожи используются в исследованиях трансдермальной доставки лекарств и других фармакологических субстанций. Также эквиваленты кожи могут быть использованы для изучения влияния различных факторов и фармакологических агентов на биогенез и реэпителизацию кожи in vitro [41], регуляцию дифференцировки кератиноцитов, абсорбцию и иммуноток-сичность, реакцию кожи на коррозирующие вещества, раздражители и аллергены [42], а также способность кератиноцитов контрактировать (сокращать) матрикс или рану [43], для оценки фототоксичности [44]. Более того, среда при культивировании на границе раздела фаз может быть собрана для анализа выбрасываемых в нее клетками медиаторов и/или метаболитов, что обеспечивает количественный и объективный результат в сравнении с такими субъективными параметрами, как эритемы и отеки in vivo.…”

Section: Discussionunclassified

The Reconstructed Human Epidermis in vitro — a Model for Basic and Applied Research of Human Skin

Beilin

Константинович²,

Леонидовна³

et al. 2020

Vestnik dermatologii i venerologii

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Background. The reconstructed human epidermis (RE) is an in vitro tissue-engineering construct similar to the native epidermis. Objective. To develop a full-layer RE. Describe its structure: determine the presence of all layers of the epidermal component, including basal, spinous and granular layers and stratum corneum of the epidermis; detect the basement membrane, the border between the epidermal and mesenchymal component. Materials and methods. Isolation of keratinocytes and fibroblasts from human donor skin. Cultivation of keratinocytes and fibroblasts in vitro under 2D conditions, cell subculturing and 3D modeling of RE, obtaining cryosections, histological staining, immunohistochemical (IHC) study with antibodies to cytokeratins 14 and 10, Ki67 protein, loricrin, laminin 5 and plectin. Results. A technique was developed for the formation of RE. Histological examination showed that the stratification of keratinocyte layers occurs during the formation of RE. Layers are formed including basal, spinous and granular layers and stratum corneum. The IHC study has shown the proliferative activity of keratinocytes of the basal layer and has detected the presence of marker proteins of keratinocytes at different stages of differentiation. RE basal keratinocytes, like native ones, form hemidesmosomes and synthesize basement membrane proteins. Conclusions. A full-layer human RE was obtained in vitro. RE meets all the characteristics of the native epidermis and it is suitable for basic and practical research in the field of skin biology, dermatology, and cosmetology.

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“…This may indicate that a more complex electrical model of the tissue, which would also take into consideration other components besides TEER, should be adopted. This can be achieved if, e.g., impedance measurements are performed instead of TEER, which can provide many more data about the analyzed tissue and its response to external changes [42]. Nevertheless, impedance measurements require a much more complex setup, a careful design of the experimental approach, as well as a more intricate extraction of the relevant parameters from the measured data which typically is done using proprietary software.…”

Section: Relationship Between Teer and Tissue Viabilitymentioning

confidence: 99%

Low-Cost Method and Biochip for Measuring the Trans-Epithelial Electrical Resistance (TEER) of Esophageal Epithelium

et al. 2020

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Trans-epithelial electrical resistance (TEER) is a good indicator of the barrier integrity of epithelial tissues and is often employed in biomedical research as an effective tool to assess ion transport and permeability of tight junctions. The Ussing chamber is the gold standard for measuring TEER of tissue specimens, but it has major drawbacks: it is a macroscopic method that requires a careful and labor intensive sample mounting protocol, allows a very limited viability for the mounted sample, has large parasitic components and low throughput as it cannot perform multiple simultaneous measurements, and this sophisticated and delicate apparatus has a relatively high cost. This paper demonstrates a low-cost home-made “sandwich ring” method which was used to measure the TEER of tissue specimens effectively. This method inspired the subsequent design of a biochip fabricated using standard soft lithography and laser engraving technologies, with which the TEER of pig epithelial tissues was measured. Moreover, it was possible to temporarily preserve the tissue specimens for days in the biochip and monitor the TEER continuously. Tissue responses after exposure tests to media of various pH values were also successfully recorded using the biochip. All these demonstrate that this biochip could be an effective, cheaper, and easier to use Ussing chamber substitute that may have relevant applications in clinical practice.

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Reepithelialization in focus: Non-invasive monitoring of epidermal wound healing in vitro

Cited by 23 publications

References 30 publications

A 3D In Vitro Model for Burn Wounds: Monitoring of Regeneration on the Epidermal Level

A 3D In Vitro Model for Burn Wounds: Monitoring of Regeneration on the Epidermal Level

The Reconstructed Human Epidermis in vitro — a Model for Basic and Applied Research of Human Skin

Low-Cost Method and Biochip for Measuring the Trans-Epithelial Electrical Resistance (TEER) of Esophageal Epithelium

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