Construction of low contracted 3D skin equivalents by genipin cross‐linking

Meng, Qin; Shen, Chong

doi:10.1111/exd.13725

Cited by 10 publications

(14 citation statements)

References 35 publications

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“…We used genipin, as the cross-link stabilator of the collagen scaffold for the fibroblast culture. Genipin was also used by Meng and Shen [ 19 ] for the stabilization of collagen for HSE production. The authors observed that 0.4 mmol/L genipin is non-toxic for the fibroblasts and greatly reduces the contraction of 3D skin equivalents from 87% to 28% ( n = 9, p < 0.05) over a 21-day follow-up period.…”

Section: Discussionmentioning

confidence: 99%

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

Szymański

Jęderka

Cios

et al. 2020

IJMS

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An important problem for researchers working in the field of dermatology is the preparation of the human skin equivalent (HSE). Here, we describe a simple and reliable protocol for preparing a skin model from the commercially available cell lines: keratinocytes, fibroblasts, and melanocytes. Importantly, in our 3D model, the keratinocytes are diverse that brings this model closer to the natural skin. For the production of HSE, we used available primary PCS-200-010, PCS-201-010, PCS-200-013, and immortalized CRL-4048 and CRL-4001 cell lines. We used genipin, which is necessary for collagen cross-linking and studied its cytotoxicity for keratinocytes and fibroblasts. The addition of 20 μM genipin reduced the shrinkage of the collagen in the constructs from 59% to 24% on day 12 of the culture of the construct. A higher concentration (80–200 µM) of genipin reduced shrinkage by 14% on average. Genipin in concentration 10 μM and below was not cytotoxic to the keratinocytes, and 150 μM and below to the fibroblasts. Hematoxylin and eosin staining showed that the morphology of HSEs was identical to that of native human skin. The immunohistochemical staining of the constructs showed the presence of vimentin-positive fibroblasts in the skin layer, while the melanocytes were in the epidermis and in the basal layer. We observed that the longer differentiation of constructs led to the higher secretion of GM-CSF, IL-10, IL-15, IL-1α, IL-6, IL-7, IL-8, and MCP-1. We also observed that the longer time of differentiation led to a more stable secretion of all analytes, which was reflected in the coefficient of variation. We described here a simple, reliable, and cost-effective production of the full-thickness human skin equivalents that can be used in the research and industry. With the global trend to decrease animal use for the research and testing, our HSE could be a useful testing tool and an alternative research model.

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Section: Discussionmentioning

confidence: 99%

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

Szymański

Jęderka

Cios

et al. 2020

IJMS

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“…3D cell cultures differ greatly from standard 2D cell cultures in cell-cell interactions, cellular mechanics and nutrient access [9]. Briefly, 3D skin equivalents are organotypic culture systems that are generated by seeding human keratinocytes onto a dermal layer with human fibroblasts [10]. The positive trend of increasing acceptance of these models in dermatological research will hopefully lead to a reduction in the number of animals used for experiments to study skin biology, wound healing, skin ageing and disease pathology [11,12].…”

Section: Introductionmentioning

confidence: 99%

“…The positive trend of increasing acceptance of these models in dermatological research will hopefully lead to a reduction in the number of animals used for experiments to study skin biology, wound healing, skin ageing and disease pathology [11,12]. In addition, 3D skin equivalents have been developed as an attractive tool for pharmacotoxicological testing of cosmetic components, for clinical applications such as artificial skin grafts and to avoid the need for sequential invasive skin biopsies from patients for standardised studies of human skin [10,13].…”

Section: Introductionmentioning

confidence: 99%

Preliminary Study of In Vitro Three-Dimensional Skin Model Using an Ovine Collagen Type I Sponge Seeded with Co-Culture Skin Cells: Submerged versus Air-Liquid Interface Conditions

2020

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Three-dimensional (3D) in vitro skin models have been widely used for cosmeceutical and pharmaceutical applications aiming to reduce animal use in experiment. This study investigate capability of ovine tendon collagen type I (OTC-I) sponge suitable platform for a 3D in vitro skin model using co-cultured skin cells (CC) containing human epidermal keratinocytes (HEK) and human dermal fibroblasts (HDF) under submerged (SM) and air-liquid interface (ALI) conditions. Briefly, the extracted OTC-I was freeze-dried and crosslinked with genipin (OTC-I_GNP) and carbodiimide (OTC-I_EDC). The gross appearance, physico-chemical characteristics, biocompatibility and growth profile of seeded skin cells were assessed. The light brown and white appearance for the OTC-I_GNP scaffold and other groups were observed, respectively. The OTC-I_GNP scaffold demonstrated the highest swelling ratio (~1885%) and water uptake (94.96 ± 0.14%). The Fourier transformation infrared demonstrated amide A, B and I, II and III which represent collagen type I. The microstructure of all fabricated sponges presented a similar surface roughness with the presence of visible collagen fibers and a heterogenous porous structure. The OTC-I_EDC scaffold was more toxic and showed the lowest cell attachment and proliferation as compared to other groups. The micrographic evaluation revealed that CC potentially formed the epidermal- and dermal-like layers in both SM and ALI that prominently observed with OTC-I_GNP compared to others. In conclusion, these results suggest that OTC_GNP could be used as a 3D in vitro skin model under ALI microenvironment.

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“…The outputs demonstrated similar findings to their previous research; higher swelling in the GNP-crosslinked group (1886 ± 56%) than EDC-crosslinked group (1,658 ± 62%) ( Fauzi et al, 2020 ). The swelling capacity of GNP-crosslinked CH-alginate (ALG)-HA polyelectrolyte composite sponges was monitored by Meng et al (2021) . They saw high swelling for all formulations after 2 h of the experiment (by using low field NMR).…”

Section: Genipin Crosslinking Effect On Biomatrix Propertiesmentioning

confidence: 99%

“…However, TG2-modified films exhibited no significant improvement in mechanical strength at low crosslinking concentration. Another comparison study of GNP and EDC crosslinking has been performed by Meng et al (2018) . They produced 3D skin equivalents from human skin cells and COL gel.…”

Section: Genipin Crosslinking Effect On Biomatrix Propertiesmentioning

confidence: 99%

Genipin-Crosslinking Effects on Biomatrix Development for Cutaneous Wound Healing: A Concise Review

Nike

Fadilah

Sallehudin

et al. 2022

Front. Bioeng. Biotechnol.

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Split skin graft (SSG), a standard gold treatment for wound healing, has numerous limitations such as lack of fresh skin to be applied, tedious process, severe scarring, and keloid formation followed by higher risks of infection. Thus, there is a gap in producing polymeric scaffolds as an alternative for wound care management. Bioscaffold is the main component in tissue engineering technology that provides porous three-dimensional (3D) microarchitecture for cells to survive. Upon skin tissue reconstruction, the 3D-porous structure ensures sufficient nutrients and gaseous diffusion and cell penetration that improves cell proliferation and vascularization for tissue regeneration. Hence, it is highly considered a promising candidate for various skin wound healing applications. To date, natural-based crosslinking agents have been extensively used to tailor the physicochemical and mechanical properties of the skin biomatrix. Genipin (GNP) is preferable to other plant-based crosslinkers due to its biological activities, such as antiinflammatory and antioxidant, which are key players to boost skin wound healing. In addition, it has shown a noncytotoxic effect and is biocompatible with human skin cells. This review validated the effects of GNP in biomatrix fabrication for skin wound healing from the last 7 years of established research articles and stipulated the biomaterial development-scale point of view. Lastly, the possible role of GNP in the skin wound healing cascade is also discussed. Through the literature output, it can be concluded that GNP has the capability to increase the stability of biomatrix and maintain the skin cells viability, which will contribute in accelerating wound healing.

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Construction of low contracted 3D skin equivalents by genipin cross‐linking

Cited by 10 publications

References 35 publications

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

A Simple Method for the Production of Human Skin Equivalent in 3D, Multi-Cell Culture

Preliminary Study of In Vitro Three-Dimensional Skin Model Using an Ovine Collagen Type I Sponge Seeded with Co-Culture Skin Cells: Submerged versus Air-Liquid Interface Conditions

Genipin-Crosslinking Effects on Biomatrix Development for Cutaneous Wound Healing: A Concise Review

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