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Skin Microcolumns as a Source of Paracrine Signaling Factors.

ABSTRACT: Objective: We recently developed the approach of using "microcolumns" of autologous full-thickness skin tissue for wound repair. The small size of these micro skin tissue columns (MSTCs, ?0.5?mm in diameter) allows donor sites to heal quickly without scarring. Treatment with MSTCs significantly accelerate wound healing, and suppled various skin cell types and skin structures to replenish the wound volume. This technology is now starting clinical use. In this study, we investigate whether MSTCs may also influence wound healing by releasing soluble signaling factors. Approach: Freshly harvested MSTCs were incubated in culture medium for 24?h. The conditioned medium was collected and tested for its effects on migration and proliferation of human dermal fibroblasts, and its ability to induce tube formation by human umbilical vein endothelial cells (HUVECs). Proteins released into the conditioned medium were characterized by multiplex enzyme-linked immunosorbent assay (ELISA), and compared with medium conditioned by an equivalent mass of intact full-thickness skin. Results: MSTC-conditioned medium increased fibroblast migration and proliferation, as well as HUVEC tube formation. MSTCs released many soluble factors known to play prominent roles in wound healing. A subset of proteins showed significantly different release profiles compared with intact full-thickness skin. Innovation: The technology for harvesting and using MSTCs to augment wound healing was recently developed as an alternative to conventional autologous skin grafting. This study shows that MSTCs could also function as "cytokine factories." Conclusion: In addition to supplying autologous cells to repopulate the wound volume, MSTCs can also function as a source of growth factors and cytokines to further enhance wound healing.

SUBMITTER: Tam J

PROVIDER: S-EPMC7047113 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Skin Microcolumns as a Source of Paracrine Signaling Factors.

Tam Joshua J Purschke Martin M Fuchs Christiane C Wang Ying Y Anderson R Rox RR

Advances in wound care 20200207 4

<b>Objective:</b> We recently developed the approach of using "microcolumns" of autologous full-thickness skin tissue for wound repair. The small size of these micro skin tissue columns (MSTCs, ∼0.5 mm in diameter) allows donor sites to heal quickly without scarring. Treatment with MSTCs significantly accelerate wound healing, and suppled various skin cell types and skin structures to replenish the wound volume. This technology is now starting clinical use. In this study, we investigate whether ...[more]

PMID: 32117581

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Project description:BackgroundIncreasing evidence has shown that mesenchymal stem cells (MSCs) yield a favorable therapeutic benefit for thermal burn skin wounds. Human amniotic MSCs (hAMSCs) derived from amniotic membrane have multilineage differentiation, immunosuppressive, and anti-inflammatory potential which makes them suitable for treating skin wounds. However, the exact effects of hAMSCs on the healing of thermal burn skin wounds and their potential mechanisms are not explored.MethodshAMSCs were isolated from amniotic membrane and characterized by RT-PCR, flow cytometry, immunofluorescence, and tumorigenicity test. We assessed the effects of hAMSCs and hAMSC conditional medium (CM) on wound healing in a deep second-degree burn injury model of mice. We then investigated the biological effects of hAMSCs and hAMSC-CM on the apoptosis and proliferation of heat stress-injured human keratinocytes HaCAT and dermal fibroblasts (DFL) both in vivo and in vitro. Next, we explored the underlying mechanisms by assessing PI3K/AKT and GSK3β/β-catenin signaling pathways in heat injured HaCAT and DFL cells after hAMSCs and hAMSC-CM treatments using PI3K inhibitor LY294002 and β-catenin inhibitor ICG001. Antibody array assay was used to identify the cytokines secreted by hAMSCs that may activate PI3K/AKT signaling pathway.ResultsOur results showed that hAMSCs expressed various markers of embryonic stem cells and mesenchymal stem cells and have low immunogenicity and no tumorigenicity. hAMSC and hAMSC-CM transplantation significantly promoted thermal burn wound healing by accelerating re-epithelialization with increased expression of CK19 and PCNA in vivo. hAMSCs and hAMSC-CM markedly inhibited heat stress-induced apoptosis in HaCAT and DFL cells in vitro through activation of PI3K/AKT signaling and promoted their proliferation by activating GSK3β/β-catenin signaling. Furthermore, we demonstrated that hAMSC-mediated activation of GSK3β/β-catenin signaling was dependent on PI3K/AKT signaling pathway. Antibody array assay showed that a panel of cytokines including PAI-1, C-GSF, periostin, and TIMP-1 delivered from hAMSCs may contribute to the improvement of the wound healing through activating PI3K/AKT signaling pathway.ConclusionOur results demonstrated that hAMSCs and hAMSC-CM efficiently cure heat stress-induced skin injury by inhibiting apoptosis of skin cells and promoting their proliferation through activating PI3K/AKT signaling pathway, suggesting that hAMSCs and hAMSC-CM may provide an alternative therapeutic approach for the treatment of skin injury.

Dataset Information

Skin Microcolumns as a Source of Paracrine Signaling Factors.

Publications

Skin Microcolumns as a Source of Paracrine Signaling Factors.

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