Wound Healing Society

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Fibroblast Activation In A Tension- Maintaining Skin Equivalent
Zoe C. Andrews, Kedriuna Townsend, Zachary Turner, Gang Xu, Melville B. Vaughan.

Background: Traditional in vitro skin equivalents lack tension, or have irregular tension based on the requirement to begin culture submerged followed by emerged culture. We developed a dermal equivalent composed of fibroblasts and collagen embedded within a plastic ring to allow isometric tension to develop. We asked whether the tension was sufficient to allow fibroblasts to proliferate during the emerged culture period, and how the epidermis was affected by this culture condition. Methods: Dermal equivalents were cultured for 4 to 7 days to allow tension generation, followed by keratinocyte plating and subsequently emerged culture for up to 21 days. Optical coherence tomography (OCT) was used to monitor the development of the epidermis during the emerged culture conditions. Skin equivalents were pulsed with ethynyl deoxyuridine to determine proliferation at day 7 and 14. Tissues were then histologically prepared and stained to determine proliferation and differentiation. Fibroblasts in the dermis entered the S phase of the cell cycle more often in tissues under tension, more so on day 7 than day 14. Results: The epidermal tissue was able to stratify and differentiate under both tension-maintaining and low-tension environments. OCT results agreed well with the histologically-prepared specimens. Conclusions: This model will be a useful in vitro model to study skin processes that require tension and fibroblast proliferation. OCT will allow the ability to monitor tissue progression while in culture, thereby reducing the need to sacrifice tissues for histology prior to end-stage assays or xenografting.

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