“Effect of Mature Adipocyte-Derived Dedifferentiated Fat (DFAT) Cells on Formation of Basement Membrane after Cultured Epithelial Autograft on Artificial Dermis”

Background: Artificial dermis (AD) is an important option for preparing full-thickness wounds for cultured epithelial autograft (CEA). Long-term fragility after CEA remains a problem, probably due to lack of basement membrane (BM) proteins. We hypothesized that treating AD with mesenchymal stem cells would promote BM protein production. We tested this using dedifferentiated fat (DFAT) cells in a porcine experimental model. Methods: This study employed four male LWD swine. Cultured epithelium (CE) and DFAT cells were prepared from skin and subcutaneous fat tissue harvested from the cervical region. Full-thickness open dorsal wounds were created and treated with AD (Pelnac®, GUNZE, Japan) to prepare a graft bed for CEA. Two groups were established. Control group: AD treated with 0.5 mL normal saline solution was applied to the wounds. DFAT group: AD treated with DFAT cells (0.5 X 105 cells) suspended in 0.5 mL of normal saline solution was sprayed to the wounds. On the 10th postoperative day, the prepared CE was grafted onto the generated dermis-like tissue. Fourteen days later, tissue specimens were harvested and histologically evaluated. Results: Light microscopy of H&E sections revealed beginning of rete ridge formation in the DFAT group. Synthesis of both Collagen IV and Laminin-5 was significantly enhanced in the DFAT group. Transmission electron microscopy revealed a nearly mature BM including anchoring fibrils in the DFAT group. Conclusion: Combined use of AD and DFAT cells promotes post-CEA production and deposition of basement membrane proteins at the dermal-epidermal junction and BM development including anchoring fibrils. Financial Disclosure Statement: This work was supported by JSPS KAKENHI Grant Number 15K10954. The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper. ACKNOWLEDGMENTS: This work was supported by JSPS KAKENHI Grant Number 15K10954. Disclosure: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper. Corresponding author: Kazutaka Soejima, MD, Department of Plastic and Reconstructive Surgery, School of Medicine, Nihon University, 30-1 Kamicho, Oyaguchi, Itabashi-ku, Tokyo 173-8610, Japan. soejima.kazutaka@nihon-u.ac.jp ©2019American Society of Plastic Surgeons

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