Effect of Mature Adipocyte-Derived Dedifferentiated Fat Cells on Formation of Basement Membrane after Cultured Epithelial Autograft on Artificial Dermis

Background: Artificial dermis is an important option for preparing full-thickness wounds for cultured epithelial autografting. Long-term fragility after cultured epithelial autografting remains a problem, probably because of the lack of basement membrane proteins. The authors hypothesized that treating artificial dermis with mesenchymal stem cells would promote basement membrane protein production. The authors tested this using dedifferentiated fat cells in a porcine experimental model. Methods: This study used four male crossbred (Landrace, Large White, and Duroc) swine. Cultured epithelium and dedifferentiated fat cells were prepared from skin and subcutaneous fat tissue harvested from the cervical region. Full-thickness open dorsal wounds were created and treated with artificial dermis to prepare a graft bed for cultured epithelial autograft. Two groups were established: the control group (artificial dermis treated with 0.5 ml of normal saline solution applied to the wounds) and the dedifferentiated fat group (artificial dermis treated with 0.5 × 10⁵ dedifferentiated fat cells suspended in 0.5 ml of normal saline solution sprayed onto the wounds). On postoperative day 10, the prepared cultured epithelium was grafted onto the generated dermis-like tissue. Fourteen days later, tissue specimens were harvested and evaluated histologically. Results: Light microscopy of hematoxylin and eosin-stained sections revealed the beginning of rete ridge formation in the dedifferentiated fat group. Synthesis of both collagen IV and laminin-5 was significantly enhanced in the dedifferentiated fat group. Transmission electron microscopy revealed a nearly mature basement membrane, including anchoring fibrils in the dedifferentiated fat group. Conclusion: Combined use of artificial dermis and dedifferentiated fat cells promotes post-cultured epithelial autograft production and deposition of basement membrane proteins at the dermal-epidermal junction and basement membrane development, including anchoring fibrils.