Plastic Surgery Research Council
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FAT GRAFT ENGINEERING BY DECELLULARIZED HUMAN ADIPOSE MATRIX AND ADIPOSE-DERIVED STEM CELLS
Presenter: Lina Wang, PhD
Co-Authors: Wang L; Johnson JA; Iyyanki TS; Zhang Q; Beahm EE
MDACC

Purpose: This study aimed to engineer human fat grafts by combining decellularized adipose matrix (DAM) and human adipose-derived stem cells (hASCs).

Methods: Human adipose tissues were treated with hypotonic solutions, Trypsin, TritonX-100, and isopropanol to remove cells and lipids, which was proved by immunohistochemistry and DNA quantification. DAM was characterized by immunohistochemistry and scanning electron microscope (SEM). hASC viability and adhesion on DAM were tested by immunostaining. DAM was implanted subcutaneously in Fisher rats to evaluate DAM-induced immunoresponse and remodeling by H&E, Masson Trichrome, CD 4, CD 8, CD 68, CD 80, CD 163, and CD31 staining. Then raw fat grafts (Coleman technique) and engineered fat grafts (DAM combined with hASCs) were implanted subcutaneously in nude rats. Both types of implants were harvested and evaluated by H&E, CD31, and HuNu staining.

Results: Negative H&E and DAPI staining and oil red O staining in DAM confirmed the absence of cells and lipids, respectively. DNA quantification further confirmed DNA removal in DAM. SEM images of DAM showed three-dimensional nanofibrous architecture. Masson Trichrome and immunohistological staining indicated that DAM was composed of collagen, laminin, and VEGF but lacked major histocompatibility complex antigen 1. hASCs were successfully integrated with DAM. hASCs established adhesion and kept proliferation on DAM in vitro. In vivo animal test demonstrated that DAM did not cause immunogenic response (few CD4+ and CD8+; completely CD68-, CD80-, and CD163-) in Fisher rats. Cell infiltration, vascularization and adipose cell-like structures were observed in DAM implanted in Fisher rats. In nude rats, implanted raw fat grafts were remodeled and replaced from the outer layer to the inner by host cells and blood vessels. Engineered fat grafts remained at week 8; hASCs contributed to adipose-like structure formation (HuNu+) while host cells formed blood vessels (CD31+/HuNu-).

Conclusion: Our study suggests engineered fat grafts (DAM combined with hASCs) may have significant potential applications in adipose tissue engineering.


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