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Adipose Derived Stem Cell to Augment Vascularization and Incorporation of Alloderm
Kenneth Fan, BS, Daniela Bueno, DDS PhD, Joyce Yuan, BS, Ginger Slack, BS, Christina Tabit, BA, Patricia Zuk, PhD, James P. Bradley, MD DMD FACS.
UCLA Division of Plastic Surgery, Los Angeles, CA, USA.

PURPOSE: Adipose-derived stem cells(ASCs) are multipotent cells with dynamic therapeutic applications, including use as growth factor biopump. ASCs may be able to augment vascularization through VEGF production. Vascularization is difficult to achieve in some instances, especially irradiated tissues. Acellular dermal matrix(ADM, AlloDerm) is widely used in breast reconstruction, hernia repair, burns, and cleft palate. The aim of our study was to determine if ASCs and/or fresh fat grafts may augment incorporation of scaffolds, in particular ADM.
METHODS: Our objectives were three part. 1)Fresh fat graft and ASCs, obtained from human lipoaspirate, were tested in vitro for VEGF secretion via ELISA. 2)After seeding of ASCs on ADM dermal surface, attachment and migration was confirmed in vitro with scanning electron microscopy(SEM) and DAPI staining. 3)ASC/cell(1*106) constructs were implanted on dorsal surfaces of athymic mice in the following conditions: a)ADM only, b)fresh fat 20 minutes before implantation, c)ASCs 20 minutes, d)ASCs 3 hours. Mice were harvested at 2, 7, 9, 14 days with 2 mice each. Surgical excision, H&E staining, mouse CD34, and DAPI staining was performed to assess cell proliferation and vascularization.
RESULTS:1)Human ASCs produced 2.19*10-3 pg/mL per ASC of VEGF 1 day in culture decreasing to 1.45*10-4 pg/mL per ASC of VEGF, 7 days in culture. Fresh fat produced less VEGF at 1.88*10-3 pg/mL per cell of VEGF 1 day in culture decreasing to 1.40*10-3 pg/mL per cell at 7 days.

2)DAPI Staining and SEM demonstrates adherence, viability, and proliferation of ASCs within ADM constructs. 3)H&E and mouse CD34 immunohistochemistry demonstrate increase vascularization by ASCs seeded 20 minutes and 3 hours compared to fresh fat seeded 20 minutes and controls. Immunoflouresence shows recruitment of local mouse cells to help augment vascularization.
CONCLUSION: In vitro findings show ASCs seeded onto AlloDerm successfully attach, migrate, and produce VEGF, promoting increased vascularization and incorporation of the graft in vivo in as early as 9 days. In the setting of devascularized tissue, as irradiated tissue, increased incorporation would be invaluable to treatment and outcome. ASCs hold immense therapetic value, which may be utilized to vascularize substrates as ADM.


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