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Ultrasound Assisted Liposuction Does Not Compromise The Regenerative Potential Of Ascs
Dominik Duscher, MD1, Anna Luan, MS2, David Atashroo, MD2, Zeshaan N. Maan, MBBS, MS2, Elizabeth R. Zielins, MD2, Kevin Paik, BA2, Alexander J. Whittam, BS2, Michelle Lin, BS2, Graham G. Walmsley, BA2, Michael S. Hu, MD, MPH, MS2, Manfred Schmidt, MD1, Georg M. Huemer, MD, MBA, MS1, Derrick C. Wan, MD2, Michael T. Longaker, MD, MBA2, Geoffrey C. Gurtner, MD, FACS2.
1Johannes Keppler University, Linz, Austria, 2Stanford University, Stanford, CA, USA.
Human adipose derived stromal cells (ASCs) have gained recent interest for their therapeutic potential in regenerative medicine. Even though liposuction is the primary method of obtaining ASCs, little is known about the effects of different liposuction methods on their regenerative abilities. Ultrasound assisted liposuction (UAL) can facilitate the process of lipoaspiration by increasing the speed and safety of the fat harvest. However, a paucity of evidence exists regarding its impact on the regenerative capacity of harvested ASCs. In this study, we evaluate the regenerative abilities of ASCs harvested with a third generation UAL device versus the industry standard suction assisted lipoaspiration (SAL).
Lipoaspirate was obtained from elective surgery patients using UAL and SAL, and processed for enzymatic isolation of the stromal vascular fraction (SVF). The SVF was then sorted using Fluorescent Assisted Cell Sorting (FACS) based on an established progenitor surface marker profile (CD34+CD31-CD45-) and live/dead gating, to obtain viable ASCs. An MTT assay was performed on cultured ASCs to compare their proliferation potential. The multlineage differentiation capacity of the ASCs was then assessed by the induction of adipogenic and osteogenic differentiation and subsequent Oil Red O, Alkaline Phosphatase and Alizarin Red staining. Additionally, qRT-PCR expression analysis of key adipogenic and osteogenic genes was performed at culture day 0, 7, and 14. Finally, the regenerative potential of the ASCs was compared in and in vivo wound healing model. Paired full thickness wounds were created on the dorsum of CD1 athymic nude mice and a hydrogel scaffold alone or seeded with 250k UAL or SAL ASCs was placed into the wound bed. Healing was assessed serially until wound re-epithelialization and histological samples were taken upon complete closure.
UAL and SAL demonstrated equivalent viable ASC yield on FACS and proliferative potential on MTT assay (*p > .05). There was no significant difference in adipogenic or osteogenic differentiation capacity (*p > .05). Similarly, qRT-PCR showed equal expression of multiple osteogenic and adipogenic genes in both ASC groups (*p > .05). Wound healing was significantly improved in both cell therapy groups compared to hydrogel alone (*p .05). This could further be confirmed on a histological level by an equally significant enhancement of neovascularization in both cell therapy groups versus the hydrogel control group.
UAL represents a successful method for obtaining ASCs for regenerative medicine. Cells harvested with this alternative approach to liposuction are suitable for potential therapeutic and tissue engineering applications.
Figure 1. Application of ASCs harvested via UAL and SAL equally improve excisional wound healing. (A) Gross appearance, (B) wound healing kinetics, and (C) closing times of humanized excisional murine wounds treated with hydrogel seeded hASCs harvested via UAL, SAL, or unseeded hydrogel. * indicates p ≤ 0.05.
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