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Sharaf, Kariem; Kleinsasser, Antonia; Schwenk-Zieger, Sabina; Gires, Olivier; Schinke, Henrik; Kohlbauer, Vera; Jakob, Mark; Canis, Martin; Haubner, Frank (2019): Molecular Characterization of Lipoaspirates Used in Regenerative Head and Neck Surgery. In: Jama Facial Plastic Surgery, Vol. 21, No. 6: pp. 526-534
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This case series study characterizes the composition of lipoaspirates and their wound-healing properties in patients receiving autologous fat injections during head and neck surgical procedures. Key PointsQuestionWhat is the composition of lipoaspirates used for lipotransfer in the head and neck, and what is their potential for wound healing in vitro? FindingsIn this case series study of lipoaspirate samples obtained from 15 patients, adipose-derived mesenchymal stem cells were found in high purity and were able to multidifferentiate. The adipose-derived stem/stromal cells and their supernatants had proliferation- and immune-modulating properties in vitro. MeaningThe findings suggest that lipoaspirates contain a concentration of adipose-derived stem/stromal cells that can be used for regenerative head and neck surgical procedures. ImportanceAdipose-derived mesenchymal stem cells (ASCs) have been used commonly in regenerative medicine and increasingly for head and neck surgical procedures. Lipoaspiration with centrifugation is purported to be a mild method for the extraction of ASCs used for autologous transplants to restore tissue defects or induce wound healing. The content of ASCs, their paracrine potential, and cellular potential in wound healing have not been explored for this method to our knowledge. ObjectiveTo evaluate the characteristics of lipoaspirates used in reconstructive head and neck surgical procedures with respect to wound healing. Design, Setting, and ParticipantsThis case series study included 15 patients who received autologous fat injections in the head and neck during surgical procedures at a tertiary referral center. The study was performed from October 2017 to November 2018, and data were analyzed from October 2017 to February 2019. Main Outcomes and MeasuresExcessive material of lipoaspirates from subcutaneous abdominal fatty tissue was examined. Cellular composition was analyzed using immunohistochemistry (IHC) and flow cytometry, and functionality was assessed through adipose, osteous, and chondral differentiation in vitro. Supernatants were tested for paracrine ASC functions in fibroblast wound-healing assays. Enzyme-linked immunosorbent assay measurement of tumor necrosis factor (TNF), vascular endothelial growth factor (VEGF), stromal-derived factor 1 alpha (SDF-1 alpha), and transforming growth factor beta 3 (TGF-beta 3) was performed. ResultsAmong the 15 study patients (8 [53.3%] male;mean [SD] age at the time of surgery, 63.0 [2.8] years), the stromal vascular fraction (mean [SE], 53.3% [4.2%]) represented the largest fraction within the native lipoaspirates. The cultivated cells were positive for CD73 (mean [SE], 99.90% [0.07%]), CD90 (99.40% [0.32%]), and CD105 (88.54% [2.74%]);negative for CD34 (2.70% [0.45%]) and CD45 (1.74% [0.28%]) in flow cytometry;and negative for CD14 (10.56 [2.81] per 300 IHC score) and HLA-DR (6.89 [2.97] per 300 IHC score) in IHC staining;they differentiated into osteoblasts, adipocytes, and chondrocytes. The cultivated cells showed high expression of CD44 (mean [SE], 99.78% [0.08%]) and CD273 (82.56% [5.83%]). The supernatants were negative for TNF (not detectable) and SDF-1 alpha (not detectable) and were positive for VEGF (mean [SE], 526.74 [149.84] pg/mL for explant supernatants;528.26 [131.79] pg/10(6) per day for cell culture supernatants) and TGF-beta 3 (mean [SE], 22.79 [3.49] pg/mL for explant supernatants;7.97 [3.15] pg/10(6) per day for cell culture supernatants). Compared with control (25% or 50% mesenchymal stem cell medium), fibroblasts treated with ASC supernatant healed the scratch-induced wound faster (mean [SE]: control, 1.000 [0.160];explant supernatant, 1.369 [0.070];and passage 6 supernatant, 1.492 [0.094]). Conclusions and RelevanceThe cells fulfilled the international accepted criteria for mesenchymal stem cells. The lipoaspirates contained ASCs that had the potential to multidifferentiate with proliferative and immune-modulating properties. The cytokine profile of the isolated ASCs had wound healing-promoting features. Lipoaspirates may have a regenerative potential and an application in head and neck surgery. Level of EvidenceNA.