Department of Otolaryngology, University Hospital, Ludwig Maximilian University of Munich, Munich, Germany.
JAMA Facial Plast Surg. 2019 Dec 1;21(6):526-534. doi: 10.1001/jamafacial.2019.0851.
Adipose-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.
To evaluate the characteristics of lipoaspirates used in reconstructive head and neck surgical procedures with respect to wound healing.
DESIGN, SETTING, AND PARTICIPANTS: This 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.
Excessive 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 1α (SDF-1α), and transforming growth factor β3 (TGF-β3) was performed.
Among 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-1α (not detectable) and were positive for VEGF (mean [SE], 526.74 [149.84] pg/mL for explant supernatants; 528.26 [131.79] pg/106 per day for cell culture supernatants) and TGF-β3 (mean [SE], 22.79 [3.49] pg/mL for explant supernatants; 7.97 [3.15] pg/106 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]).
The 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.
NA.
重要性：脂肪间充质干细胞（ASCs）已广泛应用于再生医学领域，并且越来越多地应用于头颈部外科手术。据称，通过离心的脂肪抽吸术是一种温和的方法，用于提取用于自体移植的 ASCs，以恢复组织缺陷或诱导伤口愈合。据我们所知，对于这种方法，其脂肪抽吸物中的 ASC 含量、旁分泌潜能和伤口愈合中的细胞潜能尚未得到探索。
目的：评估头颈部重建外科手术中使用的脂肪抽吸物在伤口愈合方面的特征。
设计、设置和参与者：本病例系列研究纳入了 2017 年 10 月至 2018 年 11 月在一家三级转诊中心接受头颈部自体脂肪注射的 15 名患者。该研究于 2017 年 10 月进行，并于 2019 年 2 月进行数据分析。
主要结果和测量：检查来自皮下腹部脂肪组织的过量脂肪抽吸物的材料。通过免疫组织化学（IHC）和流式细胞术分析细胞组成，并通过体外脂肪、骨和成软骨分化评估功能。将上清液用于成纤维细胞伤口愈合测定中的 ASC 旁分泌功能测试。通过酶联免疫吸附试验（ELISA）测量肿瘤坏死因子（TNF）、血管内皮生长因子（VEGF）、基质衍生因子 1α（SDF-1α）和转化生长因子β3（TGF-β3）。
结果：在 15 名研究患者（8 名男性[53.3%]；手术时的平均[标准差]年龄为 63.0[2.8]岁）中，基质血管部分（平均[SE]，53.3%[4.2%]）是原生脂肪抽吸物中最大的部分。培养细胞对 CD73（平均[SE]，99.90%[0.07%]）、CD90（99.40%[0.32%]）和 CD105（88.54%[2.74%]）呈阳性；CD34（2.70%[0.45%]）和 CD45（1.74%[0.28%]）呈阴性；CD14（每 300 IHC 评分 10.56[2.81]）和 HLA-DR（每 300 IHC 评分 6.89[2.97]）呈阴性；它们可分化为成骨细胞、脂肪细胞和成软骨细胞。培养细胞显示出高表达的 CD44（平均[SE]，99.78%[0.08%]）和 CD273（82.56%[5.83%]）。上清液对 TNF（不可检测）和 SDF-1α（不可检测）呈阴性，对 VEGF（外植体上清液的平均[SE]为 526.74[149.84]pg/mL；细胞培养上清液的 528.26[131.79]pg/106 每天）和 TGF-β3（外植体上清液的平均[SE]为 22.79[3.49]pg/mL；细胞培养上清液的 7.97[3.15]pg/106 每天）呈阳性。与对照（25%或 50%间充质干细胞培养基）相比，用 ASC 上清液处理的成纤维细胞更快地愈合划痕诱导的伤口（平均[SE]：对照，1.000[0.160]；外植体上清液，1.369[0.070]；和第 6 代上清液，1.492[0.094]）。
结论和相关性：这些细胞符合国际上公认的间充质干细胞标准。脂肪抽吸物中含有具有多向分化潜力、增殖和免疫调节特性的 ASCs。分离的 ASCs 的细胞因子谱具有促进伤口愈合的特征。脂肪抽吸物可能具有再生潜力，可应用于头颈部外科手术。
证据水平：无。
