Namgoong Sik, Lee Hyunsu, Lee Jae-Sun, Jeong Seong Ho, Han Seung-Kyu, Dhong Eun-Sang
Department of Plastic Surgery, Korea University Guro Hospital, Seoul, South Korea.
J Craniofac Surg. 2019 May/Jun;30(3):947-954. doi: 10.1097/SCS.0000000000005393.
Although therapies with human amnion/chorion are used to ameliorate acute and chronic wounds, it is unclear which component of the amnion/chorion tissue promotes wound healing. To characterize the comparative effects of amnion and chorion in wound healing, we used human adipose-derived stromal cells to assess cell viability, migration, and gel contraction after treatment with amnion membrane extract (AME) or chorion membrane extract (CME). We then correlated the possible effectors via AME and CME protein profiling, and compared them by enzyme-linked immunosorbent assay (ELISA), western blotting, and immunocytochemistry. Cell viability was significantly increased with 50 and 100 μg/mL AME treatment, but with CME treatment, a significant increase was only observed with 100 μg/mL. With CME treatment, cell migration was 2.22-fold greater than the control, and collagen gels showed 20% greater contraction. Compared to control, the expression levels of α-smooth muscle actin (SMA) and smooth muscle protein 22-alpha (SM22α) increased both with AME and CME treatments, whereas calponin expression decreased. Protein profiling revealed significantly higher tissue inhibitor of metalloproteinase-1 (TIMP-1), interleukin-8, exotoxin, and adiponectin levels in CME than in AME, and ELISA revealed 8-fold higher adiponectin levels in cells treated with CME than those treated with AME. Immunocytochemistry revealed that α-SMA, SM22α, and calponin were significantly higher in CME- than AME-treated cells; however, adiponectin treatment did not enhance α-SMA, SM22α, or calponin expression. In conclusion, amnion and chorion membrane extracts exerted differential effects on proliferation and contraction of human adipose-derived stromal cells. Amnion extract was superior at inducing cell proliferation and migration, whereas CME was superior at inducing cell contraction.
尽管使用人羊膜/绒毛膜疗法来改善急慢性伤口，但尚不清楚羊膜/绒毛膜组织的哪个成分能促进伤口愈合。为了表征羊膜和绒毛膜在伤口愈合中的比较效果，我们用人脂肪来源的基质细胞评估用羊膜膜提取物（AME）或绒毛膜膜提取物（CME）处理后的细胞活力、迁移和凝胶收缩情况。然后，我们通过AME和CME蛋白质谱分析来关联可能的效应物，并通过酶联免疫吸附测定（ELISA）、蛋白质印迹法和免疫细胞化学进行比较。用50和100μg/mL AME处理时，细胞活力显著增加，但用CME处理时，仅在100μg/mL时观察到显著增加。用CME处理时，细胞迁移比对照高2.22倍，胶原凝胶收缩增加20%。与对照相比，AME和CME处理后α-平滑肌肌动蛋白（SMA）和平滑肌蛋白22-α（SM22α）的表达水平均升高，而钙调蛋白表达降低。蛋白质谱分析显示，CME中金属蛋白酶组织抑制剂-1（TIMP-1）、白细胞介素-8、外毒素和脂联素水平显著高于AME，ELISA显示用CME处理的细胞中脂联素水平比用AME处理的细胞高8倍。免疫细胞化学显示，CME处理的细胞中α-SMA、SM22α和钙调蛋白显著高于AME处理的细胞；然而，脂联素处理并未增强α-SMA、SM22α或钙调蛋白的表达。总之，羊膜和绒毛膜膜提取物对人脂肪来源的基质细胞的增殖和收缩具有不同的影响。羊膜提取物在诱导细胞增殖和迁移方面更具优势，而CME在诱导细胞收缩方面更具优势。
