Shan Xing, Choi Jong Hyeon, Kim Ki Joo, Lee Yoon Jae, Ryu Yeon Hee, Lee Su Jin, Moon Suk-Ho, Rhie Jong Won
1Department of Plastic and Reconstructive Surgery, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul, 06591 Republic of Korea.
2Department of Molecular Biomedicine, College of Medicine, The Catholic University of Korea, 222 Banpo-daero, Seocho-gu, Seoul, 06591 Republic of Korea.
Tissue Eng Regen Med. 2018 Jan 16;15(1):49-61. doi: 10.1007/s13770-017-0105-7. eCollection 2018 Feb.
This study was to investigate the effect of subcutaneous injection of the adipose stem cells (ASCs) with conditioned media (CM) in the treatment of acne vulgaris scar. We used Adult male New Zealand white rabbit ears as an animal model and induced acne formation by Kignman method. Adipose tissue was isolated and harvested from the scapula of rabbits, and ASCs were cultured and expanded until passage 1. There have four groups in our experiment, include phosphate buffered saline (PBS), ASCs with PBS (ASC + PBS), CM, and ASCs with CM (ASC + CM) group. This solution of 0.6 ml injected to subcutaneous in each group. ASC + PBS and ASC + CM groups were containing ASCs of 5.0 × 10 cells/ml. We analyzed the treatment of 4 groups to scar tissue after 2 and 4 weeks by hematoxylin and eosin stain, immunohistochemistry, and RNA expression level of tumor necrosis factor-α (TNF-α), interleukin-1α (IL-1α), and matrix metalloproteinase-2 (MMP-2). Also, the expression of keratin 16 (K16) was detected by western blot analysis. H&E stain showed that infiltration of inflammation cells was significantly reduced at 2 and 4 weeks, as well as re-epithelialization was improved in the ASC + CM group. The ASC + CM gourp was reduced both expression levels of TNF-α, IL-1α, and MMP-2 and K16 protein level. In conclusion, the ASCs with CM has a significant curative effect on acne vulgaris scar, more to the point, the CM has a key role on treatment. It could be applied to a therapeutic approach to regenerate to treat acne vulgaris scar.
本研究旨在探讨皮下注射脂肪干细胞(ASCs)与条件培养基(CM)治疗寻常痤疮瘢痕的效果。我们以成年雄性新西兰白兔耳朵作为动物模型,采用Kligman法诱导痤疮形成。从兔肩胛骨分离并获取脂肪组织,培养并扩增ASCs至第1代。我们的实验有四组,包括磷酸盐缓冲盐水(PBS)、ASCs与PBS(ASC + PBS)、CM以及ASCs与CM(ASC + CM)组。每组皮下注射0.6 ml该溶液。ASC + PBS组和ASC + CM组含有5.0×10⁶个细胞/ml的ASCs。我们在2周和4周后通过苏木精和伊红染色、免疫组织化学以及肿瘤坏死因子-α(TNF-α)、白细胞介素-1α(IL-1α)和基质金属蛋白酶-2(MMP-2)的RNA表达水平分析四组对瘢痕组织的治疗效果。此外,通过蛋白质印迹分析检测角蛋白16(K16)的表达。苏木精和伊红染色显示,在2周和4周时炎症细胞浸润显著减少,ASC + CM组的再上皮化也得到改善。ASC + CM组降低了TNF-α、IL-1α和MMP-2的表达水平以及K16蛋白水平。总之,ASCs与CM对寻常痤疮瘢痕有显著疗效,更重要的是,CM在治疗中起关键作用。它可应用于再生治疗寻常痤疮瘢痕的治疗方法。