Department of Plastic and Reconstructive Surgery, Guangdong Second Provincial General Hospital, Guangzhou 510317, China.
The Graduate School, Southern Medical University, Guangzhou 510515, China.
J Dermatol Sci. 2018 Jan;89(1):67-76. doi: 10.1016/j.jdermsci.2017.07.020. Epub 2017 Oct 26.
There is a lack of proper animal models to study keloid formation.
To create three-dimensional poly lactic-co-glycolic acid (PLGA) scaffolds containing autologous platelet-rich plasma (PRP) as an in vitro culture environment for keloid fibroblasts (KL), and to study their implantation into nude mice to mimic the process of keloid formation.
Normal fibroblasts (FB) and KL cells were isolated from surgical specimens and transduced with lentivirus loaded with green fluorescent protein (GFP) and luciferase genes. The FB and KL cells were three-dimensionally cultured for 14-18days in PLGA scaffolds containing PRP. Ten mice were implanted with KL cells in their left forelimbs(KL), and FB-scaffolds (FB+PLGA) in their right forelimbs. An additional ten mice were implanted with PLGA scaffolds without cells (PLGA) in their left forelimbs, and KL-scaffolds (KL+PLGA) in their right forelimbs. Graft volume and collagen content were analyzed 120days after the implantation.
in vivo luminescence cell imaging showed that the FB cells proliferated in the PLGA scaffolds within 60days after implantation, and reached a plateau afterwards until 120days after implantation. The KL cells continuously proliferated in the PLGA scaffolds until 120days after implantation. The KL+PLGA group showed higher graft volumes than the FB+PLGA group 120days after the implantation (median volume, 166.95 vs. 63.34mm); however, the difference is not statistically significant (P=0.743), due to a large variation of the graft volume within each group. Furthermore, Sirius red staining revealed increased collagen I deposition, and immunohistochemistry showed large-scale accumulation of α-smooth muscle actin (α-SMA), collagen I, and collagen III in the KL+PLGA grafts.
The three-dimensional PLGA scaffold containing PRP supports keloid fibroblast growth and contributes to keloid formation in a nude mouse model.
目前缺乏合适的动物模型来研究瘢痕疙瘩的形成。
构建三维聚乳酸-羟基乙酸共聚物（PLGA）支架，内含自体富血小板血浆（PRP）作为体外培养环境，用于培养瘢痕疙瘩成纤维细胞（KL），并将其植入裸鼠体内以模拟瘢痕疙瘩形成的过程。
从手术标本中分离正常成纤维细胞（FB）和 KL 细胞，并转导携带绿色荧光蛋白（GFP）和荧光素酶基因的慢病毒。将 FB 和 KL 细胞在含有 PRP 的 PLGA 支架中进行三维培养 14-18 天。10 只小鼠的左前肢植入 KL 细胞（KL），右前肢植入 FB 支架（FB+PLGA）。另外 10 只小鼠的左前肢植入无细胞 PLGA 支架（PLGA），右前肢植入 KL 支架（KL+PLGA）。植入后 120 天分析移植物体积和胶原含量。
体内发光细胞成像显示，FB 细胞在植入后 60 天内在 PLGA 支架中增殖，并在随后的 120 天内达到平台期。KL 细胞在 PLGA 支架中持续增殖，直至植入后 120 天。植入后 120 天，KL+PLGA 组的移植物体积大于 FB+PLGA 组（中位数体积，166.95 比 63.34mm3）；然而，差异无统计学意义（P=0.743），这是由于每组内移植物体积的差异较大。此外，天狼星红染色显示胶原蛋白 I 沉积增加，免疫组织化学显示 KL+PLGA 移植物中大量α-平滑肌肌动蛋白（α-SMA）、胶原蛋白 I 和胶原蛋白 III 积聚。
含 PRP 的三维 PLGA 支架支持瘢痕疙瘩成纤维细胞的生长，并有助于裸鼠模型中瘢痕疙瘩的形成。
