Wang Ning, Liu Hanping, Li Xuefeng, Zhang Qilei, Chen Maosheng, Jin Ying, Deng Xiaoyuan
MOE Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, China.
College of Life Science, South China Normal University, Guangzhou, China.
Cell Physiol Biochem. 2017;42(2):623-639. doi: 10.1159/000477872. Epub 2017 Jun 9.
BACKGROUND/AIMS: Bone marrow Mesenchymal stem cells (MSCs) are promising for promoting cutaneous wound healing through reinforcing cellular processes. We evaluated the effect of GFP-tagged MSCs transplantation on skin regeneration in excisional wounds in mice.
MSCs from GFP-labeled transgenic mice were co-cultured with acellular dermal matrix (ADM) scaffolds, and MSC-ADM scaffolds were transplanted into surgical skin wounds of BALB/c mice. After implantation, the survival and behavior of MSCs were examined by second harmonic generation and two-photon excitation fluorescence imaging, western blotting and DNA amplification and sequencing.
GFP-tagged MSCs were retained inside the regenerating skin until day 14 post-transplantation. Alpha-smooth muscle actin (α-SMA) and vimentin (VIM) were detected at 3, 5, 7, and 14 days post-transplantation by immunofluorescence double labeling. Although the GFP+/α-SMA+- and GFP+/VIM+-cell numbers decreased gradually with healing time, α-SMA+- and VIM+-cell numbers significantly increased, most of them were endogenous functional cells which were related to angiogenesis and collagen fiber structural remodeling.
Therefore, in the initial stage of wound healing, transplanted MSCs differentiated into functional cells and played paracrine roles to recruit more endogenous cells for tissue remodeling. With the disappearance of exogenous cells, endogenous cells were responsible for the latter stage of cutaneous wound healing.
背景/目的：骨髓间充质干细胞（MSCs）有望通过加强细胞过程促进皮肤伤口愈合。我们评估了绿色荧光蛋白（GFP）标记的间充质干细胞移植对小鼠切除伤口皮肤再生的影响。
将来自GFP标记转基因小鼠的间充质干细胞与脱细胞真皮基质（ADM）支架共培养，然后将间充质干细胞-ADM支架移植到BALB/c小鼠的手术皮肤伤口中。植入后，通过二次谐波产生和双光子激发荧光成像、蛋白质免疫印迹以及DNA扩增和测序来检测间充质干细胞的存活和行为。
GFP标记的间充质干细胞在移植后14天内一直保留在再生皮肤内。通过免疫荧光双标记在移植后第3、5、7和14天检测到α-平滑肌肌动蛋白（α-SMA）和波形蛋白（VIM）。尽管GFP+/α-SMA+和GFP+/VIM+细胞数量随着愈合时间逐渐减少，但α-SMA+和VIM+细胞数量显著增加，其中大多数是与血管生成和胶原纤维结构重塑相关的内源性功能细胞。
因此，在伤口愈合的初始阶段，移植的间充质干细胞分化为功能细胞并发挥旁分泌作用，招募更多内源性细胞进行组织重塑。随着外源性细胞的消失，内源性细胞负责皮肤伤口愈合的后期阶段。
