Effect of Human Corneal Mesenchymal Stromal Cell-derived Exosomes on Corneal Epithelial Wound Healing

被引:206
作者
Samaeekia, Ravand [1 ]
Rabiee, Behnam [1 ]
Putra, Ilham [1 ]
Shen, Xiang [1 ]
Park, Young Jae [1 ]
Hematti, Peiman [2 ,3 ]
Eslani, Medi [1 ]
Djalilian, Ali R. [1 ]
机构
[1] Univ Illinois, Dept Ophthalmol & Visual Sci, 1855 W Taylor St,EEI 3164, Chicago, IL 60612 USA
[2] Univ Wisconsin, Sch Med & Publ Hlth, Dept Med, Madison, WI USA
[3] Univ Wisconsin, Sch Med & Publ Hlth, Univ Wisconsin Carbone Canc Ctr, Madison, WI USA
关键词
exosome; corneal wound healing; mesenchymal stromal cells; corneal epithelium; wound healing; OCULAR SURFACE RECONSTRUCTION; STEM-CELLS; PHYSICAL-CHARACTERIZATION; EXTRACELLULAR VESICLES; NANOFIBER SCAFFOLDS; LIGHT-SCATTERING; INHIBITION; INJURY; RNAS;
D O I
10.1167/iovs.18-24803
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
PURPOSE. Mesenchymal stromal cells (MSCs) have been used therapeutically to modulate inflammation and promote repair. Extracellular vesicles, including exosomes, have been identified as one of the important mediators. This study investigated the effect of human corneal MSC-derived exosomes on corneal epithelial wound healing. METHODS. Corneal MSCs (cMSCs) were isolated from human cadaver corneas. The secretome was collected after 72 hours and exosomes were isolated using differential ultracentrifugation. Morphology and size of exosomes were examined by electron microscopy and dynamic light scattering. Expression of CD9, CD63, and CD81 by cMSC exosomes was evaluated by western blotting. Cellular uptake of exosomes was studied using calcein-stained exosomes. The effect of exosome on wound healing was measured in vitro using a scratch assay and in vivo after 2-mm epithelial debridement wounds in mice. RESULTS. cMSC exosomes were morphologically round and main population ranged between 40 and 100 nm in diameter. They expressed CD9, CD63, and CD81, and did not express GM130, Calnexin, and Cytochrome-C. Stained cMSC exosomes were successfully taken up by human cMSCs, human corneal epithelial cells (HCECs), and human macrophages in vitro and by corneal epithelium in vivo. In scratch assay, after 16 hours, cMSC exosome treated HCECs had 30.1%+/- 14% remaining wound area compared to 72.9%+/- 8% in control (P < 0.005). In vivo, after 72 hours, cMSC exosome-treated corneas had 77.5%+/- 3% corneal wound healing compared to 41.6%+/- 7% in the control group (P < 0.05). CONCLUSIONS. Human cMSC exosomes can accelerate corneal epithelial wound healing, and thus, may provide a therapeutic approach for ocular surface injuries.
引用
收藏
页码:5194 / 5200
页数:7
相关论文
共 36 条
[1]   Mesenchymal stem cell secreted vesicles provide novel opportunities in (stem) cell-free therapy [J].
Baglio, Serena Rubina ;
Pegtel, D. Michiel ;
Baldini, Nicola .
FRONTIERS IN PHYSIOLOGY, 2012, 3
[2]   Effects of Mesenchymal Stem Cell-Derived Exosomes on Experimental Autoimmune Uveitis [J].
Bai, Lingling ;
Shao, Hui ;
Wang, Hongxing ;
Zhang, Zhihui ;
Su, Chang ;
Dong, Lijie ;
Yu, Bo ;
Chen, Xiteng ;
Li, Xiaorong ;
Zhang, Xiaomin .
SCIENTIFIC REPORTS, 2017, 7
[3]   Exosomes secreted by nematode parasites transfer small RNAs to mammalian cells and modulate innate immunity [J].
Buck, Amy H. ;
Coakley, Gillian ;
Simbari, Fabio ;
McSorley, Henry J. ;
Quintana, Juan F. ;
Le Bihan, Thierry ;
Kumar, Sujai ;
Abreu-Goodger, Cei ;
Lear, Marissa ;
Harcus, Yvonne ;
Ceroni, Alessandro ;
Babayan, Simon A. ;
Blaxter, Mark ;
Ivens, Alasdair ;
Maizels, Rick M. .
NATURE COMMUNICATIONS, 2014, 5
[4]   Suppression of alkali-induced oxidative injury in the cornea by mesenchymal stem cells growing on nanofiber scaffolds and transferred onto the damaged corneal surface [J].
Cejkova, Jitka ;
Trosan, Peter ;
Cejka, Cestmir ;
Lencova, Anna ;
Zajicova, Alena ;
Javorkova, Eliska ;
Kubinova, Sarka ;
Sykova, Eva ;
Holan, Vladimir .
EXPERIMENTAL EYE RESEARCH, 2013, 116 :312-323
[5]   Extrinsic and Intrinsic Mechanisms by Which Mesenchymal Stem Cells Suppress the Immune System [J].
Coulson-Thomas, Vivien J. ;
Coulson-Thomas, Yvette M. ;
Gesteira, Tarsis F. ;
Kao, Winston W. -Y. .
OCULAR SURFACE, 2016, 14 (02) :121-134
[6]   Cornea-Derived Mesenchymal Stromal Cells Therapeutically Modulate Macrophage Immunophenotype and Angiogenic Function [J].
Eslani, Medi ;
Putra, Ilham ;
Shen, Xiang ;
Hamouie, Judy ;
Tadepalli, Asha ;
Anwar, Khandaker N. ;
Kink, John A. ;
Ghassemi, Samaneh ;
Agnihotri, Gaurav ;
Reshetylo, Sofiya ;
Mashaghi, Alireza ;
Dana, Reza ;
Hematti, Peiman ;
Djalilian, Ali R. .
STEM CELLS, 2018, 36 (05) :775-784
[7]   Corneal Mesenchymal Stromal Cells Are Directly Antiangiogenic via PEDF and sFLT-1 [J].
Eslani, Medi ;
Putra, Ilham ;
Shen, Xiang ;
Hamouie, Judy ;
Afsharkhamseh, Neda ;
Besharat, Soroush ;
Rosenblatt, Mark I. ;
Dana, Reza ;
Hematti, Peiman ;
Djalilian, Ali R. .
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 2017, 58 (12) :5507-5517
[8]   Potential role of corneal epithelial cell-derived exosomes in corneal wound healing and neovascularization [J].
Han, Kyu-Yeon ;
Tran, Jennifer A. ;
Chang, Jin-Hong ;
Azar, Dimitri T. ;
Zieske, James D. .
SCIENTIFIC REPORTS, 2017, 7
[9]   A Comparative Study of the Therapeutic Potential of Mesenchymal Stem Cells and Limbal Epithelial Stem Cells for Ocular Surface Reconstruction [J].
Holan, Vladimir ;
Trosan, Peter ;
Cejka, Cestmir ;
Javorkova, Eliska ;
Zajicova, Alena ;
Hermankova, Barbora ;
Chudickova, Milada ;
Cejkova, Jitka .
STEM CELLS TRANSLATIONAL MEDICINE, 2015, 4 (09) :1052-1063
[10]   Mesenchymal Stem Cells, Nanofiber Scaffolds and Ocular Surface Reconstruction [J].
Holan, Vladimir ;
Javorkova, Eliska .
STEM CELL REVIEWS AND REPORTS, 2013, 9 (05) :609-619