The anti-scarring effect of corneal stromal stem cell therapy is mediated by transforming growth factorβ3  

作　　者：Lin Weng James L.Funderburgh Irona Khandaker Moira L.Geary Tianbing Yang Rohan Basu Martha L.Funderburgh Yiqin Du Gary Hin-Fai Yam 

机构地区：[1]Department of Ophthalmology,University of Pittsburgh School of medicine,203 Lothrop Street,Pittsburgh,PA 15213,USA [2]Shanghai Lanhe Optometry and Ophthalmology Clinic,Shanghai 200032,People’s Republic of China

出　　处：《Eye and Vision》2020年第1期512-525,共14页眼视光学杂志（英文）

基　　金：This work was supported by the Department of Defence Grant W81WH-19-1-0778(JLF,YD),NIH Grants RO1 EY016415(JLF)and P30 EY008098(JLF),Stein Innovator Award from Research to Prevent Blindness(JLF),Eye and Ear Foundation of Pittsburgh,and Louis J Fox Centre for Vision Restoration.

摘　　要：Background:Corneal stromal stem cells(CSSC)reduce corneal inflammation,prevent fibrotic scarring,and regenerate transparent stromal tissue in injured corneas.These effects rely on factors produced by CSSC to block the fibrotic gene expression.This study investigated the mechanism of the scar-free regeneration effect.Methods:Primary human CSSC(hCSSC)from donor corneal rims were cultivated to passage 3 and co-cultured with mouse macrophage RAW264.7 cells induced to M1 pro-inflammatory phenotype by treatment with interferonγand lipopolysaccharides,or to M2 anti-inflammatory phenotype by interleukin-4,in a Transwell system.The timecourse expression of human transforming growth factorβ3(hTGFβ3)and hTGFβ1 were examined by immunofluorescence and qPCR.TGFβ3 knockdown for>70%in hCSSC[hCSSC-TGFβ3(si)]was achieved by small interfering RNA transfection.Naïve CSSC and hCSSC-TGFβ3(si)were transplanted in a fibrin gel to mouse corneas,respectively,after wounding by stromal ablation.Corneal clarity and the expression of mouse inflammatory and fibrosis genes were examined.Results:hTGFβ3 was upregulated by hCSSC when co-cultured with RAW cells under M1 condition.Transplantation of hCSSC to wounded mouse corneas showed significant upregulation of hTGFβ3 at days 1 and 3 post-injury,along with the reduced expression of mouse inflammatory genes(CD80,C-X-C motif chemokine ligand 5,lipocalin 2,plasminogen activator urokinase receptor,pro-platelet basic protein,and secreted phosphoprotein 1).By day 14,hCSSC treatment significantly reduced the expression of fibrotic and scar tissue genes(fibronectin,hyaluronan synthase 2,Secreted protein acidic and cysteine rich,tenascin C,collagen 3a1 andα-smooth muscle actin),and the injured corneas remained clear.However,hCSSC-TGFβ3(si)lost these anti-inflammatory and anti-scarring functions,and the wounded corneas showed intense scarring.Conclusion:This study has demonstrated that the corneal regenerative effect of hCSSC is mediated by TGFβ3,inducing a scar-free tissue response.

关 键 词：Cornea wound healing Corneal stromal stem cells TGFΒ3 Inflammation FIBROSIS 

分 类 号：R772.2[医药卫生—眼科]