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Jennifer H. Ho, Hsiang-Yin Pao, Yun-Chuang Chang, Chieh-Feng Cheng; Human Orbital Fat-derived Stem Cells Promote Corneal Re-epithlialization And Attenuate Local Inflammation In Vivo . Invest. Ophthalmol. Vis. Sci. 2011;52(14):6677.
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Extensive loss of limbal epithelial cells results in recurrent corneal ulceration and corneal opacity. Replacement of stem cells by limbal transplantation is the only way for ocular surface reconstruction. However, destruction of the healthy fellow limbus tissue is inevitable while allogenic limbal graft for bilateral diseases is restricted by tissue rejection and unexpected inflammation. We have demonstrated that human orbital fat-derived stem cells (OFSCs) possess the corneal epithelial cell differentiation ability upon mix-cultured with human corneal epithelial cells. The aim of this study is to investigate the therapeutic effect and mechanisms of OFSCs on acute corneal/limbal injury in vivo.
Eight-week-old male BALB/c mice cornea (n=12) were covered by filtering paper with 0.5N NaOH for 30 seconds, and then, corneal as well as limbal epithelium was removed by surgical knife. One drop with 2 x 105 OFSCs in 5 ul PBS or 5 ul of PBS was treated twice a day. Image of corneal clearance and corneal wound stained with fluorescein strip were recorded every day. Mice were sacrificed for histopathological exam, immunohistochemistry staining, immunofluorescence staining, and western blot analysis three days after corneal injury.
Compared with PBS treatment, topical treatment with OFSCs promoted corneal re-epithelialization in vivo evidenced by significantly decreased corneal opacity and wound area. Corneal re-epithelialization were 67.2±6.24 % in PBS-treated group and 90.6±13.67 % in OFSCs-treated group (p=0.003) on the third-day after injury. Pathological examination demonstrated that OFSCs ameliorated acute alkali injury on cornea including diminishing corneal edema and decreasing stromal neutrophil and macrophage infiltration. In OFSCs treated cornea, human β2-microglobulin or human IgG expressing cells could be detected on mouse corneal epithelium, especially at limbal area and central basal layer, and some of transplanted cells expressed cytokeratin-3. OFSCs significantly decreased protein expression of inflammatory cytokines within cornea especially inducible nitric oxide synthase.
Topical OFSCs treatment was effective at promoting corneal wound healing. The therapeutic effect was achieved by both direct differentiation into corneal epithelial cells and inflammation inhibition by OFSCs.
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