| Objective: Corneal alkali burn is common clinical blinding ocular trauma. Current methods in the treatment of the disease mainly has two kinds of drugs and surgery. Although these treatments has obtained certain therapeutic effect, but in reducing after corneal alkali burns turbidity, reduce corneal inflammatory response and reduce the effect of corneal neovascularization in remains to be improved. Between bone marrow mesenchymal stem cells because of its strong ability of proliferation and multi-directional differentiation potential, combined with the potential anti-inflammatory and anti angiogenesis effect, become the cell tissue injury disease treatment of ideal sources of seed cells. Whether between bone marrow mesenchymal stem cells has a certain therapeutic effect for corneal alkali burn, between bone marrow mesenchymal stem cell culture supernatant on whether also have therapeutic effect, at home and abroad for its corneal alkali burn less research in mice. This experiment to establish and validate a stable and repeatable corneal alkali burn model in mice, and on this basis this paper discusses the application of subconjunctival injection of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cell culture supernatant points on eye between two treatment methods for the treatment of corneal alkali burn.Methods: 1.Using infiltration in Na OH solution of monolayer filter is attached with a mouse central cornea corneal alkali burn, through the slit lamp examination ocular surface after building and HE staining, Compare different concentration of Na OH solution under different filter attached with time and the effects of corneal alkali burn, so as to choose a kind of relative stability is strong, good consistency corneal alkali burn animal model making method in mice. 2. Stability of repeatable after corneal alkali burn model in mice, mice can be divided into three groups: are respectively after alkali burn ball subconjunctival injection of bone marrow mesenchymal stem cells between the treatment group;To 6 times a day after alkali burn of bone marrow mesenchymal stem cell culture supernatant point between eye group; And PBS to 6 times a day after alkali burn liquid eye group.Between subconjunctival injection of bone marrow mesenchymal stem cells in the group of injection for alkali burn after building, points 6 times a day for eye eye, until death in mice. 3. Slit lamp observation of ocular surface after building changes: after building early 1 day, 3 days, 5 days, 7 days using fluorescein staining to observe the corneal epithelial healing; Building after 3 days, 7 days, 14 days, 21 days, 28 days observation of corneal neovascularization; Building after 3 days, 7 days, 14 days, 21 days, 28 days for central corneal turbidity score, a total of 4 weeks. 4. Building after 3 days, 7 days, 14 and 28 days, by HE staining to observe the changes in corneal tissue, inflammation and angiogenesis, and the central corneal stromal cells count. 5. Building after 3 days, 7 days, 14 and 28 days, by immunohistochemical staining method detecting alpha SMA in the corneal tissue, the expression of PCNA and Ki-67.Results: 1.Using infiltration in 1 mol/L Na OH solution of monolayer filter is attached with a mouse central cornea 10 s, moderate depth, strong repeatability can be built the ideal corneal alkali burn animal models in mice. 2. After corneal alkali burn ball subconjunctival injection of bone marrow mesenchymal stem cells and corneal alkali burn after bone marrow mesenchymal stem cell culture supernatant point between eye, have promote restoration of the corneal epithelium. Building after 1 day, 3 days, five days, 7 days, application of PBS eye control corneal epithelium defect area is greater than the other two treatment group, the difference is statistically significant. 3. After corneal alkali burn ball subconjunctival injection of bone marrow mesenchymal stem cells and corneal alkali burn after bone marrow mesenchymal stem cell culture supernatant point between eye, two methods of treatment are to reduce corneal neovascularization generated. Building after 3 days, 7 days, 14 days, 21 days, 28 days, the application of PBS point control area of new blood vessels of the eye is greater than the other two treatment group, the difference is statistically significant.4. After corneal alkali burn ball subconjunctival injection of bone marrow mesenchymal stem cells and corneal alkali burn after bone marrow mesenchymal stem cell culture supernatant point between eye, two methods of treatment are to improve the degree of corneal opacity. Building after 3 days, 7 days, 14 days, 21 days, 28 days, the application of PBS corneal turbidity point of the eye control group score more than the other two treatment group, the difference is statistically significant. 5. By HE staining observation showed that compared with the two treatment groups, PBS group liquid eye corneal epithelium healing slowly, new cornea layer number is less, stromal cells arranged a disorder, inflammatory cells infiltration and generate more new blood vessels.Two treatment group and 28 days after alkali burn of cornea stroma layer collagen arrangement more rules, no inflammatory cell infiltration and less new blood vessels.Stromal cell count showed the ball subconjunctival injection of bone marrow mesenchymal stem cells between groups and between bone marrow mesenchymal stem cell culture supernatant points on eye fluid cell count is lower than the PBS point eye group, the difference is statistically significant. 6. Immunohistochemical examination showed that PBS of corneal stromal layer of ?-SMA and MMP- 9 express positive degree is higher, and this group of cortex on corneal basal cell Ki- 67 positive degree is lower than the other two groups.Conclusions: After alkali burn ball subconjunctival injection of bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cell culture supernatant point between eye both methods of mice after alkali burn of cornea have repair effect. Between the local application of bone marrow mesenchymal stem cells and their culture supernatant can improve after alkali burn of cornea transparency, and can promote healing of corneal epithelium relative inhibit the formation of corneal neovascularization. |
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