ABSTRACT
@#AIM: To study the expressions of TGFBI and microtubule-associated protein 1 light chain 3 alpha(LC3)in granular corneal dystrophy, and the influences of lithium chloride(LiCl)on corneal stromal fibroblast cell proliferation by TGFBI. <p>METHODS: Immunohistochemistry and Western-blot assays were used to detect the expression levels of TGFBI and LC3 in corneal dystrophy and normal corneal tissues. TGFBI overexpression vector was transfected into corneal stromal fibroblasts, and then the cells were treated with 5, 10, 20, 40mmol/L LiCl for different times(0, 1, 6, 12h), and Western-blot assay was performed to evaluate the expression levels of TGFBI and LC3, and CCK-8 assay was carried out to assess cell proliferation activity.<p>RESULTS: TGFBI and LC3 were highly expressed in corneal tissues of patients with corneal dystrophy. TGFBI overexpression inhibited the proliferation ability of corneal stromal fibroblasts(<i>P</i><0.05). LiCl inhibited the expression levels of TGFBI and LC3, and enhanced the cell proliferation activity in corneal stromal fibroblasts transfected with mutant TGFBI(<i>P</i><0.05).<p>CONCLUSION: LiCl promoted the proliferation and autophagy of corneal stromal fibroblasts, and its mechanism may be related to down regulated expressions of TGFBI and LC3.
ABSTRACT
AIM: To investigate the biocolonization of poly 2-hydroxyethy methacrylate (PHEMA) sponge with cornea tissue and evaluate the therapeutic effects of modified porous poly 2-hydroxyethy methacrylate-Polymethyl met-hacrylate (PHEMA-PMMA) Keratoprostheses (KPro) on rabbit and monkey corneas. METHODS:The KPro were made using two-stage polymerization combined with mechanical cutting. The experiment was divided into two groups. In the control group, ten normal rabbit eyes received lamellar implanta-tion of PHEMA sponges. The sponges were obtained 2 weeks, 1,2,3 and 4 months after operation. The cell proliferation and neovascularization inside the sponges were observed using light and transmission electron microscopy (TEM) and immunohistochemistry. In the experimental group, the porous PHEMA-PMMA KPros were inserted into the lamellar pockets of eight rabbit corneas and two monkey corneas (stage I operation). The healing process was investigated by slit-lamp microscopy. The anterior lamellar cornea tissues were removed 3 months after surgery, exposing the under-neath transparent core (stage II operation). The operated eyes were then followed up for 3-6 months.light microscope, fibroblasts started to grow into the cornea 2 weeks after operation; lots of cells, accompanied with new blood vessels, invaded into the cornea 2-3 months after surgery. Invading cells of sponge, as well as keratocytes, were positive for vimentin. Under the electron microscope, the invading cells looked healthy and were surrounded by extracellular matrix and collagen. In 8 rabbit eyes which received KPro implantation, anterior lamellar cornea melting happened in two eyes after the stage II operation. The remaining 6 corneas retained their central cores during observation after the stage II operation.Two monkey operated eyes were found no complication thoughout the whole follow-up.cornea. The modified PHEMA-PMMA KPros have obtained a relatively stable results after implantation into animal corneas.