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·AIM:To establish a simple and efficient method for the primary culture of rabbit corneal limbus stem cells. ·METHODS:Obtained the limbal tissues from rabbits, used tissue block and enzyme digestion method to culture the corneal limbus stem cells in vitro. The growth characteristics of the cultured cells in vitro were observed under inverted microscope. By means of HE, the morphology and construction features of cells were observed.And immunohistochemical method was used to identify the cultured cells. ·RESULTS:Rabbit corneal limbus stem cells could be fast and simply cultured by using tissue block and enzyme digestion method. The dynamic observation under microscope showed that rabbit corneal limbus stem cells grew well with a higher proliferative capacity. In HE staining, the morphology and structure of cells were normal.AE5 and P63 cellular immune identification were positive. · CONCLUSION: Tissue block and enzyme digestion method could be a simple and efficient mode for the primary culture of rabbit corneal limbus stem cells.
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Background Limbal stem cells (LSCs) deficiency leads to many ocular surface diseases,such as pterygium and so on.ATP-binding cassette transporter B5 (ABCB5) is a recently discovered marker of LSCs.Understanding the expression changes of ABCB5 in pterygium tissue has an important clinical significance for pterygium.Objective This study was to investigate the expression changes of ABCB5 in pterygium tissue.Methods Thirty-seven pterygium tissue specimens were collected from primary pterygium patients who received pterygium surgery,and 22 normal conjunctival tissue specimens were obtained from the strabismus patients and retinal detachment patients during surgery in 474 Hospital of Chinese PLA from January 2015 to November 2015.Immunochemistry was employed to detect the cxpression and location of ABCB5 in the specimens,and the protein and mRNA expressions of ABCB5 were detected by Western blot and real-time fluorescence quantitative PCR,respectively.The results were compared between normal conjunctival tissues and pterygium tissue.Results ABCB5 protein was expressed in the cytoplasm and nuclei of stratified squamous epithelium of 22 normal counjunctival tissue specimens,especially in the basal layer cells,and showed obvious polarity in normal conjunctiva.In pterygium group,ABCB5 protein was positively expressed in 91.89％ specimens (34/37) and the expression was absent in 8.11％ specimens (3/37).The relative expression levels of ABCB5 protein were 0.90±0.31 and 0.59±0.41,and those of ABCB5 mRNA were 1.01±0.26 and 0.65±0.32 in the normal conjunctival tissues and pterygium tissue,respectively,showing significant differences between them (protein:t =-0.266,P =0.011;mRNA:t =-4.560,P =0.000).Conclusions Down-regulation of ABCB5 in pterygium indicates the decreasing and losing of LSCs,which may play an important role in the development of pterygium.ABCB5 may be a useful indicator for the prediction of development and recurrence of pterygium and has an important implication for treating evaluation,and it may also be a target for the management of pterygium.
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@#AIM: To analyze the application value of corneal limbus stem cells transplantation in the treatment of pterygium. <p>METHODS: Totally 300 patients(318 eyes)with pterygium were divided into 2 groups according to treatment methods. Patients of the control group(70 patients with 77 eyes)were given simple surgical resection, while patients of the observation group(230 patients with 241 eyes)were given corneal limbus stem cells transplantation after surgical resection. The postoperative recurrent rate, corneal epithelium incision healing time, corneal epithelium healing degree, pain score and tear film break-up time were observed. <p>RESULTS: The corneal epithelium incision healing time of the observation group and control group were respectively 5.2±1.9d and 6.4±1.7d, and the difference had statistical significance(<i>P</i><0.05). There was no patient recurred in the observation group, while there were 7 eyes(9.1%)recurred in the control group, and the difference between groups had statistical significance(<i>P</i><0.05). At 1wk after treatment, there was no significant difference in corneal epithelium healing degree and pain score between groups(<i>P</i>>0.05). At 2wk after treatment, the corneal epithelium healing degree of the observation group was better, and the pain score of the observation group was lower than those of the control group, the differences between groups had statistical significance(<i>P</i><0.05). Before treatment, there was no significant difference in tear film break-up time between groups(<i>P</i>>0.05). At 1 and 2wk after treatment, the tear film break-up time of the observation group was longer than that of the control group(<i>P</i><0.05). <p>CONCLUSION: Corneal limbus stem cell transplantation can prevent postoperative recurrence of pterygium effectively, and its postoperative tissue repair effect is superior to simple surgery.
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To compare and analyze the clinical treatment effect of primary pterygium with amniotic membrane transplantation, autologous corneal limbus stem cell transplantation and intraoperative application of 0. 2g/L mitomycin C.METHODS: Retrospective analysis of 232 patients who were underwent primary pterygium surgery was performed. Eighty-two cases were treated with amniotic membrane transplantation, 90 with autologous corneal limbus stem cell transplantation, and 60 with intraoperative application of 0. 2g/L mitomycin C. The postoperative recurrence and complications of three operative methods were compared.RESULTS: The recurrence rate of autologous corneal limbus stem cell transplantation and intraoperative application of 0. 2g/L mitomycin C was lower than that of amniotic membrane transplantation ( P 0. 05 ). Corneal epithelium defect, tears and photophobia were higher in 0.2g/L mitomycin C group (P0. 05). Conjunctival congestion and subconjunctival hemorrhage in three groups had no statistical difference (P>0. 05).CONCLUSlON: Compared with other two operative methods, autologous corneal limbus stem cell transplantation for primary pterygium has the advantages of lower recurrence rate and less complications.
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AlM: To analyze the effect of corneal limbus stem cell transplantation and amniotic membrane transplantation in treatment of recurrent pterygium with symblepharon. METHODS:Totally 80 cases of patients with recurrent pterygium and symblepharon were randomly divided into pterygium resection combined with limbal stem cell transplantation group and pterygium excision with amniotic membrane transplantation group, 40 cases in each group. All patients were followed up for 6mo after the operation, the recurrence rate of pterygium and symblepharon were observed. RESULTS:There were 2 cases of recurrence in the group of corneal limbus stem cell transplantation (5%), 8 cases of recurrence in the group of amniotic membrane transplantation (20%), with statistical difference between two groups (P CONCLUSlON:Two kinds operation could significantly reduce the recurrence rate of recurrent pterygium, but the corneal limbus stem cell transplantation group has the lower recurrence rate.
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AIM: To evaluate the efficacy and safety of annular bubble conjunctival excision combining with amniotic membrane transplantation(AMT) for vernal keratoconjunctivitis(VKC) with gelatinous swellings(papillae) at the limbus.METHODS: Twenty eyes(10 patients) with VKC, were characterized by gelatinous swellings(papillae) at limbus and were underwent annular bubble conjunctival excision associated with AMT. The follow up ranged 5-21months.RESULTS: After operation, the symptoms were subsided remarkably: itching,discharge and foreign body sensation were almost disappear; photophobia and tearing were obvious in the first week after surgery, but were gradually alleviated, and almost disappear after one month of postoperative.Yellow-brown or filthy red gelatinous protuberances at limbus were cleaned away by operation and did not recur during the follow up. Corneal superficial punctuate keratitis(11 eyes) and corneal ulcers(3 eyes) were healed during the first week after surgery and did not recur. Conjunctival congestion was gradually lessened after surgery, which could not be noted 1months after surgery in 19 eyes, and the conjunctiva around the limbus, where AMT was performed, was whiter than the normal. But conjunctival congestion in 1 eye lasted for about 5mon accompanying with a little of itch, and this eye was treated with eyedrops with improvement but recurrence. No severe complications had been seen during the follow up.CONCLUSION: Annular bubble conjunctival excision combining with amniotic membrane transplantation is an effective and safe method for the treatment of vernal keratoconjunctivitis with gelatinous swellings (papillae) at limbus. But the candidates must be chosen with discretion.