ABSTRACT
BACKGROUND: Fibronectin(FN) plays the role of repair in the inflammation. There is no confirmed conclusion whether it can be applied to the refractory corneal epithelial defect.OBJECTIVE: To observe the repair effect of the human plasma FN for the refractory corneal epithelial defect.DESIGN: A controlled experimental study.SETTING: Guangzhou Red Cross Hospital, Guangzhou First People's Hospital, Guangzhou Children's Hospital, Guangzhou Hospital of Traditional Chinese Medicine and Guangzhou Second People's Hos pital.PARTICIPANTS: Totally 383 eyes with the refractory corneal epithelial defect were chosen, of which 309 were in the therapy group, and 74 in the control group.METHODS: The therapy group: Human plasma FN was administered by dropping it into the eyes once every two hours. The controlgroup: 10 g/L celacol M was administered by its dribbling into the eyes once every two hours. Weilesheng was taken orally in both groups, two pills once, three times per day. According to the state of illness, both groups received anti-bacterial or anti-viral treatment and reexamination was given every day or every other day after administration. 10 g/L fluorescein sodium was used to observe the changes of cornea.MAIN OUTCOSE MEASURES: The symptoms, results of staining using fluorescein sodium as well as the corneal epithelial healing of both groups.RESULTS: The symptoms, the results from staining using the fluorescein sodium and the corneal epithelial healing were used for the evaluation. In the therapy group, 309 eyes were followed up and the cure rate was 69.9%. The average therapeutic period was 6.5 days, while those of the control group were 58.1% and 8.7 days respectively. The difference in the curative effect between the two groups was significantly different( P＜0.01 ).CONCLUSION: The application of FN for the refractory corneal epithelial defect displays a more significant effect than conventional treatment.
ABSTRACT
BACKGROUND: Fibronectin serves not only as the supporter for cells,but also as an important intracellular linkage, possessing opsonic-like functions. It can promote the phagocytic function of mononucleophages and the repair in inflammation and trauma.OBJECTIVE: Type O plasma was freshly obtained from healthy males in search of the optimal preparation method for lower sampling, convenient and higher-yielding of fibronectin.DESIGN: Open experiment.SETTING: Institute of Traumatic Surgery, Fourth Affiliated Hospital of Guangzhou Red Cross Hospital, Jinan University.MATERIALS: This experiment was carried out in the Institute of Traumatic Surgery of Guangzhou Red Cross Hospital between July 2000 and July 2002. The major materials consisted of type O fresh plasma from healthy males, gelatin, sepharose 4B activated with cyanogen bromide,sephadex G-25, urea, and trishydroxymethylaminomethane.METHODS: Affinity column constituted by gelatin coupling with sepharose 4B activated with cyanogen bromide was used to purify fibronectin. ① Human plasma was added: 150 mL type O plasma was freshly obtained from healthy males and added into the column once by 25 mLat an interval of 20 minutes, the speed of flow was 3.5 mL/min. ② Removing mixed protein: the column was rinsed by Tris-sodium citrate equilibrium liquid (pH 7.5) until the absorbency of flow-out fluid decreased to ＜ 0.02 at 280 nm. Again lmol/L NaCl containing Tris-sodium citrate was used to wash off other mixed proteins. ③ Collecting fibronectin: the column was rinsed by urea-Trispurge Fluid (3 mol/L) to collect fibronectin.④ Removing urea from fibronectin collection: fibronectin collection liquid was filtrated by Sephade G-25 to remove urea. ⑤Disinfection: 0.22 μmgermtight filter was used for disinfection.MAIN OUTCOME MEASURES: ①Results of sodium dodecyl benzene sulfonate-polyacrylamide gel electrophoresis. ② The influence of retention time on the yield of purified fibronectin. ③ The influence of plasma quantity on the yield of purified fibronectin.RESULTS: ① Results of sodium dodecyl benzene sulfonate-polyacrylamide gel electrophoresis: The density of separation gel and condensed gel was 7% and 3%, respectively; fibronectin was electrophoresized into single protein lane. ② The influence of retention time on the yield of purified fibronectin: The yield of fibronectin was (65.24±3.45) %, (74.77±4.05) %,(86.99±4.10) %, and (80.47±3.75) %, respectively, when the loading amount of fibronectin was 150 mL with non-retention time and retention time of 10, 20 and 30 minutes. ③ The influence of plasma quantity on the yield of purified fibronectin: The yield of fibronectin was (72.56±3.63) %,(77.61±3.14) %, (86.99±4.12) %, and (74.67±3.05) % when the column retention time was controlled at minute 20 with the loading amount of 100,130, 150 and 180 mL, respectively.CONCLUSION: In a given column volume of gelatin, the quantity of purified fibronectin was closely related with the plasma retention time in column and the total loading amount of plasma. As a result, the optimal loading amount of plasma was 150 mL, and the retention time was 20 minutes.The preparation method, herein, has been proved to require small amounts of plasma and yield large amounts of fibronectin.