Objective To investigate the controlled release effect and the anti-cancer cell ability of a 5-FU loaded poly-L-lactic acid?（PLLA） nanofibers membrane blending with keratin. Methods Making PLLA and keratin mix together and crosslinking to generate blending solution. Then the anti-cancer drug 5-FU was added into the solution to fabricate nanofibers membrane by high voltage electrospinning method. The micro morphology was observed by scanning electron microscope （SEM）. The controlled release effect of 5-FU from the nanofibers membrane was measured by high performance liquid chromatography （HPLC）. The cytotoxicity of 5-FU/PLLA keratin nanofibers membrane was evaluated by using 3-（4, 5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （MTT） assay on HCT116 cell lines. At the meantime, cell growth morphology of HCT116 in experiment group were observed by microscope and transmission electron microscope. Results 5-FU could be dispersed homogeneous in the PLLA/keratin nanofibers membrane through SEM. HPLC suggested that 5-FU could be diffused out from the fibers slowly and uniformly, which corresponded the zero order kinetics basically. After different treatment, the longer time the 5-FU/PLLA keratin nanofibers （experiment group） immerse in the medium, the much more swelling, apoptosis, and necrocytosis of the cells were observed. The cell viability for experiment group was （47.5±2.8）% by MTT, while the PLLA keratin nanofibers without 5-FU had no significant impact on cell viability （93.9±2.8）%, which was statistic significance （P＜0.01）. Conclusion 5-FU/PLLA keratin nanofibers membrane owns good controlled release effect and satisfies cell inhibitory effect against HCT116 cells in vitro，which suggested that it has a promising prospect for clinical therapy.
Objective To assess the effect of topical appl ication of 5-fluorouracil (5-FU) on intimal hyperplasia in rabbit vein graft. Methods Sixty-four male New Zealand white rabbits, aged 5 months and weighing 2.8-3.0 kg, were randomly divided into group A, B, C, and D (n=16 rabbits per group). Artery defect model was establ ished by cutting about 1 cm artery from the middle part of the dissociated left common carotid artery. A section about 3 cm was cut from the right external jugular vein, and the harvested vein was inverted and end-to-end anastomosed to the artery defect with 9-0 non-traumatic suture. After anastomosis, the extima of the grafted veins in group A, B, and C was completely wrapped with cotton sheet (12 mm × 30 mm × 1 mm in size) immersed by 5-FU at a concentration of 50.0, 25.0, and 12.5 mg/mL, respectively, and eachvein was treated 5 times (1 minute at a time). In group D, the extima of the graft veins was treated with normal sal ine instead of 5-FU. The grafted veins were obtained 1, 2, 4, and 6 weeks after operation, HE staining and Masson staining were preformed for histological changes of grafted vein wall, prol iferating cell nuclear antigen (PCNA) immunohistochemistry staining and TUNEL label ing staining were conducted for prol iferation and apoptosis of smooth muscle cell of the grafted vein, and transmission electron microscope observation was performed for cellular ultrastructure. Results The HE staining, Masson staining, and PCNA immunohistochemistry staining showed that the thickness of intima in group A and B was obviously less than that in group C and D at 1, 2, 4, and 6 weeks after operation, and the prol iferation cells in group A and B were less than that in group C and D at 1, 2, and 4 weeks after operation. The thickness of the intima, the degree of intima hyperplasia, the degree of vessel lumen stenosis of four groups at different time points were as follows: at 1 week after operation, group A [(12.69 ± 1.68) μm, 0.73 ± 0.05, 0.025 ± 0.003], group B [(17.52 ± 2.01) μm, 0.86 ± 0.06, 0.027 ± 0.004], group C [(21.92 ± 1.85) μm, 1.06 ± 0.09, 0.036 ± 0.006] and group D [(26.45 ± 3.86) μm, 1.18 ± 0.08, 0.041 ± 0.005]; at 2 weeks after operation, group A [(24.61 ± 2.91) μm, 0.86 ± 0.06, 0.047 ± 0.003], group B [(37.28 ± 2.78) μm, 1.17 ± 0.09, 0.060 ± 0.004], group C [(46.52 ± 2.25) μm, 1.44 ± 0.08, 0.073 ± 0.003], and group D [(52.07 ± 3.29) μm, 1.45 ± 0.05, 0.081 ± 0.006]; at 4 weeks after operation, group A [(61.09 ± 6.84) μm, 1.38 ± 0.08, 0.106 ± 0.007], group B [(63.61 ± 8.25) μm, 1.40 ± 0.07, 0.107 ± 0.010], group C [(80.04 ± 7.65) μm, 1.64 ± 0.07, 0.129 ± 0.011], and group D [(84.45 ± 9.39) μm, 1.68 ± 0.10, 0.139 ± 0.014]; at 6 weeks after operation, group A [(65.27 ± 5.25) μm, 1.46 ± 0.07, 0.113 ± 0.005], group B [(65.82 ± 7.12) μm, 1.45 ± 0.05, 0.112 ± 0.011], group C [(84.45 ± 9.39) μm, 1.69 ± 0.09, 0.135 ± 0.007], and group D [(87.27 ± 8.96) μm, 1.76 ± 0.05, 0.140 ± 0.012]. Group A and B were inferior to group C and D in terms of the above three parameters and cell prol iferation index 1, 2 and 4 weeks after operation (P lt; 0.05). Group A and B were superior to group C and D in terms of cell apoptosis index of intima and media 1 and 2 weeks after operation (P lt; 0.05). Transmission electron microscope observation showed that the synthetic cell organelles such as rough endoplasmic reticulum, golgi apparatus, and ribosome in group A and B were obviously less than those in group C and D (P lt; 0.05). Conclusion Topicalappl ication of 5-FU can effectively inhibit intima hyperplasia of the vein grafts.
To evaluate the effect of 5-fluorouracil (5-FU) appl ied topically on preventing adhesion andpromoting functional recovery after tendon repair. Methods From August 2003 to June 2007, 48 patients with flexor tendonrupture of the fingers by sharp instrument were treated and randomly divided into two groups. In 5-FU group, 39 fingers of 26 patients included 17 males and 9 females, aged (29.3 ± 9.8) years; the locations were zone I in 19 fingers and zone II in 20 fingers; single finger was involved in 12 cases and more than 2 fingers were involved in 14 cases; and the time from injury to operation was (2.4 ± 1.6) hours. In control group, 36 fingers of 22 patients included 14 males and 8 females; aged (26.1 ± 8.7) years; the locations were zone I in 16 fingers and zone II in 20 fingers; single finger was involved in 10 cases and more than 2 fingers were involved in 12 cases; and the time from injury to operation was (2.1 ± 1.8) hours. No statistically significant difference was found in constituent ratio of age, gender, injured fingers and their zones, between two groups (P gt; 0.05). The repair site in 5-FU group was given 5-FU at a concentration of 25 mg/mL with a soaked sponge, and the synovial sheath of the repaired site was wrapped with the 5-FU-soaked sponge for 1 minute for 4 times after the tendons were repaired; normal sal ine was used in the control group. Results Wound healed by first intention and no infection and tendon rupture occurred in two groups. The patients were followed up for 3-8 months (mean 4.1 months) and 3-8 months (mean 3.9 months) in 5-FU group and in control group respectively. The functional recovery degrees of the fingers were evaluated with total active movement (TAM) evaluation system. In 5-FU group, the results were excellent in 22 fingers, good in 13 fingers, fair in 3 fingers and poor in 1 finger; the excellentand good rate was 89.7%. In control group, the results were excellent in 11 fingers, good in 15 fingers, fair in 9 fingers andpoor in 1 finger; the excellent and good rate was 72.2%. There was statistically significant difference in the functional recovery degrees of fingers between two groups (P lt; 0.05). The 2 fingers which had a poor result in 5-FU group and control group were served with tenolysis was performed in 2 cases having poor results after 6 months of operation and had an excellent result at last. Conclusion 5-FU appl ied topically can reduce tendon adhesions after the ruptured tendon repair.
Objective To assess the effect of basic fibroblast growth factor (bFGF) and 5-fluorouracil (5-FU) appl ied topically on the tendon adhesion and the heal ing process after the flexor tendon repair in Leghorn chickens. Methods Ninety male Leghorn chickens (weighing 3.0-3.5 kg) were randomly divided into 3 groups, with 30 chickens in each group. The flexordigitorum profundus tendons of the third right toes were transected and sutured directly. The repair site in group A was given 0.6 μL fibrin sealant (FS). In group B, the repair site was given 0.6 μL FS containing 500 ng bFGF. In group C, before the tendons were transected, they had been soaked in 5-FU solution, and then the same treatment as group B was given. Six specimens of the third toe were harvested to perform the macroscopical and histological examinations at 1, 2, 4, and 8 weeks, respectively, and to perform the biomechanical test at 8 weeks. Results All animals survived until the experiment was completed. All incisions healed smoothly. No rupture occurred in the reparied tendon. At 8 weeks, the adhesion degree was l ighter in group C than in group B (P lt; 0.05), but there was no significant difference in the adhesion degree between group A and groups B, C (P gt; 0.05). At 1, 2, and 4 weeks after operation, the number of fibroblast cells of group A was significantly less than that of group B (P lt; 0.05), and the number of fibroblast cells of group C was significantly less than that of group A and group B in the tendon sheath and epitenon (P lt; 0.05); however, it was significantly more than that of group A in the tendon parenchyma (P lt; 0.05), and no significant difference was observed when compared with that of group B (P gt; 0.05). At 8 weeks, no difference was found among 3 groups (P gt; 0.05). The collagen fiber content of group A was significantly less than that of group B at 4 and 8 weeks (P lt; 0.05). In the sheath and epitenon, the collagen fiber content of group A was significantly more than that of group C at 4 weeks (P lt; 0.05); however, no significant difference was found between 2 groups at 8 weeks (P gt; 0.05). The collagen fiber content of group A wassignificantly less than that of group C in the parenchyma at 4 and 8 weeks (P lt; 0.05). At all time points, the collagen fiber content of group B was significantly more than that of group C in the sheath and epitenon (P lt; 0.05), but no significant difference in the parenchyma was observed between 2 groups (P gt; 0.05). The biomechanical tests showed that the gl iding excursion of the tendon in groups A, B, and C was (3.51 ± 0.56), (2.84 ± 0.42), and (4.56 ± 0.59) mm, respectively; the work of flexion was (14.08 ± 1.85), (20.62 ± 3.52), and (10.91 ± 1.53) N.mm, respectively; and the ultimate tensile strength of the tendon was (11.26 ± 1.83), (15.02 ± 2.20), and (14.40 ± 1.57) N, respectively. There were significant differences in the gl iding excursion of the tendon and the work of flexion among 3 groups (P lt; 0.05) and in the ultimate tensile strength of the tendon between group A and groups B, C (P lt; 0.05), but there was no significant difference in the ultimate tensile strength of the tendon between group B and group C (P gt; 0.05). Conclusion Local single-use bFGF and 5-FU can not only effectively promote the heal ing of flexor tendon, but also significantly reduce tendon adhesion.