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The effective concentration and exposure time of mitomycin-C for the inhibition of lens epithelial cell proliferation in rabbit eyes.

The proliferation of residual lens epithelial cells following cataract surgery is assumed to be a major cause of posterior capsular opacification. To assess the efficacy of mitomycin-C in preventing posterior capsular opacification, we determined the effective concentration and exposure time of mitomycin-C in inhibiting rabbit lens epithelial cell proliferation. The fourth-passaged rabbit lens epithelial cells were maintained for one day and then exposed to mitomycin-C for 1, 2, 3, and 5 minutes, respectively. There were 9 different plating concentrations of mitomycin-C with two-fold serial dilution. The maintenance of the phenotypic properties of lens epithelial cells was confirmed by continuous transcription of lambda-crystalline mRNA determined by reverse transcription-polymerase chain reaction and the polymorphism of the restriction fragment. Cell proliferation was assayed with 3H-thymidine incorporation into DNA. The fourth-passaged cells maintained the expression of lambda-crystalline mRNA, suggesting that they are phenotypically authentic lens epithelial cells. The effective concentrations and exposure time of mitomycin-C were 0.1 mg/ml for 1 minute and 2 minutes, and 0.025 mg/ml for 2 minutes. By these results, we postulated that mitomycin-C at relatively short incubation times could be clinically used for prevention of posterior capsular opacification after cataract surgery.

Animals↗

Mitomycin in the treatment of hypertrophic conjunctival scars after strabismus surgery.

A recognized complication of strabismus surgery is the development of hypertrophic conjunctival scars over the operated-on muscle. Our standard treatment of these scars has been excision followed by corticosteroid eye drops, which results in a scar recurrence rate of 33%. Antimetabolic chemotherapeutic agents, such as mitomycin, can reduce posttraumatic scarring and vascularization of the ocular surface. We studied the use of mitomycin eye drops as an adjunct to standard treatment of hypertrophic conjunctival scars after strabismus surgery. Four patients were treated with adjunct mitomycin eye drops (0.2 to 0.4 mg/ml). Postoperative follow up ranged from 16 to 56 weeks. None of these cases showed significant scar recurrence. Mitomycin in the higher dosage caused mild superficial punctate keratitis after 8 days of use. Despite a lamellar sclerectomy in a patient treated with 0.2 mg/ml of mitomycin, there was no further clinically apparent loss of scleral tissue. The results of this pilot study suggest that topical mitomycin is a safe and effective adjunct to excision and corticosteroid eyedrops in the treatment of hypertrophic conjunctival scars after strabismus surgery.

Administration, Topical↗

Effectiveness of mitomycin C in reducing reformation of adhesions following surgery for restrictive strabismus.

PURPOSE: To evaluate the efficacy of mitomycin C in reducing reformation of adhesions following surgery for restrictive strabismus. METHODS: Sixteen patients with restrictive strabismus resulting from trauma, retinal detachment surgery, and multiple squint surgeries were randomized to either a mitomycin group (8 patients) or a control group (8 patients). Mitomycin C (0.2 mg/mL) was applied intraoperatively for 5 minutes after release of adhesions. Range of passive ductions was scored for comparison. Postoperative follow-up ranged from 3-18 months. RESULTS: Compared to controls, the mitomycin C group had better range of passive ductions. There were no complications related to the cornea, intraocular pressure, uveitis, or muscle disinsertion in the mitomycin C group. CONCLUSION: Intraoperative application of mitomycin C (0.2 mg/mL for 5 minutes) is a safe and effective adjunct to surgery in the treatment of restrictive strabismus.

Adolescent↗

Long-term results of trabeculectomy with different concentrations of mitomycin C in refractory developmental glaucoma.

PURPOSE: To evaluate the long-term results of trabeculectomy with two different concentrations of mitomycin C in refractory developmental glaucoma. PATIENTS AND METHODS: A total of 48 eyes of 37 patients with previous failure of congenital glaucoma surgery underwent mitomycin C-augmented trabeculectomy. Data were analyzed retrospectively, and patients were divided into two groups. Group 1 consisted of 18 patients (25 eyes) who underwent trabeculectomy with a 0.4 mg/mL mitomycin C solution and group 2 consisted of 19 patients (23 eyes) who underwent trabeculectomy with a 0.2 mg/mL mitomycin C solution. Pre- and postoperative intraocular pressure (IOP), best-corrected visual acuity, bleb characteristics, success rate, and complications were compared for both groups. RESULTS: Mean time between primary surgery and mitomycin C-augmented trabeculectomy was 19.3 months (range, 11 months to 7 years). Mean preoperative IOP was 28.4 +/- 2.9 mm Hg in group 1 and 29.0 +/- 2.4 mm Hg in group 2. Final postoperative IOP was 17.0 +/- 1.6 for group 1 and 17.1 +/- 1.4 mm Hg for group 2. There were no significant differences between the groups for IOP, success rate, and complications (P > .05). The most common postoperative complication was shallow anterior chamber (11 [22.9%] of 48 eyes). CONCLUSIONS: A 0.4 or 0.2 mg/mL mitomycin C solution for 4 minutes may be used during trabeculectomy in refractory developmental glaucoma.

Child, Preschool↗

Pharmacokinetic of intraarterial mitomycin C with extra corporeal detoxification in humans.

PURPOSE: A study has been performed in 15 patients with liver metastases from colorectal cancer to evaluate the pharmacokinetic of mitomycin C and the effectiveness of drug removal techniques during high-dose locoregional chemotherapy. PATIENTS AND METHODS: Haemofiltration and/or haemodialysis of post-hepatic venous blood were performed during 22 intra-arterial infusions of mitomycin C by the use of a double lumen catheter surgically introduced in the inferior vena cava. Mitomycin C levels were measured by high performance liquid chromatography in the extracorporeal circuit blood, in the peripheral venous blood and ultrafiltrate. RESULTS: The mean reduction of mitomycin C bioavailability in the extracorporeal circuit blood was 42.3 per cent using haemofiltration, 58.9 per cent using haemofiltration plus haemodialysis and 59.3 per cent with haemodialysis alone. The resulting mean mitomycin C plasmatic half-life was 40.2 minutes using haemofiltration, 24.9 minutes using haemofiltration plus haemodialysis and 24.1 minutes using haemodialysis alone. CONCLUSIONS: The detoxification of posthepatic venous blood during intra-arterial hepatic chemotherapy is an effective method to increase mitomycin C concentrations and oncological response and limit extra hepatic toxicity.

Antibiotics, Antineoplastic↗

[Effects of mitomycin C on haze after photorefractive keratectomy for myopia in rabbits].

OBJECTIVE: To investigate the effects of mitomycin C on haze after photorefractive keratectomy. METHODS: Forty-five rabbits underwent bilateral 193 nm excimer laser photorefractive keratectomy to correct 8 diopters of myopia. All eyes were allocated randomly to be treated with 0.008% mitomycin C during operation, or 0.1% dexamethasone after operation, or no medical treatment as control. Clinical and histopathologic examinations were made with slit-lamp microscope, specular microscope, light microscope and transmission electron microscope. RESULTS: At 4, 8 weeks after operation, the corneal haze was significantly less in mitomycin C group than that in the control group or in dexamethasone group, and the corneal haze was significantly less in dexamethasone group than that in the control group. At 1, 4, 8 weeks, the number of keratocyte in the anterior stroma of ablation area was significantly less in mitomycin C group than in the control group or in dexamethasone group. The difference between dexamethasone group and the control group was insignificant. The changes in time of reepithelialization, thickness of epithelium and keratocyte number of anterior stroma in ablation area were statistically insignificant among all groups. CONCLUSION: Mitomycin C can reduce corneal haze by inhibiting the proliferation of keratocyte and has no toxicity on cornea. Mitomycin C is more effective than dexamethasone on haze.

Animals↗

Role of NAD(P)H:(quinone acceptor) oxidoreductase (DT-diaphorase) in activation of mitomycin C under hypoxia.

The role of the two-electron reducing enzyme DT-diaphorase in the activation of mitomycin C under hypoxic conditions was investigated. Mitomycin C activity was compared in L5178Y murine lymphoblasts, which have low levels of DT-diaphorase activity, and L5178Y/HBM10 cells, which have elevated levels of enzyme activity. The cytotoxic and DNA cross-linking activities of mitomycin C were greater in L5178Y/HBM10 cells than in L5178Y cells. In L5178Y/HBM10 cells, dicoumarol, an inhibitor of DT-diaphorase, decreased cell kill and DNA cross-linking by mitomycin C in air but had no significant effect on these activities under hypoxia. By comparison, in L5178Y cells, dicoumarol had no effect on drug activity under either aerobic or hypoxic conditions. A model for the activation of mitomycin C by both one-electron and two-electron reduction is proposed. Our findings suggest that two-electron reduction by DT-diaphorase has only a limited role in the activation of mitomycin C under hypoxic conditions, although this enzyme appears to be an important contributor to drug activation under aerobic conditions.

Animals↗

Mitomycin C-induced colitis in rats: a new animal model of acute colonic inflammation implicating reactive oxygen species.

The mechanism of the tissue damage induced by colonic inflammation in ulcerative colitis is not established. We therefore developed and characterized a simple new rat model of acute colonic inflammation induced by a single systemic injection of mitomycin C. After an intraperitoneal injection of mitomycin-C, colon histologic examination revealed transient (3 to 14 days) diffuse, colonic inflammation and injury that, like human ulcerative colitis, was limited to the mucosal layer. The rest of the gastrointestinal tract was spared. Gut permeability, as measured by urinary excretion of orally administered lactulose and mannitol, was unchanged 3 days after injection, when inflammation was already present; permeability was increased at 7 days, when inflammation was maximal. Mitomycin C did not produce inflammation in experimentally bypassed segments of small bowel despite the presence of colonic-type bacteria, suggesting that lack of intraluminal bacteria was not responsible for the absence of inflammation in the small intestine. Chemiluminescence, a means of estimating levels of reactive oxygen species, was greater in the intact, inflamed colon of mitomycin C-treated rats than in bypassed segments. Moreover, inflamed mucosal scrapings produced more in vitro luminol-enhanced chemiluminescence. Furthermore, the reactive oxygen species scavengers allopurinol, catalase, and WR-2721 decreased inflammation severity. We therefore conclude: (1) the mitomycin C-treated rat is a novel, easy to prepare animal model of acute inflammation of colonic mucosa, with morphologic similarities to the acute phase of ulcerative colitis in human beings; (2) increased gut permeability in mitomycin C-treated rats is the result, not the cause, of the inflammation; and (3) reactive oxygen species play an important role in colonic inflammation and tissue injury in this model, and possibly in human ulcerative colitis.

Animals↗

Postoperative concurrent chemoradiotherapy with mitomycin in advanced squamous cell carcinoma of the head and neck: results from three prospective randomized trials.

UNLABELLED: Recent prospective randomized trials have shown concurrent chemoradiotherapy improves locoregional control in postoperative patients with squamous cell carcinoma of the head and neck using cisplatin-based regimes. This report presents data pooled from three randomized trials employing mitomycin, selecting those patients treated postoperatively, to evaluate the long-term benefit of mitomycin in the postoperative setting and to compare these results with those of two other recently published randomized trials. METHODS AND MATERIALS: Between 1980 and 1999, a total of 331 patients with squamous cell carcinoma of the head and neck from the three prospective trials were enrolled. Of the 205 postoperative patients in these trials, 103 were randomized to receive mitomycin and radiation, while 102 received radiation alone or radiation with porfiromycin in the third trial. Patients were treated with standard daily radiotherapy to a total median dose of 60 Gy over 47 days. Patients who were randomized to mitomycin C received 15 mg/m2 of the drug on days 5 and 47 (or last day). RESULTS: The 5-year rate of locoregional control was higher in the mitomycin arms. There was no statistically significant difference in the rates of overall survival or distant metastasis. Patients had a lower percentage of high-risk factors in both arms of the study, compared with patients in the large prospective trials, including positive margins, two or more positive lymph nodes, or oropharynx primary tumors. The gains in locoregional control realized with mitomycin were similar to the improvements in the recently published randomized trials using platinum. CONCLUSIONS: These results confirm significant gains in locoregional control using concurrent chemoradiotherapy in the postoperative setting for patients with squamous cell carcinoma of the head and neck. The lack of consensus over a benefit in the rates of overall survival and distant metastasis emphasizes the need for further prospective trials in the postoperative management of squamous cell carcinoma of the head and neck.

Antibiotics, Antineoplastic↗

[In vitro interaction of mezlocillin with fluorouracil and mitomycin].

The interaction of mezlocillin (Baypen) with either fluorouracil (5-fluorouracil, 5-FU) or mitomycin was tested in vitro by incubating colorectal carcinoma cell line HT 29 with varying concentrations of mezlocillin, 5-FU, mitomycin and the combinations of mezlocillin + 5-FU and mezlocillin + mitomycin. Inhibition of colony growth in soft agar of treated compared with untreated cells was used as an in vitro response parameter. Concentration ranges were representative for regional chemotherapy with mitomycin or 5-FU and perioperative antibiotic therapy with mezlocillin. Mezlocillin, 5-FU and mitomycin showed dose-dependent inhibition of colony growth. Mezlocillin produced a maximal toxicity of 16% at 1000 micrograms/ml. Toxicity of 5-FU was reduced by addition of mezlocillin. The increase of colony growth after coincubation of mezlocillin with 5-FU (compared to cells treated with 5-FU alone) at a mezlocillin concentration of 10 micrograms/ml was 47% to 116%, at 100 micrograms/ml 44% to 132%, depending on the 5-FU test concentrations. The 5-FU toxicity in some instances was nearly abolished. There was no interaction between mezlocillin and mitomycin. The interference of mezlocillin with 5-FU-toxicity should be respected in case of combined perioperative antitumor and antibiotic therapy with 5-FU mezlocillin. Similar interactions with related antibiotics cannot be excluded.

Cell Survival↗

Animal experiments as a model for clinical trials of a new dosage form of mitomycin C for peritoneal carcinomatosis.

Rabbits with peritoneal carcinomatosis induced by VX2 carcinoma were used as an experimental model to study a new dosage form of mitomycin C, MMC-CH, comprising activated carbon adsorbing mitomycin C. Rabbits revived an intraperitoneal injection of mitomycin C (1 mg/kg in the form of MMC-CH or 0.185 mg/kg in the form of mitomycin C solution). The two doses were equal in toxicity. MMC-CH extended survival time measured in days to 139% as compared to mitomycin C solution. Autopsy findings showed peritoneal carcinomatosis to be much less developed with MMC-CH than with mitomycin C solution. Hematological and blood biochemical analyses showed no remarkable abnormality.

Animals↗

Preferential activation of mitomycin C to cytotoxic metabolites by hypoxic tumor cells.

Mitomycin C, a bioreductive alkylating agent with clinical utility against several human tumors, was found to be selectively toxic at a relatively low concentration (1.5 micro M) to EMT6 tumor cells made chronically hypoxic by preincubation in 95% N2-5% CO2 for 4 hr prior to drug exposure. This selective cytotoxicity correlated well with the preferential activation and metabolism of mitomycin C by sonicated cell preparations. The bioactivation of mitomycin C to an alkylating agent by EMT6 and Sarcoma 180 cell sonicates required hypoxic conditions and a reduced nicotinamide adenine dinucleotide phosphate-generating system. Furthermore, the formation of reactive drug metabolites and the disappearance of mitomycin C from the reaction mixture were inhibited by carbon monoxide. The presence of potassium cyanide in the incubation mixture did not affect either the rate of overall metabolism or the rate of formation of reactive metabolites. A high rate of disappearance of mitomycin C from the medium of intact cultures of EMT6 cells was found only in those cultures which were made chronically hypoxic. These data suggest that bioreductive alkylating agents like mitomycin C have the potential to attack selectively the chemotherapeutically resistant hypoxic cell component of solid tumors. Thus, agents capable of bioreductive alkylation should be useful adjuncts to existing therapeutic regimens which are effective against well-oxygenated cells.

Alkylating Agents↗

Effects of topically applied mitomycin-C on intraocular pressure, facility of outflow, and fibrosis after glaucoma filtration surgery in clinically normal dogs.

The effects of mitomycin-C on intraocular pressure (IOP), facility of outflow (C), and Tenon's capsule fibrosis were studied over 60 days in 10 clinically normal dogs. A-1-piece, silicone glaucoma implant was surgically implanted into both eyes; the filtration site of one eye was treated with a single, 5-minute intraoperative application of mitomycin (0.5 mg/ml), and the fellow eye was treated in a similar manner with balanced salt solution. There were no significant differences in preoperative IOP or C-values between treatment groups. Mean IOP in eyes of both groups initially decreased from the preoperative value, but returned to the baseline value by day 21. Mean facility of aqueous outflow (C-value) increased in all eyes during the first 14 days (mitomycin-C-value = 2.26 +/- 0.72; control C-value = 2.38 +/- 0.81), then reached a plateau that was significantly higher than the baseline value in mitomycin (P = 0.039) and control (P = 0.041) eyes. Histologic evaluation revealed all implants surrounded by a connective tissue capsule composed of regular dense collagen and fibroblasts that was significantly (P = 0.003) thinner in the mitomycin-treated (scleral side = 167 +/- 62 microns; conjunctival side = 122 +/- 41 microns) than the control (scleral side = 261 +/- 92 microns; conjunctival side = 180 +/- 48 microns) group. There were, however, no significant differences in IOP or C-values between groups at any postoperative time interval. Results of this study indicate that intraoperative treatment with mitomycin suppresses, but does not prevent fibrosis around silicone filtering implants.

Animals↗

Formation of 8-hydroxyguanine in DNA during mitomycin C activation.

DNA damage caused indirectly via reactive oxygen species generated during reductive activation of mitomycin C was evaluated. This oxidative DNA damage was measured by determining the formation of 8-hydroxyguanine in DNA exposed to chemically or enzymatically activated mitomycin C. The level of 8-hydroxyguanine was measured indirectly by determining formamidopyrimidine-DNA glycosylase-sensitive sites induced in plasmid DNA exposed to mitomycin C and directly by a 32P-postlabeling assay for the modified base. Activation of mitomycin C by sodium borohydride in air, by H2/Pt, or xanthine oxidase in N2 caused increases in the level of 8-hydroxyguanine. The extent of the increase varied according to the incubation conditions with the greatest increase being observed in DNA exposed to mitomycin C activated under hypoxic conditions. These results support a possible indirect mechanism for DNA damage caused by mitomycin C that is mediated by reactive oxygen species.

Anaerobiosis↗

Role of NAD(P)H:(quinone acceptor) oxidoreductase (DT-diaphorase) in activation of mitomycin C under acidic conditions.

The quinone antitumor agent mitomycin C is preferentially toxic to some cells under hypoxic and acidic conditions. The two-electron reducing enzyme DT-diaphorase may be a major contributor to mitomycin C activation under aerobic conditions, but its role in drug activation under hypoxic and acidic conditions is unclear. In this study, we observed that mitomycin C produced increased DNA cross-linking and cytotoxicity in Chinese hamster ovary cells at pH 6.6, compared with pH 7.2, under aerobic conditions, but drug activity was similar at these pH values under hypoxic conditions. The DT-diaphorase inhibitor dicoumarol completely inhibited the enhanced activity of mitomycin C at acidic pH under aerobic conditions but had no effect on DNA cross-linking or cytotoxicity under hypoxic conditions. These finding suggest that the enhanced activity of mitomycin C at acidic pH, in air, is due to increased drug activation by DT-diaphorase. However, the role of DT-diaphorase in activating mitomycin C under hypoxic conditions appears to be limited, even at acidic pH.

Animals↗

[Acute bronchospasm due to periwinkle alkaloid and mitomycin association].

We report six cases in which patients presented with acute dyspnoea following injections of either vindesine or vinorelbin. These patients were receiving chemotherapy in association with cisplatin, mitomycin, and vindesine or vinorelbin, for inoperable bronchial cancer. Three of the patients had evidence of airflow obstruction before these incidents. The clinical picture suggested bronchospasm and appeared in the two hours following an injection of the vinca alkaloid and a significant time away from the administration of the mitomycin. Additional respiratory support was necessary in one patient, the bronchial spasm stopped spontaneously in three cases, and following bronchodilator in two. The respiratory toxicity of vinca alkaloids (vindesine, vinblastin) was observed in 4% of the cases, uniquely when they were associated with mitomycin. Vinorelbin seems to possess the same respiratory toxicity. The bronchospasm, sometimes very severe, seems to occur in the two hours following the injection in the case of the cytotoxics and some time after the administration of mitomycin. The recurrence of the bronchospasm is a constant feature when the vinca alkaloid is readministered. This side effect is different to the pulmonary fibrosis due to mitomycin. Clinical follow up and spirometry is thus necessary in those patients receiving chemotherapy in which vinca alkaloids and mitomycin are associated and the regime should be followed after each administration of a vinca derivative. After the first episode of dyspnoea, it is probably wise to stop the administration of these anti-mitotics to prevent any further respiratory side-effects which could be more severe.

Acute Disease↗

Update on the Dutch Cooperative Trial: mitomycin versus bacillus Calmette-Guérin-Tice versus bacillus Calmette-Guérin RIVM in the treatment of patients with pTA-pT1 papillary carcinoma and carcinoma in situ of the urinary bladder. Dutch South East Cooperative Urological Group.

Prophylactic intravesical treatment with chemotherapeutic agents or one of the different strains of the immunomodulator bacillus Calmette-Guérin (BCG) reduces tumor recurrence and may prevent progression after transurethral resection (TUR) of superficial bladder cancer. Clinical and pathological prognostic factors are used to categorize different groups at risk for disease recurrence and progression. Solitary low-grade (G1, 2A), low-stage (pTa) tumors have less than a 5% risk of progression, while pT1 G3 tumors with concomitant carcinoma in situ (CIS) have a considerably higher risk of progression and metastases. Thus, prospective clinical trials should consider the different risks associated with different prognostic groups and stratify patients accordingly. The Dutch Cooperative Trial evaluated mitomycin versus BCG-Tice versus BCG-RIVM in 469 patients with pTA/pT1 carcinoma and CIS of the urinary bladder after TUR. Of 437 evaluable patients, 50 had CIS, 254 had pTA tumors, and 133 had pT1 tumors. No statistical differences were observed in toxicity between the two strains of BCG, but local and systemic side effects were more frequent in the BCG groups versus the mitomycin group. No differences in response rate were observed between the 3 treatment groups in patients with CIS. A statistically significant difference in favor of mitomycin was seen, however, in patients with papillary tumors. Mitomycin and BCG-RIVM were equally effective, and mitomycin proved significantly more effective than BCG-Tice. No significant difference in efficacy was observed between the two strains of BCG. Disease recurrence during the study in patients with papillary tumors was seen in 43% of mitomycin-treated patients, 64% of the BCG-Tice group, and 46% of patients treated with BCG-Rivm. However, a subgroup analysis for time to first recurrence for pTa versus pT1 and for grade 1 and 2 versus grade 3 papillary tumors showed no statistically significant between-group difference. Further studies comparing identical regimens, doses, and patient groups are needed to define more clearly which patient groups and tumors are more likely to respond to intravesical therapy.

Adjuvants, Immunologic↗

Low-dose mitomycin C trabeculectomy in patients with advanced glaucoma.

PURPOSE: The purpose of this study was to analyze the efficacy and safety of low-dose (0.2 mg/ml) intraoperative mitomycin C in primary trabeculectomy. METHODS: Twenty-eight eyes of 28 patients with advanced primary open-angle glaucoma undergoing primary trabeculectomy were randomized to either mitomycin C (0.2 mg/ml) or saline solution intraoperatively for 3 min. Intraocular pressure was measured at 1 day; 1 week; 1, 3, and 6 months postoperatively; and at the final visit. RESULTS: Mean follow-up was 17.0 +/- 5.6 months for the mitomycin C group and 15.7 +/- 5.1 months for the control group. Mean intraocular pressures were significantly lower in the treated group on the first postoperative day (p = 0.021), at the 6-month interval (p = 0.001), and at the final visit (p = 0.001). At the last follow-up examination, intraocular pressure was < or = 15 mm Hg in 12 (85.7%) of the mitomycin C-treated eyes and in four (28.6%) of the control eyes (p = 0.002). Life table analysis showed a significantly higher probability of intraocular pressure control in the mitomycin C group than in the control group (p = 0.0065). Choroidal effusion was observed in five (35.7%) treated eyes and two (14.3%) control eyes, whereas shallow anterior chamber were present in five (35.7%) treated eyes and one control eye (7.1%). CONCLUSION: Despite inducing a higher short-term complication rate, low-dose mitomycin C may be an alternative in the treatment of eyes with advanced glaucomatous damage requiring low final intraocular pressures.

Aged↗