METHODS: 3c-induced inhibition of proliferation was measured in the absence and presence NAC using MTT in HT-29 and SW620 cells and xCELLigence RTCA DP instrument. 3c-induced apoptotic studies were performed using flow cytometry. 3c-induced redox alterations were measured by ROS production using fluorescence plate reader and flow cytometry and mitochondrial membrane potential by flow cytometry; NADPH and GSH levels were determined by colorimetric assays. Bcl2 family protein expression and cytochrome c release and PARP activation was done by western blotting. Caspase activation was measured by ELISA. Cell migration assay was done using the real time xCELLigence RTCA DP system in SW620 cells and wound healing assay in HT-29.
RESULTS: Many anticancer therapeutics exert their effects by inducing reactive oxygen species (ROS). In this study, we demonstrate that 3c-induced inhibition of cell proliferation is reversed by the antioxidant, N-acetylcysteine, suggesting that 3c acts via increased production of ROS in HT-29 cells. This was confirmed by the direct measurement of ROS in 3c-treated colorectal cancer cells. Additionally, treatment with 3c resulted in decreased NADPH and glutathione levels in HT-29 cells. Further, investigation of the apoptotic pathway showed increased release of cytochrome c resulting in the activation of caspase-9, which in turn activated caspase-3 and -6. 3c also (i) increased p53 and Bax expression, (ii) decreased Bcl2 and BclxL expression and (iii) induced PARP cleavage in human colorectal cancer cells. Confirming our observations, NAC significantly inhibited induction of apoptosis, ROS production, cytochrome c release and PARP cleavage. The results further demonstrate that 3c inhibits cell migration by modulating EMT markers and inhibiting TGFβ-induced phosphorylation of Smad2 and Samd3.
CONCLUSIONS: Our findings thus demonstrate that 3c disrupts redox balance in colorectal cancer cells and support the notion that this agent may be effective for the treatment of colorectal cancer.
AIM OF THE STUDY: To investigate the effects of E. guineensis leaf on wound healing activity in rats.
METHODS: A phytochemical screening was done to determine the major phytochemicals in the extract. The antimicrobial activity of the extract was examined using the disk diffusion technique and broth dilution method. The wound healing activity of leaves of E. guineensiswas studied by incorporating the methanolic extract in yellow soft paraffin in concentration of 10% (w/w). Wound healing activity was studied by determining the percentage of wound closure, microbial examination of granulated skin tissue and histological analysis in the control and extract treated groups.
RESULTS: Phytochemical screening reveals the presence of tannins, alkaloids, steroids, saponins, terpenoids, and flavonoids in the extract. The extract showed significant activity against Candida albicans with an MIC value of 6.25 mg/mL. The results show that the E. guineensis extract has potent wound healing capacity, as evident from better wound closure, improved tissue regeneration at the wound site, and supporting histopathological parameters pertaining to wound healing. Assessment of granulation tissue every fourth day showed a significant reduction in microbial count.
CONCLUSIONS: E. guineensis accelerated wound healing in rats, thus supporting this traditional use.
METHOD: This is prospective and randomized clinical trial. Our study population included 30 eyes undergoing pars plana vitrectomy that required near total corneal debridement intra-operatively for surgical view. We compared the residual wound and wound healing rate in between 3 groups: 10 diabetic eyes (DMV) on topical SH 0.18%; 10 diabetic eyes (DMC) and 10 non-diabetic eyes (NDM) not treated with topical SH 0.18%. The corneal epithelial wound was measured at 12, 24, 36, 48, 60, 72 and 120 h after the vitrectomy surgery.
RESULTS: DMC group had corneal wounds that reepithelialization significantly more slowly than in NDM and DMV groups at 12, 24, 36 and 48 h (Mann-Whitney test p wound and wound healing were detected in between NDM and DMV groups. The mean for epithelial closure in DMC group was delayed 87.6 ± 28.31 h, compared with DMV group (64.8 ± 21.31) and NDM group (56.4 ± 9.88). All groups were followed up 1 month beyond completed wound closure. No recurrent corneal epithelial wound, corneal melting or corneal neovascularization was noted.
CONCLUSION: Diabetic patients on SH 0.18% four times daily for epithelial defect had similar corneal wounds healing rate as non-diabetics. This treatment significantly improved corneal wound healing and accelerated complete corneal wound resurfacing in diabetic patients.
DESIGN: A double blind randomized controlled hospital-based study involving diabetic patients with postoperative corneal epithelial defect after vitreoretinal surgery.
METHODS: Diabetic patients were randomized to 3 different concentrations of topical insulin (DTI 0.5, DTI 1, and DTI 2) or placebo in the control group (DNS). Primary outcome measure was the rate of corneal epithelial wound healing (mm² per hour) over pre-set interval and time from baseline to minimum size of epithelial defect on fluorescein stained anterior segment digital camera photography. Secondary outcome measure was any adverse effect of topical insulin. Follow-up was 1 month.
RESULTS: Thirty-two eyes of 32 patients undergoing intraoperative corneal debridement with resultant epithelial defect (8 eyes per group) were analyzed. DTI 0.5 was superior to other concentrations achieving 100% healing rate within 72 hours of treatment compared with 62.5% in DNS, 75% in DTI 1, and 62.5% in DTI 2. Statistically, DTI 0.5 achieved significant results (P = 0.036) compared with the diabetic control group (DNS) in terms of mean rate of corneal epithelial wound healing from maximum to minimum defect size. No adverse effect of topical insulin was reported.
CONCLUSIONS: Topical insulin 0.5 units QID is most effective for healing corneal epithelial defect in diabetic patients after vitrectomy surgery compared with placebo and higher concentrations. Topical insulin is safe for human ocular usage.