Mekanisme Kematian Sel Epitel Kornea Oleh Karena Infeksi Pseudomonas Aeruginosa Melalui Analisis Ekspresi Caspase-1, Tnfα, Ripk1, Ripk3, Caspase-3 Pada Model Tikus (Rattus Norvegicus)

Background: Pseudomonas aeruginosa is one of the leading causes of severe keratitis that may cause in progressive inflammation with rapid onset, resulting in permanent tissue destruction, then eventually, blindness and damage to corneal integrity. It is important to know the virulence factor of Pseudomonas aeruginosa mechanism and modes of regulation in corneal epithelial cells to prevent progressivity, increasing chance of recovery, and decreasing perforation corneal complication. Objective: The aim of this study to investigate mechanism of corneal epithelial cell death by Pseudomonas aeruginosa infection through the analysis of expressions of caspase-1, TNFα, RIPK1, RIPK3, and caspase-3. Methods: The study design was randomized post-test only with control group. Fifty-three Wistar rats are divided into 2 groups, each of them with 6 treatments. One control group and one experimental group. In the experimental group, three epithelial abrasions were produced on the left cornea with a 26-gauge needle and inoculated with 2x106 CFU/ml in 5μl of bacterial suspension, while in the control group only three epithelial abrasions. In both groups use waiting time 1 hour, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours after that the eyeball was enucleated and rat was terminated thus examine for histology and immunohistochemical staining examination. Result: Pseudomonas aeruginosa was shown to cause increasing expression of caspase-1 (p = 0.006), TNFα (p = 0.000), caspase-3 (p = 0.061), and decreasing expression of RIPK3 (p = 0.047). TNFα showed to cause increasing expression of RIPK1 (p = 0.000), RIPK1 showed to cause increasing expression of RIPK3 (p = 0.000), but TNFα showed not to cause change of expression of caspase-3 (p = 0.141). Conclusion: This study demonstrated infection of Pseudomonas aeruginosa in corneal rats model showed to cause increasing expression of caspase-1, TNFα, caspase-3, increasing RIPK1 through TNFα, and decreasing expression of RIPK3 in 1 hour until 72 hours after inoculation as prime biomarkers in mechanism of corneal epithelial cell death

The Effect of Limbal Mesenchymal Stem Cell Secretome on Corneal Neovascularization in Experimental Model of Corneal Alkali Injury

Abstract Background: To investigate the effect of limbal mesenchymal stem cell (L-MSC) secretome in inhibition of corneal neovascularization in animal model of alkali chemical injury Methods: Secretome was collected from limbal mesenchymal stem cells of corneoscleral tissue of healthy male rabbit. Rabbits were exposed to NaOH. The research subjects consisted of 3 groups, normal control group, Balance salt solution (BSS) group, and (L-MSC) secretome group. Clinical corneal neovascularization level, number of corneal neovascularization histopathologically with hematoxylin-eosin staining and VEGF expression by immunohistochemical staining were analyzed. Results: After seven days of administration of topical instillation of L-MSC secretome, neovascularization level in L-MSC group was lower than BSS group even though the difference was not statistically significant (p>0.05). The mean histopathological neovascular count in the L-MSC secretome group was lower than that BSS group even though the difference was not statistically significant (p>0.05) Conclusions: In conclusion, topical instillation of limbal mesenchymal stem secretome has a potential effect on inhibiton of corneal neovascularization even though not significant at day-7 in experimental alkali injury model

Case Report Awareness to Prolapse of the Eye in Baby

Corneal perforation may be associated with prolapse of ocular tissue. We present a rare case reportan11 months-old female presented prolapsed of right eye. She was earlier diagnosedwithright corneal ulcer anddescemetocele. Physicalexamination revealed perforatedcornea.On ultrasound examination showedvitreous hazzinessand membranous opacity assumed due to inflammation. Based on examinationswe suspectedmalignancy process and performed enucleation withautogenous Dermis Fat Graft (DFG) which harvested onthigh region.Histopathology examination revealed chronic inflammation process.Hence, determining the causative factors of the prolapsed eye is the key for better management and prognosis

Phlyctenule Conjunctivitis of Tuberculosis (A Case Report)

Phlyctenule conjunctivitis of tuberculosis is a very rare case. The diagnosis is very difficult because always not considered in conjunctivitis unless which is not improved with a number of treatments, contact history with tuberculosis patient is not always clear and culture material is not available to confirm the diagnosis

Matrix Metalloproteinase-3 Down Regulation and Cell Migration Inhibition in Human Pterygium Fibroblasts by Mitomycin-C, Curcumin and Fibrin Glue

BACKGROUND: Pterygium is an ocular surface disease that often occurs in tropical countries with a high recurrence rate. Matrix metalloproteinase-3 (MMP-3) play a key role in the inflammatory process of pterygium. This study aims to investigate the ability of curcumin and fibrin glue (FG) in suppressing the expression of MMP-3, and whether can be expected as adjuvant therapy to reduce pterygium recurrence.METHODS: Human pterygium fibroblasts (HPF) obtained from primary cultured of pterygium were treated with no treatment, curcumin, mitomycin-C (MMC), and FG. MMP-3 expression was analyzed using immunocytochemistry and the intensity measurement was done using ImageJ software. Cell migration was measured by scratching and stratification of fibroblast culture after cell confluence, and assessed for 48 hours.RESULTS: The expression of MMP-3 were lower in the HPF treated with 100 mol/mL curcumin, 200 mol/mL, and FG (2205.84±86.1 pg/mL, 1002.51±25.22 pg/mL, 1131.55±17.71 pg/mL, respectively) in comparison with untreated HPF (4703.49±108.9 pg/mL). The expression of MMP-3 were significantly different between groups (p<0.001). Cell migration of HPF after scratching with curcumin intervention at 200 mol/mL decrease from 178.67±2.85 (24 hours) to 88.83±1.48 (48 hours). Meanwhile the migration in FG group also decrease from 180.4±2.56 (24 hours) to 72.45±1.25 (48 hours).CONCLUSION: Curcumin and FG able to reduce the expression of MMP-3 and inhibit the migration of HPF cells.KEYWORDS: curcumin, mitomycin C, fibrin glue, human pterygium fibroblast, MMP-

Curcumin as Potential Adjuvant Therapy to Prevent Recurrence in Pterygium Excision

Abstract Pterygium is a wing-shaped growth from the conjunctiva and fibrovascular tissue onto the surface of the cornea. It has a high postoperative recurrence rate (which can be as high as 89% and its severity may vary according to the approach adopted and the preoperative conditions). To reduce the postoperative recurrence rate, several surgical techniques have been developed. The pathophysiology of pterygium development is diverse, one of the main factors is the proliferative process initiated by the TGF-? mediator. One of the therapies developed to prevent pterygium recurrence is by targeting therapy on the proliferative pathway by adding pterygium surgery with adjuvant MMC therapy. MMC plays a role in inhibiting DNA synthesis, thereby reducing TGF-? expression and ultimately inhibiting cell proliferation. Although it can reduce the recurrence rate quite well, MMC has numbers of complications. Therefore, an agent is needed to suppress pterygium proliferation which is relatively safer. Curcumin is a compound derived from the tuber of Curcuma longa L. which includes polyphenols. Curcumin modulates proliferation by stimulating the formation of TGIF which is a negative regulator of TGF-? so that it can modulate proliferation

Matrix Metalloproteinase-3 Down Regulation and Cell Migration Inhibition in Human Pterygium Fibroblasts by Mitomycin-C, Curcumin and Fibrin Glue

Abstract BACKGROUND: Pterygium is an ocular surface disease that often occurs in tropical countries with a high recurrence rate. Matrix metalloproteinase-3 (MMP-3) play a key role in the inflammatory process of pterygium. This study aims to investigate the ability of curcumin and fibrin glue (FG) in suppressing the expression of MMP-3, and whether can be expected as adjuvant therapy to reduce pterygium recurrence. METHODS: Human pterygium fibroblasts (HPF) obtained from primary cultured of pterygium were treated with no treatment, curcumin, mitomycin-C (MMC), and FG. MMP-3 expression was analyzed using immunocytochemistry and the intensity measurement was done using ImageJ software. Cell migration was measured by scratching and stratification of fibroblast culture after cell confluence, and assessed for 48 hours. RESULTS: The expression of MMP-3 were lower in the HPF treated with 100 mol/mL curcumin, 200 mol/mL, and FG (2205.84±86.1 pg/mL, 1002.51±25.22 pg/mL, 1131.55±17.71 pg/mL, respectively) in comparison with untreated HPF (4703.49±108.9 pg/mL). The expression of MMP-3 were significantly different between groups (p<0.001). Cell migration of HPF after scratching with curcumin intervention at 200 mol/mL decrease from 178.67±2.85 (24 hours) to 88.83±1.48 (48 hours). Meanwhile the migration in FG group also decrease from 180.4±2.56 (24 hours) to 72.45±1.25 (48 hours). CONCLUSION: Curcumin and FG able to reduce the expression of MMP-3 and inhibit the migration of HPF cells

Cryotherapy Has No Significant Effect on MMP-9 and TGF-β1 Expression in Fungal Corneal Ulcer

BACKGROUND: Usually, surgical intervention is needed to eradicate the fungal microorganism that cause fungal corneal ulcers. However, since surgical intervention is invasive, the latest technology uses cryotherapy in treating it. Cryotherapy plays a vital role in the wound healing process. We aimed to evaluate is to study the decreased expression of matrix metalloproteinase 9 (MMP-9) and transforming growth factor β1 (TGF-β1) in fungal corneal ulcers after the administration of cryotherapy.METHODS: Aspergillus flavus fungus was injected to the intrastromal corneas of all Sprague Dawley rats. The rats were divided into four groups, the first group was not given any therapy, the second group was given topical natamycin therapy, the third group was given cryotherapy, and the fourth group was given a combination between cryotherapy and topical natamycin therapy. Therapy was given after five days of follow up on the formation of a corneal ulcer. After four days of therapy, the eyes were enucleated to determine MMP-9 and TGF-β1 expression.RESULTS: The result in the third group showed lower MMP-9 expression (20.0±10.0% cells per field of view) compared to the second group (40.0±20.0% cells per field of view) and the fourth group (30.0±25.0% cells per field of view), but had the same MMP-9 expression value as the first group. There was no significant difference in MMP-9 expression between the four groups (p=0.356). The third group reduced more TGF-β1 expression (10.0±12.50% cells per field of view) compared to the fourth group (30±27.5% cells per field of view) and the first group (30±32.5% cells per field of view). There was also no significant difference in TGF-β1 between the four groups (p=0.315).CONCLUSION: There is no significant difference in the expression of TGF-β1 and MMP-9 after the cryotherapy treatment.KEYWORDS: corneal ulcer, cryotherapy, MMP-9, TGF-β

Mitomycin C, curcumin, and fibrin glue inhibit the cell proliferation and expression of TGF-β in human pterygium fibroblast

Introduction: High recurrence of a fibrotic disease, pterygium, following the surgical procedure is perceived as the primary challenge of its management. As the standard procedure, adjuvant therapy of using mitomycin C could dramatically reduce the recurrence of pterygium but could cause multiple serious complications. Our study aimed to investigate curcumin and fibrin glue as alternative candidates for adjuvant therapy in pterygium surgery. Methods: Human pterygium fibroblast (HPF) was isolated from the patient and cultured in-vitro. The HPF culture was then exposed with mitomycin C (0.4 mg/mL), curcumin (200 μmol/L), and fibrin glue, respectively, for 48 hours. The outcomes were determined by the proliferation of HPF and the expression of transforming growth factor-beta (TGF-β) which were obtained from 2,5-diphenyl-2H-tetrazolium bromide assay and immunofluorescence staining analysis with TGF-β antibody. Results: The experiment revealed that mitomycin C, curcumin, and fibrin glue could significantly inhibit the proliferation of HPF (p<0.05) suggesting their antifibrotic effect. Further analysis with immunofluorescence staining showed that mitomycin C, curcumin, and fibrin glue could significantly reduce the level of TGF-β as compared with control group (p<0.05). Conclusion: Mitomycin C was the most potent adjuvant agent to reduce the recurrence of pterygium, followed by curcumin and fibrin glue. Taken altogether, curcumin and fibrin glue have role as adjuvant therapy to prevent recurrence in pterygium surgery

Mitomycin C, curcumin, and fibrin glue inhibit the cell proliferation and expression of TGF-β in human pterygium fibroblast

Abstract Introduction: High recurrence of a fibrotic disease, pterygium, following the surgical procedure is perceived as the primary challenge of its management. As the standard procedure, adjuvant therapy of using mitomycin C could dramatically reduce the recurrence of pterygium but could cause multiple serious complications. Our study aimed to investigate curcumin and fibrin glue as alternative candidates for adjuvant therapy in pterygium surgery. Methods: Human pterygium fibroblast (HPF) was isolated from the patient and cultured in-vitro. The HPF culture was then exposed with mitomycin C (0.4 mg/mL), curcumin (200 μmol/L), and fibrin glue, respectively, for 48 hours. The outcomes were determined by the proliferation of HPF and the expression of transforming growth factor-beta (TGF-β) which were obtained from 2,5-diphenyl-2H-tetrazolium bromide assay and immunofluorescence staining analysis with TGF-β antibody. Results: The experiment revealed that mitomycin C, curcumin, and fibrin glue could significantly inhibit the proliferation of HPF (p<0.05) suggesting their antifibrotic effect. Further analysis with immunofluorescence staining showed that mitomycin C, curcumin, and fibrin glue could significantly reduce the level of TGF-β as compared with control group (p<0.05). Conclusion: Mitomycin C was the most potent adjuvant agent to reduce the recurrence of pterygium, followed by curcumin and fibrin glue. Taken altogether, curcumin and fibrin glue have role as adjuvant therapy to prevent recurrence in pterygium surgery