RESUMO
PURPOSE: We aimed to examine the effectiveness of mother milk exosomes in treating corrosive esophageal burns. MATERIALS AND METHODS: 32 rats were separated into four equal groups and weighed individually before the procedure. A corrosive esophageal burn model was created with 12.5% sodium hydroxide by a 3F Fogarty catheter. Group 1 did not apply any process or treatment, Group 2 was burned, and no treatment was performed. Group 3 was burned, and then 0.5 cc/day of mother milk exosome extract was given. Group 4 was not applied any process, and 0.5 cc/day mother milk exosome extract was given. All rats were weighed again and sacrificed. Biopsy samples were sent to the pathology laboratory for histopathological examination (in terms of inflammation, fibrosis, and necrosis).Kindly check and confrm all email ids.The e-mail addresses and affiliation of all authors were checked. Affiliation departments are as stated on the title page. There is no change. RESULTS: A significant difference was found in the results of inflammation and fibrosis. There was a meaningful difference in fibrosis between the 2nd and 3rd groups. There was weight gain in groups 1, 3 and 4. Statistical evaluations for each group were significant. CONCLUSION: It was observed that breast milk exosomes may be effective in inflammation and fibrosis formation in treating corrosive esophageal burns. This suggested that breast milk exosomes reduce stricture formation due to esophageal corrosion.Please confirm if the author names are presented accurately and in the correct sequence (given name, middle name/initial, family name). Author 1 Given name: [specify authors given name] Last name [specify authors last name]. Also, kindly confirm the details in the metadata are correct.The names and affiliation of all authors were checked. Affiliation departments are as stated on the title page. There is no change. Also we confirm the details in the metadata.
Assuntos
Queimaduras Químicas , Modelos Animais de Doenças , Exossomos , Animais , Ratos , Queimaduras Químicas/terapia , Esofagite/induzido quimicamente , Esofagite/patologia , Cáusticos/toxicidade , Leite Humano , Feminino , Hidróxido de Sódio/toxicidade , Esôfago/patologia , MasculinoRESUMO
PURPOSE: The current study used various techniques to develop a rabbit animal model of lacrimal gland damage caused by scarring conjunctivitis in the periglandular area. METHODS: Left eyes of New Zealand white rabbits were injected with 0.1 ml of 1M NaOH subconjunctivally around superior and inferior lacrimal gland orifices (Group 1, n = 4), touched with 1M NaOH for 100 s to the superior and inferior fornices with conjunctival denuding (Group 2; n = 4), and electrocauterization to the ductal opening area (Group 3; n = 4). The ocular surface staining, Schirmer I, lacrimal gland, and conjunctival changes were observed at baseline,1, 4, 8, and 12 weeks. The degree of glandular inflammation, conjunctival fibrosis (Masson Trichrome), and goblet cell density (PAS) were also assessed. RESULTS: At 12 weeks, the lacrimal glands of group 1 rabbits with periglandular injection showed severe inflammation with mean four foci/10HPF and a significant mean reduction in the Schirmer values by 7.6 mm (P = 0.007). Lacrimal glands had diffuse acinar atrophy, loss of myoepithelial cells, and ductular dilatation. The overlying conjunctiva showed fibrosis, goblet cell loss, and corneal vascularization in the inferotemporal quadrant. No lacrimal gland or ocular surface changes were observed in groups 2 and 3 at 12 weeks, except for localized subconjunctival fibrosis. CONCLUSION: Periglandular injection of 0.1 ml of 1M NaOH induced extensive lacrimal gland damage with reduced secretion and scarring in the subconjunctival plane compared to direct cauterization or direct NaOH contact to the ductal orifices of the rabbit lacrimal gland.
Assuntos
Cicatriz , Túnica Conjuntiva , Conjuntivite , Modelos Animais de Doenças , Síndromes do Olho Seco , Células Caliciformes , Lágrimas , Animais , Coelhos , Síndromes do Olho Seco/metabolismo , Cicatriz/patologia , Células Caliciformes/patologia , Túnica Conjuntiva/patologia , Lágrimas/metabolismo , Conjuntivite/patologia , Aparelho Lacrimal/patologia , Hidróxido de Sódio/toxicidade , Fibrose , Masculino , Contagem de Células , Feminino , EletrocoagulaçãoRESUMO
INTRODUCTION: In some cases, the tongue and oesophagus tissues are damaged by the corrosive burn. Surgical interventions may cause scar formation, and severe burns treatment methods are limited. This study aims to investigate bromelain, a phytotherapeutic product, on the corrosive burn as a non-surgical option and as an adjunctive therapy, insofar as the treatment of corrosive wounds is not limited only to the treatment of oxidative stress and inflammatory reactions. METHODS: On the tongues of Wistar albino rats, chemically produced oral ulcers were created by topical application of NaOH (40%) solution, and in the distal oesophagus same mixture was applied to produce a corrosive oesophageal burn. For a week, they were treated orally by bromelain (100 mg/kg/day) or saline solution. At the end of seven days, animals were decapitated to remove the tongue and oesophagus, and blood samples were collected to obtain serum. Myeloperoxidase (MPO) activity, malondialdehyde (MDA), glutathione (GSH), interleukin-1 beta (IL-1ß) and tumour necrosis factor-alpha (TNF-α) concentrations were measured in serum, and luminol and lucigenin chemiluminescence (CL) were measured in tissue samples. RESULTS: MDA and CL values were significantly increased, and GSH levels in tissue significantly decreased due to the corrosive burns. Saline treated corrosive burn group measured higher in the serum cytokines in according to the control group. CONCLUSIONS: Bromelain administration decreased oxidant and inflammatory parameters and increased antioxidant levels in NaOH-induced corrosive burns. Thus, we concluded that bromelain may protect the tongue and oesophagus tissues with its anti-inflammatory and antioxidant effects.
Assuntos
Bromelaínas , Queimaduras , Cáusticos , Esôfago/lesões , Animais , Antioxidantes , Bromelaínas/uso terapêutico , Queimaduras/tratamento farmacológico , Cáusticos/toxicidade , Glutationa , Interleucina-1beta , Malondialdeído , Peroxidase , Ratos , Ratos Wistar , Hidróxido de Sódio/toxicidade , Fator de Necrose Tumoral alfaRESUMO
Potassium bromate (KBrO3) present in consumed ozonised water was recently documented to exacerbate experimental gastric ulcer. Information, however, is vague as regards its effects in the colon where water reabsorption occurs. In this study, we observed the possible effects of KBrO3 on oxidative stress and inflammatory biomarkers in sodium hydroxide (NaOH) - induced Crohn's colitis (CC). Wistar rats (180-200 g) were divided into six groups (n = 10): (i) control; (ii) untreated CC (induced by 1.4% NaOH; intra-rectal administration); and (iii-vi) CC treated with vitamin E, KBrO3, vitamin E+KBrO3, and sulphazalazine, respectively, for 7 days. Body weight and stool score were monitored daily. By day 3 and 7, excised colon was evaluated for ulcer scores and biochemical and histological analysis. Blood samples collected on days 3 and 7 were assayed for haematological indices using standard methods. Data were subjected to analysis of variance (ANOVA) and p ≤ 0.05 considered significant. Platelet/lymphocyte ratio, colonic ulcer score, malondialdehyde, and mast cells were significantly decreased while colonic sulfhydryl, and Ca2+- and Na+/K+-ATPase activities were increased following KBrO3 treatment compared with untreated CC. These findings suggest that KBrO3 may mitigate against NaOH-induced CC via inhibiting mast cell population and oxidative and inflammatory content but stimulating colonic sulfhydryl and Ca2+- and Na+/K+-ATPase activities.
Assuntos
Bromatos/farmacologia , Colite/tratamento farmacológico , Doença de Crohn/tratamento farmacológico , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Biomarcadores/metabolismo , Colite/induzido quimicamente , Colite/metabolismo , Colite/patologia , Doença de Crohn/induzido quimicamente , Doença de Crohn/metabolismo , Doença de Crohn/patologia , Interações Medicamentosas , Aditivos Alimentares/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Oxirredução , Ratos , Ratos Wistar , Hidróxido de Sódio/toxicidadeRESUMO
Limbal Stem Cell Deficiency (LSCD), caused due to corneal injury, primarily by chemical/alkali burns, leads to compromised vision. Recently, several animal models of corneal alkali burn injury have become available. The majority of the studies with these animal models start interventions soon after the injury. However, in the clinical setting, there is a considerable delay before the intervention is initiated. Detailed knowledge of the molecular, histopathological, and clinical parameters associated with the progression of the injury leading to LSCD is highly desirable. In this context, we set out to investigate clinical, histopathological parameters of ocular surface alkali burn over a long period of time, post-injury. Limbal stem cell-deficient animal models of rabbits were created by alkali burn using sodium hydroxide, which was then assessed for their progression towards LSCD by grading the alkali burn, corneal haze, and vascularization. Additionally, cells present on the corneal surface after the burn was investigated by histology and immunophenotyping. Grading of rabbit eyes post-alkali burn had shown complete conjunctivalization in 80% (n = 12/15) of the rabbits with the alkali burn grade score of 3.88 ± 0.29 in three months and remained stable at four months (4.12 ± 0.24). However, ocular surface showed self-healing in 20% (n = 3/15) of the rabbits with a score of 1.67 ± 0.34 in four months irrespective of similar alkali injury. These self-healing corneas exhibited decreased opacity score from 2.51 ± 0.39 to 0.66 ± 0.22 (p = 0.002) and regressed vascularity from 1.66 ± 0.41 to 0.66 ± 0.33 in one to nine months, respectively. Restoration of the corneal phenotype (CK3+) was observed in central and mid-peripheral regions of the self-healing corneas, and histology revealed the localization of inflammatory cells to the peripheral cornea when compared to conjunctivalized and scarred LSCD eyes. Our study shows the essentiality to consider the time required for surgical intervention after the corneal alkali injury in rabbit models as evident from their tendency to self-heal and restore corneal phenotype without therapy. Such information on the possibility of self-healing should be useful in further studies as well as determining interventional timings and strategy during clinical presentation of corneal alkali burns.
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Queimaduras Químicas/fisiopatologia , Lesões da Córnea/fisiopatologia , Neovascularização da Córnea/fisiopatologia , Opacidade da Córnea/fisiopatologia , Queimaduras Oculares/induzido quimicamente , Recuperação de Função Fisiológica/fisiologia , Hidróxido de Sódio/toxicidade , Animais , Cáusticos/toxicidade , Túnica Conjuntiva/fisiopatologia , Córnea/fisiopatologia , Modelos Animais de Doenças , Queimaduras Oculares/fisiopatologia , Seguimentos , Limbo da Córnea/citologia , Coelhos , Transplante de Células-Tronco , Cicatrização/fisiologiaRESUMO
Alkali burn to the cornea is one of the most intractable injuries to the eye due to the opacity resulting from neovascularization (NV) and fibrosis. Numerous studies have focused on studying the effect of drugs on alkali-induced corneal injury in mouse, but fewer on the involvement of alkali-induced DNA methylation and the PI3K/AKT/mTOR signaling pathway in the mechanism of alkali-induced corneal injury. Thus, the aim of this study was to determine the involvement of DNA methyltransferase 3 B-madiated DNA methylation and PI3K/AKT/mTOR signaling modulation in the mechanism of alkali-induced corneal injury in a mouse model. To this end, we used bisulfite sequencing polymerase chain reaction and Western blot analysis, to study the effects of 5-aza-2'-deoxycytidine and 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one, which inhibit methyltransferase and PI3K respectively, on DNA methylation and expression of downstream effectors of PI3K related to corneal NV, including TSC1 and mTOR genes. The results showed that, after an intraperitoneal injection of rapamycin (2 mg/kg/day) for seven days, the alkali-induced opacity and NV were remarkably decreased mainly by suppressing the infiltration of immune cells into injured corneas, angiogenesis, VEGF expression and myofibroblasts differentiation; as well as by promoting corneal cell proliferation and PI3K/AKT/mTOR signaling. More significantly, these findings showed that epigenetic regulatory mechanisms by DNA methylation played a key role in corneal NV, including in corneal alkali burn-induced methylation modification and rapamycin-induced DNA demethylation which involved the regulation of the PI3K/AKT/mTOR signaling pathway at the protein level. The precise findings of morphological improvement and regulatory mechanisms are helpful to guide the use of rapamycin in the treatment of corneal angiogenesis induced by alkaline-burn.
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Queimaduras Químicas/prevenção & controle , Lesões da Córnea/prevenção & controle , Queimaduras Oculares/induzido quimicamente , Imunossupressores/uso terapêutico , Sirolimo/uso terapêutico , Serina-Treonina Quinases TOR/genética , Proteína 1 do Complexo Esclerose Tuberosa/genética , Actinas/genética , Animais , Western Blotting , Queimaduras Químicas/genética , Queimaduras Químicas/patologia , Cromonas/farmacologia , Lesões da Córnea/genética , Lesões da Córnea/patologia , Metilação de DNA , Modelos Animais de Doenças , Técnica Indireta de Fluorescência para Anticorpo , Regulação da Expressão Gênica/fisiologia , Masculino , Camundongos , Morfolinas/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Inibidores de Fosfoinositídeo-3 Quinase/farmacologia , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-akt/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Hidróxido de Sódio/toxicidade , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
Corneal calcification is a vision-threatening manifestation of calcium containing agents in ocular burn. As we previously reported, our interest was sparked by a particular discrepancy of a case: A patient treated for a non-calcium containing agent in eye burn from exposure to an alkaline mixture of NaOH and KOH, who unexpectedly developed corneal calcification. This current study aims to elucidate whether the 2min lasting irrigation with a phosphate-buffered saline itself, regardless of rinsing regimen, triggers corneal calcification. The Ex Vivo Eye Irritation Test (EVEIT) system was used on rabbit corneas to replicate the very same phosphate-buffered saline solution the patient was treated with. The rabbit corneas were first burned with 1 M NaOH, rinsed with 4.9% phosphate-buffered saline for 2 min, and were then moisturized with an artificial tear solution for 48 h. All corneas were fluorescein-stained for photo documentation, snap-frozen, lyophilizated, and the electrolyte content was analyzed by Energy-Dispersive X-ray spectroscopy (EDX). The EDX analysis revealed pathological phosphorous in corneal stroma after a single rinsing with phosphate-buffered saline. Ongoing application of artificial tears containing physiological 14.581 mmol Ca2+ /l led to macroscopically visible calcification, but only in areas of induced corneal erosion. Regardless of the rinsing protocol neither 2 or 15 min of eye rinsing with phosphate containing rinsing solutions, we have given proof that corneal calcification is a foreseeable effect of the phosphate-buffered saline rinsing of mechanically epithelial damaged and chemically burnt eyes. Thus, it is crucial to legally restrict the formulations of phosphate-buffered salines in the medical treatment of eye burns, corneal erosions or chemical splashes of the eye.
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Queimaduras Químicas/terapia , Calcinose/induzido quimicamente , Córnea/efeitos dos fármacos , Queimaduras Oculares/terapia , Fosfatos/farmacologia , Solução Salina/química , Irrigação Terapêutica/métodos , Adulto , Animais , Soluções Tampão , Queimaduras Químicas/etiologia , Calcinose/patologia , Córnea/diagnóstico por imagem , Queimaduras Oculares/induzido quimicamente , Primeiros Socorros , Humanos , Técnicas In Vitro , Masculino , Fosfatos/efeitos adversos , Coelhos , Hidróxido de Sódio/toxicidade , Espectrometria por Raios XRESUMO
BACKGROUND: Angiogenesis is tightly linked to inflammation. Cytokines of interleukin 1 (IL-1) family are key mediators in modulating inflammatory responses. METHODS: In this study, we examined the role of IL-38, a member of the IL-1 family, in mediating inflammation-induced angiogenesis. RESULTS: The results showed that the angiogenesis was attenuated by topical administration of IL-38 to the injured corneas in a mouse model of alkali-induced corneal neovascularization (CNV). Further study showed that the expression of inflammatory cytokines TNF-α, IL-6, IL-8 and IL-1ß was decreased in the IL-38-treated corneas. Moreover, the angiogenic activities including the proliferation, migration and tube formation of human retinal endothelial cells were reduced by IL-38 treatment in vitro. CONCLUSION: The data indicate that IL-38 modulates inflammation-induced angiogenesis.
Assuntos
Neovascularização da Córnea/tratamento farmacológico , Interleucina-1/fisiologia , Administração Tópica , Animais , Movimento Celular , Células Cultivadas , Neovascularização da Córnea/etiologia , Citocinas/biossíntese , Citocinas/genética , Células Endoteliais/efeitos dos fármacos , Humanos , Interleucina-1/antagonistas & inibidores , Interleucina-1/farmacologia , Interleucina-1/uso terapêutico , Ceratite/induzido quimicamente , Ceratite/complicações , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/uso terapêutico , Retina/citologia , Hidróxido de Sódio/toxicidade , Fator A de Crescimento do Endotélio Vascular/farmacologiaRESUMO
Limbal stem cell deficiency (LSCD) is one of the serious cause of visual impairment and blindness with loss of corneal clarity and vascularization. Factors such as ocular burns (acids, lime, thermal), genetic disorders or infections results in the loss of limbal stem cells leading to LSCD. Reliable animal models of LSCD are useful for understanding the pathophysiology and developing novel therapeutic approaches. The purpose of the present study was to validate small and large animal models of LSCD by immunohistochemcal, clinical and histopathological comparison with human. The animal models of LSCD were created by topical administration of sodium hydroxide on the ocular surface of C57BL/6 mice (m, nâ¯=â¯12) and New Zealand white rabbits (r, nâ¯=â¯12) as per the standard existing protocol. Human corneal specimens (h, nâ¯=â¯12) were obtained from tissue bank who had chemical burn-induced LSCD. All samples were either paraffin embedded or frozen in cryogenic medium and the sections were processed for Hematoxylin-Eosin and Periodic Acid-Schiff staining to analyse the morphology and histopathological features of the corneal surface such as vascularization, inflammation, presence of goblet cells, epithelial hyperplasia and keratinization. Immunofluorescence was performed to distinguish between corneal (CK3+), conjunctival (CK19+) and epidermal (CK10+) epithelial phenotype. Histological analysis of corneal specimens from the three groups showed the presence of goblet cells (h:83%, m:50%, r:50%, p = 0.014), epithelial hypertrophy (h:92%, m:50%, r:66.6%, p = 0.04), epithelial hyperplasia (h:50%, m:17%, r:17%, p = 0.18), intra epithelial edema (h:42%, m:33%, r:100%, p = 0.02), stromal inflammation (h:100%, m:67%, r:67%, p = 0.01) and stromal vascularization (h:100%, m:50%, r:67%), in varying proportions. Immunostaining showed presence of total LSCD (CK19 + and/or CK10+, CK3-) in 92% of human and 50% of animal specimens. While partial LSCD (CK19 + and/or CK10+, CK3+) was seen in 8% of human and 50% of animal specimens. Our study shows the significant differences in the extent of vascularization, inflammation, epithelial thickness and goblet cell formation in mice and rabbit models of LSCD when compared to post-chemical burn LSCD in human corneas. In both mice and rabbit models complete LSCD developed in only 50% of cases and this important fact needs to be considered when working with animal models of LSCD.
Assuntos
Queimaduras Químicas/patologia , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Queimaduras Oculares/induzido quimicamente , Células Caliciformes/patologia , Ceratite/patologia , Limbo da Córnea/patologia , Animais , Queimaduras Químicas/metabolismo , Doenças da Córnea/metabolismo , Doenças da Córnea/patologia , Neovascularização da Córnea/metabolismo , Células Epiteliais/metabolismo , Epitélio Corneano , Queimaduras Oculares/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Células Caliciformes/metabolismo , Humanos , Imunofenotipagem , Inflamação/metabolismo , Inflamação/patologia , Queratina-19/metabolismo , Queratina-3/metabolismo , Ceratite/metabolismo , Limbo da Córnea/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucinas/metabolismo , Coelhos , Hidróxido de Sódio/toxicidadeRESUMO
Effective methods to detect viable Mycobacterium tuberculosis, the main causative agent of tuberculosis (TB), are urgently needed. To date, cultivation of M. tuberculosis is the gold standard, which depends on initial sample processing with N-acetyl-l-cysteine-sodium hydroxide (NALC-NaOH), chemicals that compromise M. tuberculosis viability and, consequently, the performance of downstream tests. We applied culture and the novel molecular bacterial load assay (MBLA) to measure the loss of M. tuberculosis viability following NALC-NaOH treatment of M. tuberculosis H37Rv pure culture and clinical sputum samples from pulmonary TB patients. Compared to the bacterial loads of untreated controls, NALC-NaOH treatment of M. tuberculosis reduced the MBLA-detectable bacillary load (estimated number of CFU [eCFU] per milliliter) by 0.66 ± 0.21 log10 at 23°C (P = 0.018) and 0.72 ± 0.08 log10 at 30°C (P = 0.013). Likewise, NALC-NaOH treatment reduced the viable count on solid culture by 0.84 ± 0.02 log10 CFU/ml at 23°C (P < 0.001) and 0.85 ± 0.01 log10 CFU/ml at 30°C (P < 0.001), respectively. The reduction in the viable count was reflected by a corresponding increase in the time to positivity of the mycobacterial growth indicator tube (MGIT) liquid culture: 1.2 days at 23°C (P < 0.001) and 1.1 days at 30°C (P < 0.001). This NaOH-induced M. tuberculosis viability loss was replicated in clinical sputum samples, with the bacterial load dropping by 0.65 ± 0.17 log10 from 5.36 ± 0.24 log10 eCFU/ml to 4.71 ± 0.16 log10 eCFU/ml for untreated and treated sputa, respectively. Applying the model of Bowness et al. (R. Bowness, M. J. Boeree, R. Aarnoutse, R. Dawson, et al., J Antimicrob Chemother 70:448-455, 2015, https://doi.org/10.1093/jac/dku415) revealed that the treated MGIT time to culture positivity of 142 ± 7.02 h was equivalent to 4.86 ± 0.28 log10 CFU, consistent with the MBLA-measured bacterial load. Our study confirms the contribution of NALC-NaOH treatment to the loss of viable bacterial counts. Tests that obviate the need for decontamination may offer an alternative option for the accurate detection of viable M. tuberculosis and treatment response monitoring.
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Carga Bacteriana/métodos , Mycobacterium tuberculosis/isolamento & purificação , Hidróxido de Sódio/toxicidade , Tuberculose/microbiologia , Contagem de Colônia Microbiana , DNA Bacteriano/genética , Testes Diagnósticos de Rotina , Viabilidade Microbiana/efeitos dos fármacos , Técnicas de Diagnóstico Molecular , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crescimento & desenvolvimento , RNA Ribossômico 16S/genética , Manejo de Espécimes , Escarro/microbiologia , Temperatura , Fatores de Tempo , Tuberculose/diagnósticoRESUMO
Purpose: To compare the therapeutic effects of human derivatives in a mouse alkali burn model. Methods: The right eyes of mice were injured using NaOH. After alkali injury, one of the following agents was topically administered for 7 d: human amniotic membrane (hAM) suspension, human umbilical cord serum (hUCS), and human peripheral blood serum (hPBS), or saline. The epithelial defect areas on days 1, 2, and 3 degrees of opacity on days 2, 3, and 7, and corneal neovascularization (NV) areas on day 7 were evaluated. Histologic examination and mRNA expression levels of tumour necrosis factor (TNF)-α, interleukin (IL)-6, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2, MMP-8, and MMP-9 were also evaluated on day 7. Results: The epithelial defect areas in the hUCS group were smaller than those in the control and hPBS groups on day 3 (p < .05, respectively). The epithelial defect areas in the hAM suspension group showed smaller than those in the control and hPBS groups on days 1 and 2 (p < .05, respectively). The degrees of opacity were lower in all treatment groups than that of the saline control group on day 7 (p < .05, respectively). Corneal NV areas were not different among groups on day 7 (p = 0.20). The expression levels of TNF-α, IL-6, MMP-8, and MMP-9 mRNA and the infiltration of the inflammatory cells in all treatment groups were lesser than those in the control group on day 7 (p< .05, respectively). Conclusions: All treatments reduced inflammatory reactions and corneal opacity development. Corneal reepithelialization was faster in the hUCS group.
Assuntos
Âmnio , Queimaduras Químicas/terapia , Neovascularização da Córnea/terapia , Opacidade da Córnea/terapia , Queimaduras Oculares/terapia , Soro , Hidróxido de Sódio/toxicidade , Animais , Queimaduras Químicas/patologia , Córnea/efeitos dos fármacos , Córnea/patologia , Neovascularização da Córnea/patologia , Opacidade da Córnea/patologia , Queimaduras Oculares/induzido quimicamente , Queimaduras Oculares/patologia , Humanos , Masculino , Camundongos Endogâmicos BALB CRESUMO
Purpose: To investigate the preventive effects of topical sunitinib, sunitinib-hesperetin and sunitinib-doxycycline combinations on corneal neovascularization (CNV), apoptosis and fibrosis in a corneal alkali burn model. Materials and Methods: The corneas of 32 Wistar albino rats were cauterized with silver nitrate to induce CNV. Four groups were created receiving artificial tears (sham), sunitinib (0.5 mg/ml), sunitinib-hesperetin (0.5 mg/ml-0.2 mg/ml), and sunitinib-doxycycline (0.5 mg/ml-20 mg/ml) treatments. Corneal photographs were taken on days 0, 7 and 15 . Photographs of the cornea were digitally analyzed to measure the size of the neovascularization area in comparison to the total corneal surface area. On the 15th day, the animals were euthanized, and the eyes were enucleated for immunohistochemical staining to investigate neovascularization, apoptosis, and fibrosis. Results: CNV areas on the 7th day in the sunitinib (4.8% ± 0.07%) and sunitinib-hesperetin (1.1% ± 0.03%) groups were smaller than those in the sham group (33.9% ± 0.12%) (p = 0.001 and, p < 0.001 respectively). On the 15th day, the CNV area in the sunitinib-hesperetin (20.8% ± 0.37%) group was significantly smaller than that of the sham group (74.6% ± 0.32%) (p = 0.039). The combination groups had lower levels of VEGF, TUNEL and α-SMA positivity than the sunitinib monotherapy group. TUNEL positivity was lowest in the sunitinib-hesperetin and sunitinib-doxycycline groups, and α-SMA positivity was lowest in the sunitinib-hesperetin group. Conclusion: Topical sunitinib-hesperetin was more effective than sunitinib alone and the sunitinib-doxycycline combination in the treatment of CNV. The combination of sunitinib and hesperetin seems to be a promising treatment for preventing corneal fibrosis and apoptosis.
Assuntos
Inibidores da Angiogênese/uso terapêutico , Antibacterianos/uso terapêutico , Neovascularização da Córnea/prevenção & controle , Doxiciclina/uso terapêutico , Hesperidina/uso terapêutico , Sunitinibe/uso terapêutico , Actinas/metabolismo , Administração Oftálmica , Animais , Apoptose/efeitos dos fármacos , Queimaduras Químicas/prevenção & controle , Modelos Animais de Doenças , Quimioterapia Combinada , Queimaduras Oculares/induzido quimicamente , Marcação In Situ das Extremidades Cortadas , Masculino , Soluções Oftálmicas , Ratos Wistar , Hidróxido de Sódio/toxicidade , Resultado do Tratamento , Fator A de Crescimento do Endotélio Vascular/sangueRESUMO
Toll-like receptors (TLRs) play an important role in inflammatory and immunological responses, which are intimately related to neovascularization. However, the precise mode of action of TLR3 in neovascularization still remains ambiguous. In this study, we sought to investigate the role of TLR3 in pathological corneal neovascularization (CNV) using a mouse model of alkali-induced CNV. CNV was attenuated in TLR3-deficient mice, and the absence of TLR3 led to decreased production of stromal cell-derived factor 1 (SDF-1), a well-characterized cytokine that regulates the recruitment of endothelial progenitor cells (EPCs) to the sites of neo-angiogenic niches in the injured tissues. Topical administration of polyinosinic-polycytidylic acid [poly (I:C)], a synthetic ligand for TLR3, to the injured cornea promoted CNV in wild type (WT) mice but not in TLR3-deficient mice. In addition, the effect of poly (I:C) on WT mice was abolished by addition of SDF-1 receptor antagonist AMD 3100. Furthermore, poly (I:C) treatment in vitro enhanced the migration of EPCs, whereas the enhanced migration was abolished by AMD 3100. These results indicate an essential role of TLR3 signalling in CNV that involves upregulating SDF-1 production and recruiting EPCs to the sites of injury for neovascularization. Thus, targeting the TLR3 signalling cascade may constitute a novel therapeutic approach for treating neovascularization-related diseases.
Assuntos
Quimiocina CXCL12/metabolismo , Neovascularização da Córnea/metabolismo , Células Progenitoras Endoteliais/citologia , Transdução de Sinais/fisiologia , Receptor 3 Toll-Like/metabolismo , Administração Oftálmica , Animais , Queimaduras Químicas/metabolismo , Movimento Celular/fisiologia , Córnea/efeitos dos fármacos , Neovascularização da Córnea/induzido quimicamente , Neovascularização da Córnea/patologia , Modelos Animais de Doenças , Queimaduras Oculares/induzido quimicamente , Técnica Indireta de Fluorescência para Anticorpo , Camundongos , Camundongos Endogâmicos C57BL , Poli I-C/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Hidróxido de Sódio/toxicidadeRESUMO
The aim of the present study was to evaluate the effect and the mechanism of action of the conditioned medium from human uterine cervical stem cells (CM-hUCESC) on corneal wound healing in a rabbit dry eye model. To do this, dry eye and corneal epithelial injuries were induced in rabbits by topical administration of atropine sulfate and NaOH. Hematoxylin-Eosin (H&E) and Ki-67 immunostaining were carried out to evaluate corneal damage and cell proliferation, and real-time PCR was used to evaluate proinflammatory cytokines in the cornea. In addition, in order to investigate possible factors involved in corneal regeneration, primary cultures of rat corneal epithelial cells (rCECs) were used to evaluate cell migration, proliferation, and apoptosis before and after immunoprecipitation of specific factors from the CM-hUCESC. Results showed that CM-hUCESC treatment significantly improved epithelial regeneration in rabbits with dry eye induced by atropine and reduced corneal pro-inflammatory TNF-α, MCP-1, MIP-1α and IL-6 cytokines. In addition, metalloproteinase inhibitors TIMP-1 and TIMP-2, which are present at high levels in CM-hUCESC, mediated corneal regenerative effects by both inducing corneal epithelial cell proliferation and inhibiting apoptosis. In summary, CM-hUCESC induces faster corneal regeneration in a rabbit model of dry eye induced by atropine than conventional treatments, being TIMP-1 and TIMP-2 mediators in this process. The results indicate that an alternative CM-based treatment for some corneal conditions is achievable, although future studies would be necessary to investigate other factors involved in the multiple observed effects of CM-hUCESC.
Assuntos
Colo do Útero/citologia , Meios de Cultivo Condicionados/farmacologia , Síndromes do Olho Seco/tratamento farmacológico , Epitélio Corneano/fisiologia , Regeneração/fisiologia , Células-Tronco/citologia , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Animais , Apoptose , Atropina/toxicidade , Western Blotting , Movimento Celular , Proliferação de Células , Citocinas/genética , Modelos Animais de Doenças , Síndromes do Olho Seco/induzido quimicamente , Síndromes do Olho Seco/metabolismo , Feminino , Antígeno Ki-67/metabolismo , Masculino , Coelhos , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Hidróxido de Sódio/toxicidade , Espectrometria de Massas em Tandem , Cicatrização/efeitos dos fármacosRESUMO
Las sustancias cáusticas son productos químicos capaces de provocar lesiones de diferente gravedad, según sea la concentración del producto, el tiempo de contacto y la vía de ingreso. La exposición es habitual por tratarse de productos utilizados en el hogar como destapacañerías y/o limpiahornos. Los cáusticos alcalinos producen necrosis por licuefacción de los tejidos. La ingesta causa edema, ulceraciones, sangrado, placas blanquecinas e intensa actividad fibroblástica con cicatrización en tres semanas. El esófago se afecta más que el estómago, en especial cuando se trata de productos sólidos o en escamas. La secuela observada es la estenosis esofágica, la cual puede requerir desde procedimientos de dilatación hasta cirugía de reemplazo, con alta morbilidad. El objetivo del trabajo es presentar tres casos clínicos y sus complicaciones a largo plazo; recordar el manejo inicial del paciente que ingiere cáusticos alcalinos, su seguimiento multidisciplinario y resaltar las medidas de prevención para evitar este tipo de accidentes graves.
Caustic are chemical substances capable of causing different degree of lesions, according to the product concentration, the time and the route of contact. The usual exposure is because of their use as household products such as drain openers and oven cleaners. Caustic alkalis produce tissue liquefaction necrosis. Ingestion causes edema, ulceration, bleeding, whitish plaques and intense fibroblastic activity with healing in three weeks. The esophagus is more affected than the stomach, especially when solids are involved. The observed sequel is esophageal stricture, requiring treatments as dilation or replacement surgery, with high morbilidad. The aim of the paper is to report three clinical cases and their long-term complications; review the initial management of patients who ingested caustic alkali, highlighting its multidisciplinary monitoring and prevention measures to avoid such serious accidents.
Assuntos
Humanos , Pré-Escolar , Hidróxido de Sódio/efeitos adversos , Hidróxido de Sódio/toxicidade , Queimaduras Químicas , Esôfago/lesões , Cáusticos/toxicidade , Estenose Esofágica/induzido quimicamenteRESUMO
The aim of this study was to examine the effect of molecular hydrogen (H2) on the healing of alkali-injured cornea. The effects of the solution of H2 in phosphate buffered saline (PBS) or PBS alone topically applied on the alkali-injured rabbit cornea with 0.25 M NaOH were investigated using immunohistochemical and biochemical methods. Central corneal thickness taken as an index of corneal hydration was measured with an ultrasonic pachymeter. Results show that irrigation of the damaged eyes with H2 solution immediately after the injury and then within next five days renewed corneal transparency lost after the injury and reduced corneal hydration increased after the injury to physiological levels within ten days after the injury. In contrast, in injured corneas treated with PBS, the transparency of damaged corneas remained lost and corneal hydration elevated. Later results-on day 20 after the injury-showed that in alkali-injured corneas treated with H2 solution the expression of proinflammatory cytokines, peroxynitrite, detected by nitrotyrosine residues (NT), and malondialdehyde (MDA) expressions were very low or absent compared to PBS treated injured corneas, where NT and MDA expressions were present. In conclusion, H2 solution favorably influenced corneal healing after alkali injury via suppression of oxidative stress.
Assuntos
Lesões da Córnea/etiologia , Hidrogênio/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Hidróxido de Sódio/toxicidade , Actinas/metabolismo , Animais , Córnea/metabolismo , Córnea/patologia , Lesões da Córnea/metabolismo , Lesões da Córnea/patologia , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Expressão Gênica/efeitos dos fármacos , Imuno-Histoquímica , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Queratina-12/metabolismo , Queratina-3/metabolismo , Malondialdeído/metabolismo , Ácido Peroxinitroso/metabolismo , Coelhos , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
The aim of this study was to examine whether nanofiber scaffolds seeded with rabbit bone marrow mesenchymal stem cells (MSCs nanofibers) transferred onto the damaged corneal surface and covered with cyclosporine A (CsA)-loaded nanofiber scaffolds (CsA nanofibers) enable healing of the rabbit cornea injured with 1N NaOH. The healing of damaged corneas was examined morphologically, immunohistochemically and biochemically on day 24 after the injury. Compared to untreated injured corneas, where corneal ulceration or large corneal thinning or even perforation were developed, injured corneas treated with drug free nanofibers healed without profound disturbances in a majority of cases, although with fibrosis and scar formation. In injured corneas treated with CsA nanofibers or MSCs nanofibers, the development of scar formation was reduced. Best healing results were obtained with a combination of MSCs and CsA nanofibers (MSCs-CsA nanofibers). Corneas healed with highly restored transparency. Neovascularization highly expressed in untreated injured corneas and reduced in corneas treated with CsA nanofibers or MSCs nanofibers, was suppressed in corneas treated with MSCs-CsA nanofibers. The levels of matrix metalloproteinase 9, inducible nitric oxide synthase, interleukin 6, α-smooth muscle actin, tumor growth factor ß and vascular endothelial growth factor were significantly decreased in these corneas as compared to untreated corneas, where the levels of the above mentioned markers were high. In conclusion, MSCs-CsA nanofibers were effective in the treatment of severe alkali-induced corneal injury.
Assuntos
Lesões da Córnea/terapia , Neovascularização da Córnea/prevenção & controle , Ciclosporina/administração & dosagem , Imunossupressores/administração & dosagem , Transplante de Células-Tronco Mesenquimais/métodos , Animais , Cáusticos/toxicidade , Cicatriz/prevenção & controle , Modelos Animais de Doenças , Portadores de Fármacos , Sistemas de Liberação de Medicamentos/métodos , Feminino , Nanofibras , Coelhos , Hidróxido de Sódio/toxicidade , Alicerces Teciduais , Cicatrização/efeitos dos fármacosRESUMO
A superfusion apparatus (SA) was developed to maintain isolated human corneas ex vivo under conditions which mimic the natural eye environment in vivo, including controlled temperature, tear flow and intraocular pressure. The SA was designed, developed and tested for use in ophthalmic pre-clinical research and to test new pharmaceutical formulations. Corneas undergo an equilibration process in the new physiological environment for one day. The test was then initiated by the application of the test substance, incubation, and temporal assessment of corneal damage using various parameters. The effects of mild and severe irritant concentrations of NaOH (2% and 8%, respectively) on corneal opacity, swelling and epithelial integrity were studied, and the inflammatory status assessed using F4/80 and MPO as macrophages and neutrophils markers, respectively. The SA was then used to test new artificial tear formulations supplemented with silver ions as an active constituent, showing different degrees of inflammatory responses as indicated by the migration of MPO and F4/80 positive cells towards the epithelium. The human cornea superfusion apparatus was proposed as a model for acute eye irritation research.
Assuntos
Alternativas aos Testes com Animais , Cáusticos/toxicidade , Córnea/efeitos dos fármacos , Irritantes/toxicidade , Hidróxido de Sódio/toxicidade , Antígenos de Diferenciação/metabolismo , Ácido Ascórbico/toxicidade , Córnea/patologia , Opacidade da Córnea , Humanos , Técnicas In Vitro , Lubrificantes Oftálmicos/toxicidade , Soluções Oftálmicas , Peroxidase/metabolismo , Nitrato de Prata/toxicidadeRESUMO
AIMS: To investigate whether allogeneic limbal mesenchymal stem cell (LMSC) therapy affects corneal healing after a severe chemical burn and whether the route of administration of LMSCs differs in its therapeutic effect in this respect. METHODS: A total of 60 Sprague-Dawley rats with clinically proven alkali injury were divided into four equal groups (n = 15) as follows: group 1: 2 × 10(5) cells/drop LMSCs, topically applied 6 times a day for 2 days; group 2: 2.4 × 10(6) cells in 0.5 ml LMSCs, subconjunctivally applied; group 3: 2.4 × 10(6) cells in 1 ml LMSCs, intraperitoneally applied, and group 4: no LMSC treatment. The groups were compared according to grades of corneal opacity (CO), corneal neovascularization (CNV) and corneal fluorescein staining (CFS). The migration of LMSCs into the cornea and the inflammatory characteristics of the groups were evaluated with BrdU (5-bromo-2'-deoxyuridine bromodeoxyuridine) immunostaining and histopathologically in a 4-week follow-up. RESULTS: There were statistically significant differences between the LMSC-treated and control groups in each week regarding mean CO scores and in the 3rd week regarding the mean CNV and CFS scores (p < 0.05). The statistical significance was due to the differences between the topical and the control group and between the subconjunctival and the control group. BrdU+ LMSCs were seen in the corneal epithelium of the all LMSC-administered rats, and fewer inflammatory changes were observed in these rats. CONCLUSION: Allogeneic LMSC treatment, especially topical and subconjunctival administration, seems to be helpful in affecting corneal healing after a severe corneal burn.
Assuntos
Queimaduras Químicas/terapia , Doenças da Córnea/terapia , Queimaduras Oculares/induzido quimicamente , Limbo da Córnea/citologia , Transplante de Células-Tronco Mesenquimais/métodos , Cicatrização/fisiologia , Animais , Bromodesoxiuridina/metabolismo , Técnicas de Cultura de Células , Modelos Animais de Doenças , Células-Tronco Mesenquimais/fisiologia , Ratos , Ratos Sprague-Dawley , Hidróxido de Sódio/toxicidade , Transplante HomólogoRESUMO
BACKGROUND: Corrosive esophageal stricture is a major cause of morbidity among Nigerians. In most cases, this follows accidental or parasuicidal ingestion of caustic sodium hydroxide solution (NaOH) often used in the local production of soaps. Various treatment modalities have been advocated for the treatment of esophageal stricture. In this study, we review the results of our adopted technique in the past 10 years for pedicled colonic interposition. METHODS: This is a retrospective study of 21 patients who underwent substernal isoperistaltic pedicled colonic interposition graft for management of corrosive esophageal stricture. The right colon was pulled up into the neck in all the patients without resecting the strictured esophagus. RESULTS: Long segment strictures and multiple strictures were the main indications for the procedure. The mean duration of the procedure was 339.6 ± 71.1 min. The average intraoperative blood loss was 673.1 ± 398.1 mL. There were two (9.5 %) hospital mortalities. Graft infarction (9.5 %), cervical fistulae (19.0 %), and reflux neo-esophagitis (14.3 %) were the main non-fatal complications. In the mid-term, dysphagia was completely relieved in a little over 84 % (16/19) of patients, while one patient (4.8 %) still experienced reflux neo-esophagitis requiring treatment. There was no case of gross regurgitation or nocturnal aspiration in the mid-term. CONCLUSIONS: Although the use of pedicled colonic interposition graft offers a good mid-term functional outcome with relief of dysphagia, early postoperative morbidity is high. Graft infarction is the single most important factor for poor functional outcome and every effort must be made to prevent its occurrence.