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Maackia amurensis lectins serve as research and botanical agents that bind to sialic residues on proteins. For example, M. amurensis seed lectin (MASL) targets the sialic acid modified podoplanin (PDPN) receptor to suppress arthritic chondrocyte inflammation, and inhibit tumor cell growth and motility. However, M. amurensis lectin nomenclature and composition are not clearly defined. Here, we sought to definitively characterize MASL and its effects on tumor cell behavior. We utilized SDS-PAGE and LC-MS/MS to find that M. amurensis lectins can be divided into two groups. MASL is a member of one group which is composed of subunits that form dimers, evidently mediated by a cysteine residue in the carboxy region of the protein. In contrast to MASL, members of the other group do not dimerize under nonreducing conditions. These data also indicate that MASL is composed of 4 isoforms with an identical amino acid sequence, but unique glycosylation sites. We also produced a novel recombinant soluble human PDPN receptor (shPDPN) with 17 threonine residues glycosylated with sialic acid moieties with potential to act as a ligand trap that inhibits OSCC cell growth and motility. In addition, we report here that MASL targets PDPN with very strong binding kinetics in the nanomolar range. Moreover, we confirm that MASL can inhibit the growth and motility of human oral squamous cell carcinoma (OSCC) cells that express the PDPN receptor. Taken together, these data characterize M. amurensis lectins into two major groups based on their intrinsic properties, clarify the composition of MASL and its subunit isoform sequence and glycosylation sites, define sialic acid modifications on the PDPN receptor and its ability to act as a ligand trap, quantitate MASL binding to PDPN with KD in the nanomolar range, and verify the ability of MASL to serve as a potential anticancer agent.
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Antineoplásicos , Carcinoma de Células Escamosas , Neoplasias de Cabeça e Pescoço , Neoplasias Bucais , Humanos , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas de Cabeça e Pescoço , Ácido N-Acetilneuramínico/metabolismo , Maackia/química , Maackia/metabolismo , Neoplasias Bucais/patologia , Cromatografia Líquida , Ligantes , Espectrometria de Massas em Tandem , Lectinas/farmacologia , Antineoplásicos/farmacologia , Análise de Sequência , Movimento CelularRESUMO
PURPOSE: This study aims to elucidate the tear proteome and understand the underlying molecular mechanisms involved in the ocular complications following Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN). METHODS: Mass spectrometry (MS) was performed to quantify the tear fluid proteins from chronic SJS/TEN patients (n = 22 eyes) and age- and gender-matched controls (n = 22 eyes). The candidate proteins were validated using ELISA (n = 80 eyes) in tear samples and immunohistochemistry (IHC; n = 12) in eyelid margin specimens. These proteins were compared for significant differences based on age, gender, disease duration, and ocular severity. RESULTS: A total of 1692 tear fluid proteins were identified, of which 470 were significantly differentially regulated in chronic SJS/TEN. The top 10 significantly upregulated proteins were neutrophil secretions including neutrophil elastase (p < .0001), defensin (p < .0001), and matrix metalloproteinase 8 (p < .0001). The presence of neutrophils was confirmed by the upregulation of IL-8 (p < .001) in tears, a key cytokine known for recruiting neutrophils. Additionally, positive expression of myeloperoxidase was observed in the keratinized eyelid margins of SJS/TEN to validate the presence of neutrophils. Among 41 unique proteins identified by MS, IL-36γ (p < .01) was expressed in three SJS/TEN patients and was confirmed in SJS/TEN tears and eyelid margins by ELISA and IHC, respectively. IL-36γ was specifically expressed in the superficial layers of eyelid margin keratinized conjunctiva. The majority of the significantly downregulated proteins were lacrimal gland secretions such as lacritin (p < .0001) and opiorphin (p < .002). Neutrophil elastase (p < .02) was significantly elevated in patients with severe eyelid margin keratinization. CONCLUSION: Our observations indicate a clear correlation between eyelid margin keratinization and the expression of IL-36γ, potentially mediated by neutrophils recruited via IL-8. Future experimental studies are needed to test the role of therapies targeting IL-8 and/or IL-36γ in reducing eyelid margin keratinization and its associated ocular complications in SJS/TEN.
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Interleucina-1 , Neutrófilos , Síndrome de Stevens-Johnson , Lágrimas , Humanos , Feminino , Masculino , Neutrófilos/metabolismo , Neutrófilos/imunologia , Interleucina-1/metabolismo , Síndrome de Stevens-Johnson/metabolismo , Síndrome de Stevens-Johnson/patologia , Adulto , Lágrimas/metabolismo , Pessoa de Meia-Idade , Doença Crônica , Inflamação/metabolismo , Idoso , Adulto JovemRESUMO
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.
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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
Keratoconus (KC) is a degenerative disorder resulting from the degradation of the stromal collagen fibril network in the cornea, leading to its thinning and conical deformation. Various studies have established animal models of KC by using the collagenase type II enzyme to gain a better understanding of the pathogenesis, however, long-term monitoring or follow-up of the models have not been reported so far. This study evaluates the long-term stability of collagenase type II-induced KC in a rabbit model. Six New Zealand rabbits were divided into 4 study groups with 3 eyes per group. The groups were control (group 1), 0.5% proparacaine + 5 min collagenase treatment on day 0 and day 30 (group 2), 0.5% proparacaine + 10 min collagenase treatment on day 0 (group 3) and, mechanical debridement + 2 min collagenase treatment on day 0 (group 4). Inflammation was observed in group 4 till week 10. Significant decrease in the central corneal thickness was observed in group 3 by week 4 (p < 0.001) however, the thickness was regained in the subsequent follow-ups in all the groups. Keratography results showed no changes in Km values but an increased astigmatic power in all groups. Scanning electron microscopy images revealed thinner collagen fibrils arranged in a mesh-like pattern above the uniform layer of the collagen lamellae in the central part of the treated corneas. Similarly, histological staining revealed loosely packed stromal fibrils in the anterior portion of the cornea which corroborates with the immunofluorescent staining results. This study revealed the remodeling of the corneal structure by eight weeks of collagenase treatment. Consequently, the possibility of creating a rabbit keratoconus model induced by collagenase may warrant further consideration.
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Ceratocone , Propoxicaína , Coelhos , Animais , Ceratocone/induzido quimicamente , Ceratocone/tratamento farmacológico , Ceratocone/metabolismo , Córnea/metabolismo , Colágeno/metabolismo , Colagenases , Progressão da DoençaRESUMO
Dry eye disease (DED) is an emerging health problem affecting millions of individuals every year. The current treatments for DED include lubricating eye drops and anti-inflammatory agents. These agents have to be used frequently and contain preservatives, which can damage the ocular surface. A substantially long-acting treatment with better bioavailability on the ocular surface might reduce the frequency of drug use and its side effects. This review summarizes the current state of different biomaterials-nanosystems, hydrogels, and contact lenses used as drug delivery systems in DED. The explored drugs in biomaterial formulation are cyclosporin, ocular lubricants, and topical steroids. Most of the data is from animal models where increased drug delivery and desired therapeutic effects could be obtained; however, trials involving human participants are yet to happen. There is no published study comparing the different types of biomaterials for DED use. Long-term studies evaluating their ocular toxicity and biocompatibility would enhance their transition to human use. Overall they look promising for DED treatment, but they are still in the stage of technological advancement and clinical studies.
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Lentes de Contato , Síndromes do Olho Seco , Animais , Humanos , Síndromes do Olho Seco/tratamento farmacológico , Anti-Inflamatórios/uso terapêutico , Doença Iatrogênica , Disponibilidade Biológica , LágrimasRESUMO
A simple and reproducible method is necessary to generate reliable animal models of limbal stem cell deficiency (LSCD) for assessing the safety and efficacy of new therapeutic modalities. This study aimed to develop and validate a rabbit model of LSCD through mechanical injury. The corneal and limbal epithelium of New Zealand White rabbits (n = 18) were mechanically debrided using an ophthalmic burr (Algerbrush II) with a 1.0-mm rotating head after 360° conjunctival peritomy. The debrided eyes were serially evaluated for changes in corneal opacity, neo-vascularization, epithelial defect and corneal thickness using clinical photography, slit lamp imaging, fluorescein staining, and anterior segment optical coherence tomography scanning (AS-OCT). Following this, an assessment of histopathology and phenotypic marker expression of the excised corneas was conducted. The experimental eyes were grouped as mild (n = 4), moderate (n = 10), and severe (n = 4) based on the grade of LSCD. The moderate group exhibited abnormal epithelium, cellular infiltration in the stroma, and vascularization in the central, peripheral, and limbal regions of the cornea. The severe group demonstrated central epithelial edema, peripheral epithelial thinning with sparse goblet cell population, extensive cellular infiltration in the stroma, and dense vascularization in the limbal region of the cornea. A significant decrease in the expression of K12 and p63 (p < 0.0001) was observed, indicating the loss of corneal epithelium and limbal epithelial stem cells in the LSCD cornea. This study demonstrates that the Alger brush-induced mechanical debridement model provides a reliable model of LSCD with comprehensive clinic-pathological features and that is well suited for evaluating novel therapeutic and regenerative approaches.
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Doenças da Córnea , Epitélio Corneano , Limbo da Córnea , Coelhos , Animais , Limbo da Córnea/metabolismo , Desbridamento , Células-Tronco do Limbo , Córnea/metabolismo , Epitélio Corneano/metabolismo , Doenças da Córnea/patologiaRESUMO
BACKGROUND: Keratoconjunctivitis sicca or dry eye disease (DED) is a multifactorial disorder underpinned by a complex inflammatory cycle. Introduction of topical cyclosporine has been a significant advance in the management of DED. In recent years advancements in formulation technology have led to development of micellar nano-particulate (MNP) cyclosporine formulations that promise better penetration into ocular target tissues and potential for reduced ocular surface irritation. METHODS: We compared two dosing regimes of a proprietary MNP cyclosporine emulsion with the widely marketed topical cyclosporine formulation Restasis™ in a multicenter parallel-group randomised trial in patients with DED. Patients were randomised to one of 3 treatment groups with 90 patients eligible for the per protocol analysis: 30 in the higher dose test arm A; 32 in the lower dose test arm B; and 28 in the Restasis™ control arm C. All scored efficacy endpoints were tested for significance by comparing the mean change in scores from baseline in the test groups with that in the control group at 12 weeks, using the Student's t test. Wilcoxon's rank sum test was used to test individual symptom scores and clinician's global evaluation of treatment grades. RESULTS: Corneal fluorescein staining score, the primary efficacy endpoint, decreased by 6.8 ± 4.0, 5.7 ± 3.9, and 4.6 ± 3.6 points in the 3 groups respectively, indicating superior efficacy in test arm A in comparison to control arm C (p = 0.0026). Schirmer's tear test, conjunctival lissamine staining score, ocular surface disease index, and individual dry eye symptom scores also favoured higher dose MNP cyclosporine over Restasis™. The study failed to differentiate the treatment arms in terms of clinician's global evaluation of treatment, use of tear substitutes, best corrected visual acuity or safety and toleration. CONCLUSION: The results indicate that the dose of 1 drop of a 0.05% w/v ophthalmic emulsion of MNP cyclosporine administered topically twice daily yields better outcomes at 12 weeks than the lower dose tested in the study, and is more efficacious than an equivalent dose of Restasis™, the active control used in the study. TRIAL REGISTRATION: This trial was registered in the Clinical Trials Registry of India on 29/03/2019, and was assigned registration number CTRI/2019/03/018319.
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Síndromes do Olho Seco , Ceratoconjuntivite Seca , Humanos , Ceratoconjuntivite Seca/tratamento farmacológico , Ceratoconjuntivite Seca/induzido quimicamente , Micelas , Emulsões/uso terapêutico , Soluções Oftálmicas , Síndromes do Olho Seco/tratamento farmacológico , Ciclosporina/uso terapêutico , Lágrimas , Método Duplo-Cego , Resultado do TratamentoRESUMO
PURPOSE: The purpose of this study was to assess the long-term outcomes of severe cicatricial entropion repair with mucous membrane grafting in patients with chronic cicatrizing conjunctivitis and report histopathological changes in the eyelid margin area. METHODS: Prospective interventional study included 19 patients with severe cicatricial entropion with trichiasis (N = 20 eyelids; 19 upper and 1 lower eyelid) who underwent anterior lamellar recession (with back cuts) and mucous membrane grafting cover for bare anterior tarsus, lid margin, and 2 mm of marginal tarsus, and had a minimum 6 months of follow-up. The anterior lamella and metaplastic eyelid margins were sent for routine Haematoxylin and Eosin and special staining with Masson trichrome stain. RESULTS: The etiologies were chronic Stevens-Johnson syndrome (N = 6), chemical injury (N = 11), and drug-induced pseudopemphigoid (N = 2). Five eyes had undergone entropion correction in the past, and 9 had electroepilation for trichiasis. Entropion was well corrected (without residual trichiasis) in 85% of eyelids with primary surgery. The etiology-wise success rates were 100% for Stevens-Johnson syndrome, 72.7% for chemical injury, and 100% for drug-induced pseudopemphigoid. Three eyelids with failure belonged to chemical injury, and trichiasis in these eyes could be managed with subsequent interventions except in 1 case. All eyelids had no entropion at a mean follow-up of 10.8 months (range, 6-18). Histopathological evaluation of anterior lamella (N = 10) and eyelid margins revealed significant fibrosis in subepithelial, perimysium (muscle of Riolan), and perifollicular areas. CONCLUSION: Anterior lamellar recession combined with mucous membrane grafting achieves good cicatricial entropion correction except in eyes with chemical injury. The eyelid margins in these eyes have persistent inflammation, and fibrosis involving lash follicles.
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Conjuntivite , Entrópio , Síndrome de Stevens-Johnson , Triquíase , Humanos , Entrópio/etiologia , Entrópio/cirurgia , Síndrome de Stevens-Johnson/complicações , Síndrome de Stevens-Johnson/diagnóstico , Síndrome de Stevens-Johnson/cirurgia , Estudos Prospectivos , Cicatriz/complicações , Cicatriz/diagnóstico , Resultado do TratamentoRESUMO
Histone protein modifications control the inflammatory state of many immune cells. However, how dynamic alteration in histone methylation causes endothelial inflammation and apoptosis is not clearly understood. To examine this, we explored two contrasting histone methylations; an activating histone H3 lysine 4 trimethylation (H3K4me3) and a repressive histone H3 lysine 27 trimethylation (H3K27me3) in endothelial cells (EC) undergoing inflammation. Through computer-aided reconstruction and 3D printing of the human coronary artery, we developed a unique model where EC were exposed to a pattern of oscillatory/disturbed flow as similar to in vivo conditions. Upon induction of endothelial inflammation, we detected a significant rise in H3K4me3 caused by an increase in the expression of SET1/COMPASS family of H3K4 methyltransferases, including MLL1, MLL2, and SET1B. In contrast, EC undergoing inflammation exhibited truncated H3K27me3 level engendered by EZH2 cytosolic translocation through threonine 367 phosphorylation and an increase in the expression of histone demethylating enzyme JMJD3 and UTX. Additionally, many SET1/COMPASS family of proteins, including MLL1 (C), MLL2, and WDR5, were associated with either UTX or JMJD3 or both and such association was elevated in EC upon exposure to inflammatory stimuli. Dynamic enrichment of H3K4me3 and loss of H3K27me3 at Notch-associated gene promoters caused ADAM17 and Jagged-1 derepression and abrupt Notch activation. Conversely, either reducing H3K4me3 or increasing H3K27me3 in EC undergoing inflammation attenuated Notch activation, endothelial inflammation, and apoptosis. Together, these findings indicate that dynamic chromatin modifications may cause an inflammatory and apoptotic switch of EC and that epigenetic reprogramming can potentially improve outcomes in endothelial inflammation-associated cardiovascular diseases.
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Histonas , Lisina , Proteína ADAM17/metabolismo , Células Endoteliais/metabolismo , Histonas/genética , Histonas/metabolismo , Humanos , Inflamação/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteína Jagged-1/genética , Proteína Jagged-1/metabolismo , Lisina/metabolismoRESUMO
Conjunctival swabs (CS) are the major source of sampling for ocular microbiome studies, however collecting CS from the diseased eyes is difficult and painful. In this study, as an alternative to CS, a less invasive approach of tear collection was used to establish the bacterial microbiome in healthy eyes. Tear bacterial microbiome was generated from the DNA of tears (n = 24; male = 16 and female = 8) of healthy volunteers aged from 20 to 52 years. Sequencing of V3-V4 region of 16S rRNA gene was performed on the Illumina platform. Reads were processed in QIIME to assign the taxa. Statistical analysis of the tear microbiome was done in R to assess the alphadiversity and betadiversity indices. Tear microbiome was generated in all the 24 tear samples. Eight out of the top 10 predominant bacterial genera remained same in both tear and CS microbiomes, which include genera such as Corynebacterium, Staphylococcus, Streptococcus, Mycobacterium, Escherichia-Shigella, Lactobacillus, Bacillus and Acinetobacter. The similarity network analysis indicates that 144 out of 145 genera of tear cohort matched with conjunctival swabs. However, tear and CS microbiomes differed in the abundance of the predominant bacterial genera. The bacterial microbiome of tears in adults appears to be stable and is comparable with that of CS microbiome.
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Microbiota , Bactérias/genética , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Microbiota/genética , RNA Ribossômico 16S/genética , Lágrimas , Adulto JovemRESUMO
PURPOSE: To study the cytokeratin profile and keratinization-related gene expression in keratinized lid margins of chronic Stevens-Johnson syndrome (SJS) patients. METHODS: Posterior eyelid margins from 24 chronic SJS patients undergoing mucous membrane grafting and six healthy margins (orbital exenteration, fresh body donors) were studied using immunofluorescence staining (CK10, CK1, filaggrin, transglutaminase 1 (TGM1), (CK19, MUC5AC)) and quantitative PCR (keratinization-related genes-HBEGF, KGF, EGF, TGFα, TGFß, and TNFα). The staining and gene expression were studied separately in the lid margin epidermis (LME) and lid margin conjunctiva (LMC). RESULTS: The expression of CK 1/10, filaggrin, and TGM1 in the LMC was similar to the LME in SJS patients. CK19 was expressed only in the basal epithelial layer of the LMC with loss of MUC5AC expression. Increased expression of KGF (p ≤ 0.056), TNFα (p ≤ 0.02), and TGFα (p ≤ 0.01) was observed in the LME of SJS patients compared to normal LME. LMC of SJS patients showed an increased expression of HBEGF (p ≤ 0.002), EGF (p ≤ 0.0002), KGF (p ≤ 0.02), TNFα (p ≤ 0.04), TGFα (p ≤ 0.003), and TGFß (p ≤ 0.001) compared to normal LMC. Significant differences were observed in the expression of these genes between LME and LMC of SJS patients. These genes were validated using String analysis, which revealed the positive regulation of keratinization. CONCLUSION: In lid margins of SJS, there is an increased expression of keratinization-related genes compared to the normal lid margin. Keratinized LMC shares similar cytokeratin profile and keratinization gene expression as seen in cutaneous epithelium of SJS patients, indicating the possibility of the cutaneous epithelium as a source for keratinized LMC.
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Síndrome de Stevens-Johnson , Fator de Crescimento Epidérmico , Pálpebras , Expressão Gênica , Humanos , Queratinócitos , Queratinas , Fator de Crescimento Transformador alfa , Fator de Crescimento Transformador beta , Fator de Necrose Tumoral alfaRESUMO
Limbus-derived stromal/mesenchymal stem cells (LMSCs) are vital for corneal homeostasis and wound healing. However, despite multiple pre-clinical and clinical studies reporting the potency of LMSCs in avoiding inflammation and scarring during corneal wound healing, the molecular basis for the ability of LMSCs remains unknown. This study aimed to uncover the factors and pathways involved in LMSC-mediated corneal wound healing by employing RNA-Sequencing (RNA-Seq) in human LMSCs for the first time. We characterized the cultured LMSCs at the stages of initiation (LMSC-P0) and pure population (LMSC-P3) and subjected them to RNA-Seq to identify the differentially expressed genes (DEGs) in comparison to native limbus and cornea, and scleral tissues. Of the 28,000 genes detected, 7800 DEGs were subjected to pathway-specific enrichment Gene Ontology (GO) analysis. These DEGs were involved in Wnt, TGF-ß signaling pathways, and 16 other biological processes, including apoptosis, cell motility, tissue remodeling, and stem cell maintenance, etc. Two hundred fifty-four genes were related to wound healing pathways. COL5A1 (11.81 ± 0.48) and TIMP1 (20.44 ± 0.94) genes were exclusively up-regulated in LMSC-P3. Our findings provide new insights involved in LMSC-mediated corneal wound healing.
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Lesões da Córnea , Células-Tronco Mesenquimais , Córnea/metabolismo , Lesões da Córnea/metabolismo , Humanos , Células-Tronco Mesenquimais/metabolismo , Transcriptoma , Cicatrização/genéticaRESUMO
Dry eye disease (DED) is emerging as an eye health pandemic, affecting millions worldwide. The development of novel drugs, drug delivery systems, and targeted therapies for addressing the inflammation in DED necessitates progress in experimental models of DED. Animal models of DED have been created for simulating the two clinically described forms of DED: lacrimal insufficiency and the evaporative DED models. Although most DED models have relied upon rodents, the larger eye size and longer life span of rabbits and the closer resemblance to human lacrimal glands, render rabbits a promising near-ideal model for studying DED. Since the first rabbit DED model was described, numerous modifications including the use of topical epitheliotoxic drugs, neural abolition, activated lymphocytes injection, and surgical dacryoadenectomy have been introduced. The stability of these models, whether short-term or long-term, accordingly guides their experimental or therapeutic utility. A rabbit autoimmune dacryoadenitis model has successfully simulated DED signs and features of lacrimal gland inflammation, as observed in Sjogren's syndrome, that improved with mesenchymal stem cell therapy. This review summarizes the comparative microscopic anatomy of rabbit and human lacrimal glands, various existing rabbit DED models and their respective suitability for understanding pathogenetic mechanism of DED or for experimental drug testing. Also, the insights gained from animal models in dry eye management is described along with the future perspectives. There is still a pressing need of developing rabbit models for studying the pathogenesis of complex ocular surface changes in evaporative and aqueous deficiency DED other than autoimmune dacryoadenitis.
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Síndromes do Olho Seco/terapia , Aparelho Lacrimal/diagnóstico por imagem , Lágrimas/metabolismo , Pesquisa Translacional Biomédica/métodos , Animais , Modelos Animais de Doenças , Síndromes do Olho Seco/metabolismo , Humanos , Aparelho Lacrimal/metabolismo , CoelhosRESUMO
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.
Assuntos
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
OBJECTIVE: To describe the clinical profile of pterygium in patients presenting to a multi-tier ophthalmology hospital network in India. METHODS: This cross-sectional hospital-based study included 1,610,843 new patients presenting between 2010 and 2019. Patients with a clinical diagnosis of pterygium in at least one eye were included as cases. The data were collected using an electronic medical record system. Multiple logistic regression analysis with odds ratios (OR) was performed to identify the associated risk factors. RESULTS: Overall, 168,807 (10.5%) new patients were diagnosed with pterygium, of which 43,692 (26%) patients complained about the lesion. The prevalence rates were 0.7% in children and 12.6% in adults. Majority of patients were female (54.5%) and had unilateral (57%) affliction. Among the 241,631 affected eyes, the pterygia were primary in 99.6%, nasally located in 94%, and were grade I-II in 84.8%. Four in 5 eyes did not have any cylindrical refractive error, and 44% had coexistent cataract. Pterygium surgery was indicated in 10.3% eyes. Female sex (OR 1.37), increasing age (OR 19.5), rural residence (OR 1.21), agriculture work (OR 2.19), manual labor (OR 2.05), low socioeconomic status (OR 2.14) and geographical location closer to the equator (OR 3.4) were identified as the risk factors for developing pterygium. CONCLUSION: About one-tenth of individuals seeking eye care in India have pterygium in at least one eye. It rarely impacts vision, is commonly unilateral and nasal and usually does not require surgery. It is associated with increasing age, females, outdoor work, low income and geographical location closer to the equator.
Assuntos
Gerenciamento Clínico , Registros Eletrônicos de Saúde/estatística & dados numéricos , Hospitais/estatística & dados numéricos , Pterígio/epidemiologia , População Rural/estatística & dados numéricos , Adulto , Distribuição por Idade , Estudos Transversais , Ciência de Dados , Feminino , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Pterígio/terapia , Estudos Retrospectivos , Fatores de Risco , Distribuição por SexoRESUMO
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
PURPOSE: To assess the outcome of aspiration and sclerotherapy with sodium tetradecyl sulfate in the management of conjunctival inclusion cysts. METHODS: Retrospective interventional case series of 6 patients with clinical diagnosis of conjunctival inclusion cysts treated with cyst aspiration and foam sclerotherapy with 3% sodium tetradecyl sulfate. The volume of the sclerosant was 20% of the aspirated cyst volume. RESULTS: Four patients had an inclusion cyst in anophthalmic sockets and 2 patients in sighted eyes. Average time lag between primary surgery and cyst formation was 14.6 months (range 2-30 months). Average amount of fluid aspirated from cyst was 3.07 ml (range 1-9 ml). Average volume of sclerosant injected was (20% of the aspirated volume) 0.55 ml (range 0.2-1.1 ml). All 6 patients showed complete resolution of cyst at a mean follow-up period of 15.6 months (range 9-24 months). All but one showed complete resolution of cyst with single injection sclerosant. Only 1 patient required a second sclerosant injection. There was no ocular surface or implant-related complications in this cohort. CONCLUSIONS: Cyst aspiration and sodium tetradecyl sulfate foam sclerotherapy is a minimally invasive procedure for the management of conjunctival inclusion cysts in anophthalmic sockets and sighted eyes. The injection of sodium tetradecyl sulfate in a dose of 20% of the aspirate is effective in the management of conjunctival inclusion cysts over a follow-up period of 13 months. The procedure is safe, with insignificant inflammation and without ocular surface or implant complications.
Assuntos
Túnica Conjuntiva/patologia , Doenças da Túnica Conjuntiva/terapia , Cistos/terapia , Paracentese/métodos , Tetradecilsulfato de Sódio/uso terapêutico , Adolescente , Adulto , Biópsia , Criança , Pré-Escolar , Túnica Conjuntiva/efeitos dos fármacos , Doenças da Túnica Conjuntiva/diagnóstico , Cistos/diagnóstico , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Soluções Esclerosantes/uso terapêutico , Escleroterapia/métodos , Resultado do Tratamento , Adulto JovemAssuntos
Conjuntivite , Entrópio , Humanos , Entrópio/etiologia , Entrópio/cirurgia , Túnica Conjuntiva/patologia , Cicatriz/etiologiaRESUMO
AIM: To determine whether evisceration with optic nerve disinsertion and nonporous implant placement increases the risk of implant migration. METHODS: This was a single-center, retrospective consecutive comparative interventional case series including patients undergoing evisceration with nonporous implant between January and December 2014. Patients were grouped into 2 groups: group I where the optic nerve was not disinserted (n = 37) and group II with optic nerve disinsertion (n = 50). Implant migration was assessed clinically and on patient photographs. Migration was subclassified as decentration that did not affect the prosthetic outcome and displacement that affected the prosthetic outcome. The secondary outcome measures were the mean implant diameter, volume of the custom ocular prosthesis, and implant-related complications like exposure and extrusion between the 2 groups. RESULTS: At a mean follow up of 12.5 months, none of the sockets in group I and 3 (6%) sockets in group II (p = 0.35) had evidence of implant decentration. There were no cases of implant displacement in both groups. The mean implant diameter in group I was 16.97 mm ± 0.65 mm and in group II 19.2 mm ± 0.83 mm (p = 0.0001). Implant extrusion was not different between the 2 groups. The mean custom ocular prosthesis volume in group I was 3.86 ml ± 0.52 ml and in group II 2.50 ml ± 0.68 ml (p < 0.0001). CONCLUSIONS: The rate of nonporous implant migration due to optic nerve disinsertion is not statistically or clinically significant in evisceration with optic nerve disinsertion, allowing placement of a larger implant and fabrication of a custom ocular prosthesis with an ideal weight.