RESUMO
The trabecular meshwork (TM) is an ocular tissue that maintains intraocular pressure (IOP) within a physiologic range. Glaucoma patients have reduced TM cellularity and, frequently, elevated IOP. To establish a stem cell-based approach to restoring TM function and normalizing IOP, human adipose-derived stem cells (ADSCs) were induced to differentiate to TM cells in vitro. These ADSC-TM cells displayed a TM cell-like genotypic profile, became phagocytic, and responded to dexamethasone stimulation, characteristic of TM cells. After transplantation into naive mouse eyes, ADSCs and ADSC-TM cells integrated into the TM tissue, expressed TM cell markers, and maintained normal IOP, outflow facility, and extracellular matrix. Cell migration and affinity results indicated that the chemokine pair CXCR4/SDF1 may play an important role in ADSC-TM cell homing. Our study demonstrates the possibility of applying autologous or allogeneic ADSCs and ADSC-TM cells as a potential treatment to restore TM structure and function in glaucoma.
Assuntos
Células-Tronco Adultas/citologia , Células-Tronco Adultas/transplante , Glaucoma/terapia , Malha Trabecular/citologia , Tecido Adiposo/citologia , Células-Tronco Adultas/efeitos dos fármacos , Animais , Câmara Anterior/citologia , Câmara Anterior/imunologia , Apoptose , Humor Aquoso/fisiologia , Diferenciação Celular , Movimento Celular , Células Cultivadas , Quimiotaxia , Dexametasona/farmacologia , Modelos Animais de Doenças , Glaucoma/patologia , Glaucoma/fisiopatologia , Xenoenxertos , Humanos , Técnicas In Vitro , Pressão Intraocular/fisiologia , Camundongos , Fagocitose , Medicina Regenerativa , Malha Trabecular/fisiologiaRESUMO
PURPOSE: To compare the outcomes and complications of topical difluprednate 0.05% and loteprednol gel 0.5% after routine cataract surgery. METHODS: Subjects received either difluprednate emulsion 0.05% (n=30 eyes) or loteprednol gel 0.5% (n=30 eyes) after routine cataract surgery. Topical steroid drops were initiated 3 days before cataract surgery and continued for 2 weeks postoperatively. Anterior chamber (AC) cell grade, corneal edema, corneal pachymetry, visual acuity, ocular surface quality (Oxford scale), and intraocular pressure (IOP) were evaluated at 1 day, 1 week, and 1 month postoperatively. RESULTS: Patients treated with difluprednate or loteprednol had statistically similar resolution of their AC cell grade and corneal edema at 1 day, 1 week, and 1 month postoperatively (P>0.05 at each study visit). Difluprednate-treated and loteprednol-treated eyes achieved a mean best-corrected visual acuity of at least 20/25 by 1 week postoperatively (0.055 and 0.061 logarithm of the minimum angle of resolution, respectively; P=0.82). The nasal ocular surface quality at 1 week had improved in loteprednol-treated eyes compared with difluprednate-treated eyes (1.0 vs. 1.9 Oxford score, respectively; P<0.001), but similar at all other visits. There was no statistical difference between IOP levels between both treatment groups (P>0.05). In the difluprednate-treated group, one patient developed rebound inflammation and two patients developed cystoid macular edema at their 1-month postoperative visit. CONCLUSIONS: The anti-inflammatory effect, visual recovery, and IOP of patients using topical difluprednate or loteprednol gel after cataract surgery are equivalent. There may be an additional short-term benefit of loteprednol gel in protecting the ocular surface after cataract surgery.
Assuntos
Anti-Inflamatórios/uso terapêutico , Extração de Catarata , Oftalmopatias/tratamento farmacológico , Fluprednisolona/análogos & derivados , Glucocorticoides/uso terapêutico , Inflamação/tratamento farmacológico , Etabonato de Loteprednol/uso terapêutico , Complicações Pós-Operatórias/tratamento farmacológico , Idoso , Idoso de 80 Anos ou mais , Câmara Anterior/citologia , Extração de Catarata/efeitos adversos , Córnea/patologia , Edema da Córnea/tratamento farmacológico , Edema da Córnea/fisiopatologia , Oftalmopatias/etiologia , Oftalmopatias/fisiopatologia , Feminino , Fluprednisolona/uso terapêutico , Géis/uso terapêutico , Humanos , Inflamação/fisiopatologia , Pressão Intraocular , Masculino , Pessoa de Meia-Idade , Acuidade Visual/fisiologiaRESUMO
To investigate the accuracy of the measurements of Nidek AL-Scan by comparing with Sirius (CSO, Florence, Italy), a corneal tomography which also employs the Scheimpflug principle, and a commonly used device, ultrasound biometry (UB) (Aviso A/B, Quantel Medical, MT, USA). Right eyes of 85 healthy volunteers (58 women 27 men) with a mean age of 39.24 ± 14.37 years (range 15-68) were enrolled into this comparative prospective study. Average K 2.4, average K 3.3, CCT (central corneal thickness), WTW (white to white distance), ACD (anterior chamber depth) and AL (axial length) were obtained from the AL-Scan and compared with average SimK, CCT, WTW (horizontal anterior chamber diameter) and ACD obtained from Sirius and also compared with ACD and AL obtained from UB. The statistically significant difference was found between all of the measurements (p < 0.001) except the average keratometry values (K2.4, K3.3, SimK) (p = 0.083). There was a perfect correlation between keratometry, CCT and AL measurements of the devices (ICC = 0.977, 0.954, 0.923, respectively) and there was a strong correlation between the WTW measurements of AL-Scan and Sirius (ICC = 0.865). While ACD parameter of AL-Scan and UB showed a perfect correlation (ICC = 0.977), there was a moderate correlation between AL-Scan and Sirius and also between UB and Sirius (ICC = 0.608 and 0.664, respectively). There was a high correlation between the all measurements, besides ACD, of AL-Scan and Sirius and they can be used interchangeably for average keratometry and WTW confidently. However, ACD and CCT have a broader 95 % LoA (-0.039 to 0.744 and -24.985 to 3.691, respectively). In addition, AL-Scan and UB were in good agreement regarding ACD, while differences in AL measurements of UB and AL-Scan were clinically important (95 % LoA = -0.091 to 0.703). Furthermore, UB and Sirius have a moderate agreement regarding ACD (95 % LoA = -0.047 to 0.680).
Assuntos
Biometria/instrumentação , Córnea/citologia , Topografia da Córnea/instrumentação , Adolescente , Adulto , Idoso , Câmara Anterior/citologia , Desenho de Equipamento , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Curva ROC , Reprodutibilidade dos Testes , Adulto JovemRESUMO
AIMS/HYPOTHESIS: In vivo imaging of the developing pancreas is challenging due to the inaccessibility of the tissue. To circumvent this, on embryonic day 10.5 (E10.5) we transplanted a mouse developing pancreatic bud into the anterior chamber of the eye (ACE) to determine whether the eye is a useful transplant site to support pancreas development. METHODS: We transplanted an E10.5 dorsal pancreatic bud into the ACE of a syngeneic recipient mouse. Using a mouse insulin promoter-green fluorescent protein (MIP-GFP) mouse as the tissue donor, we non-invasively imaged the pancreatic bud as it develops at single beta cell resolution across time. RESULTS: The transplanted pancreatic bud rapidly engrafts and vascularises when transplanted into the ACE. The pancreatic progenitor cells differentiate into exocrine and endocrine cells, including cells expressing insulin, glucagon and somatostatin. The morphology of the transplanted pancreatic bud resembles that of the native developing pancreas. Beta cells within the transplanted pancreatic bud respond to glucose in a manner similar to that of native fetal beta cells and superior to that of in vitro developed beta cells. Unlike in vitro grown pancreatic explants, pancreatic tissue developing in the ACE is vascularised, providing the developing pancreatic tissue with a milieu resembling the native situation. CONCLUSIONS/INTERPRETATION: Altogether, we show that the ACE is able to support growth, differentiation and function of a developing pancreatic bud across time in vivo.
Assuntos
Câmara Anterior/citologia , Ilhotas Pancreáticas/embriologia , Pâncreas/embriologia , Animais , Câmara Anterior/metabolismo , Diferenciação Celular/fisiologia , Feminino , Insulina/metabolismo , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/metabolismo , Masculino , Camundongos , GravidezRESUMO
PURPOSE: To describe live mouse, anterior chamber constant-pressure perfusion by an approach using feedback-controlled coupling of pressure and flow to maintain a preset pressure. METHODS: We established a microperfusion system that maintains a constant preset pressure in the anterior chamber of live mice by automatically regulating the microsyringe pump flow rate with a computer-controlled voltage feedback loop. Perfusion was by single-needle cannulation. We characterized the following in C57BL/6 mice aged 3-4 months in vivo: (i) pressure stability, (ii) pressure and flow rate reproducibility, (iii) total outflow facility, and (iv) anterior segment histology after perfusion. RESULTS: Twenty live mice underwent perfusion. Constant pressure was quickly attained and stably maintained. The coefficient of pressure variation over time during perfusion at a preset pressure was <0.001. The average coefficient of variation for repeat pressure and flow rate measurements was 0.0005 and 0.127, respectively. The relationship between flow rate and pressure was linear for perfusions between 15 and 35 mmHg. The total outflow facility was 0.0066 µl/min/mmHg. Perfusion system resistance (0.5 mmHg/min/µl) was negligible relative to the ocular outflow resistance (147 mmHg/min/µl) at physiologically relevant perfusion pressures of 15-35 mmHg. No histological disruption of the drainage tissue was seen following perfusion. CONCLUSIONS: Predetermined pressure was stably maintained during constant-pressure perfusion of live mouse eyes by a method using feedback-controlled coupling of pressure and flow along with single-needle anterior chamber cannulation. Perfusion measurements were reproducible. This approach is potentially useful for exploring aqueous drainage tissue biology, physiology, and pharmacology in live mice.
Assuntos
Câmara Anterior/fisiologia , Retroalimentação Fisiológica , Pressão Intraocular/fisiologia , Animais , Câmara Anterior/citologia , Drenagem , Camundongos , Camundongos Endogâmicos C57BL , Agulhas , Perfusão , Reprodutibilidade dos Testes , ReologiaRESUMO
Intravital imaging emerged as an indispensible tool in biological research, and a variety of imaging techniques have been developed to noninvasively monitor tissues in vivo. However, most of the current techniques lack the resolution to study events at the single-cell level. Although intravital multiphoton microscopy has addressed this limitation, the need for repeated noninvasive access to the same tissue in longitudinal in vivo studies remains largely unmet. We now report on a previously unexplored approach to study immune responses after transplantation of pancreatic islets into the anterior chamber of the mouse eye. This approach enabled (i) longitudinal, noninvasive imaging of transplanted tissues in vivo; (ii) in vivo cytolabeling to assess cellular phenotype and viability in situ; (iii) local intervention by topical application or intraocular injection; and (iv) real-time tracking of infiltrating immune cells in the target tissue.
Assuntos
Câmara Anterior/citologia , Ilhotas Pancreáticas/citologia , Microscopia Confocal/métodos , Linfócitos T/citologia , Amidas/farmacologia , Animais , Câmara Anterior/metabolismo , Câmara Anterior/cirurgia , Antagonistas dos Receptores CCR5 , Quimiocinas/farmacologia , Diabetes Mellitus Experimental/terapia , Ensaio de Imunoadsorção Enzimática , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Interferon gama/metabolismo , Interleucina-2/metabolismo , Ilhotas Pancreáticas/metabolismo , Transplante das Ilhotas Pancreáticas/métodos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Microscopia de Vídeo/métodos , Compostos de Amônio Quaternário/farmacologia , Receptores CCR5/metabolismo , Análise de Célula Única/métodos , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , Fatores de TempoRESUMO
Systemic immunological tolerance to Ag encountered in the eye restricts the formation of potentially damaging immune responses that would otherwise be initiated at other anatomical locations. We previously demonstrated that tolerance to Ag administered via the anterior chamber (AC) of the eye required Fas ligand-mediated apoptotic death of inflammatory cells that enter the eye in response to the antigenic challenge. Moreover, the systemic tolerance induced after AC injection of Ag was mediated by CD8(+) regulatory T cells. This study examined the mechanism by which these CD8(+) regulatory T cells mediate tolerance after AC injection of Ag. AC injection of Ag did not prime CD4(+) T cells and led to increased TRAIL expression by splenic CD8(+) T cells. Unlike wild-type mice, Trail(-/-) or Dr5(-/-) mice did not develop tolerance to Ag injected into the eye, even though responding lymphocytes underwent apoptosis in the AC of the eyes of these mice. CD8(+) T cells from Trail(-/-) mice that were first injected via the AC with Ag were unable to transfer tolerance to naive recipient wild-type mice, but CD8(+) T cells from AC-injected wild-type or Dr5(-/-) mice could transfer tolerance. Importantly, the transferred wild-type (Trail(+/+)) CD8(+) T cells were also able to decrease the number of infiltrating inflammatory cells into the eye; however, Trail(-/-) CD8(+) T cells were unable to limit the inflammatory cell ingress. Together, our data suggest that "helpless" CD8(+) regulatory T cells generated after AC injection of Ag enforce systemic tolerance in a TRAIL-dependent manner to inhibit inflammation in the eye.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/transplante , Olho/imunologia , Olho/patologia , Tolerância Imunológica , Ligante Indutor de Apoptose Relacionado a TNF/biossíntese , Animais , Câmara Anterior/citologia , Câmara Anterior/imunologia , Câmara Anterior/metabolismo , Antígenos Virais/administração & dosagem , Antígenos Virais/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Células Cultivadas , Chlorocebus aethiops , Olho/virologia , Herpesvirus Humano 1/imunologia , Tolerância Imunológica/genética , Imunidade Celular/genética , Mediadores da Inflamação/administração & dosagem , Mediadores da Inflamação/imunologia , Mediadores da Inflamação/metabolismo , Injeções Intraoculares , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Baço/citologia , Baço/imunologia , Baço/transplante , Ligante Indutor de Apoptose Relacionado a TNF/deficiência , Ligante Indutor de Apoptose Relacionado a TNF/genética , Células VeroRESUMO
Optical coherence tomography (OCT) is a noninvasive method that can quickly and accurately examine the eye at the cellular level. Several studies have used OCT for analysis of anterior chamber cells. However, these studies have several limitations. This study was performed to supplement existing reports of automated analysis of anterior chamber cell images using spectral domain OCT (SD-OCT) and to compare this method with the Standardization of Uveitis Nomenclature (SUN) grading system. We analyzed 2398 anterior segment SD-OCT images from 34 patients using code written in Python. Cell density, size, and eccentricity were measured automatically. Increases in SUN grade were associated with significant cell density increases at all stages (p < 0.001). Significant differences were observed in eccentricity in uveitis, post-surgical inflammation, and vitreous hemorrhage (p < 0.001). Anterior segment SD-OCT is reliable, fast, and accurate means of anterior chamber cell analysis. This method showed a strong correlation with the SUN grade system. Also, eccentricity could be helpful as a supplementary evaluation tool.
Assuntos
Câmara Anterior/citologia , Rastreamento de Células/métodos , Processamento de Imagem Assistida por Computador/métodos , Software , Idoso , Algoritmos , Contagem de Células , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tomografia de Coerência ÓpticaRESUMO
Primary congenital glaucoma (PCG) is a severe disease characterized by developmental defects in the trabecular meshwork (TM) and Schlemm's canal (SC), comprising the conventional aqueous humor outflow pathway of the eye. Recently, heterozygous loss of function variants in TEK and ANGPT1 or compound variants in TEK/SVEP1 were identified in children with PCG. Moreover, common variants in ANGPT1and SVEP1 have been identified as risk alleles for primary open angle glaucoma (POAG) in GWAS studies. Here, we show tissue-specific deletion of Angpt1 or Svep1 from the TM causes PCG in mice with severe defects in the adjacent SC. Single-cell transcriptomic analysis of normal and glaucomatous Angpt1 deficient eyes allowed us to identify distinct TM and SC cell populations and discover additional TM-SC signaling pathways. Furthermore, confirming the importance of angiopoietin signaling in SC, delivery of a recombinant ANGPT1-mimetic promotes developmental SC expansion in healthy and Angpt1 deficient eyes, blunts intraocular pressure (IOP) elevation and RGC loss in a mouse model of PCG and lowers IOP in healthy adult mice. Our data highlight the central role of ANGPT1-TEK signaling and TM-SC crosstalk in IOP homeostasis and provide new candidates for SC-targeted glaucoma therapy.
Assuntos
Humor Aquoso/metabolismo , Comunicação Celular/fisiologia , Glaucoma de Ângulo Aberto/patologia , Glaucoma de Ângulo Aberto/terapia , Angiopoietina-1/administração & dosagem , Angiopoietina-1/genética , Angiopoietina-1/metabolismo , Animais , Câmara Anterior/irrigação sanguínea , Câmara Anterior/citologia , Câmara Anterior/metabolismo , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Moléculas de Adesão Celular/genética , Moléculas de Adesão Celular/metabolismo , Comunicação Celular/efeitos dos fármacos , Modelos Animais de Doenças , Células Endoteliais/citologia , Células Endoteliais/metabolismo , Perfilação da Expressão Gênica , Glaucoma de Ângulo Aberto/genética , Glaucoma de Ângulo Aberto/metabolismo , Pressão Intraocular/efeitos dos fármacos , Pressão Intraocular/genética , Camundongos , Camundongos Knockout , Crista Neural/citologia , Crista Neural/metabolismo , Proteínas/genética , Proteínas/metabolismo , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transdução de Sinais/efeitos dos fármacos , Análise de Célula Única , Malha Trabecular/citologia , Malha Trabecular/metabolismoRESUMO
P-selectin glycoprotein ligand-1 (PSGL-1) is constitutively expressed on leukocytes and was thought to be down-regulated with cell activation. However, this work shows the surprising finding of functional PSGL-1 up-regulation during acute inflammation. PSGL-1 function was studied in our autoperfusion assay, in which blood from a mouse carotid flows through a microchamber coated with a fixed density of P-selectin. Under the inflammatory conditions--uveitis induced by systemic lipopolysaccharide injection--we recorded significantly reduced leukocyte rolling velocity, which suggests PSGL-1 up-regulation; however, flow cytometry showed reduced PSGL-1. When bound leukocytes were released from the vasculature by PSGL-1 blockade, a large peripheral blood leukocyte (PBL) population showed elevated PSGL-1, which could account for the reduced PSGL-1 in the remaining unbound population. In the eye, systemic blockade of PSGL-1 with a monoclonal antibody or recombinant soluble PSGL-1 drastically reduced the severe manifestations of uveitis. Furthermore, PSGL-1 blockade was significantly more effective in reducing retinal leukostasis than was P-selectin blockade. Our results provide surprising evidence for functional PSGL-1 up-regulation in PBLs during acute inflammation. The temporal overlap between PSGL-1 and P-selectin up-regulation reveals an as yet unrecognized collaboration between this receptor-ligand pair, increasing efficiency of the first steps of the leukocyte recruitment cascade.
Assuntos
Endotoxinas/toxicidade , Glicoproteínas de Membrana/metabolismo , Regulação para Cima/fisiologia , Uveíte/induzido quimicamente , Animais , Câmara Anterior/citologia , Inflamação/metabolismo , Leucócitos/fisiologia , Masculino , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Técnicas Analíticas Microfluídicas , Retina/citologia , Uveíte/metabolismoRESUMO
Imaging beta cells is a key step towards understanding islet transplantation. Although different imaging platforms for the recording of beta cell biology have been developed and utilized in vivo, they are limited in terms of allowing single cell resolution and continuous longitudinal recordings. Because of the transparency of the cornea, the anterior chamber of the eye (ACE) in mice is well suited to study human and mouse pancreatic islet cell biology. Here is a description of how this approach can be used to perform continuous longitudinal recordings of grafting and revascularization of individual human islet grafts. Human islet grafts are inserted into the ACE, using NOD.(Cg)-Gt(ROSA)26Sortm4-Rag2-/-mice as recipients. This allows for the investigation of the expansion of recipient versus donor cells and the contribution of recipient cells in promoting the encapsulation and vascularization of the graft. Further, a step-by-step approach for image analysis and quantification of the islet volume or segmented vasculature and islet capsule forming recipient cells is outlined.
Assuntos
Câmara Anterior/citologia , Imageamento Tridimensional , Transplante das Ilhotas Pancreáticas , Ilhotas Pancreáticas/citologia , Animais , Humanos , Células Secretoras de Insulina/citologia , Ilhotas Pancreáticas/cirurgia , Camundongos Endogâmicos NOD , Microscopia de Fluorescência por Excitação Multifotônica , Neovascularização FisiológicaRESUMO
Background CD8+ regulatory T cells (Treg) have been considered to be involved in a model of ocular-induced tolerance, known as anterior chamber-associated immune deviation (ACAID). The mechanisms of suppression by CD8+ T cells in ACAID remain only poorly understood. TGF-beta1 is considered as an inhibitory cytokine for immunosuppression in some models. The production of TGF-beta1 by CD8+ T cells in ACAID, and whether CD8+ T cells exert suppression through TGF-beta1, is unknown. Methods The suppressive effect of CD8+ T cells in ACAID mice was determined by a local adoptive transfer (LAT) assay. The production of TGF-beta1 by CD8+ T cells was measured by enzyme-linked immunosorbent assay (ELISA). Anti-TGF-beta1 antibodies were used in the LAT assay to test if they could block the inhibitory effect of CD8+ T cells. Results CD8+ T cells from ACAID mice were shown to block the delayed-type hypersensitivity (DTH) response in an antigen-specific manner in a LAT assay. These CD8+ T cells secreted TGF-beta1, and their suppression could partially be blocked by anti-TGF-beta1 antibodies. Conclusions Our study confirms that CD8+ T cells from ACAID mice possess inhibitory properties. This population exerts part of its suppressive function via the production of TGF-beta1.
Assuntos
Câmara Anterior/imunologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Hipersensibilidade Tardia/imunologia , Tolerância Imunológica/imunologia , Fator de Crescimento Transformador beta1/metabolismo , Transferência Adotiva , Animais , Câmara Anterior/citologia , Feminino , Hipersensibilidade Tardia/metabolismo , Interleucina-10/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Baço/citologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismoRESUMO
Purpose: Corneal endothelial dysfunction leads to corneal edema, pain, and vision loss. Adequate animal models are needed to study the safety and efficacy of novel cell therapies as an alternative to corneal transplantation. Methods: Primary human corneal endothelial cells (HCECs) were isolated from cadaveric donor corneas, expanded in vitro, transduced to express green fluorescent protein (GFP), loaded with superparamagnetic nanoparticles, and injected into the anterior chamber of adult rabbits immediately after endothelial cell or Descemet's membrane stripping. The same volume of balanced salt solution plus (BSS+) was injected in control eyes. We compared different models for inducing corneal edema in rabbits, and examined the ability of transplanted HCECs to reduce corneal edema over time by measuring central corneal thickness and tracking corneal clarity. GFP-positive donor cells were tracked in vivo using optical coherence tomography (OCT) fluorescence angiography module, and the transplanted cells were confirmed by human nuclei immunostaining. Results: Magnetic HCECs integrated onto the recipient corneas with intact Descemet's membrane, and donor identity was confirmed by GFP expression and immunostaining for human nuclei marker. Donor HCECs formed a monolayer on the posterior corneal surface and expressed HCEC functional markers of tight junction formation. No GFP-positive cells were observed in the trabecular meshwork or on the iris, and intraocular pressure remained stable through the length of the study. Conclusions: Our results demonstrate magnetic cell-based therapy efficiently delivers HCECs to restore corneal transparency without detectable toxicity or adverse effect on intraocular pressure. Magnetic delivery of HCECs may enhance corneal function and should be explored further for human therapies.
Assuntos
Transplante de Células/métodos , Doenças da Córnea/cirurgia , Sistemas de Liberação de Medicamentos , Endotélio Corneano/transplante , Magnetoterapia/métodos , Nanopartículas de Magnetita/química , Animais , Câmara Anterior/citologia , Sobrevivência Celular/fisiologia , Células Cultivadas , Doenças da Córnea/patologia , Portadores de Fármacos , Endotélio Corneano/metabolismo , Endotélio Corneano/cirurgia , Proteínas de Fluorescência Verde/metabolismo , Humanos , Pressão Intraocular , Substâncias Luminescentes/metabolismo , Modelos Animais , Coelhos , Doadores de Tecidos , TransfecçãoRESUMO
PURPOSE: The iridocorneal angle in the mammalian eye including the trabecular meshwork (TM) develops from undifferentiated mesenchyme/neural crest between the iris root and cornea. The precise mechanisms underlying anterior angle development are unclear, and the contribution of cell death and phagocytic resorption by macrophages in angle development is controversial. In this study, we examined the human anterior chamber angle during various stages of development for evidence of cell death and phagocytic resorption. METHODS: Eyes from the human fetus (F) of 7, 8, 9, 10, 11, 13, 15, 18, 19, 21, 22, 23, and 27 weeks as well as eyes from 5- and 11-month-old children and donors 24, 48, and 67 years of age were obtained. Formalin-fixed and paraffin-embedded sections were examined by hematoxylin and eosin (H&E) staining. Immunohistochemistry was performed using polyclonal antibodies against CD68. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) labeling was also performed to evaluate cell death. RESULTS: By light microscopy, the development of human angle structures appeared to progress as previously described. Histological evidence of cellular death or resorption by macrophages was not observed. Furthermore, the chamber angle tissues did not stain with CD68 at any stage of development. Few CD68 positive cells were observed in the iris stroma and the anterior ciliary body between fetal weeks 10 and 18 (F10w and F18w). TUNEL labeled nuclei were not detected in the anterior chamber angle in any fetal or infant eyes. By contrast, TUNEL positive nuclei in TM cells were observed in the examined adult donor specimens. CONCLUSIONS: The results suggest that at the time points examined, neither cell death nor phagocytic resorption with macrophages appear to play a role in the development of the human anterior chamber angle.
Assuntos
Câmara Anterior/citologia , Câmara Anterior/embriologia , Macrófagos/citologia , Adulto , Idoso , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/metabolismo , Morte Celular , Núcleo Celular/metabolismo , Feto/citologia , Feto/embriologia , Fluoresceínas , Humanos , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Indóis , Macrófagos/metabolismo , Pessoa de Meia-IdadeRESUMO
PURPOSE: To describe the distribution of anterior chamber depth (ACD), anterior chamber volume (ACV), and anterior chamber angle (ACA) and establish the associated factors in the pediatric population in Shanghai, China. MATERIALS AND METHODS: In this cross-sectional study, children aged 6 to 18 years from 9 primary and middle schools in Shanghai were enrolled. The Pentacam Scheimpflug camera was used to measure anterior eye chamber parameters. The distribution of ACD, ACV, ACA, and their associations with age, sex, body mass index, cycloplegic refractive error, axial length, intraocular pressure, and other parameters were analyzed. RESULTS: A total of 1321 children were included, with a mean age of 9.65±2.95 years. The mean ACA, ACD, and ACV values were 37.95±7.96 degrees, 3.22±0.23 mm and 194.89±28.95 mm, respectively, and were higher in boys than in girls. ACV and ACD had similar growth trend curves with age, whereas ACA was stable. Overall, 5% of the tested children had ACA values ≤24.91 degrees. Greater ACV, deeper ACD, shorter pupil diameter, shorter axial length, and thinner apex corneal thickness were the independent factors associated with wider ACA (R=13.0%, P<0.001). CONCLUSIONS: As one of diagnostic indicators of angle closure, ACA was stable with age. The results of this study should improve the current understanding of the distribution of anterior chamber parameters and the main factors affecting their variation.
Assuntos
Câmara Anterior/anatomia & histologia , Câmara Anterior/diagnóstico por imagem , Povo Asiático , Adolescente , Fatores Etários , Câmara Anterior/citologia , Câmara Anterior/fisiologia , Índice de Massa Corporal , Criança , China , Estudos Transversais , Feminino , Humanos , Pressão Intraocular/fisiologia , Masculino , Tamanho do Órgão , Estudos Prospectivos , Tonometria OcularRESUMO
PURPOSE: The purpose of the study was to evaluate tissue reaction to polycaprolactone (PCL) nanofiber patches in the cornea, conjunctiva, and anterior chamber (AC) in rabbit eyes and to assess their biocompatibility for use as patch grafts. METHODS: Two 100 µ PCL patches were implanted under the conjunctiva and in the corneal stroma of one albino New Zealand rabbit, and pathologic evaluation was done after 3 weeks. In the next step, two PCL patches were implanted; one in the corneal stroma and the other in the AC of two rabbits followed by pathologic evaluation after 3 months. RESULTS: On slit-lamp examination, there was minimum inflammation in all cases. Pathologic examination showed that the contact and probably merging between the host tissue and PCL fibers were achieved with minimal tissue reaction. CONCLUSION: As a biocompatible material, PCL nanofibers seem to be a promising modality for the repair of different tissue defects including melting, thinning, and perforation. They may also be a suitable material for manufacturing keratoprostheses.
Assuntos
Câmara Anterior/efeitos dos fármacos , Túnica Conjuntiva/efeitos dos fármacos , Doenças da Córnea/cirurgia , Substância Própria/efeitos dos fármacos , Transplante de Córnea/métodos , Nanofibras/administração & dosagem , Poliésteres/administração & dosagem , Animais , Câmara Anterior/citologia , Curativos Biológicos , Células Cultivadas , Túnica Conjuntiva/citologia , Túnica Conjuntiva/cirurgia , Substância Própria/citologia , Substância Própria/cirurgia , Modelos Animais de Doenças , Oftalmologia/métodos , Complicações Pós-Operatórias/prevenção & controle , Coelhos , Microscopia com Lâmpada de FendaRESUMO
A 14-year-old, female dog was presented for a recheck following intracapsular lens removal 1 year earlier. On examination, epithelial downgrowth was identified in the anterior chamber, and an evisceration was performed. The intraocular contents were submitted for histopathologic examination, which confirmed the presence of epithelial downgrowth.
Assuntos
Doenças da Córnea/veterinária , Doenças do Cão/patologia , Epitélio Corneano/patologia , Lentes Intraoculares , Animais , Câmara Anterior/citologia , Câmara Anterior/patologia , Câmara Anterior/cirurgia , Doenças da Córnea/patologia , Doenças da Córnea/cirurgia , Doenças do Cão/cirurgia , Cães , Células Epiteliais , Feminino , Complicações Pós-Operatórias/veterináriaRESUMO
PURPOSE: To test the effect of ambient illumination scaling on the reproducibility and reliability anterior chamber metrics using the Visante time domain optical coherence tomography (TD-OCT) instrument. MATERIALS AND METHODS: The inferior irido-corneal angles of 25 normal, healthy eyes were imaged twice with the Zeiss Visante TD-OCT under five strictly controlled ambient light conditions (foot candles (fc) measured with a light meter at camera/eye interface). Each eye was imaged 10 times totaling 250 assessments. Angle opening distance (AOD500/750), trabecular iris space area (TISA500/750), and scleral spur (SS) angle were graded twice by masked, trained graders at the Doheny Imaging Reading Center using the Visante's intrinsic tools. Lighting effects on measurements, intra-/inter-grader and acquisition analyses, and Bland-Altman plots were computed using Statistical Package for Social Science (SPSS Inc. version 18.0, Armonk, NY). RESULTS: With a near linear relationship of angle metrics to lights levels (R2 = 0.8-0.95), the analysis examines the differences from the brightest to darkest light levels. Decreasing ambient light levels from 1.0 to 0.0 fc decreased the average AOD500 measurement from 407 ± 136 µm to 315 ± 114 µm (mean percent difference (MPD) 29%, p < 0.001), AOD750 from 587 ± 184 µm to 496 ± 155 µm (MPD 18%, p < 0.001), TISA500 from 136 ± 43 µm2 to 101 ± 37 µm2 (MPD 35%, p < 0.001), TISA750 from 269 ± 81 µm2 to 212 ± 68 µm2 (MPD 27%, p < 0.001), and SS angle from 38.3% ± 9% to 32.1% ± 9% (MPD 19%, p < 0.001). Intra-/inter-grader results showed good reproducibility for each grader (MPD = 0.7-3%; coefficient of variation (CV) = 3.2-8.3%; R2 = 0.8-0.95; p < 0.001 for all metrics) and between graders (MPD = 1.4-5.9%; CV = 6.7-14.2%; R2 = 0.81-0.89; Pearson Correlation Coefficient (PCC) = 0.8-0.97 (p<0.001)). Bland-Altman plots did not demonstrate any apparent bias, with similar repeatability and agreement. CONCLUSIONS: The results of this study show the high sensitivity of the anterior chamber to changes in the illumination. The slight decrease in light had a corresponding large decrease in Anterior Chamber Angle (ACA) metrics. With clinical diagnoses and treatments of eye diseases relying on these angle measurements, these findings emphasize the importance of strictly controlling light conditions in order to obtain reproducible measurements of anterior chamber geometry.
Assuntos
Câmara Anterior/citologia , Tomografia de Coerência Óptica/métodos , Idoso , Feminino , Glaucoma/diagnóstico , Voluntários Saudáveis , Humanos , Iluminação , Masculino , Reprodutibilidade dos TestesRESUMO
Polyvinylalcohol (PVA) hydrogel cross-linked by gamma irradiation was assessed as a possible vitreous substitute. From a series of experiments, rise of intraocular pressure and inflammatory changes in the vitreous cavity after operation were observed in some cases. Crab-eating macaques were used for this experiment. PVA gels were injected into vitreous cavity after vitrectomy and followed clinically by opthalmoscopy, tonometry, fundus photography, electroretinogram (ERG), chemotaxis, and flare cell meter. Histopathologic examination by light and electron microscopy was performed after 3 months. As a result, there were no significant changes in ophthalmoscopic findings. No abnormal rising of intraocular pressure (IOP) was recognized. ERG did not show meaningful amplitude weakness. From the photon counting of flare cell meter, significant break of blood-aqueous barrier and blood-retinal barrier was not observed. Histopathologic examination revealed that all layers of the retina were intact and no loss of tissue was evident. However, in PVA gel-injected eyes, some vacuolations of the inner retina were found in some specimens. To conclude, PVA gel was well tolerated in these experiments. The gel with a network similar to the vitreous body showed the best biocompatibility, though it is necessary to investigate the biocompatibility for the long-term. PVA gel is a good candidate for a vitreous substitute.
Assuntos
Materiais Biocompatíveis/farmacologia , Teste de Materiais , Álcool de Polivinil/farmacologia , Corpo Vítreo , Animais , Câmara Anterior/citologia , Câmara Anterior/efeitos dos fármacos , Barreira Hematoaquosa/efeitos dos fármacos , Modelos Animais de Doenças , Eletrorretinografia , Géis , Técnicas In Vitro , Injeções , Pressão Intraocular/efeitos dos fármacos , Macaca fascicularis , Masculino , Microscopia Eletrônica , Coelhos , Retina/efeitos dos fármacos , Retina/fisiologia , Retina/ultraestrutura , VitrectomiaRESUMO
The fetal environment plays a decisive role in modifying the risk for developing diabetes later in life. Developing novel methodology for noninvasive imaging of ß-cell development in vivo under the controlled physiological conditions of the host can serve to understand how this environment affects ß-cell growth and differentiation. A number of culture models have been designed for pancreatic rudiment but none match the complexity of the in utero or even normal physiological environment. Speier et al. recently developed a platform of noninvasive in vivo imaging of pancreatic islets using the anterior chamber of the eye where islets get vascularized, grow and respond to physiological changes. The same methodology was adapted for the study of pancreatic development. E13.0, still undifferentiated rudiments with fluorescent lineage tracing were implanted in the AC of the eye, allowing the longitudinal study of their growth and differentiation. Within 48 h the anlages get vascularized and grow but their mesenchyme displays a selective growth advantage. The resulting imbalance leads to alteration in the differentiation pattern of the progenitors. Reducing the mesenchyme to its bare minimum before implantation allows the restoration of a proper balance and a development that mimics the normal pancreatic development. These groundbreaking observations demonstrate that the anterior chamber of the eye provides a good system for noninvasive in vivo fluorescence imaging of the developing pancreas under the physiology of the host and can have important implications for designing strategies to prevent or reverse the deleterious effects of hyperglycemia on altering ß-cell function later in life.