Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 19 de 19
Filtrar
Mais filtros











Base de dados
Intervalo de ano de publicação
1.
Stem Cells Transl Med ; 8(10): 1030-1040, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31179638

RESUMO

Mesenchymal stromal cells (MSCs) are multipotent stem cells that participate in tissue repair and possess considerable immunomodulatory potential. MSCs have been shown to promote allograft survival, yet the mechanisms behind this phenomenon have not been fully defined. Here, we investigate the capacity of MSCs to suppress the allogeneic immune response by secreting the pleiotropic molecule hepatocyte growth factor (HGF). Using an in vivo mouse model of corneal transplantation, we report that MSCs promote graft survival in an HGF-dependent manner. Moreover, our data indicate that topically administered recombinant HGF (a) suppresses antigen-presenting cell maturation in draining lymphoid tissue, (b) limits T-helper type-1 cell generation, (c) decreases inflammatory cell infiltration into grafted tissue, and (d) is itself sufficient to promote transplant survival. These findings have potential translational implications for the development of HGF-based therapeutics. Stem Cells Translational Medicine 2019;8:1030-1040.


Assuntos
Córnea/patologia , Transplante de Córnea/métodos , Fator de Crescimento de Hepatócito/metabolismo , Transplante de Células-Tronco Mesenquimais/métodos , Transplante Homólogo/métodos , Animais , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Transfecção
2.
Mol Ther ; 25(8): 1881-1888, 2017 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-28502469

RESUMO

Corneal injuries are among the major causes of ocular morbidity and vision impairment. Optimal epithelial wound healing is critical for the integrity and transparency of the cornea after injury. Hepatocyte growth factor (HGF) is a mitogen and motility factor that primarily regulates epithelial cell function. Herein, we investigate the effect of HGF on proliferation of corneal epithelial cells (CECs) in inflamed conditions both in vitro and in vivo. We demonstrate that HGF not only promotes CEC proliferation in homeostatic conditions but also reverses the anti-proliferative effect of the inflammatory environment on these cells. Furthermore, using a mouse model of ocular injury, we show that HGF treatment suppresses ocular inflammation and actively augments CEC proliferation, leading to improved and accelerated corneal epithelial repair. These findings have potential translational implications and could provide a framework for the development of novel HGF-based therapies for corneal epithelial defects.


Assuntos
Lesões da Córnea/genética , Lesões da Córnea/metabolismo , Epitélio/metabolismo , Fator de Crescimento de Hepatócito/genética , Cicatrização/genética , Animais , Biomarcadores , Antígeno CD11b/metabolismo , Proliferação de Células , Lesões da Córnea/patologia , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Epitélio/patologia , Expressão Gênica , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Inflamação/genética , Inflamação/metabolismo , Inflamação/terapia , Mediadores da Inflamação/metabolismo , Camundongos
3.
Sci Rep ; 7(1): 971, 2017 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-28428556

RESUMO

Antigen-presenting cells (APCs) play an important role in transplant rejection and tolerance. In high-risk corneal transplantation, where the graft bed is inflamed and vascularized, immature APCs in the donor corneal stroma quickly mature and migrate to lymphoid tissues to sensitize host T cells. In this study, using a mouse model of corneal transplantation, we investigated whether enrichment of tolerogenic APCs (tolAPCs) in donor corneas can enhance graft survival in corneal allograft recipients with inflamed graft beds. Treatment of donor corneas with interleukin-10 (IL-10) and transforming growth factor-ß1 (TGFß1) altered the phenotype and function of tissue-residing APCs. Transplantation of these tolAPC-enriched corneas decreased frequencies of interferon gamma (IFNγ)+ effector T cells (Teffs), as well as allosensitization in the hosts, diminished graft infiltration of CD45+ and CD4+ cells, and significantly improved corneal allograft survival compared to saline-injected controls. These data provide a novel approach for tolAPC-based immunotherapy in transplantation by direct cytokine conditioning of the donor tissue.


Assuntos
Córnea/efeitos dos fármacos , Transplante de Córnea/métodos , Citocinas/farmacologia , Sobrevivência de Enxerto , Animais , Células Apresentadoras de Antígenos/imunologia , Córnea/imunologia , Interleucina-10/farmacologia , Camundongos , Modelos Animais , Fenótipo , Doadores de Tecidos , Fator de Crescimento Transformador beta1/farmacologia
4.
Stem Cell Reports ; 7(4): 583-590, 2016 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-27693426

RESUMO

Transparency of the cornea is indispensable for optimal vision. Ocular trauma is a leading cause of corneal opacity, leading to 25 million cases of blindness annually. Recently, mesenchymal stem cells (MSCs) have gained prominence due to their inflammation-suppressing and tissue repair functions. Here, we investigate the potential of MSCs to restore corneal transparency following ocular injury. Using an in vivo mouse model of ocular injury, we report that MSCs have the capacity to restore corneal transparency by secreting high levels of hepatocyte growth factor (HGF). Interestingly, our data also show that HGF alone can restore corneal transparency, an observation that has translational implications for the development of HGF-based therapy.


Assuntos
Córnea/fisiologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Regeneração , Animais , Linhagem Celular , Córnea/citologia , Córnea/efeitos dos fármacos , Lesões da Córnea/genética , Lesões da Córnea/metabolismo , Lesões da Córnea/patologia , Lesões da Córnea/terapia , Fibroblastos , Expressão Gênica , Fator de Crescimento de Hepatócito/genética , Fator de Crescimento de Hepatócito/metabolismo , Fator de Crescimento de Hepatócito/farmacologia , Humanos , Interleucina-1beta/farmacologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Modelos Animais , RNA Interferente Pequeno/genética , Cicatrização
5.
Invest Ophthalmol Vis Sci ; 57(4): 1643-8, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27054516

RESUMO

PURPOSE: To compare the effects of post-penetrating keratoplasty (PK) and post-keratoprosthesis (KPro) surgery-related inflammation on the posterior segment of the eye and to assess inhibition of tumor necrosis factor alpha (TNFα) and interleukin-1 beta (IL-1ß) on these effects. METHODS: BALB/C (syngeneic) or C57BL/6 (allogeneic) corneas were transplanted onto BALB/C host beds as part of PK or miniature KPro (m-KPro) implantation. Intraocular pressure (IOP) was measured via an intracameral pressure sensor; tissues were harvested and analyzed 8 weeks after surgery. Expression of TNFα and IL-1ß in the retina was analyzed using real-time quantitative (q)PCR. Optic nerve degeneration (axon count, circularity, and area) was assessed quantitatively using ImageJ software. After m-KPro implantation, mice were treated with saline, anti-TNFα, or anti-IL-1ß antibody, and axonal loss was assessed after 10 weeks. RESULTS: Mean IOP was within normal limits in the operated and fellow eyes in all groups. The mRNA expression of TNFα and IL-1ß was highest in m-KPro groups with either syngeneic or an allogeneic carrier. We observed optic nerve degeneration in both allogeneic PK and m-KPro implanted eyes with an allogeneic carrier. However, TNFα blockade significantly reduced axonal loss by 35%. CONCLUSIONS: Allogeneic PK and m-KPro implants with an allogeneic carrier lead to chronic inflammation in the posterior segment of the eye, resulting in optic nerve degeneration. In addition, blockade of TNFα prevents axonal degeneration in this preclinical model of allogeneic m-KPro (alloKPro) implantation.


Assuntos
Órgãos Artificiais , Bioprótese/efeitos adversos , Córnea , Ceratoplastia Penetrante/efeitos adversos , Degeneração Neural/etiologia , Doenças do Nervo Óptico/etiologia , Segmento Posterior do Olho/patologia , Animais , Axônios/patologia , Expressão Gênica/fisiologia , Interleucina-1beta/antagonistas & inibidores , Interleucina-1beta/genética , Pressão Intraocular , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Degeneração Neural/metabolismo , Degeneração Neural/patologia , Doenças do Nervo Óptico/metabolismo , Doenças do Nervo Óptico/patologia , Segmento Posterior do Olho/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Retina/metabolismo , Transplante Homólogo , Transplante Isogênico , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética
6.
Transplantation ; 99(4): 678-86, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25606789

RESUMO

BACKGROUND: Graft failure because of immune rejection remains a significant problem in organ transplantation, and lymphatic and blood vessels are important components of the afferent and efferent arms of the host alloimmune response, respectively. We compare the effect of antihemangiogenic and antilymphangiogenic therapies on alloimmunity and graft survival in a murine model of high-risk corneal transplantation. METHODS: Orthotopic corneal transplantation was performed in hemevascularized and lymph-vascularized high-risk host beds, and graft recipients received subconjunctival vascular endothelial growth factor (VEGF)-trap, anti-VEGF-C, sVEGFR-3, or no treatment, beginning at the time of surgery. Fourteen days after transplantation, graft hemeangiogenesis and lymphangiogenesis were evaluated by immunohistochemistry. The frequencies of Th1 cells in regional lymphoid tissue and graft-infiltrating immune cells were evaluated by flow cytometry. Long-term allograft survival was compared using Kaplan-Meier curves. RESULTS: VEGF-trap significantly decreased graft hemangiogenesis as compared to the control group and was most effective in reducing the frequency of graft-infiltrating immune cells. Anti-VEGF-C and sVEGFR3 significantly decreased graft lymphangiogenesis and lymphoid Th1 cell frequencies as compared to control. VEGF-trap (72%), anti-VEGF-C (25%), and sVEGFR-3 (11%) all significantly improved in the 8-week graft survival compared to control (0%), although VEGF-trap was significantly more effective than both anti-VEGF-C (P < 0.05) and sVEGFR-3 (P < 0.05). CONCLUSION: In a clinically relevant model of high-risk corneal transplantation in which blood and lymphatic vessels are present and treatment begins at the time of transplantation, VEGF-trap is significantly more effective in improving long-term graft survival as compared to anti-VEGF-C and sVEGFR-3, but all approaches improve survival when compared to untreated control.


Assuntos
Inibidores da Angiogênese/farmacologia , Transplante de Córnea/efeitos adversos , Rejeição de Enxerto/prevenção & controle , Sobrevivência de Enxerto/efeitos dos fármacos , Receptores de Fatores de Crescimento do Endotélio Vascular/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Animais , Anticorpos Monoclonais/farmacologia , Anticorpos Neutralizantes/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/metabolismo , Rejeição de Enxerto/fisiopatologia , Linfangiogênese/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Modelos Animais , Neovascularização Fisiológica/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th1/metabolismo , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator C de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator C de Crescimento do Endotélio Vascular/imunologia , Fator C de Crescimento do Endotélio Vascular/metabolismo , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/farmacologia
7.
Invest Ophthalmol Vis Sci ; 55(10): 6631-8, 2014 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-25228546

RESUMO

PURPOSE: To investigate whether systemically injected syngeneic mesenchymal stem cells (MSCs) can home to the transplanted cornea, suppress induction of alloimmunity, and promote allograft survival. METHODS: Mesenchymal stem cells were generated from bone marrow of wild-type BALB/c or GFP (green fluorescent protein)+ C57BL/6 mice, and 1×10(6) cells were intravenously injected to allografted recipients 3 hours after surgery. Mesenchymal stem cells homing to the cornea were examined at day 3 post transplantation by immunohistochemistry. MHC (major histocompatibility complex) II+CD11c+ cells were detected in the cornea and lymph nodes (LNs) 14 days post transplantation using flow cytometry. Cytokine expression of bone marrow-derived dendritic cells (BMDCs) was determined using real-time PCR. ELISPOT assay was used to assess indirect and direct host T cell allosensitization, and graft survival was evaluated by slit-lamp biomicroscopy weekly up to 8 weeks. RESULTS: Intravenously injected GFP+ MSCs were found in abundance in the transplanted cornea, conjunctiva, and LNs, but not in the ungrafted (contralateral) tissue. The frequencies of mature MHC II+CD11c+ antigen-presenting cells (APCs) were substantially decreased in the corneas and draining LNs of MSC-injected allograft recipients compared to control recipients. Maturation and function of in vitro cultured BMDCs were decreased when cocultured with MSCs. Draining LNs of MSC-injected allograft recipients showed lower frequencies of IFNγ-secreting Th1 cells compared to the control group. Allograft survival rate was significantly higher in MSC-injected recipients compared to non-MSC-injected recipients. CONCLUSIONS: Our data demonstrate that systemically administered MSCs specifically home to the inflamed ocular surface and promote allograft survival by inhibiting APC maturation and induction of alloreactive T cells.


Assuntos
Transplante de Córnea/métodos , Células Dendríticas/imunologia , Rejeição de Enxerto/imunologia , Complexo Principal de Histocompatibilidade/imunologia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/imunologia , Células Th1/imunologia , Animais , Autoimunidade , Células Cultivadas , Citocinas/biossíntese , Citocinas/genética , Modelos Animais de Doenças , Queimaduras Oculares/induzido quimicamente , Queimaduras Oculares/patologia , Queimaduras Oculares/cirurgia , Citometria de Fluxo , Regulação da Expressão Gênica , Rejeição de Enxerto/patologia , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Microscopia de Fluorescência , RNA/genética , Reação em Cadeia da Polimerase em Tempo Real , Transplante Homólogo
8.
Graefes Arch Clin Exp Ophthalmol ; 252(11): 1755-62, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25091513

RESUMO

PURPOSE: To investigate the effect of VEGF-C and VEGF-D blockade via soluble VEGFR-3 (sVEGFR-3) on T cell allosensitization, corneal neovascularization, and transplant survival. METHODS: Corneal intrastromal suture placement and allogeneic transplantation were performed on BALB/c mice to evaluate the effect of sVEGFR-3 on corneal neovascularization. Soluble VEGFR-3 trap was injected intraperitoneally to block VEGF-C/D (every other day starting the day of surgery). Immunohistochemical staining of corneal whole mounts was performed using anti-CD31 (PECAM-1) and anti-LYVE-1 antibodies to quantify the levels of hem- and lymphangiogenesis, respectively. Mixed lymphocyte reaction (MLR) was performed to assess indirect and direct host T cell allosensitization and the frequencies of IFN-γ-producing T cells in the draining lymph nodes were assessed using flow cytometry. Graft opacity and survival was evaluated by slit-lamp biomicroscopy. RESULTS: Treatment with sVEGFR-3 resulted in a significant blockade of lymphangiogenesis 2 weeks post-transplantation and significantly prolonged corneal allograft survival compared to the control group at 8 weeks post-transplantation (87.5 % vs. 50 %), and this was associated with significant reduction in the frequencies of allosensitized T cells and decreased frequencies of IFN-γ-producing CD4 T cells. CONCLUSIONS: Soluble VEGFR-3 suppresses corneal lymphangiogenesis and allograft rejection and may offer a viable therapeutic modality for corneal neovascularization and corneal transplantation.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Córnea/imunologia , Neovascularização da Córnea/prevenção & controle , Sobrevivência de Enxerto/efeitos dos fármacos , Ceratoplastia Penetrante , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/farmacologia , Animais , Técnicas de Cocultura , Neovascularização da Córnea/imunologia , Citometria de Fluxo , Glicoproteínas/metabolismo , Sobrevivência de Enxerto/imunologia , Isoantígenos/imunologia , Linfangiogênese/efeitos dos fármacos , Teste de Cultura Mista de Linfócitos , Masculino , Proteínas de Membrana Transportadoras , Camundongos , Camundongos Endogâmicos BALB C , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Transplante Homólogo , Fator C de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator D de Crescimento do Endotélio Vascular/antagonistas & inibidores
9.
Invest Ophthalmol Vis Sci ; 55(6): 3681-5, 2014 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-24833747

RESUMO

PURPOSE: To establish a murine model for keratoprosthesis. METHODS: A miniature keratoprosthesis (m-KPro) device was created consisting of a poly[methyl methacrylate] front part and a titanium back plate, designed after the Boston KPro, which is in widespread clinical use. BALB/c mice were used and a 2 mm in diameter donor cornea was punched out. After 2-mm trepanation of the syngeneic recipient cornea, extracapsular crystalline lens extraction was performed. The m-KPro was assembled onto the cornea button in a similar manner to human KPro implantation. The cornea-device complex was secured to the recipient bed with eight interrupted 11-0 sutures. All mice (n = 10) were followed up for 8 weeks postoperatively. RESULTS: All m-KPros were successfully implanted and retained in all 10 animals. There were no critical complications such as endophthalmitis, corneal melting, device extrusions, leakage, extensive inflammation, or weight loss in the animals. We observed mild to moderate donor and host corneal neovascularization in all cases throughout the follow-up period. CONCLUSIONS: We have established a novel murine model of KPro implantation that we anticipate will serve as a good experimental system for evaluating host responses after KPro surgery.


Assuntos
Órgãos Artificiais , Doenças da Córnea/cirurgia , Próteses e Implantes , Animais , Modelos Animais de Doenças , Seguimentos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Desenho de Prótese , Resultado do Tratamento
10.
PLoS One ; 7(11): e47716, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23152758

RESUMO

BACKGROUND: Peripheral nerve damage of the cornea is a complication following surgery or infection which may lead to decreased visual function. We examined the efficacy of the semaphorin 3A inhibitor, SM-345431, in promoting regeneration of peripheral nerves in a mouse corneal transplantation model. METHODOLOGY/PRINCIPAL FINDINGS: P0-Cre/Floxed-EGFP mice which express EGFP in peripheral nerves cells were used as recipients of corneal transplantation with syngeneic wild-type mouse cornea donors. SM-345431 was administered subconjunctivally every 2 days while control mice received vehicle only. Mice were followed for 3 weeks and the length of regenerating nerves was measured by EGFP fluorescence and immunohistochemistry against ßIII tubulin. Cornea sensitivity was also measured by the Cochet-Bonnet esthesiometer. CD31 staining was used to determine corneal neovascularization as a possible side effect of SM-345431. Regeneration of ßIII tubulin positive peripheral nerves was significantly higher in SM-345431 treated mice compared to control. Furthermore, corneal sensitivity significantly improved in the SM-345431 group by 3 weeks after transplantation. Neovascularization was limited to the peripheral cornea with no difference between SM-345431 group and control. CONCLUSIONS/SIGNIFICANCE: Subconjunctival injections of SM-345431 promoted a robust network of regenerating nerves as well as functional recovery of corneal sensation in a mouse keratoplasty model, suggesting a novel therapeutic strategy for treating neurotrophic corneal disease.


Assuntos
Transplante de Córnea , Regeneração Nervosa/efeitos dos fármacos , Nervos Periféricos/efeitos dos fármacos , Nervos Periféricos/fisiologia , Semaforina-3A/antagonistas & inibidores , Xantonas/farmacologia , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Córnea/inervação , Córnea/patologia , Córnea/cirurgia , Neovascularização da Córnea , Células Epiteliais/efeitos dos fármacos , Camundongos , Camundongos Transgênicos , Modelos Animais , Neovascularização Fisiológica/efeitos dos fármacos , Semaforina-3A/metabolismo
11.
Invest Ophthalmol Vis Sci ; 53(7): 3638-44, 2012 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-22562508

RESUMO

PURPOSE: Bone marrow-derived mesenchymal stem cells (MSCs) hold great promise for wound healing and tissue regeneration. In the present study, we investigated the impact of corneal injury on the homeostasis of endogenous MSCs, and the potential of MSCs to home to injured tissue and promote corneal repair. METHODS: Corneal injury in mice was induced by thermal cauterization. Circulating MSCs were quantified by flow cytometric analysis. Ex vivo expanded red Q-dot-labeled or GFP+ bone marrow-derived MSCs were intravenously injected after injury and detected using epifluorescence microscopy. Corneal fluorescein staining was performed to evaluate epithelial regeneration. RESULTS: Following the induction of corneal injury in mice, a 2-fold increase in the frequency of circulating endogenous MSCs was observed within 48 hours of injury, which was accompanied by increased levels of the stem cell chemoattractants, substance P and SDF-1, in both the injured cornea and blood. Systemically administered MSCs homed to the injured cornea, but not to the normal cornea, and showed long-term survival. In addition, in the setting of corneal injury, MSC administration showed significant and rapid corneal epithelial regeneration. CONCLUSIONS: These findings provide novel evidence that corneal injury causes significant mobilization of endogenous MSCs into blood, and that MSCs home specifically to the injured cornea and promote regeneration, highlighting the therapeutic implications of MSC-mediated tissue repair in corneal injury.


Assuntos
Movimento Celular/fisiologia , Doenças da Córnea/patologia , Lesões da Córnea , Traumatismos Oculares/patologia , Células-Tronco Mesenquimais/fisiologia , Transplante de Células-Tronco , Cicatrização/fisiologia , Animais , Células da Medula Óssea/citologia , Diferenciação Celular , Sobrevivência Celular , Córnea/patologia , Doenças da Córnea/cirurgia , Modelos Animais de Doenças , Traumatismos Oculares/cirurgia , Citometria de Fluxo , Seguimentos , Masculino , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos C57BL
12.
Mol Vis ; 16: 2727-32, 2010 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-21179238

RESUMO

PURPOSE: To determine whether epithelial-mesenchymal transition is involved in the development of corneal subepithelial fibrosis (pannus). METHODS: Frozen samples of pannus tissue removed from human corneas with a diagnosis of total limbal stem cell deficiency were characterized by immunostaining for both epithelial and mesenchymal markers. We selected transformation-related protein 63 (p63) and pancytokeratin as epithelial markers and vimentin and α-smooth muscle actin (α-SMA) as mesenchymal markers. Immunostaining for ß-catenin and E-cadherin was performed to determine wingless-Int (Wnt)-pathway activation. RT-PCR analysis was also performed on epithelial tissue obtained from pannus samples after dispase digestion. RESULTS: Immunohistochemistry revealed strong nuclear expression of p63 and weak intercellular expression of E-cadherin in epithelial basal cells of pannus tissue. Furthermore, translocation of ß-catenin from intercellular junctions to the nucleus and cytoplasm was also observed. Double-positive cells for both p63 and α-SMA were observed in the subepithelial stroma of pannus tissue, which was supported by RT-PCR and cytospin analysis. CONCLUSIONS: Epithelial-mesenchymal transition may be partially involved in the development of subepithelial corneal fibrosis due to total limbal stem cell deficiency.


Assuntos
Transição Epitelial-Mesenquimal , Epitélio Corneano/patologia , Actinas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Caderinas/metabolismo , Criança , Demografia , Epitélio Corneano/enzimologia , Feminino , Fibrose , Regulação da Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transativadores/metabolismo , Fatores de Transcrição , Proteínas Supressoras de Tumor/metabolismo , Vimentina/metabolismo , beta Catenina/metabolismo
13.
Jpn J Ophthalmol ; 54(6): 537-43, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21191713

RESUMO

PURPOSE: To report the efficacy of simultaneous keratolimbal allograft (KLAL) surgery and deep anterior lamellar keratoplasty (DALK) for limbal stem cell deficiency (LSCD). METHODS: We conducted a retrospective, interventional case series of six consecutive eyes of five patients with LSCD and stromal opacity due to gelatinous drop-like dystrophy (two eyes), Stevens-Johnson syndrome (SJS, two eyes), or aniridia (two eyes). Only patients with normal lid anatomy and Schirmer test values greater than 3 mm were enrolled. DALK was performed by viscodissection followed by a thin, 360° KLAL designed by using an artificial anterior chamber. KLAL sutures were removed after 2 weeks. RESULTS: DALK and KLAL were successfully performed in all eyes, which were followed for an average of 17.2 ± 10.8 months. All eyes recovered a smooth corneal epithelium, although one SJS patient developed a persistent epithelial defect (PED) leading to opacification of the central cornea. Visual acuity improved by more than 2 lines in all eyes except that of the SJS patient with PED. No other complications were observed. CONCLUSION: Simultaneous DALK and thin-section KLAL is an effective treatment for ocular surface disease in patients with residual tear function and normal lid anatomy.


Assuntos
Doenças da Córnea/cirurgia , Transplante de Córnea , Epitélio Corneano/transplante , Limbo da Córnea/citologia , Transplante de Células-Tronco , Células-Tronco/patologia , Adulto , Idoso , Doenças da Córnea/patologia , Células Epiteliais/transplante , Feminino , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos , Tomografia de Coerência Óptica , Transplante Homólogo , Acuidade Visual/fisiologia
14.
Mol Vis ; 15: 2751-61, 2009 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-20019883

RESUMO

PURPOSE: To establish a novel murine intraocular lens (IOL) implantation model to study the protective effects of colored-IOLs against retinal phototoxicity. METHODS: Two-millimeter diameter IOL buttons were created from IOLs for clinical use. Extra-capsular crystalline lens extraction and IOL implantation were performed in BALB/c mice using a technique similar to human cataract surgery. For light exposure experiments, mice were exposed to 5,000 LUX of white light for 24 h on the day after surgery. To investigate the protective effects of yellow IOL against light exposure, ERG measurements were conducted in vivo, followed by TdT-mediated dUTP Nick-End Labeling (TUNEL) and outer nuclear layer (ONL) thickness measurement of retinal tissue in yellow or clear IOL-implanted mice and control mice without surgery. RESULTS: IOLs were successfully implanted in all animals, and IOL buttons without haptics were well stabilized in the capsular bag. Murine eyes developed posterior capsule opacification (PCO) after IOL implantation by postoperative day 5 at the latest. In contrast to the clear IOL-implanted animals stimulated by light exposure, the yellow IOL-implanted animals had significantly reduced numbers of TUNEL-positive cells and retained thickness of the ONL. The ERG showed that yellow IOL implantation prevents a decrease of amplitude in both the a-wave and b-wave compared with clear IOL implantation. CONCLUSIONS: We established a new animal model of IOL implantation and demonstrated the protective effects of colored-IOL against retinal phototoxicity after cataract surgery.


Assuntos
Lentes Intraoculares , Luz/efeitos adversos , Implantação de Prótese , Retina/patologia , Retina/efeitos da radiação , Animais , Catarata/complicações , Catarata/patologia , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Retina/cirurgia , Degeneração Retiniana/complicações , Degeneração Retiniana/patologia
15.
Invest Ophthalmol Vis Sci ; 50(5): 2109-15, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19136703

RESUMO

PURPOSE: To report the efficacy of human bone marrow-derived mesenchymal stem cells as a source of feeder cells for the cultivation of transplantable corneal epithelial cell sheets. METHODS: Human mesenchymal stem cells (marrow adherent stem cells; MASCs) were cultured in alpha-modified Eagle's medium with 10% serum and were treated with mitomycin C. Expression of cytokines in MASCs was confirmed by reverse transcription-polymerase chain reaction. Human limbal epithelial cells were cocultured with MASCs or 3T3 feeder cells to compare colony-forming efficiency (CFE). Limbal epithelial cells were cultured on MASCs or 3T3 feeder cells at the air-liquid interface to allow stratification, and stratified epithelial sheets were analyzed by immunohistochemistry against cytokeratin 3 (K3), K15, p63alpha, and ABCG2. Rabbit limbal epithelial cell sheets were cultivated with MASC feeder cells and transplanted to the ocular surface of the limbal-deficient rabbits. Epithelial grafts were observed by slit lamp microscopy for 4 weeks and then evaluated by histology and immunohistochemistry against K3 and K4. RESULTS: MASC feeder cells expressed keratinocyte growth factor, hepatocyte growth factor, and N-cadherin. The CFE of human limbal epithelial cells was similar in MASC and 3T3 feeder groups. Stratified cell sheets were successfully cultivated with MASC feeder cells expressing K3, K15, p63alpha, and ABCG2. Transplanted epithelial sheets regenerated the corneal phenotype in limbal-deficient rabbits. CONCLUSIONS: MASC-derived feeder cells are suitable for the engineering of epithelial sheets, avoiding the use of potentially hazardous xenologic feeder cells.


Assuntos
Transplante de Células , Doenças da Córnea/cirurgia , Células Epiteliais/citologia , Epitélio Corneano/citologia , Células-Tronco Mesenquimais/citologia , Membro 2 da Subfamília G de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Antígenos CD/metabolismo , Células da Medula Óssea , Caderinas/metabolismo , Técnicas de Cultura de Células , Técnicas de Cocultura , Ensaio de Unidades Formadoras de Colônias , Doenças da Córnea/metabolismo , Doenças da Córnea/patologia , Citocinas/genética , Células Epiteliais/transplante , Epitélio Corneano/metabolismo , Fator 7 de Crescimento de Fibroblastos/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Queratina-15/metabolismo , Queratina-3/metabolismo , Limbo da Córnea/citologia , Proteínas de Membrana/metabolismo , Células-Tronco Mesenquimais/metabolismo , Camundongos , Células NIH 3T3 , Proteínas de Neoplasias/metabolismo , RNA Mensageiro/metabolismo , Coelhos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
16.
Nippon Ganka Gakkai Zasshi ; 111(5): 391-6, 2007 May.
Artigo em Japonês | MEDLINE | ID: mdl-17536498

RESUMO

PURPOSE: To demonstrate the efficacy and safety of deep lamellar keratoplasty(DLKP) in the treatment of corneal opacity in severe ocular surface disease. METHODS: A total of 12 eyes of 11 patients were analyzed in this retrospective case series. The original diseases were Stevens Johnson syndrome (SJS : 3 eyes), ocular ciccatricial pemphigoid(OCP : 2 eyes), thermal burns (2 eyes), limbal deficiency due to unknown cause (2 eyes), and corneal scarring due to trachoma (3 eyes). Cases with total limbal deficiency (6 eyes) were also treated with secondary or simultaneous limbal transplantation. Cataract surgery was also performed in 4 eyes following DLKP. RESULTS: DLKP was successfully done in all cases. One case with SJS experienced an immunological rejection against the limbal graft. Two eyes with SJS eventually developed ulcers that required therapeutic penefrating keratoplasty. Visual improvement was observed in 9 out of 12 eyes, of which 2 cases maintained a corrected visual acuity of 0.1 despite conjunctivalization of the ocular surface. CONCLUSION: DLKP is an effective means to treat stromal opacity in patients with ocular surface disease.


Assuntos
Doenças da Córnea/cirurgia , Transplante de Córnea/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Opacidade da Córnea/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
17.
J Cataract Refract Surg ; 31(9): 1699-701, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16246770

RESUMO

This study reports a technique in which phacoemulsification and intraocular lens (IOL) insertion are performed in conjunction with a microkeratome flap in a patient with ocular surface disease. Microkeratome-assisted phacoemulsification was performed in a 72-year-old woman with a history of trachoma during childhood. A 130 microm corneal flap was made using a microkeratome prior to phacoemulsification and IOL insertion. Lifting the flap during surgery allowed a clear view of the anterior chamber through the smooth lamellar interface created by the microkeratome blade. The flap was washed and repositioned without sutures at the end of surgery. There were no complications associated with the microkeratome flap following surgery. Visual acuity improved from hand motion to 20/200. Microkeratome-assisted phacoemulsification is a safe technique for cataract patients with diseases of the ocular surface.


Assuntos
Implante de Lente Intraocular/métodos , Facoemulsificação/métodos , Retalhos Cirúrgicos , Idoso , Feminino , Humanos
18.
J Cataract Refract Surg ; 31(5): 895-902, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15975453

RESUMO

PURPOSE: To prospectively assess the clinical outcome after implantation of the 1CU accommodating intraocular lens (IOL) and a foldable acrylic IOL (AcrySof, Alcon). SETTING: Department of Ophthalmology, Tokyo Dental College, Ichikawa Hospital, Ichikawa, and Minami Aoyama Eye Clinics, Tokyo, Yokohama, Japan. METHODS: Twenty-two eyes of 16 patients with cataract had phacoemulsification implantation of 1CU accommodating IOL. Twenty eyes of 10 age-matched and sex-matched patients with cataract had the same surgery but with a foldable acrylic IOL. All patients had assessments of the amplitude of accommodation, refraction, uncorrected and best corrected distance and near visual acuity, and distance corrected near visual acuity before surgery up to 12 months after surgery. Contrast visual acuities were measured 1 year after surgery. Anterior segment photography, intraocular pressure measurements, specular microscopy, and computerized topography were also performed. RESULTS: The final best corrected distance visual acuity was above 20/25 in all eyes with the 1CU and the AcrySof IOLs. The mean distance corrected near visual acuity was significantly higher in the 1CU IOL group than in the acrylic IOL group after 3 months. None of the eyes with the AcrySof IOL implants displayed an accommodative response at any examination. The peak mean amplitude of accommodation with the 1CU IOLs was observed at 3 months and was 0.5 diopters +/- 0.44 (SD). Accommodation amplitude declined after 6 months. CONCLUSION: The 1CU IOL provided additional near acuity postoperatively, but the benefit disappeared at 12 months with a concomitant decrease in accommodation amplitude owing to an increase in anterior and posterior capsular opacities.


Assuntos
Acomodação Ocular , Implante de Lente Intraocular , Lentes Intraoculares , Facoemulsificação , Acuidade Visual/fisiologia , Resinas Acrílicas , Idoso , Contagem de Células , Topografia da Córnea , Endotélio Corneano/patologia , Feminino , Humanos , Pressão Intraocular , Cápsula do Cristalino/patologia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/patologia , Estudos Prospectivos , Desenho de Prótese , Resultado do Tratamento
19.
J Cataract Refract Surg ; 30(3): 558-65, 2004 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15050249

RESUMO

PURPOSE: To prospectively assess the clinical and visual outcomes of phacoemulsification and implantation of a rollable intraocular lens (IOL) with a thin optic and compare the results with those of implantation of a foldable acrylic IOL. SETTING: Department of Ophthalmology, Tokyo Dental College, Ichikawa General Hospital, Ichikawa, Chiba, Japan. METHODS: Sixteen consecutive eyes of 8 patients (4 women, 4 men) with corticonuclear cataract had small-incision clear corneal phacoemulsification with implantation of a rollable ThinOptX IOL (ThinOptX Inc.) in the capsular bag. Twenty eyes of 10 age- and sex-matched patients (5 women, 5 men) with the same diagnosis had phacoemulsification and intracapsular implantation of an AcrySof foldable acrylic IOL (MA60BM, Alcon). The patients' refractive status and uncorrected and best corrected distance visual acuities were assessed preoperatively and 1 week and 1, 3, and 6 months after surgery. The uncorrected and best corrected near acuities were measured before and 6 months after surgery. Contrast visual acuity was measured with variable contrast charts 1, 3, and 6 months after surgery, and the results in the 2 IOL groups were compared. Anterior segment photography, intraocular pressure (IOP) measurement, specular microscopy, and fundoscopy were done before surgery and at 1, 3, and 6 months. RESULTS: The final best corrected distance acuity was better than 20/25 in all eyes with a ThinOptX IOL and 18 eyes (90%) with an AcrySof IOL. The best corrected near acuity was better than 20/40 in 12 eyes (75%) and 14 eyes (70%), respectively. The mean contrast acuity with charts 2 and 3 was significantly higher in the ThinOptX group than in the AcrySof group at all examinations (P<.05). The final mean postoperative induced astigmatism was 0.06 diopter (D) +/- 0.50 (SD) and 0.25 +/- 0.68 D, respectively (P>.05). There were no differences in IOP or corneal endothelial cell density between the 2 groups at any examination. No intraoperative or postoperative complications occurred. CONCLUSIONS: ThinOptX IOL implantation provided best corrected near and distance visual acuities comparable to those provided by the AcrySof IOLs. The significantly higher contrast acuities attained after implantation of the ThinOptX lens may be attributable to its ultrathin properties.


Assuntos
Implante de Lente Intraocular/métodos , Lentes Intraoculares , Facoemulsificação/métodos , Acuidade Visual/fisiologia , Resinas Acrílicas , Idoso , Contagem de Células , Sensibilidades de Contraste/fisiologia , Endotélio Corneano/citologia , Feminino , Humanos , Pressão Intraocular , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Minimamente Invasivos , Estudos Prospectivos , Desenho de Prótese
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA