Reduction in corneal haze and apoptosis by amniotic membrane matrix in excimer laser photoablation in rabbits

Wang, Ming X.; Gray, Trevor; Park, Woo Chan; Prabhasawat, Pinnita; Culbertson, William W.; Forster, Richard K.; Hanna, Khalil; Tseng, Scheffer C.G.

doi:10.1016/s0886-3350(00)00467-3

Cited by 114 publications

(72 citation statements)

References 38 publications

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“…In a murine model of HSV-1 necrotizing keratitis, application of human AM as a temporary graft for 2 days induces rapid regression of corneal stromal edema and inflammation, and reduces infiltration of lymphocytes and macrophages in the corneal stroma (Heiligenhaus et al, 2001). In a rabbit model of excimer laser ablation, human AM as a temporary graft for 24 hr reduces infiltration of polymorphonuclear (PMN) cells in the ablated stroma while PMNs adherent to AM stroma undergo apoptosis (Park and Tseng, 2000;Wang et al, 2001). Clinically, Shimmura et al (2001) removed AM one week after being placed as a temporary AM graft for treating persistent corneal epithelial defects or in conjunction with keratolimbal allograft, and disclosed that most monocytes and macrophages trapped by AM exhibit characteristics of apoptosis.…”

Section: Introductionmentioning

confidence: 99%

Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro

Kawakita

et al. 2006

Experimental Eye Research

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Amniotic membrane (AM) used as a temporary or permanent graft for ocular surface reconstruction has a potent anti-inflammatory effect. We would like to investigate the mechanism whereby AM induces macrophage apoptosis in vitro. Mouse macrophages, Raw 264.7 cells, were cultured on plastic, type I collagen, corneal stromal slice or AM stromal matrix in serum-free medium with or without interferon-γ (IFN-γ). Cells were stained by LIVE/DEAD assay, Hoechst-33342, and TUNEL assay for cell death and apoptosis. Cell lysates and conditioned media were analysed by Cell Death Detection ELISA assay for quantitation of apoptosis. Conditioned media were also analysed by Griess assay for the nitrite concentration and ELISA assay for tumour necrosis factor alpha (IFN-α) concentration. Lysates of cells were subjected to Western blot analyses of IKK-α, IKK-β, p65 (RelA) subunit of nuclear factor κB (NF-κB), total Akt, phospho-Akt (Ser473), and phospho-FKHR (Thr24)/phosphor-FKHRL1 (Thr32). At 48 hr after cultivation, cells showed a low level of apoptosis when cultured on plastic, type I collagen and corneal stromal slice with or without IFN-γ and on AM without IFN-γ. Nevertheless, cells showed a significant increase of apoptosis when cultured on AM with IFN-γ activation, and this phenomenon became apparent only after 48 hr. IFN-γ-activated macrophages on plastic continuously produced nitric oxide (NO) and IFN-α during 72 hr culturing. In contrast, there was no NO and IFN-α production after 48 hr culture on AM. NO inhibitors, L-NMMA and L-NIL, attenuated NO production of IFN-γ-activated macrophages on AM, while apoptosis was not decreased accordingly. Expression of IKK-α, IKK-β, p65 (RelA) subunit of NF-κB total Akt, phosopho-Akt (Ser473), and phospho-FKHR (Thr24)/FKHRL1 (Thr32) was all down-regulated in IFN-γ-activated macrophages cultured on AM. In conclusion, AM stromal matrix induces apoptosis of IFN-γ activated, but not non-activated macrophages, not through the generation of NO, but instead by down-regulating anti-apoptotic NF-κB and Akt-FKHR signalling pathways.

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Section: Introductionmentioning

confidence: 99%

Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro

Kawakita

et al. 2006

Experimental Eye Research

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“…Já no século XVIII, Quengsy, em 1789, citado por Guimarães (1979), propôs a utilização de placas de vidro em substituição às córneas lesadas. A partir dessa época, vários estudos envolvendo técnicas de ceratoplastia foram desenvolvidos, com ênfase, nos últimos anos, para a utilização de membranas fetais (Matthews et al, 1982;Wang et al, 2001;Woo et al, 2001). …”

Section: Introductionunclassified

Aspectos clínicos e imunopatológicos da ceratoplastia com membrana amniótica xenógena fresca e conservada em glicerina: estudo experimental em coelhos

Sampaio

Ranzani

Júnior

et al. 2006

Arq. Bras. Med. Vet. Zootec.

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Avaliaram-se, por meio da análise clínica, histopatologia e imunoistoquímica, os efeitos da aplicação da membrana amniótica xenógena fresca e conservada em glicerina, sobre os mecanismos imunológicos da superfície ocular. Para tal, utilizaram-se 40 coelhos, distribuídos em dois grupos experimentais, os quais foram avaliados por 21 dias. A avaliação clínica revelou que a membrana amniótica xenógena conservada em glicerina estimulou uma resposta inflamatória aguda maior que a membrana aplicada fresca. A análise histopatológica indicou que ambas se comportaram de forma semelhante a partir da primeira semana de pós-operatório, apresentando as alterações clássicas da resposta inflamatória da córnea, com o predomínio de infiltrado do tipo polimorfonuclear. A análise imunoistoquímica indicou que, ainda aos 21 dias, a resposta imune local é inespecífica, permitindo concluir que a resposta imune específica na córnea é tardia e que a córnea é um sítio privilegiado para aplicação de enxertos com características imunológicas diferentes, visto que não houve o estímulo para o desenvolvimento de uma resposta mais específica nos grupos avaliados durante toda a execução do experimento.

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“…Corneal haze is more common after PRK than after LASIK, because the corneal wound healing response and the stimulus for the fibrotic response are stronger (1) . Many substances have been used to prevent haze formation in PRK: amniotic membrane, vitamins A and E, collagenase inhibitors, and ubiquinone Q10 (19)(20)(21)(22) , but no controlled clinical trials have been reported, except for mitomicin C (23) . MMC is a drug that has been used frequently to reduce haze formation owing, presumably, to its inhibition of proliferation of activated keratocytes (1) .…”

Section: Discussionmentioning

confidence: 99%

Effects of mitomycin C on infiltration of polymorphonuclear leukocytes after epithelial scrape injury in the mouse cornea

Escarião¹,

Nagasaki²,

Braunstein³

2008

Arq. Bras. Oftalmol.

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Purpose: To determine whether mitomycin C (MMC) alters appearance and disappearance of polymorphonuclear leucocytes (PMN) in the cornea stroma, using an epithelial scrape injury in eye mouse model. Methods: Twenty-mice underwent mechanical epithelium debridement in the central cornea using 20% ethanol. After the scrape, the right eye received 0.02% MMC for one minute, while the left eye received physiological saline. The animals were sacrificed on days 1, 2, 5, and 14 after surgery, and corneal whole mounts were prepared for histology. PMN distribution was analyzed in digitized microscope images. Cell division in the cornea was determined by immunohistochemical detection of bromodeoxyuridine (BrdU), which was injected intraperitoneally before the mice were sacrificed. Results: Epithelial scrape injury triggered infiltration of PMNs into the corneal stroma. An analysis of PMN distribution revealed that there was no difference between eyes treated with and without MMC at all time points. BrdU labeling showed that 0.02% MMC for one minute blocked keratocyte proliferation completely. Conclusion: MMC treatment regimen, which is common in clinical practice, inhibits keratocyte proliferation during wound healing, but when used at 0.02% for one minute, it does not affect PMN infiltration into the corneal stroma, and subsequent movement toward the injury site, or the disappearance of PMNs from the stroma, in the mouse epithelial injury model. ABSTRACT INTRODUCTION

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Reduction in corneal haze and apoptosis by amniotic membrane matrix in excimer laser photoablation in rabbits

Cited by 114 publications

References 38 publications

Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro

Amniotic membrane induces apoptosis of interferon-γ activated macrophages in vitro

Aspectos clínicos e imunopatológicos da ceratoplastia com membrana amniótica xenógena fresca e conservada em glicerina: estudo experimental em coelhos

Effects of mitomycin C on infiltration of polymorphonuclear leukocytes after epithelial scrape injury in the mouse cornea

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