Yazarlar laboratuvarda mesleki ve mükemmel teknik çalışmaları için Sayın Dalia Sela ve Bay Emi Sharon müteşekkiriz.

Hayvanlar araştırma hayvanların kullanımına ilişkin ARVO kararına uygun olarak tedavi edildi. Çalışma protokolü Laboratuar Hayvanları Araştırma kurumsal Komitesi (.; 021211 onay numarası 022-4598-2) tarafından onaylandı. Cerrahi 1. 1. Hazırlık <ol><li> Tartılır ve ketamin hidroklorür 100 mg / ml (100 mg / kg) ve ksilazin hidroklorür 25 mg / ml (12.5 mg / kg) içeren bir intramüsküler enjeksiyon ile 13 günlük bir Yeni Zelanda beyaz tavşanı anestetize. Bu dozlar nedeniyle, cerrahi prosedürün süresi için kullanılmaktadır. Bir ağrı refleksi eksikliği ile anestezi uygun seviyede sağlamak. </li><li> Steril bir pamuklu çubuk kullanarak, kurumasını kornea önlemek için sigara ameliyat gözle oftalmik merhem küçük bir miktar uygulanır. kurumasını kornealar önlemek için işlem sırasında normal tuzlu su işletilen göz kornea üzerine damla% 0.9 uygulayın. </li><li> oxybuprocain (topikal anestezi sonrası her gözün üç aksiyel uzunluk ölçümleri gerçekleştirmeksonra e hidroklorür% 0.4) bir ultrason A-tarayıcı kullanarak ve ölçümler ortalama. merkezi kornea dik probu uygulayın. </li></ol> 2. Cerrahi # 1 - Ön Çapraz Bağlama Adım <ol><li> bir göz ameliyat mikroskobu altında cerrahi göz forseps ve makas kullanılarak, cerrahi göz forseps ve makas ile 360 ​​° konjonktiva kesi yapmak. </li><li> açılı forseps veya kas kanca dört ekstraoküler rektus kasları tanımlamak ve 2-0 örgülü ipek olmayan Needled dikişlerle onları izole kullanma. NOT: ipek dikişler istenilen yönde dünyayı hareket ile yardımcı olacaktır. gözün dört rektus kaslarının (dört kadran) arasındaki kadranın ayrılmıştır. <ol><li> Bir cilt işaretleyici her çeyrekte iki bölge, ekvator sklera az bir ve posterior sklera az bir Mark. Bunlar ışınlama bölgeleri bulunmaktadır. </li></ol></li><li> % 0.1 dekstran-ücretsiz riboflavin-5-fosfat içeren 5ml şırınga 3 ml hazırlayıne. 26 G lakrimal kanül veya 25 G konik hydrodelineator şırınga bağlayın. </li><li> aşağı eğimli ısmarlama fiber optik bağlı bir UV A (370 nm) ışık kaynağı içeren ışınlama cihazını hazırlayın. Enerji gücü ve kalibrasyon ölçümü takiben (aşağıda Not bakınız), 57 mW / cm² şekilde ayarlayabilirsiniz. NOT: Kalibrasyon üretici tarafından ticari gerçekleştirildi. Bu yöntem, 57 J / cm² toplam UVA ışık dozu içerir. (0.2 cm 2 57 mW / cm2 2.2 J 0.2 cm2 nokta kümülatif yük 200 saniye boyunca 11.4 mW / 0.2 cm 2 kullanılarak 11.4 mW / 0.2 cm 2 olduğunu) </li></ol> 3. Cerrahi # 1 - Çapraz Bağlama <ol><li> Seçilen çeyreğin ters yönde ipek sütür çekerek gözün taşımak tedavi edilecek (alt temporal kadranda tedavi ederken, örneğin, yukarı çekin ve burun). 2-0 örgülü ipek olmayan iğneli sütür kullanarak küresi hareketini gerçekleştirmek hangi iHer rektus kas solates. </li><li> ışınlama başlamadan önce ışınlama bölgesi 20 sn üzerine% 0.1 dekstran-ücretsiz riboflavin-5-fosfat içeren foto çözümü uygulayın. </li><li> 200 sn bir ışınlama süre için aşağı eğimli özel yapılmış fiber optik kullanarak ışınlama bölgesi ışın tedavisi. 4 mm bir çapa sahip 0.2 cm² her bölgenin alanı ölçülür. </li><li> 200 sn ışınlama süresince ışınlama bölgesi üzerine her 20 saniyede% 0.1 dekstran içermeyen riboflavin-5-fosfat ihtiva eden foto-çözeltisi uygulanır. NOT: ışınlama ve iki cerrah tarafından aynı anda riboflavin düşmesini gerçekleştirin. </li><li> Tekrarlayın dört gözün kadranın her biri için 3.1-3.4 adımları. </li></ol> 4. Cerrahi # 1 - Gönder Çapraz Bağlama Adım <ol><li> Göz çevresinde kürk tıraş. Cerrahi oftalmik makas kullanılarak kapak kenar boşluklarını Trim ve daha sonra yavaşça üst ve 4-0 örgülü ipek fildişi rengi kullanarak düşük göz kapakları sütür - 3/8 daire yenidenayetin demircilik 13 mm uzunluk C-3 iğne (tarsorrhaphy). prosedür (kloramfenikol% 5) sonunda kapak kenarındaki göz merhem küçük bir miktar uygulayın. </li></ol> 5. Cerrahi # 1 - Gönder Operatif Bakım <ol><li> kurtarma sırasında bir ısıtma lambası altında hayvanların tutun. sternal yatma korumak için yeterli bilinci yerine kadar gözetimsiz bir hayvan bırakmayın. </li><li> tamamen iyileşti kadar yeni yatak ile temiz bir kafeste hayvan yerleştirin. İyileştikten sonra, hayvan, hayvan odasına iade edilecektir. sadece uyanık annesi hayvan dönün. </li><li> İlk 48 saat boyunca, genel durumunu ve hastalık veya enfeksiyon belirtileri ile ilgili hayvanları inceler. Tavşan davranış değişikliği gözlenmesi halinde gerektiğinde bölüm 1. Yönet analjezi başına analjezi yönetme. (Metamizol 50 mg / kg her 6 saat düşer). </li></ol> 6. Cerrahi 2. <ol><li> Elli beş gün yıllardan sonraurgery 1., ağırlık ve ketamin hidroklorür 100 mg / ml (100 mg / kg) ve ksilazin hidroklorür 25 mg / ml (12.5 mg / kg) içeren bir intramüsküler enjeksiyon ile anestezi tavşan. </li><li> bir göz ameliyat mikroskobu altında cerrahi göz makas kullanarak, tarsorrhaphy çıkarın. topikal anestezi (oksibuprokain hidroklorür% 0.4) bir ultrason A-tarayıcı kullandıktan sonra her gözün üç aksiyel uzunluk ölçümleri gerçekleştirmek ve sonra ölçümler ortalama. merkezi kornea dik probu uygulayın. </li><li> intraperitonal fenobarbital sodyum (200 mg / 1.5 kg vücut ağırlığı) ile tavşan öldürülür. </li></ol>

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Yazarlar hiçbir rakip mali çıkarları olduğunu beyan ederim.

Bu riboflavin ve UVA ışıması ile çapraz bağlama teknolojisini kullanan bir tavşan modelinde, eksenel miyopi önlenmesi vivo çalışmasında ilk sunulmaktadır. Farklı deney hayvanları bu tip çalışmalar kullanılabilir olsa da, çoğunlukla göz boyutu ve sklera yüzeyi üzerinde çapraz bağlanmasını gerçekleştirmek için ihtiyaç tavşan seçtik. Biz tavşan sklera açığa üst ve alt göz kapakları dikilmesi zorlu prosedürler olduğu bulundu. Biz kapak...

Miyopi kırılma bozukluklarının en yaygın olanıdır. ABD&#39;de ve Avrupa&#39;da miyopi prevalansı% 30 civarında olduğu bildirilmektedir ve Asya ülkelerinde genel nüfus 1,2% 60 kadar etkiler. Miyop ilerlemesi, genellikle iki yıllık bir aralık üzerinde 3 yaklaşık -0.5 diyoptri arasında bir oranda, miyoplara% 50 kadar gerçekleşir. Miyopi dayattığı sağlık maliyetleri gözlük masrafları, kontakt lens ve refraktif cerrahi ve glokom, katarakt, retina dekolmanı ve görme bozukluğu 4-6 artmış sağlık risklerine ilişkin masraflar dahil olmak üzere, önemli bulunmaktadır. Miyopi Hayvan çalışmalarında, görme azalması göz ve kornea dövme 11 kısa mesafede gözkapağı sütür 7-10, bir tıkayıcı yerleştirme indüklenir. Yapay miyopi Bu çalışmaların gerçekleştirilmesi için, ancak, oklüzyon sürecinin hiçbir görüş yoksunluğu deneyleri Carr gibi, çok küçük hayvanlar üzerinde gerçekleştirilmelidirYetişkin numuneler üzerinde ied başarılı olmuştur. Şiddetli miyopi önemli özelliklerinden biri gibi Ehlers gibi bağlı sklera bazı metabolik bozukluk nedeniyle görsel yoksunluğu 12 veya daha sonra emmetropization bir rahatsız geri bildirim mekanizması muhtemelen sklera ilerici incelme ile sklera bir patolojik değişiklik Danlos sendromu 13. Sonuçta, her iki mekanizma nedeniyle böyle fibrillogenesis 16 bir azalma kollajen lif çapı 14,15 ve bozuklukları gibi miyop sklera yapısal anormallikler kadar uzanan ve sklera, retina ve koroid incelmesi yol açar. Çeşitli çalışmalar bozulmuş kollajen çapraz bağlama miyop sklera 17-18 zayıflaması sürecinde önemli bir faktör olduğunu göstermiştir. WOLLENSAK ve ark., 19-21 riboflavin ve ultraviyole-A (UVA) ışınlama foto uygulayarak kollajen çapraz bağlanmasını uyarılan (370 nm)ve in vitro 19 domuz ve insan sklera sertlik önemli bir,% 157 artış ve in vivo tavşan sklera sertliğinde bir 465% artış (Young modülü) 20 kaydetti. Sertliği 4 ay sonra, 3 gün sonra 320,4 göre% 277.6% arttı ve 8 ay (Young modülü) 22 sonra 502%: Çapraz bağlanma, aynı zamanda in vivo tavşan sklera üzerinde uzun süreli bir etkisi vardı. Miyop ilerlemesi tutuklama terapötik girişimler 23-26 yayınlandı ancak bu yöntemlerin başarısı tartışmalıdır. progresif miyopi önlenmesi Resim etkili bir araç bugüne kadar tespit edilmiştir. miyopi etiyolojisi hala tartışmalıdır ve tedavisi bir sorun teşkil etmektedir. Bu bulgulara dayanarak, skleral çapraz miyop ilerlemesi sklera tabanlı tedavi için bir araç olarak hizmet verebilir varsayılmaktadır. Bu çalışmanın amacı, skleral kollajen crosslinki incelemekgörsel eksen tıkanması ile uyarılan eksenel miyopi gelişmesi üzerinde etkisi ng.

<table><tbody><tr><td>2-0 braided silk non-needled sutures&amp;nbsp;</td><td>ETHICON</td><td>W193</td><td></td></tr><tr><td>4-0 braided silk ivory color&amp;nbsp;</td><td>ETHICON</td><td>W816</td><td></td></tr><tr><td>0.1% dextran-free riboflavin-5-phosphate 1 mg:1 ml</td><td>Concept for Pharmacy Ltd</td><td>D2-5025</td><td></td></tr><tr><td>UV A (370 nm) light source&amp;nbsp;</td><td>O/E LAND Inc</td><td>NCSU033B</td><td></td></tr><tr><td>Beveled down custom made fiber optic&amp;nbsp;</td><td>Prizmatix Ltd</td><td></td><td></td></tr><tr><td>26 G lacrimal cannula&amp;nbsp;</td><td>Beaver-visitec International Ltd.&amp;nbsp;</td><td>REF581276</td><td></td></tr><tr><td>25 G tapered hydrodelineator [Blumenthal]&amp;nbsp;</td><td>Beaver-visitec International Ltd.&amp;nbsp;</td><td>REF585107</td><td></td></tr><tr><td>13 days old rabbits</td><td>Harlan&amp;nbsp;</td><td>1NZWR40</td><td></td></tr><tr><td>Ultrasonic biometer</td><td>Allergan-Humphrey</td><td>820-519</td><td></td></tr><tr><td>Skin marker</td><td>Devon</td><td>4237101664X</td><td></td></tr></tbody></table>

scleral cross-linking using riboflavin and ultraviolet-a radiation for prevention of axial myopia in a rabbit model

Miyop kişiler, şiddetli miyopi ile özellikle, katarakt, glokom, retina dekolmanı ve korioretinal anormalliklerin yüksek normalden risk altındadır. Buna ek olarak, patolojik miyopi görme bozukluğu ve körlük 1-3 ortak bir geri dönüşü olmayan bir nedenidir. Çalışmamız bir tavşan modelinde eksenel miyopi gelişimine riboflavin ve ultraviyole-A ışınları kullanılarak skleral çapraz bağlanma etkisini gösterir. gözün aksiyel uzunluk 13 gün (erkek ve dişi) yaş Yeni Zelanda beyaz tavşanlarda A-tarama ultrason ile ölçüldü. Sonra göz tarsorrhaphy tarafından takip sklera çapraz bağlanma, 360 ° konjonktiva kesi uygulandı. Eksenel uzama onların sağ göz kapaklarını (tarsorrhaphy) dikilmesi ile 13 gün eski Yeni Zelanda tavşanlarda oluşturuldu. gözler çeyrek ayrıldı ve her kadran 0.2 cm² bir alanı ve 4 mm çapa sahip, iki sklera ışınlama bölgeleri, her biri vardı. Çapraz bağlanma,% 0.1 bırakarak gerçekleştirildidekstran içermeyen morötesi A radyasyondan önce radyasyon bölgeleri 20 saniye ve 200 saniye ışınlama süresi boyunca her 20 saniyede üzerine riboflavin-5-fosfat. UVA radyasyonu (370 nm) 57 mW / cm² (toplam UVA ışık dozu, 57 J / cm²) de sklera dik uygulandı. Tarsorrhaphies tekrar eksenel uzunlukta ölçümleri, ardından günde 55 ayrılmıştır. Bu çalışma, riboflavin ve ultraviyole-Radyasyonla skleral çapraz bağlama etkin bir tavşan modelinde oklüzyon kaynaklı eksenel uzamaya önlediğini göstermektedir.

Şekiller 1 ve 2, grafiksel iki grup eksenel uzunluğu ölçümleri göstermektedir. Sol göz (Şekil 1) ameliyat değil iken Grup 1 tavşan sağ göz skleral çapraz bağlanma ve tarsorrhaphy uygulandı. Sol göz (Şekil 2) ameliyat değil iken Grup 2 tavşan sağ gözüne sadece kesi ve tarsorrhaphy uygulandı. skleral çapraz bağlanma ve tarsorrhaphy uygul...

Watch this Scientific Journal Video about Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model at JoVE.com

Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model

Bu eksenel uzama tavşan göz riboflavin ve UVA sklera çapraz bağlanma etkisini göstermektedir. Eksenel uzama onların sağ göz kapaklarını (tarsorrhaphy) dikilmesi yoluyla (erkek ve dişi) 13 gün eski Yeni Zelanda tavşanlarda oluşturuldu.

Skleral bir Tavşan Modelinde Aksiyel Miyopide Önleme Riboflavin ve Ultraviyole-A Radyasyon Kullanımı Çapraz bağlama

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal ...

scleral-cross-linking-using-riboflavin-ultraviolet-radiation-for

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

8.8K Görüntüleme.  Rabin Medical Center, Beilinson Campus.  Bu eksenel uzama tavşan göz riboflavin ve UVA sklera çapraz bağlanma etkisini göstermektedir. Eksenel uzama onların sağ göz kapaklarını (tarsorrhaphy) dikilmesi yoluyla (erkek ve dişi) 13 gün eski Yeni Zelanda tavşanlarda oluşturuldu. 

Video: Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes

Photoacoustic ocular imaging is an emerging ophthalmic imaging technology that can noninvasively visualize ocular tissue by converting light energy into sound waves and is currently under intensive investigation. However, most reported work to date is focused on the imaging of the posterior segment of the eyes of small animals, such as rats and mice, which poses challenges for clinical human translation due to small eyeball sizes. This manuscript describes a novel photoacoustic microscopy (PAM) and optical coherence tomography (OCT) dual-modality system for posterior segment imaging of the eyes of larger animals, such as rabbits. The system configuration, system alignment, animal preparation, and dual-modality experimental protocols for in vivo, noninvasive, label-free chorioretinal imaging in rabbits are detailed. The effectiveness of the method is demonstrated through representative experimental results, including retinal and choroidal vasculature obtained by the PAM and OCT. This manuscript provides a practical guide to reproducing the imaging results in rabbits and advancing photoacoustic ocular imaging in larger animals.

This manuscript describes the novel setup and operating procedure of a photoacoustic microscopy and optical coherence tomography dual-modality system for noninvasive, label-free chorioretinal imaging of larger animals, such as rabbits.

Yaşam tavşan gözünde roman Photoacoustic mikroskobu ve optik Koherens tomografi çift-modality Chorioretinal görüntüleme

Photoacoustic ocular imaging is an emerging ophthalmic imaging technology that can noninvasively visualize ocular tissue by converting light energy into ...

Mühendislik

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

Sodyum Hyaluronat subretinal Enjeksiyon Kemirgenler Retina Dekolmanı Modeli

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

Tıp

Three Different Protocols of Corneal Collagen Crosslinking in Keratoconus: Conventional, Accelerated and Iontophoresis

Keratoconus is a bilateral and progressive corneal ectasia. In order to slow down its progression, corneal collagen cross-linking (CXL) has recently been introduced as an efficient treatment option. In biological and chemical sciences, crosslinking refers to new chemical bonds formed between reactive molecules. Hence, the aim of corneal collagen CXL is to synthetically increase the formation of crosslinks between collagen fibrils in the corneal stroma. Despite the fact that the efficiency of the conventional CXL (C-CXL) protocol has already been shown in several clinical studies, it might benefit from improvements in duration of the procedure and removal of corneal epithelium. Hence, in order to provide a coherent evaluation of two new and optimized CXL protocols, we studied keratoconus patients who had undergone one of the three CXL treatments: iontophoresis (I-CXL), accelerated CXL (A-CXL), and conventional CXL (C-CXL). A-CXL is a 6 time faster CXL procedure using a ten time higher UVA irradiance but still including an epithelium removal. Iontophoresis is a transepithelial non-invasive technique in which a small electric current is applied to improve riboflavin penetration throughout the cornea. Using anterior segment optical coherence tomography (AS OCT) and in vivo confocal microscopy (IVCM), we conclude that regarding the depth of treatment penetration, conventional CXL protocol remains the standard for treating progressive keratoconus. Accelerated CXL seems to be a quick, effective and safe alternative to treat thin corneas. The use of iontophoresis is still being investigated and should be considered with greater caution.

Corneal collagen cross-linking (CXL) is the only conservative treatment currently available to halt keratoconus progression by improving the biomechanical rigidity of the corneal stroma. The aim of this manuscript is to highlight the methods of three different protocols of CXL: conventional CXL (C-CXL), accelerated CXL (A-CXL), and iontophoresis CXL (I-CXL).

Keratokonus Kornea Kollajen çapraz bağlama Üç Farklı Protokoller: Konvansiyonel, Hızlandırılmış ve İyontoforez

Keratoconus is a bilateral and progressive corneal ectasia. In order to slow down its progression, corneal collagen cross-linking (CXL) has recently been ...

A Step by Step Protocol for Subretinal Surgery in Rabbits

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in adults in industrially developed countries. RPE transplantation appears to be a promising therapy, as it may replace dysfunctional RPE, restore its function, and thereby vision.
Here we describe a method for transplanting a cultured RPE monolayer on a scaffold into the subretinal space (SRS) of rabbits. After vitrectomy xenotransplants were delivered into the SRS using a custom made shooter consisting of a 20-gauge metallic nozzle with a polytetrafluoroethylene (PTFE) coated plunger. The current technique evolved in over 150 rabbit surgeries over 6 years. Post-operative follow-up can be obtained using non-invasive and repetitive in vivo imaging such as spectral domain optical coherence tomography (SD-OCT) followed by perfusion-fixed histology.
The method has well-defined steps for easy learning and high success rate. Rabbits are considered a large eye animal model useful in preclinical studies for clinical translation. In this context rabbits are a cost-efficient and perhaps convenient alternative to other large eye animal models.

Retinal pigment epithelium (RPE) replacement strategies and gene-based therapy are considered for several retinal degenerative conditions. For clinical translation, large eye animal models are required to study surgical techniques applicable in patients. Here we present a rabbit model for subretinal surgery geared towards RPE transplantation, which is versatile and cost-efficient.

Tavşanlarda Subretinal Cerrahi Adım Protokolle Bir Adım

Age related macular degeneration (AMD), retinitis pigmentosa, and other RPE related diseases are the most common causes for irreversible loss of vision in ...

In vivo Structural Assessments of Ocular Disease in Rodent Models using Optical Coherence Tomography

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable tool to evaluate and characterize changes in a variety of retinal and ocular disease and injury models. In light induced retinal degeneration models, SD-OCT can be used to track thinning of the photoreceptor layer over time. In glaucoma models, SD-OCT can be used to monitor decreased retinal nerve fiber layer and total retinal thickness and to observe optic nerve cupping after inducing ocular hypertension. In diabetic rodents, SD-OCT has helped researchers observe decreased total retinal thickness as well as decreased thickness of specific retinal layers, particularly the retinal nerve fiber layer with disease progression. In mouse models of myopia, SD-OCT can be used to evaluate axial parameters, such as axial length changes. Advantages of SD-OCT include in vivo imaging of ocular structures, the ability to quantitatively track changes in ocular dimensions over time, and its rapid scanning speed and high resolution. Here, we detail the methods of SD-OCT and show examples of its use in our laboratory in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia. Methods include anesthesia, SD-OCT imaging, and processing of the images for thickness measurements.

Here, we describe the use of spectral-domain optical coherence tomography (SD-OCT) to visualize retinal and ocular structures in vivo in models of retinal degeneration, glaucoma, diabetic retinopathy, and myopia.

Optik Koherens Tomografi Kullanılarak Kemirgen Modellerinde Oküler Hastalığın İn vivo Yapısal Değerlendirmeleri

Spectral-domain optical coherence tomography (SD-OCT) is useful for visualizing retinal and ocular structures in vivo. In research, SD-OCT is a valuable ...

Biyomühendislik

Second Harmonic Generation Signals in Rabbit Sclera As a Tool for Evaluation of Therapeutic Tissue Cross-linking (TXL) for Myopia

Methods to strengthen tissue by introducing chemical bonds (non-enzymatic cross-linking) into structural proteins (fibrillar collagens) for therapy include photochemical cross-linking and tissue cross-linking (TXL) methods. Such methods for inducing mechanical tissue property changes are being employed to the cornea in corneal thinning (mechanically weakened) disorders such as keratoconus as well as the sclera in progressive myopia, where thinning and weakening of the posterior sclera occurs and likely contributes to axial elongation. The primary target proteins for such tissue strengthening are fibrillar collagens which constitute the great majority of dry weight proteins in the cornea and sclera. Fortuitously, fibrillar collagens are the main source of second harmonic generation signals in the tissue extracellular space. Therefore, modifications of the collagen proteins, such as those induced through cross-linking therapies, could potentially be detected and quantitated through the use of second harmonic generation microscopy (SHGM). Monitoring SHGM signals through the use of a laser scanning microscopy system coupled with an infrared excitation light source is an exciting modern imaging method that is enjoying widespread usage in the biomedical sciences. Thus, the present study was undertaken in order to evaluate the use of SHGM microscopy as a means to measure induced cross-linking effects in ex vivo rabbit sclera, following an injection of a chemical cross-linking agent into the sub-Tenon&#39;s space (sT), an injection approach that is standard practice for causing ocular anesthesia during ophthalmologic clinical procedures. The chemical cross-linking agent, sodium hydroxymethylglycinate (SMG), is from a class of cosmetic preservatives known as formaldehyde releasing agents (FARs). Scleral changes following reaction with SMG resulted in increases in SHG signals and correlated with shifts in thermal denaturation temperature, a standard method for evaluating induced tissue cross-linking effects.

Second Harmonic Generation Signals in Rabbit Sclera As a Tool for Evaluation of Therapeutic Tissue Cross-linking TXL for Myopia

This protocol describes techniques for evaluating chemical cross-linking of the rabbit sclera using&#160;second harmonic generation imaging and differential scanning calorimetry.

Tavşan sklera bir araç olarak ikinci harmonik üretimi sinyalleri tedavi doku (TXL) için miyopi Cross-linking değerlendirilmesi için

Methods to strengthen tissue by introducing chemical bonds (non-enzymatic cross-linking) into structural proteins (fibrillar collagens) for therapy ...

Establishment of a Severe Dry Eye Model Using Complete Dacryoadenectomy in Rabbits

Dry eye disease (DED) is a complex disease with multiple etiologies and variable symptoms, having ocular surface inflammation as its key pathophysiologic step. Despite advances in our understanding of DED, significant knowledge gaps remain. Advances are limited in part due to the lack of informative animal models. The authors recently reported on a method of DED induced by injecting all orbital lacrimal gland (LG) tissues with the lectin concanavalin A. Here, we report a novel model of aqueous-deficient DED based on the surgical resection of all orbital LG (dacryoadenectomy) tissues. Both methods use rabbits because of their similarity to human eyes in terms of the size and structure of the ocular surface. One week after removal of the nictitating membrane, the orbital superior LG was surgically removed under anesthesia, followed by removal of the palpebral superior LG, and finally removal of the inferior LG. Dacryoadenectomy induced severe DED, evidenced by a marked reduction in the tear break up time test and the Schirmer&#39;s tear test, and significantly increased tear osmolarity and rose bengal staining. Dacryoadenectomy-induced DED lasted at least eight weeks. There were no complications and animals tolerated the procedure well. The technique can be mastered relatively easily by those with adequate surgical experience and appreciation of the relevant rabbit anatomy. Since this model recapitulates the features of human aqueous-deficient DED, it is suitable for studies of ocular surface homeostasis, DED, and candidate therapeutics.

A novel approach is presented to induce chronic dry eye disease in rabbits by surgically removing all orbital lacrimal glands. This method, distinct from those previously reported, produces a stable, reproducible model of aqueous deficient dry eye well suited to study tear physiology and pathophysiology and ocular therapeutics.

Tavşanlarda Komple Dakriyoadenektomi Kullanarak Şiddetli Kuru Göz Modelinin Oluşturulması

Dry eye disease (DED) is a complex disease with multiple etiologies and variable symptoms, having ocular surface inflammation as its key pathophysiologic ...

Ex Vivo and In Vivo Animal Models for Mechanical and Chemical Injuries of Corneal Epithelium

Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, and result in a large, persistent corneal epithelial defect. Epithelial defect with the following corneal opacity and peripheral neovascularization result in irreversible visual impairment and hinder future management, especially keratoplasty. Since the animal model can be used as an effective drug development platform, models of corneal injury to the mouse and alkali burn to rabbit corneal epithelium are developed here. New Zealand white rabbit is used in the alkali burn model. Different concentrations of sodium hydroxide can be applied onto the central circular area of the cornea for 30 s under intramuscular and topical anesthesia. After copious isotonic normal saline irrigation, residual loose corneal epithelium was removed with corneal burr deep down to the Bowman's layer within this circular area. Wound healing was documented by fluorescein staining under Cobalt blue light. C57BL/6 mice were used in the traumatic model of murine corneal epithelium. The murine central cornea was marked using a skin punch, 2 mm in diameter, and then debrided by a corneal rust ring remover with a 0.5 mm burr under a stereomicroscope. These models can be prospectively used to validate the therapeutic effect of eye drops or mixed agents such as stem cells, which potentially facilitate corneal epithelial regeneration. By observing corneal opacity, peripheral neovascularization, and conjunctival congestion with stereomicroscope and imaging software, therapeutic effects in these animal models can be monitored.

Ex Vivo and In Vivo Animal Models for Mechanical and Chemical Injuries of Corneal Epithelium

Here, animal models based on mouse and rabbit are developed for mechanical and chemical injury of corneal epithelium to screen new therapeutics and the underlying mechanism.

Kornea Epitelinin Mekanik ve Kimyasal Yaralanmaları için Ex Vivo ve In Vivo Hayvan Modelleri

Corneal injury to the ocular surface, including chemical burn and trauma, may cause severe scarring, symblepharon, corneal limbal stem cells deficiency, ...

Subretinal Transplantation of Human Embryonic Stem Cell Derived-retinal Pigment Epithelial Cells into a Large-eyed Model of Geographic Atrophy

Geographic atrophy (GA), the late stage of dry age-related macular degeneration is characterized by loss of the retinal pigment epithelial (RPE) layer, which leads to subsequent degeneration of vital retinal structures (e.g., photoreceptors) causing severe vision impairment. Similarly, RPE-loss and decrease in visual acuity is seen in long-term follow up of patients with advanced wet age-related macular degeneration (AMD) receiving intravitreal anti-vascular endothelial growth factor (VEGF) treatment. Therefore, on the one hand, it is fundamental to efficiently derive RPE cells from an unlimited source that could serve as replacement therapy. On the other hand, it is important to assess the behavior and integration of the derived cells in a model of the disease entailing surgical and imaging methods as close as possible to those applied in humans. Here, we provide a detailed protocol based on our previous publications that describes the generation of a preclinical model of GA using the albino rabbit eye, for evaluation of the human embryonic stem cell derived retinal pigment epithelial cells (hESC-RPE) in a clinically relevant setting. Differentiated hESC-RPE are transplanted into naive eyes or eyes with NaIO3-induced GA-like retinal degeneration using a 25 G transvitreal pars plana technique. Evaluation of degenerated and transplanted areas is performed by multimodal high-resolution non-invasive real-time imaging.

Retinal pigment epithelial cells could serve as a cell-replacement therapy for the advanced form of dry age-related macular degeneration. This protocol describes the generation of a large-eyed model of geographic atrophy and the subretinal transplantation of human embryonic stem cell-derived retinal pigment epithelial cells into this model of disease.

Büyük gözlü manken coğrafi Atrofi, içine insan embriyonik kök hücre elde edilen retina Pigment epitel hücrelerinin subretinal nakli

Geographic atrophy (GA), the late stage of dry age-related macular degeneration is characterized by loss of the retinal pigment epithelial (RPE) layer, ...

Biyoloji

Establishing a Severe Corneal Inflammation Model in Rats Based on Corneal Epithelium Curettage Combined with Corneal Sutures

Corneal inflammation, especially severe corneal inflammation, plays a significant role in the development of corneal limbal stem cell dysfunction. Constructing appropriate animal models can help us focus on the effects of severe inflammation on corneal limbal stem cells. A 2 mm rust remover&#160;was&#160;used to remove the central corneal epithelium of Sprague Dawley (SD) rats to create an injury. Then, the central stroma of the cornea was sutured with nylon sutures to induce persistent inflammation. In this way, a corneal inflammation model with central corneal epithelium abrasion and central stroma suturing was constructed, which induced severe corneal inflammation. The changes in corneal inflammation and the condition of the limbal stem cells at 1, 3, and 7 days post-modeling were observed. On the 3rd day after modeling, the rats&#39; corneal limbus was severely edematous, with obvious neovascularization and local hyperplasia, which are typical signs of limbal stem cell deficiency. By the 7th day, the corneal edema gradually worsened, and the neovascularization continued to increase. Through quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence staining, we found that the corneal epithelial inflammatory factors were significantly upregulated, the corneal epithelial differentiation was abnormal, the corneal epithelial stem cells were significantly reduced, and the cell proliferation and stemness had also decreased. Therefore, this model demonstrates that severe inflammation can induce limbal stem cell damage without directly damaging the limbal stem cells. The model is beneficial for observing the effects of severe inflammation on the biological mechanisms of stem cells and provides an ideal platform for studying the mechanisms of corneal epithelial stem cell dysfunction induced by inflammation.

We developed a rat model of severe corneal inflammation through corneal epithelium curettage combined with corneal sutures. The study evaluated corneal inflammation patterns, epithelial proliferation, and changes in limbal stem cells under inflammatory conditions.