Ocular Surface最新文献_第3页

Readership awareness series – Paper 13: Key concepts of translational research 读者意识系列--论文 13：转化研究的关键概念。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-30 DOI: 10.1016/j.jtos.2024.08.013

Mohammad Javed Ali MD, PhD, FRCS, Ali Djalilian MD

引用次数: 0

Development and characterization of a preclinical mouse model of alkali-induced limbal stem cell deficiency 碱性诱导的睑缘干细胞缺乏症临床前小鼠模型的开发和特征描述。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-29 DOI: 10.1016/j.jtos.2024.08.015

Lina Sprogyte, Mijeong Park, Lamia Nureen, Nicodemus Tedla, Alexander Richardson, Nick Di Girolamo

Purpose

Limbal stem cell deficiency (LSCD) secondary to ocular surface alkali burn is a blinding condition that features corneal conjunctivalization. Mechanistic insights into its pathophysiology are lacking. Here, we developed a mouse model that recapitulates human disease to comprehensively delineate the clinicopathological features of a conjunctivalized cornea.

Methods

LSCD was induced in the right eyes of 6-8-week-old C57BL/6 male and female mice (n = 151) by topical administration of 0.25N sodium hydroxide on the cornea. Uninjured left eyes served as controls. Clinical, histological, phenotypic, molecular, and immunological assessments were performed at multiple time-points over 6-months.

Results

Clinically, alkali burn caused persistent corneal opacity (p = 0.0014), increased punctate staining (p = 0.0002), and reduced epithelial thickness (p = 0.0082) compared to controls. Total LSCD was confirmed in corneal whole mounts by loss of K12 protein (p < 0.0001) and mRNA expression (p = 0.0090). Instead, K8⁺, K13⁺, K15⁺ and MUC5AC⁺ conjunctival epithelia prevailed. 20 % of injured corneas developed islands of K12⁺ epithelia, suggesting epithelial transdifferentiation. Squamous metaplasia was detected in 50 % of injured corneas. Goblet cell density peaked early post-injury but decreased over time (p = 0.0047). Intraepithelial corneal basal nerve density remained reduced even at 6-months post-injury (p = 0.0487).

Conclusions

We developed and comprehensively characterized a preclinical mouse model of alkali-induced LSCD. Understanding the pathophysiological processes that transpire on the ocular surface in LSCD is key to discovering, testing, and advancing biological and pharmacological interventions that can be dispensed prior to or in conjunction with stem cell therapy to rehabilitate the cornea and restore vision.

目的：继发于眼表碱烧伤的角膜缘干细胞缺乏症（LSCD）是一种以角膜结膜化为特征的致盲性疾病。目前还缺乏对其病理生理学机制的深入了解。在此，我们建立了一个能再现人类疾病的小鼠模型，以全面描述角膜结膜化的临床病理特征：方法：在 6-8 周大的 C57BL/6J 雌雄小鼠（n=151）的右眼角膜上局部注射 0.25N 氢氧化钠，诱发 LSCD。未受伤的左眼作为对照组。在 6 个月内的多个时间点进行了临床、组织学、表型、分子和免疫学评估：结果：与对照组相比，碱烧伤在临床上导致持续性角膜混浊（p=0.0014）、点状染色增加（p=0.0002）和上皮厚度减少（p=0.0082）。在角膜全切片中，K12 蛋白的缺失证实了角膜全层缺损（以 p+、K13+、K15+ 和 MUC5AC+ 结膜上皮为主）。20% 的受伤角膜出现了 K12+ 上皮细胞岛，表明上皮细胞发生了转分化。50%的受伤角膜检测到鳞状化生。上皮细胞密度在受伤后早期达到峰值，但随着时间的推移逐渐下降（p=0.0047）。角膜上皮内基底神经密度在损伤后6个月仍有所降低（p=0.0487）：我们建立并全面描述了碱诱导的角膜缺损临床前小鼠模型。了解LSCD眼表的病理生理过程是发现、测试和推进生物和药物干预的关键，这些干预可在干细胞疗法之前或与干细胞疗法结合使用，以修复角膜和恢复视力。

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引用次数: 0

Distinctive small molecules blend: Promotes lacrimal gland epithelial cell proliferation in vitro and accelerates lacrimal gland injury repair in vivo 独特的小分子混合物：体外促进泪腺上皮细胞增殖，体内加速泪腺损伤修复

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-28 DOI: 10.1016/j.jtos.2024.08.014

Baihui Zeng , Lina Xu , Guoliang Wang , Ruize Shi , Kerui Wang , Shurong Wang , Cheng Li

Purpose

This study aims to develop a novel serum-free culture strategy containing only two small molecules, Y27632 and SB431542 (2C), for in vitro expansion of mouse lacrimal gland epithelial cells (LGECs) and investigate an innovative therapeutic approach for lacrimal gland (LG) injury.

Methods

LGECs proliferative capacity was assessed by cell counting, crystal violet staining, qRT-PCR and immunofluorescence. Cell differentiation was achieved by manipulating culture conditions and assessed by qRT-PCR and AQP5 immunofluorescence. LGECs were seeded in Matrigel for three-dimensional culture and assessed by qRT-PCR and immunofluorescence. Secretory function of the cultures was assayed by ELISA. In vivo, 2C injection verified its reparative capacity in a mouse LG injury model. Corneal fluorescein staining, phenol red cotton thread, H&E, immunofluorescence and Western blot were used to assess LG injury repair.

Results

LGECs cultured with 2C exhibited high expression of stemness/proliferation markers and maintained morphology and proliferative capacity even after the tenth passage. Removal of 2C was efficacious in achieving LGECs differentiation, characterized by the increased AQP5 expression and LTF secretion. 3D spheroids cultured with 2C demonstrated differentiation potential, forming microglandular structures containing multiple LG cell types with secretory functions after 2C removal. In vivo, 2C improved the structural integrity and function of the injured LG.

Conclusions

We present a small molecule combination, 2C, that promotes LGECs expansion and differentiation in vitro and accelerates LG injury repair in vivo. This approach has potential applications for providing a stable source of seed cells for tissue engineering applications, providing new sights for LG-related diseases treatment.

目的本研究旨在开发一种新型无血清培养策略，该策略仅含有两种小分子 Y27632 和 SB431542 (2C)，用于小鼠泪腺上皮细胞（LGECs）的体外扩增，并研究一种治疗泪腺（LG）损伤的创新方法。方法通过细胞计数、水晶紫染色、qRT-PCR 和免疫荧光评估 LGECs 的增殖能力。通过调节培养条件实现细胞分化，并通过 qRT-PCR 和 AQP5 免疫荧光进行评估。将 LGECs 接种到 Matrigel 中进行三维培养，并通过 qRT-PCR 和免疫荧光进行评估。培养物的分泌功能通过 ELISA 进行检测。在小鼠 LG 损伤模型中，注射 2C 验证了其体内修复能力。用角膜荧光素染色、酚红棉线、H&E、免疫荧光和 Western blot 评估 LG 损伤修复情况。去除 2C 能有效实现 LGECs 分化，其特征是 AQP5 表达和 LTF 分泌增加。用 2C 培养的三维球体具有分化潜能，去除 2C 后可形成包含多种具有分泌功能的 LG 细胞类型的微腺体结构。结论我们提出了一种小分子组合 2C，它能促进 LGECs 在体外的扩增和分化，并加速 LG 损伤在体内的修复。这种方法有望为组织工程应用提供稳定的种子细胞来源，为 LG 相关疾病的治疗提供新的视角。

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引用次数: 0

β-Catenin gain of function mutant in mouse periocular neural crest-derived mesenchymal cells impairs embryonic eyelid morphogenesis and leads to blepharophimosis syndrome in mice 小鼠眼周神经嵴衍生间充质细胞中的β-Catenin功能增益突变体会损害胚胎眼睑形态发生并导致小鼠睑外翻综合征

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-26 DOI: 10.1016/j.jtos.2024.08.012

Yen-Chiao Wang, Yong Yuan, Jianhua Zhang, Yujin Zhang, Winston W.-Y. Kao, Chia-Yang Liu

Purpose

The aberrant canonical Wnt-β-catenin signaling can cause devastating outcomes of tissue morphogenesis and tumor formation. In this study, we examined the impact of overexpression of constitutive active β-catenin in mouse periocular neural crest-derived mesenchymal cells during embryonic eyelid morphogenesis.

Methods

We expressed a stabilized β-catenin in which the exon 3 of the Ctnnb1 gene was deleted in periocular neural crest (PONC)-derived eyelid stromal cells (Ctnnb1^Δex3-PONC). Histopathological examinations were performed to examine the eyelid morphogenetic alterations in Ctnnb1^Δex3-PONC mice. Immunohistochemical investigations for cell proliferation, apoptosis, and differentiation were also assessed.

Results

We discovered that nuclear accumulation of β-catenin resulted in a reduction of nuclear Ki-67 and phospho-Erk1/2 expression levels and elevation of apoptosis in PONC cells during embryonic eyelid closure morphogenesis. Interestingly, however, the eyelid epithelial migration was not affected, which resulted in only eyelid epidermal closure but lacked underneath dermal formation at embryonic (E) day 16.5. The sequelae of Ctnnb1^Δex3-PONC revealed the malformation of the eyelid margin and Meibomian gland and deficiency of Muller's smooth muscle fibers formation. Consequently, Ctnnb1^Δex3-PONC mice manifested blepharophimosis syndrome at P21.

Conclusion

Our data suggested that aberrant expression of β-catenin gain of function in PONC interrupts the interplay between epithelium and stroma for the morphogenesis of eyelid closure during embryonic development.

目的正常 Wnt-β-catenin 信号的异常可导致组织形态发生和肿瘤形成的破坏性结果。方法我们在眼周神经嵴（PONC）衍生的眼睑基质细胞（Ctnnb1Δex3-PONC）中表达了稳定的β-catenin，其中Ctnnb1基因的第3外显子被删除。对Ctnnb1Δex3-PONC小鼠的眼睑形态发生变化进行了组织病理学检查。结果我们发现，在胚胎眼睑闭合形态发生过程中，β-catenin的核积累导致PONC细胞的核Ki-67和phospho-Erk1/2表达水平降低，细胞凋亡增加。但有趣的是，眼睑上皮的迁移并没有受到影响，这导致在胚胎（E）16.5天时只有眼睑表皮闭合，而缺乏下面真皮的形成。Ctnnb1Δex3-PONC的后遗症表现为眼睑边缘和睑板腺畸形以及穆勒平滑肌纤维形成不足。结论我们的数据表明，β-catenin功能增益在PONC中的异常表达中断了胚胎发育过程中眼睑闭合的上皮和基质之间的相互作用。

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引用次数: 0

Serum-derived extracellular vesicles for the treatment of severe ocular surface disease 用于治疗严重眼表疾病的血清衍生细胞外囊泡

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-17 DOI: 10.1016/j.jtos.2024.08.009

Namita Saraf , Rajalakshmy Ayilam Ramachandran , Mou Cao , Andrew Lemoff , Hamid Baniasadi , Danielle M. Robertson

Purpose

Autologous serum is widely used for the treatment of severe ocular surface disease with mixed efficacy. Extracellular vesicles (EVs) are small membrane bound structures present in all body fluids, including serum. This study compared the proteomic, metabolomic, and inflammatory cytokine composition of serum-derived EVs (SDEVs) to that of the soluble free protein fraction and the subsequent capacity of SDEVs to induce corneal epithelial cell migration and inflammation.

Methods

SDEVs were isolated from human serum using size exclusion chromatography. SDEVs were analyzed using nanoparticle tracking analysis, transmission electron microscopy, and western blotting. The effects of SDEVs on corneal epithelial cell migration were tested using a standard scratch assay. Inflammatory cytokines in SDEVs and the free protein fraction were quantified using a microarray. A mutli-omics approach was further used to define SDEV cargo. The ability of SDEVs to modulate inflammation in corneal epithelial cells was quantified using ELISAs.

Results

Western blot and TEM confirmed the presence of SDEVs. Proinflammatory cytokines, along with complement proteins and TGF-β, were decreased in SDEVs compared to serum. Metabolites present in SDEVs were mostly involved in amino acid biosynthesis, the TCA cycle and oxidative phosphorylation. SDEVs exhibited pro-migratory effects similar to serum however, SDEVs did not induce secretion of IL-6 or IL-8.

Conclusions

SDEVs exhibit reduced levels of pro-inflammatory cytokines while retaining the beneficial wound healing properties of serum. Unlike serum, SDEVs do not induce inflammation. SDEVs may represent an alternative option for patients with severe ocular surface disease where traditional autologous serum has failed.

目的：自体血清被广泛用于治疗严重的眼表疾病，但疗效参差不齐。细胞外囊泡（EVs）是存在于包括血清在内的所有体液中的小型膜结合结构。本研究比较了血清衍生EVs（SDEVs）与可溶性游离蛋白部分的蛋白质组、代谢组和炎症细胞因子组成，以及SDEVs随后诱导角膜上皮细胞迁移和炎症的能力：方法：使用尺寸排阻色谱法从人血清中分离出 SDEVs。方法：采用尺寸排阻色谱法从人类血清中分离出 SDEV，并利用纳米粒子跟踪分析、透射电子显微镜和 Western 印迹法对 SDEV 进行分析。使用标准划痕试验测试了 SDEV 对角膜上皮细胞迁移的影响。使用芯片对 SDEV 和游离蛋白部分中的炎性细胞因子进行了量化。此外，还采用了一种突变组学方法来确定 SDEV 的载体。使用 ELISAs 对 SDEVs 调节角膜上皮细胞炎症的能力进行了量化：Western印迹和TEM证实了SDEV的存在。与血清相比，SDEVs 中的促炎细胞因子、补体蛋白和 TGF-β 均有所减少。SDEVs 中的代谢物主要参与氨基酸的生物合成、TCA 循环和氧化磷酸化。SDEVs 表现出与血清相似的促迁移作用，但 SDEVs 不会诱导 IL-6 或 IL-8 的分泌：结论：SDEV 降低了促炎细胞因子的水平，同时保留了血清有益的伤口愈合特性。与血清不同，SDEVs 不会诱发炎症。对于传统自体血清治疗失败的严重眼表疾病患者来说，SDEV 可能是一种替代选择。

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引用次数: 0

Tear neuropeptide Y as a non-invasive marker of peripheral microvascular complications in type 1 diabetes 将泪液神经肽 Y 作为 1 型糖尿病患者外周微血管并发症的非侵入性标记物。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-15 DOI: 10.1016/j.jtos.2024.08.011

Alexis Ceecee Britten-Jones , Mengliang Wu , Leslie J. Roberts , Richard J. MacIsaac , Haihan Jiao , Jennifer P. Craig , Holly R. Chinnery , Laura E. Downie

Aims

To investigate tear neuropeptide Y (NPY) and substance P concentrations in individuals with type 1 diabetes, comparing those with and without both diabetic retinopathy (DR) and peripheral neuropathy.

Methods

This cross-sectional study involved 41 participants with type 1 diabetes and none to moderate DR, and 22 healthy controls. Assessments included clinical ocular surface parameters, quantification of corneal nerve attributes (based on in vivo confocal microscopy imaging), DR grading, and evaluation for small and large fibre neuropathy. Concentrations of NPY and substance P in tear samples were measured using enzyme-linked immunosorbent assay.

Results

Mean (± standard deviation) tear NPY concentrations in participants with type 1 diabetes and length-dependent small fibre neuropathy (SFN) was lower than in controls (10.84 ± 4.10 ng/mL vs 14.72 ± 3.12 ng/mL; p=0.004), but not significantly different from type 1 diabetes participants without SFN (13.39 ± 4.66 ng/mL; p=0.11). Tear NPY levels were lower in individuals with type 1 diabetes and mild/moderate non-proliferative DR (10.44 ± 3.46 ng/mL) compared to none/minimal DR (13.79 ± 4.76 ng/mL; p=0.0005) and controls. In separate linear regression models, both the presence of SFN (β = −0.75, p=0.02) and the presence of mild/moderate DR (β = −0.84, p=0.009) were significantly associated with tear NPY levels relative to controls, after adjusting for participant age, sex, and dry eye disease. There were no inter-group differences for tear substance P concentrations.

Conclusions

Tear NPY has potential utility as an indicator of peripheral microvascular complications associated with type 1 diabetes.

目的：研究 1 型糖尿病患者的泪液神经肽 Y (NPY) 和 P 物质浓度，并对同时患有和未患有糖尿病视网膜病变 (DR) 和周围神经病变的患者进行比较：这项横断面研究涉及 41 名患有 1 型糖尿病且无中度 DR 的患者和 22 名健康对照者。评估内容包括临床眼表参数、角膜神经属性量化（基于体内共聚焦显微镜成像）、DR 分级以及小纤维和大纤维神经病变评估。采用酶联免疫吸附法测定了泪液样本中 NPY 和 P 物质的浓度：结果：1 型糖尿病和长度依赖性小纤维神经病变（SFN）患者的泪液 NPY 浓度平均值（± 标准偏差）低于对照组（10.84±4.10 ng/mL vs 14.72±3.12 ng/mL；p=0.004），但与无 SFN 的 1 型糖尿病患者（13.39±4.66 ng/mL；p=0.11）无显著差异。与无/轻度DR（13.79±4.76 ng/mL；p=0.0005）和对照组相比，1型糖尿病和轻度/中度非增生性DR患者的泪液NPY水平较低（10.44±3.46 ng/mL）。在单独的线性回归模型中，在对参与者的年龄、性别和干眼症进行调整后，SFN（β=-0.75，p=0.02）和轻度/中度 DR（β=-0.84，p=0.009）的存在与对照组的泪液 NPY 水平显著相关。泪液物质P浓度没有组间差异：结论：泪液NPY可作为与1型糖尿病相关的外周微血管并发症的指标。

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引用次数: 0

The clinical outcomes of minor salivary gland transplantation for severe dry eye disease secondary to chronic Stevens-Johnson syndrome 小唾液腺移植治疗慢性史蒂文斯-约翰逊综合征继发性严重干眼症的临床疗效。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-14 DOI: 10.1016/j.jtos.2024.08.010

Namrata Sharma, Vishal Kumar, Aafreen Bari, Renu Venugopal, Shivam Sharma, Tushar Agarwal, Tanuj Dada, Neelam Pushker

Purpose

To study the outcomes of minor salivary gland transplantation for severe dry eye disease secondary to chronic Steven Johnson Syndrome.

Methods

It was an ambispective, interventional case series conducted at Rajendra Prasad Centre for Ophthalmic Sciences, Delhi, India from 2022 to 2023 evaluating the outcomes of minor salivary gland transplantation with anchorage of the minor salivary glands to superior rectus muscle in twenty cases of severe dry eye disease secondary to chronic Steven-Johnson Syndrome. The pre-operative clinical parameters were compared to those at post-operative 1 year follow-up.

Results

At 1 year follow-up, there was an improvement in mean Schirmer-1 value (p = 0.0004), hyperemia score (p = 0.0004), keratinization score (p = 0.04), corneal epithelial defect score (p = 0.0004), corneal opacification score (p = 0.001), corneal neovascularization score (p = 0.001), palisades of Vogt score (p = 0.007), corneal keratinization score (p = 0.04) and corneal conjunctivalization score (p = 0.08).

Conclusion

The minor salivary gland transplantation is a viable management option for cases with severe dry eye disease secondary to chronic Steven Johnson Syndrome with clinical improvement in corneal and conjunctival parameters of the ocular surface.

目的：研究小唾液腺移植治疗继发于慢性史蒂文-约翰逊综合征的严重干眼症的效果：这是 2022 年至 2023 年期间在印度德里 Rajendra Prasad 眼科科学中心进行的一项前瞻性、介入性病例系列研究，评估了小唾液腺移植手术的效果，并将小唾液腺固定在上直肌上，用于治疗 20 例继发于慢性史蒂芬-约翰逊综合征的严重干眼症。将术前的临床参数与术后 1 年随访时的临床参数进行了比较：随访 1 年时，Schirmer-1 平均值（p=0.0004）、充血评分（p=0.0004）、角质化评分（p=0.04）、角膜上皮缺损评分（p=0.0004）、角膜不透明评分（p=0.001）、角膜新生血管评分（p=0.001）、Vogt 宫评分（p=0.007）、角膜角质化评分（p=0.04）和角膜结膜化评分（p=0.08）：小唾液腺移植对于继发于慢性史蒂文-约翰逊综合征的严重干眼症病例是一种可行的治疗方案，可改善眼表角膜和结膜的临床参数。

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引用次数: 0

Punctal cautery in dry eye disease: A systematic review 干眼症的眼球穿孔烧灼术：系统回顾。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-09 DOI: 10.1016/j.jtos.2024.08.006

Ashish Ranjan , Sayan Basu , Swati Singh

Purpose

To critically appraise the evidence on the efficacy and recanalization rates of permanent punctal occlusion via thermal or surgical means in managing dry eye disease (DED).

Methods

In PubMed, Scopus, and Cochrane databases, two authors systematically reviewed the literature for prospective studies on punctal cautery or surgical occlusion (excluding punctal plugs) for DED. The studied outcomes were changes in tear volume, tear film stability, punctal recanalization rates, and patient symptomatology.

Results

Nine studies (all single-arm) had 150 subjects (96 females). Five studies were on thermal punctal cauterization, and four used surgical occlusion techniques. One hundred eighty puncta were operated for eyes not responding to maximal lubricants or recurrent plug extrusion. DED etiologies were Sjogren's syndrome (78), cicatricial ADDE (27), graft-versus-host disease (12), and non-SS DED (50). Follow-up ranged from 3 to 24 months. At the final follow-up, improvements in Schirmer I and TBUT were 2.5 mm and 0.8s with thermal and 2.1 mm and 0.6s with surgical methods, respectively (P = 0.17 for Schirmer, P = 0.18 for TBUT). Punctal recanalization rates varied between thermal (0–38.7 %) and surgical (5–9%) techniques (p = 0.22). Different cautery devices show different recanalization rates; disposable thermal cautery tips directly inserted into the punctum had lesser recanalization than radiofrequency monopolar cautery. Most patients reported subjective improvement following the procedure, but no quantification measure was given in the studies. None of the published studies had a comparison group for performing a meta-analysis.

Conclusion

Based on non-comparative studies, thermal or surgical punctal occlusion improves tear volume in DED with similar recanalization rates; however, randomized controlled trials are needed to ascertain the real effects of punctal cautery on DED.

目的：对通过热敷或手术方法永久性阻塞穿刺点治疗干眼症（DED）的疗效和再通率的证据进行严格评估：在 PubMed、Scopus 和 Cochrane 数据库中，两位作者系统地查阅了有关穿刺烧灼或手术闭塞（不包括穿刺栓）治疗 DED 的前瞻性研究文献。研究结果包括泪液量变化、泪膜稳定性、穿刺再通率和患者症状：九项研究（均为单臂研究）共有 150 名受试者（96 名女性）。其中五项研究采用了热穿刺烧灼法，四项采用了手术闭塞技术。对最大润滑剂无效和复发性塞子挤出的眼睛进行了 180 例穿刺手术。DED的病因包括：Sjogren综合征（78例）、卡他性ADE（27例）、移植物抗宿主病（12例）和非SS DED（50例）。随访时间从 3 个月到 24 个月不等。在最后的随访中，热疗法的 Schirmer I 值和 TBUT 值分别提高了 2.5 毫米和 0.8 秒，手术法分别提高了 2.1 毫米和 0.6 秒（Schirmer 值 P=0.17，TBUT 值 P=0.18）。热灼法（0-38.7%）和手术法（5-9%）的穿孔再通率各不相同（P=0.22）。不同的烧灼设备显示出不同的再通率；直接插入穿刺孔的一次性热烧灼刀头的再通率低于射频单极烧灼。大多数患者在术后都表示主观症状有所改善，但研究中并未给出量化指标。所有已发表的研究都没有为进行荟萃分析设立对比组：结论：根据非比较性研究，热疗或手术穿刺闭塞可改善 DED 的泪液量，且再通率相近；但要确定穿刺烧灼术对 DED 的实际效果，还需要进行随机对照试验。

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引用次数: 0

Duloxetine enhances PAX6 expression and suppresses innate immune responses in murine LPS-induced corneal inflammation 度洛西汀能增强 PAX6 的表达，并抑制 LPS 诱导的小鼠角膜炎症中的先天性免疫反应。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-08 DOI: 10.1016/j.jtos.2024.08.008

Petros Moustardas , Mojdeh Abbasi , Dina Javidjam , Cindy Saah Asamoah , Arnaud Schweitzer-Chaput , Salvatore Cisternino , Dominique Bremond-Gignac , Daniel Aberdam , Neil Lagali

Background-aim

PAX6 is a key regulator of eye development and epithelial homeostasis in the cornea. When deficient, chronic corneal inflammation, neovascularization and limbal stem cell deficiency can occur. Here we investigated the potential of duloxetine, a generic serotonin reuptake inhibitor that can upregulate PAX6 in vitro, for its in vivo activity in the context of corneal inflammation.

Methods

Duloxetine tolerance was tested in a human limbal stem cell line and isogenic CRISPR-knockout PAX6^+/− cells. C57BL/6-Wildtype mice were administered duloxetine eye drops at concentrations of 1 μM - 2 mM and tested for toxicity and corneal PAX6 expression. In LPS-induced corneal inflammation in mice, duloxetine's effect on PAX6 expression, corneal opacification and inflammatory responses were evaluated by in vivo corneal imaging, immunostaining, and whole-transcriptome microarray analysis.

Results

No toxicity was observed in vitro for duloxetine concentrations up to 10μΜ. In vivo, duloxetine drops were well-tolerated up to 50 μM. Duloxetine drops at 10μΜ significantly upregulated PAX6 protein levels in the cornea by 30 % within 2 days. In the LPS model, duloxetine resulted in a sustained 33 % PAX6 protein upregulation in the cornea at 7 days, and in reduced opacity within 2 days, accompanied by a significant dampening of IL-17A signaling, neutrophil degranulation, microglial activation, macrophage markers, and MMP expression, despite non-significant changes in total inflammatory cell infiltration.

Conclusion

Short-term administration of a repurposed generic drug, duloxetine, upregulates PAX6 protein levels in the cornea of mice and exerts an anti-inflammatory activity by dampening innate immune responses.

背景-目的：PAX6 是眼睛发育和角膜上皮稳态的关键调节因子。当PAX6缺乏时，会出现慢性角膜炎症、新生血管和角膜缘干细胞缺乏症。杜洛西汀是一种普通的血清素再摄取抑制剂，可在体外上调 PAX6，我们在此研究了其在角膜炎症中的体内活性潜力：方法：在人类角膜缘干细胞系和同源 CRISPR 基因敲除 PAX6+/- 细胞中测试度洛西汀耐受性。给 C57BL/6 野生型小鼠滴入浓度为 1μM - 2mM 的度洛西汀眼药水，并测试其毒性和角膜 PAX6 表达。在 LPS 诱导的小鼠角膜炎中，通过体内角膜成像、免疫染色和全转录组芯片分析，评估了度洛西汀对 PAX6 表达、角膜翳和炎症反应的影响：结果：在体外，度洛西汀浓度达到10μΜ时未观察到毒性。在体内，度洛西汀滴剂的耐受性高达 50μM。10μΜ的度洛西汀滴眼液可在2天内使角膜中的PAX6蛋白水平显著提高30%。在LPS模型中，度洛西汀可使角膜中的PAX6蛋白水平在7天内持续上调33%，并在2天内降低角膜翳，同时显著抑制IL-17A信号传导、中性粒细胞脱颗粒、小胶质细胞活化、巨噬细胞标志物和MMP表达，尽管炎症细胞浸润总量无明显变化：结论：短期服用一种再利用的普通药物度洛西汀可上调小鼠角膜中的 PAX6 蛋白水平，并通过抑制先天性免疫反应发挥抗炎活性。

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引用次数: 0

Pipeline: Wiley Chambers, MD, FDA Ophthalmology regulator 管道：Wiley Chambers, MD, FDA 眼科监管者。

IF 5.9 1区医学 Q1 OPHTHALMOLOGY

Ocular Surface

Pub Date : 2024-08-08 DOI: 10.1016/j.jtos.2024.08.004

Gary D. Novack

引用次数: 0