Experimental eye research最新文献_第4页

Commentary on “A sustainable tissue-specific hydrogel from decellularized corneal ECM for 3D in vitro ocular models” by Santos et al. Santos等人对“一种来自脱细胞角膜ECM的可持续组织特异性水凝胶用于体外3D眼模型”的评论。

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-28 DOI: 10.1016/j.exer.2025.110833

Abhisarika Maduri , Rohit Saxena , Varshini Vadhithala , Sachin Kumar

引用次数: 0

From risk markers to treatable traits in retinal oculomics 从视网膜眼组学的风险标记到可治疗的特征

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-26 DOI: 10.1016/j.exer.2025.110832

Henry Bair

引用次数: 0

Layer-by-layer coated chitosan-CRISPR/Cas9 mTOR nanoparticles: A novel approach to inhibit lens epithelial cell proliferation and migration for preventing posterior capsule opacification 壳聚糖- crispr /Cas9 mTOR纳米颗粒：一种抑制晶状体上皮细胞增殖和迁移以防止后囊膜混浊的新方法

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-26 DOI: 10.1016/j.exer.2025.110828

Chen Chang , Jiaming Yang , Ziming Liu , Jiayu Chen , Biying Wang , Junqi Li , Hongling Liu

Posterior capsular opacification (PCO) is the most common complication following cataract surgery and a significant cause of vision impairment. PCO arises from the proliferation, migration, and epithelial-mesenchymal transition (EMT) of residual lens epithelial cells (LECs), driven by an activated mTOR signalling pathway. Previous research has demonstrated that inhibiting mTOR activity effectively reduces LEC proliferation and EMT in rabbit models. However, achieving sustained mTOR inhibition remains a challenge.

In this study, we encapsulated the CRISPR/Cas9 system targeting mTOR into chitosan nanoparticles (Chi-gRNA) with an average size of 135 nm. These nanoparticles exhibited resistance to DNase I digestion. To prolong release duration, we incorporated these Chi-gRNA nanoparticles onto the surface of intraocular lenses (IOLs) via layer-by-layer (LbL) assembly. The LbL coatings consisted of alternating layers of positively charged polyethyleneimine (PEI) and negatively charged heparin, interspersed with Chi-gRNA nanoparticles over five consecutive cycles.

Spectral analysis confirmed the successful integration and coating of nanoparticles, with characteristic peaks validating the electrostatic assembly of the layers. In vitro assays demonstrated that Chi-gRNA-coated IOLs significantly inhibited the proliferation, migration, and adhesion of human lens epithelial cells (hLECs).

These findings highlight the potential of LbL-coated IOLs to deliver CRISPR/Cas9 system-targeting mTOR nanoparticles as a novel and effective strategy to prevent PCO in patients undergoing cataract surgery. This approach offers a promising avenue for the long-term management of this prevalent postoperative complication.

后囊膜混浊（PCO）是白内障手术后最常见的并发症，也是视力受损的重要原因。残晶状体上皮细胞（LECs）的增殖、迁移和上皮-间质转化（EMT）由激活的mTOR信号通路驱动。先前的研究表明，抑制mTOR活性可有效降低兔模型中LEC增殖和EMT。然而，实现持续的mTOR抑制仍然是一个挑战。在这项研究中，我们将靶向mTOR的CRISPR/Cas9系统封装在平均尺寸为135 nm的壳聚糖纳米颗粒（Chi-gRNA）中。这些纳米颗粒表现出对dna酶I消化的抵抗力。为了延长释放时间，我们将这些Chi-gRNA纳米颗粒通过逐层组装（LbL）结合到人工晶状体（iol）表面。LbL涂层由带正电荷的聚乙烯亚胺（PEI）和带负电荷的肝素交替层组成，并在连续五个循环中点缀Chi-gRNA纳米颗粒。光谱分析证实了纳米粒子的成功集成和涂层，特征峰证实了层的静电组装。体外实验表明，chi - grna包被的iol可显著抑制人晶状体上皮细胞（hLECs）的增殖、迁移和粘附。这些发现强调了lbl涂层iol递送CRISPR/Cas9系统靶向mTOR纳米颗粒作为一种新的有效策略来预防白内障手术患者的PCO的潜力。这种方法为这种常见的术后并发症的长期治疗提供了一条有希望的途径。

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引用次数: 0

Role of hypoxia-related differentially expressed genes in pathogenesis of keratoconus and construction of diagnostic models 低氧相关差异表达基因在圆锥角膜发病机制中的作用及诊断模型的建立

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-24 DOI: 10.1016/j.exer.2025.110827

Mengnan Wu, Mansha He, Yaolei Du

Keratoconus is a dilated corneal disorder caused by multiple factors, which is characterized by progressive thinning and protrusion of the paracentral cornea. It may be influenced both by genetic and environmental factors. Clinically, it manifests as significant irregular astigmatism in the affected eye, which can result in irreversible vision impairment that is challenging to correct with standard spectacles. Nevertheless, the specific hypoxia-related genes implicated in keratoconus, along with their diagnostic significance, remain inadequately explored. This research intend to explore the involvement of hypoxia-related differentially expressed genes in the pathogenesis of keratoconus, aiming to explore new diagnostic methods and therapeutic strategies. By employing bioinformatics methodologies, we analyzed RNA sequencing and microarray data from the GEO database to pinpoint differentially expressed genes associated with hypoxic conditions in keratoconus. Our analysis identified 490 differentially expressed genes, encompassing 62 genes that were upregulated and 428 that were downregulated, indicating their probable role in the advancement of keratoconus. Gene Ontology and pathway enrichment analyses showed that 43 HRDEGs were significantly enriched in various biological processes, underscoring the pivotal influence of hypoxia in the pathophysiology of keratoconus. Additionally, a support vector machine model highlighted nine hypoxia-related differentially expressed genes, while least absolute shrinkage and selection operator regression identified five genes suitable for modeling, culminating in two diagnostic models exhibiting high accuracy (AUC >0.9) for the early detection of keratoconus. Furthermore, the immune cell infiltration analysis revealed a positive association between DUSP1 and follicular helper T cells, as well as a negative association with resting mast cells, indicating an immune-related aspect in the progression of keratoconus. The results of our study lay the groundwork for subsequent investigations into the molecular pathways involved in keratoconus, as well as the creation of specialized diagnostic instruments.

圆锥角膜是一种由多种因素引起的扩张性角膜疾病，以中心旁角膜逐渐变薄和突出为特征。它可能受到遗传和环境因素的影响。在临床上，它表现为受影响的眼睛出现明显的不规则散光，这可能导致不可逆的视力损害，很难用标准眼镜矫正。然而，圆锥角膜中涉及的特定缺氧相关基因及其诊断意义仍未得到充分探讨。本研究旨在探讨缺氧相关的差异表达基因在圆锥角膜发病中的作用，旨在探索新的诊断方法和治疗策略。通过采用生物信息学方法，我们分析了GEO数据库中的RNA测序和微阵列数据，以确定与圆锥角膜缺氧状况相关的差异表达基因。我们的分析确定了490个差异表达基因，其中62个基因上调，428个基因下调，表明它们可能在圆锥角膜的进展中起作用。基因本体和通路富集分析显示，43个hrdeg在多个生物过程中显著富集，提示缺氧在圆锥角膜病理生理中的关键影响。此外，一个支持向量机模型突出了9个与缺氧相关的差异表达基因，而最小绝对收缩和选择算子回归确定了5个适合建模的基因，最终形成了两个诊断模型，在圆锥角膜的早期检测中表现出较高的准确性（AUC >0.9）。此外，免疫细胞浸润分析显示DUSP1与滤泡辅助性T细胞呈正相关，而与静止肥大细胞负相关，表明圆锥角膜的进展与免疫相关。我们的研究结果为后续研究圆锥角膜的分子通路以及创建专门的诊断仪器奠定了基础。

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引用次数: 0

Single zonular fiber extraction and characterization: A platform for biomechanical, biochemical and structural analyses 单带状纤维提取和表征：生物力学、生化和结构分析的平台

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-24 DOI: 10.1016/j.exer.2025.110830

Riley Forinash , Sean Staggs , Steven Bassnett , Juan Rodriguez

Zonular fibers are essential for suspending the lens within the eye and, in primates, for transmitting forces generated by the ciliary body to drive ocular accommodation. Despite their biomechanical importance, the response of these fibers to sudden strains, and the molecular basis of their exceptional durability remain poorly understood. This is due, in part, to the absence of tools for studying them in isolation. Here, we present a custom-built, cost-effective apparatus designed to isolate, manipulate, and characterize individual zonular fibers under controlled physical and chemical conditions. The apparatus enables quantification of elastic moduli, stress-relaxation behavior, Poisson's ratio, and response to chemical or enzymatic treatments all from a single fiber, while supporting simultaneous imaging and post-experiment analyses by atomic force microscopy (AFM) or other modalities. We describe the design and calibration of the system, provide protocols for sample mounting and mechanical testing, and introduce accessories for high-resolution imaging and temperature control. Using bovine zonular fibers as a model, we demonstrate the system's ability to capture dynamic responses to enzymatic digestion, including time-resolved degradation kinetics following exposure to α-amylase and trypsin. The modular design, precision, and affordability of the apparatus make it a valuable platform for investigating the mechanics of fine elastic fibers in the eye and beyond, with relevance to connective tissue biology across organ systems.

带状纤维对于悬浮眼内晶状体至关重要，在灵长类动物中，用于传递睫状体产生的力以驱动眼部调节。尽管具有重要的生物力学意义，但这些纤维对突然应变的反应，以及其特殊耐久性的分子基础仍然知之甚少。这在一定程度上是由于缺乏单独研究它们的工具。在这里，我们提出了一种定制的，具有成本效益的设备，用于在受控的物理和化学条件下分离，操作和表征单个带状纤维。该仪器可以量化弹性模量、应力松弛行为、泊松比以及对化学或酶处理的反应，所有这些都来自一根纤维，同时支持通过原子力显微镜（AFM）或其他方式进行同步成像和实验后分析。我们描述了该系统的设计和校准，提供了样品安装和机械测试的协议，并介绍了用于高分辨率成像和温度控制的附件。使用牛带状纤维作为模型，我们证明了该系统捕捉酶消化动态响应的能力，包括暴露于α-淀粉酶和胰蛋白酶后的时间分辨降解动力学。该仪器的模块化设计、精度和可负担性使其成为研究眼部及其他部位精细弹性纤维力学的有价值的平台，与跨器官系统的结缔组织生物学相关。

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引用次数: 0

Retinal organoids in age-related macular degeneration: Promise, pitfalls, and progress 视网膜类器官在年龄相关性黄斑变性：前景，缺陷和进展

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-24 DOI: 10.1016/j.exer.2025.110831

Anwar A. Palakkan, Gopika S. Kumar

Recent advancements in organoid technology have revolutionized our understanding of organ development and disease modelling. However, despite significant progress, retinal organoids continue to face several limitations, including imperfect architecture, incomplete maturation, limited functional integration, and lack of vascularization. These challenges hinder their potential as accurate models of human retinal biology and disease. In this review, we examine the currently available models of age-related macular degeneration, evaluate the potential of retinal organoids, and discuss their respective advantages and limitations. We also explore strategies to overcome the existing challenges in retinal organoid systems, drawing parallels from progress made in other organoid fields, where similar limitations have been addressed. Implementing these strategies may enhance the structural and functional fidelity of retinal organoids, thereby improving their utility in modelling retinal disorders and advancing regenerative medicine.

最近类器官技术的进步彻底改变了我们对器官发育和疾病建模的理解。然而，尽管取得了重大进展，视网膜类器官仍然面临着一些限制，包括不完善的结构，不完整的成熟，有限的功能整合和缺乏血管化。这些挑战阻碍了它们作为人类视网膜生物学和疾病精确模型的潜力。在这篇综述中，我们研究了目前可用的年龄相关性黄斑变性模型，评估了视网膜类器官的潜力，并讨论了它们各自的优点和局限性。我们还探讨了克服视网膜类器官系统中现有挑战的策略，并从其他类器官领域取得的进展中得出了相似的结论，这些领域已经解决了类似的局限性。实施这些策略可以提高视网膜类器官的结构和功能保真度，从而提高它们在视网膜疾病建模和推进再生医学方面的应用。

引用次数: 0

ATAC-seq revealing chromatin accessibility and novel motifs linked to corneal fibrosis ATAC-seq揭示与角膜纤维化相关的染色质可及性和新基序。

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-24 DOI: 10.1016/j.exer.2025.110826

Swati Sood , Rajnish Kumar , Nishant R. Sinha , Rajiv R. Mohan

Corneal fibrosis results in loss of corneal transparency and vision impairment. Dynamic chromatin accessibility is a key regulatory mechanism driving fibrotic process, allowing transcription factors to access specific DNA sequences to activate or shut down transcription of pro- and anti-fibrotic genes. Genome-wide chromatin accessibility profiling was employed to explore the epigenomic landscape in alkali-induced fibrosis in rabbit cornea to identify key fibrotic regulators. Chromatin accessible regions (peaks) were identified using MACS2 and annotated relative to genomic features using ChIPseeker. de novo motif enrichment was evaluated by HOMER. Differentially accessible regions (DARs) were quantified with csaw and edgeR followed by pathway enrichment analysis using clusterProfiler. Of the total 121,305 peaks identified across all samples, 5890 DARs (FDR<0.05) were identified exhibiting reduced accessibility. Distinct chromatin condensation was observed at promoter region in fibrotic (5 %) vs. naïve (15 %) corneas. Functional enrichment analysis revealed that cell cycle, metabolism and stress response regulation genes were the most affected. de novo motifs analysis identified motifs with high similarities to KLF17 (p = 1e⁻⁴⁹¹), NRF2 (p = 1e⁻⁴⁸³), and ETV6 (p = 1e⁻²⁰¹) as potential regulators of corneal fibrosis. The qRT-PCR evaluation of motifs in human corneal myofibroblasts showed low expression of KLF17 (p < 0.05) and NRF2 (p = 0.23), and elevated levels of ETV6 (p = 0.64) compared to fibroblast cells. ATAC-sequences are made available at the NCBI Gene Expression Omnibus (GEO accession no. GSE305524). Fibrosis triggers extensive epigenomic reprogramming, characterized by promoter-to-enhancer regulatory shifts and selective activation of fibrosis-associated transcriptional networks providing promising avenue for therapeutic interventions managing corneal fibrosis.

角膜纤维化导致角膜透明度丧失和视力损害。动态染色质可及性是驱动纤维化过程的关键调控机制，允许转录因子进入特定的DNA序列，激活或关闭促纤维化和抗纤维化基因的转录。采用全基因组染色质可及性分析来探索兔角膜碱诱导纤维化的表观基因组景观，以确定关键的纤维化调节因子。使用MACS2识别染色质可接近区域（峰），并使用ChIPseeker根据基因组特征进行注释。用HOMER评价de novo motif富集程度。用csaw和edgeR对差异可达区（dar）进行量化，然后用clusterProfiler进行通路富集分析。在所有样本中鉴定的121305个峰中，5890个dar （FDR-491）、NRF2 （p=1e-483）和ETV6 （p=1e-201）是角膜纤维化的潜在调节因子。qRT-PCR对人角膜肌成纤维细胞中基元的评价显示KLF17的低表达(p

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引用次数: 0

Development of a multi-parametric MRI platform to evaluate steady-state water content and optical properties in multiple ex vivo bovine lens 多参数磁共振成像平台的开发，以评估稳态含水量和光学性质的多个离体牛晶状体。

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-23 DOI: 10.1016/j.exer.2025.110822

Xingzheng Pan , Yadi Chen , Beau Pontre , Catherine Morgan , Jin Jin , Chen Qiu , Courtney A. Thorne , Thomas W. White , Paul J. Donaldson

Here we describe the development of magnetic resonance imaging (MRI) protocols which when run on a standard clinical 3T MRI system can non-invasively monitor changes in the water content and refractive properties of bovine lenses maintained under organ culture. Fresh bovine lenses were incubated in artificial aqueous humour and placed in a custom holder within a hand/wrist coil. Multi-parametric mapping sequences (T1, T2∗, proton density [PD], and magnetisation transfer [MT] ratio) were optimised to quantify the total, free, and bound water compartments in distinct lens regions (outer cortex, inner cortex, and nucleus). Repeated scanning on different days demonstrated robust inter-sample reproducibility. MRI-derived T2∗ and MT ratio values were calibrated against refractive index (n) profiles calculated using laser ray tracing and subsequently applied to ZEMAX optical modelling to calculate lens power. The imaging protocol achieved high spatial resolution and reproducibility, which allowed the detection of regional variations in water composition within a practical scan time. Hypotonic lens swelling induced significant increases in total and free water across all lens regions but had no effect on bound water or the refractive index gradient. Lens swelling increased lens power (ΔP = 2.98D, p = 0.04) primarily due to a change in the surface geometry of the lens. This ability to quantify lens water states and refractive changes across multiple lens samples will facilitate the screening of novel pharmacological reagents that modulate lens water transport, allowing for testing of their efficacy as potential anti-cataract therapies.

在这里，我们描述了磁共振成像（MRI）方案的发展，当在标准的临床3T MRI系统上运行时，可以无创地监测器官培养下维持的牛晶状体含水量和折射特性的变化。新鲜的牛晶状体在人工幽默水中孵育，并放置在手/手腕线圈内的定制支架中。优化了多参数图谱序列（T1、T2*、质子密度[PD]和磁化转移[MT]比率），以量化不同晶状体区域（外皮层、内皮层和核）中的总水区、游离水区和结合水区。不同日期的重复扫描显示了样本间的可重复性。根据激光射线追踪计算的折射率(n)曲线，对mri导出的T2*和MT比值值进行校准，随后将其应用于ZEMAX光学建模以计算透镜功率。成像方案实现了高空间分辨率和再现性，从而可以在实际扫描时间内检测到水成分的区域变化。低渗晶状体肿胀导致晶状体各区域的总水和自由水显著增加，但对束缚水和折射率梯度没有影响。透镜膨胀增加了透镜的功率（ΔP = 2.98D, p = 0.04），主要是由于透镜表面几何形状的变化。这种量化晶状体水分状态和多个晶状体样品屈光变化的能力，将有助于筛选调节晶状体水分运输的新型药理试剂，从而测试其作为潜在抗白内障疗法的功效。

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引用次数: 0

Development of a microscopy-based diagnostic test for alkali injury-induced limbal stem cell deficiency through Autofluorescence Multispectral Imaging 基于自体荧光多光谱成像的碱损伤诱导的角膜缘干细胞缺乏症显微诊断方法的建立。

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-23 DOI: 10.1016/j.exer.2025.110829

Xiaohu Xu , Lina Sprogyte , Alexander Richardson , Lamia Nureen , Ewa Magdalena Goldys , Nick Di Girolamo , Abbas Habibalahi

Diagnosing limbal stem cell deficiency (LSCD) remains challenging, and this study evaluates a newly developed Autofluorescence Multispectral Imaging (AFMI) technology for its diagnostic potential. Unstained fixed corneal cross-sections from 36 mice were used in this study. The right eyes of these mice had alkali-induced LSCD, while the left eyes served as healthy controls. Autofluorescence multispectral images of the corneal epithelium were acquired across multiple spectral channels to track changes in endogenous fluorophore spectra. Big data analytics and machine learning classifiers were applied to differentiate LSCD-affected from healthy tissues. The performance of this technique was evaluated by histological evaluation. Imaging was conducted on both the Operetta CLS (PerkinElmer) for research applications and a custom-modified Olympus IX83 system designed for potential clinical translation. Significant differences in autofluorescence signal intensities (p < 0.0001) were observed between diseased and control tissues across several key fluorophores. The machine learning classifier achieved high prediction accuracy using both the Operetta CLS system (90.38 %) and a custom-modified Olympus IX83 platform (83.28 %) during 10-fold cross-validation, and the false-color prediction maps showed strong agreement with histological assessments. These results highlight new perspectives on LSCD through the characterization of autofluorescent biomarkers and demonstrate the feasibility of AFMI as a diagnostic tool in a mouse model, providing a basis for future investigations into its possible adaptation for non-invasive clinical assessment of LSCD and other ocular surface disorders.

诊断角膜缘干细胞缺乏症（LSCD）仍然具有挑战性，本研究评估了新开发的自体荧光多光谱成像（AFMI）技术的诊断潜力。本研究采用36只小鼠未染色的固定角膜横切面。这些小鼠的右眼患有碱诱导的LSCD，而左眼作为健康对照。通过多个光谱通道获得角膜上皮的自体荧光多光谱图像，以跟踪内源性荧光团光谱的变化。应用大数据分析和机器学习分类器区分受lscd影响的组织和健康组织。通过组织学评价评价该技术的性能。在用于研究应用的Operetta CLS （Perkin Elmer）和用于潜在临床翻译的定制改进的Olympus IX83系统上进行成像。自体荧光信号强度的显著差异(p

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引用次数: 0

An improved and more stable mouse model of lens-induced myopia 一种改进且更稳定的晶状体性近视小鼠模型。

IF 2.7 2区医学 Q1 OPHTHALMOLOGY

Experimental eye research

Pub Date : 2025-12-22 DOI: 10.1016/j.exer.2025.110825

Yining Shen , Jinfang Huang , Leilei Zou , Yuanyang Huang , Rui Duan , Huiling Shen , Hong Liu

The increasing prevalence of myopia is a pressing public health concern. Robust experimental models are needed to investigate its mechanisms and evaluate potential interventions. Mouse models are particularly advantageous due to their concise and well-characterized genetic background, which facilitates genetic manipulation. However, the small head and eye size, combined with high activity levels of mice, present substantial challenges for the stable establishment of myopia models. The lens-induced myopia (LIM) model, closely resembling the natural progression of human myopia, offers significant research value. Here, we present an improved and reproducible mouse LIM model incorporating a 3D-printed head-mounted spectacle frame and custom-designed high-strength, wear-resistant resin lenses. The device was securely affixed to the skull using dental resin and tissue adhesive, ensuring long-term stability while minimizing interference with feeding and grooming behavior. Four-week induction with −25 diopter lenses in four-week-old C57BL/6J mice resulted in a mean myopic shift of 5.19 D and axial elongation of 110 μm. Electroretinography confirmed preserved retinal function, and the lens detachment rate remained below 15 %. No significant differences in general health status were observed between LIM and control groups. This optimized LIM model provides a reliable, well-tolerated platform for experimental myopia studies and is well-suited for mechanistic and interventional research in mice.

近视日益普遍是一个紧迫的公共卫生问题。需要稳健的实验模型来研究其机制并评估潜在的干预措施。小鼠模型由于其简洁和特征良好的遗传背景而特别有利，这有利于遗传操作。然而，小的头和眼睛尺寸，加上小鼠的高活动水平，为稳定建立近视模型提出了实质性的挑战。透镜性近视（LIM）模型与人类近视的自然发展过程非常相似，具有重要的研究价值。在这里，我们提出了一种改进的、可重复的小鼠LIM模型，该模型结合了3d打印的头戴式眼镜框架和定制的高强度、耐磨树脂镜片。该装置使用牙科树脂和组织粘合剂牢固地固定在颅骨上，确保长期稳定性，同时最大限度地减少对喂食和梳理行为的干扰。4周龄C57BL/6J小鼠用-25屈光度透镜诱导4周后，平均近视位移5.19 D，轴向伸长110 μm。视网膜电图证实视网膜功能保留，晶状体脱离率保持在15%以下。LIM组与对照组在总体健康状况上无显著差异。优化后的LIM模型为实验近视研究提供了一个可靠、耐受性良好的平台，非常适合小鼠的机制和介入性研究。

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引用次数: 0