Karkada Sharel P, Karkada Sheril P, Bhatkal Anushree, Balaraj Srihari, Mukherji Ritica, Ghosh Arkasubhra, Khamar Pooja, Kumar Nimisha R
GROW Research Laboratory, Narayana Netralaya Foundation, Bangalore, Karnataka, India.
Department of Cornea and Refractive Surgery, Narayana Nethralaya, Bangalore, Karnataka, India.
Indian J Ophthalmol. 2025 Jun 1;73(Suppl 3):S435-S443. doi: 10.4103/IJO.IJO_2151_24. Epub 2025 May 6.
Ocular surface discomfort and dry eye disease (DED) are the most common conditions addressed by ophthalmologists worldwide. Artificial tear substitutes are used as the first line of treatment management for DED patients. The present study was performed to understand the role of artificial tear formulation namely Soha Liquigel (0.18% sodium hyaluronate with trehalose) and Soha (0.1% sodium hyaluronate) for the treatment of DED in vitro . Human corneal epithelial (HCE) cells were used in adapted cell culture conditions which induce relevant cellular and molecular modifications thus mimicking the DED.
Artificial tears containing either sodium hyaluronate (SH) (Soha 0.1%, Sun Pharma) or a combination of SH with trehalose (Soha Liquigel 0.18%, Sun Pharma) were compared with respective controls to analyze the effect on desiccation-induced stress or oxidative stress or hyperosmolarity induced stress on HCE cells. Cellular viability was evaluated using the trypan blue assay, while epithelial morphology was observed under light microscopy. Real-time polymerase chain reaction (RT-PCR) was utilized to analyze the transcriptional profile of a specific set of gene signatures, namely S100A7, FOS, SOD-2, COX2, TonEBP, IL6, MCP1, and IL10.
The response of HCE cells to desiccation stress (24 hr) was observed through alterations in their cellular morphology, which were subsequently restored by applying Soha Liquigel. Oxidative stress was induced using 100 nM of H 2 O 2 on HCE cells (short- 24 h and long-term 5 days) and measured using increased expression of S100A7, an oxidative stress-responsive gene. Oxidative-stressed HCE cells after treatment with Soha Liquigel showed reduced pro-oxidant gene and COX2 expression and elevated anti-oxidant genes, FOS, and SOD levels. HCE cells were subjected to +100mOsmol and +200mOsmol NaCl-containing media, inducing hyperosmolar stress that imitates the symptoms of DED. Further, these hyperosmolar stressed cells were treated with Soha Liquigel and Soha eye drops for 24 h and 5 days. Both eye drops rescued the cell morphology under hyperosmolar conditions in both short- and long-term treatments. Increased TonEBP levels confirm the osmotic stress in HCE cells. Reduction in IL6, MCP1, TonEBP, and elevated expression of IL10 in hyperosmotic stressed HCE cells treated with either of the artificial tears indicates their osmo-protection properties.
By using desiccation, oxidative, and hyperosmolar stress simulated in HCE cells in culture, we observed that SH-containing artificial tears provided bio-protection, osmo-protection, and anti-oxidant benefits that were further strengthened with SH and trehalose combination.
眼表不适和干眼症（DED）是全球眼科医生诊治的最常见病症。人工泪液替代品被用作干眼症患者治疗管理的一线用药。本研究旨在了解人工泪液制剂即索哈凝胶（含0.18%透明质酸钠和海藻糖）和索哈（含0.1%透明质酸钠）在体外治疗干眼症中的作用。在适应性细胞培养条件下使用人角膜上皮（HCE）细胞，这种条件可诱导相关细胞和分子改变，从而模拟干眼症。
将含透明质酸钠（SH）（0.1%索哈，太阳制药）或SH与海藻糖组合（0.18%索哈凝胶，太阳制药）的人工泪液与各自的对照进行比较，以分析其对HCE细胞干燥诱导应激、氧化应激或高渗诱导应激的影响。使用台盼蓝试验评估细胞活力，同时在光学显微镜下观察上皮形态。利用实时聚合酶链反应（RT-PCR）分析一组特定基因特征即S100A7、FOS、SOD-2、COX2、TonEBP、IL6、MCP1和IL10的转录谱。
通过观察HCE细胞形态变化来观察其对干燥应激（24小时）的反应，随后应用索哈凝胶可使其恢复。用100 nM过氧化氢对HCE细胞诱导氧化应激（短期24小时和长期5天），并通过氧化应激反应基因S100A7表达增加来测定。用索哈凝胶处理后的氧化应激HCE细胞显示促氧化基因和COX2表达降低，抗氧化基因、FOS和SOD水平升高。使HCE细胞置于含+100 mOsmol和+200 mOsmol NaCl的培养基中，诱导高渗应激，模拟干眼症症状。此外，用索哈凝胶和索哈滴眼液对这些高渗应激细胞进行24小时和5天的处理。两种滴眼液在短期和长期处理中均能在高渗条件下挽救细胞形态。TonEBP水平升高证实了HCE细胞中的渗透应激。用任一种人工泪液处理的高渗应激HCE细胞中IL6、MCP1、TonEBP降低以及IL10表达升高表明它们具有渗透保护特性。
通过在培养的HCE细胞中模拟干燥、氧化和高渗应激，我们观察到含SH的人工泪液具有生物保护、渗透保护和抗氧化益处，而SH与海藻糖组合可进一步增强这些益处。
