Purpose

Cornea epithelial-stromal scarring is related to the differentiation of fibroblasts into opaque myofibroblasts. Our study aims to assess the effectiveness of Lycium barbarum polysaccharide (LBP) solution as a pre-treatment in minimizing corneal scarring.

Methods

Human corneal fibroblasts were cultured in a three-dimensional collagen type I-based hydrogel in an eye-on-a-chip model. Fibroblasts were pre-treated with 2 mg/mL LBP for 24 h, followed by another 24-h incubation with 10 ng/mL transforming growth factor-beta 1 (TGF-β1) to induce relevant physiological events after stromal injury. Intracellular pro-fibrotic proteins, extracellular matrix proteins, and pro-inflammatory cytokines that involved in fibrosis, were assessed using immunocytochemistry and enzyme-linked immunosorbent assays.

Results

Compared to the positive control TGF-β1 group, LBP pre-treated cells had a significantly lower expression of alpha-smooth muscle actin, marker of myofibroblasts, vimentin (p < 0.05), and also extracellular matrix proteins both collagen type II and type III (p < 0.05) that can be found in scar tissues. Moreover, LBP pre-treated cells had a significantly lower secretion of pro-inflammatory cytokines interleukin-6 and interleukin-8 (p < 0.05). The cell-laden hydrogel contraction and stiffness showed no significant difference between LBP pre-treatment and control groups. Fibroblasts pretreated with LBP as well had reduced angiogenic factors expression and suppression of undesired proliferation (p < 0.05).

Conclusion

Our results showed that LBP reduced both pro-fibrotic proteins and pro-inflammatory cytokines on corneal injury in vitro. We suggest that LBP, as a natural Traditional Chinese Medicine, may potentially be a novel topical pre-treatment option prior to corneal refractive surgeries with an improved prognosis.

中文翻译：

---

枸杞多糖对角膜上皮间质损伤的抗瘢痕化作用

目的

角膜上皮间质瘢痕形成与成纤维细胞分化为不透明的肌成纤维细胞有关。我们的研究旨在评估枸杞多糖 (LBP) 溶液作为减少角膜瘢痕的预处理的有效性。

方法

人角膜成纤维细胞在基于 I 型胶原蛋白的三维水凝胶中以芯片眼模型进行培养。成纤维细胞用 2 mg/mL LBP 预处理 24 小时，然后与 10 ng/mL 转化生长因子-β1 (TGF-β1) 再孵育 24 小时，以诱导基质损伤后的相关生理事件。使用免疫细胞化学和酶联免疫吸附试验评估了参与纤维化的细胞内促纤维化蛋白、细胞外基质蛋白和促炎细胞因子。

结果

与阳性对照 TGF-β1 组相比，LBP 预处理细胞的 α-平滑肌肌动蛋白、肌成纤维细胞标志物、波形蛋白 (p < 0.05) 以及细胞外基质蛋白 II 型和 III 型胶原蛋白的表达显着降低(p < 0.05) 可以在疤痕组织中找到。此外，LBP 预处理细胞的促炎细胞因子白细胞介素 6 和白细胞介素 8 的分泌显着降低（p < 0.05）。LBP 预处理组和对照组之间的细胞负载水凝胶收缩和刚度没有显着差异。用 LBP 预处理的成纤维细胞也降低了血管生成因子的表达并抑制了不需要的增殖 (p < 0.05)。

结论

我们的结果表明，LBP在体外减少了角膜损伤的促纤维化蛋白和促炎细胞因子。我们建议 LBP 作为一种天然中药，可能是角膜屈光手术前的一种新型局部预处理选择，具有改善的预后。