Epithelial Basement Membrane Regeneration After PRK-Induced Epithelial-Stromal Injury in Rabbits: Fibrotic Versus Non-fibrotic Corneal Healing.
Epithelial Basement Membrane Regeneration After PRK-Induced Epithelial-Stromal Injury in Rabbits: Fibrotic Versus Non-fibrotic Corneal Healing.
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DOI：
10.3928/1081597x-20211007-02
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发表时间：
2022-01
期刊：
影响因子：
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通讯作者：
Wilson SE
中科院分区：
文献类型：
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作者：
de Oliveira RC;Sampaio LP;Shiju TM;Santhiago MR;Wilson SE
研究屈光性角膜切除术（PRK）后非纤维化和纤维化角膜的上皮基底膜（EBM）再生。兔子（总共 120 只）要么仅进行上皮刮擦、-4.5 屈光度 (D) PRK、-9D PRK，要么不进行手术。在未受伤至八周的时间点对冷冻固定的角膜进行免疫组织化学 (IHC)（每组每个时间点四个角膜）。对 EBM 成分进行多重 IHC，包括 IV 型胶原、层粘连蛋白 β 3、层粘连蛋白 α 5、基底膜聚糖和 nidogen-1。通过三重 IHC 研究了基质细胞组成中的角蛋白、波形蛋白和 α-平滑肌肌动蛋白 (SMA)。术后 4 天，PRK 损伤的 EBM 显着再生。然而，纤维化角膜手术后 4 至 7 天，早期 TGF β 调节基底膜聚糖与新生 EBM 的结合量下降。具有完全再生 EBM（所有五种成分均纳入 EBM）的非纤维化角膜是透明的，并且基质中几乎没有 SMA 阳性肌成纤维细胞。相反，具有缺陷的新生EBM且缺乏基底膜聚糖的角膜在术后3至4周出现许多前基质肌成纤维细胞和纤维化，并且在新生EBM和前基质中具有大量IV型胶原。肌成纤维细胞合成基底膜聚糖，但无法将硫酸肝素蛋白聚糖整合到新生的 EBM 中。即使在纤维化角膜中，角膜透明度也会在几个月内恢复，这与 EBM 基底膜蛋白的恢复、肌成纤维细胞消失和紊乱的细胞外基质的重吸收有关。基底膜蛋白聚糖通过上皮下肌成纤维细胞（可能还有其前体细胞）有缺陷地掺入再生 EBM，是 PRK 角膜损伤后基质纤维化发生和持续的基础。
To study epithelial basement membrane (EBM) regeneration in non-fibrotic and fibrotic corneas after photorefractive keratectomy (PRK). Rabbits (120 total) had either epithelial scrape alone, −4.5 diopter (D) PRK, −9D PRK, or no surgery. Immunohistochemistry (IHC) was performed on cryofixed corneas at time points from unwounded to eight weeks (four corneas at each time point in each group). Multiplex IHC was performed for EBM components, including collagen type IV, laminin beta 3, laminin alpha 5, perlecan, and nidogen-1. Stromal cellular composition was studied by triplex IHC for keratocan, vimentin, and alpha-smooth muscle actin (SMA). PRK-injured EBM significantly regenerated by 4 days after surgery. However, early TGF beta-regulating perlecan incorporation into the nascent EBM declined 4 to 7 days after surgery in fibrotic corneas. Non-fibrotic corneas that had fully-regenerated EBM (with all five components incorporated into the EBM) were transparent, and had few SMA-positive myofibroblasts in the stroma. Conversely, corneas with defective nascent EBM that lacked perlecan developed many anterior stromal myofibroblasts and fibrosis at 3 to 4 weeks after surgery and had large amounts of collagen type IV in the nascent EBM and anterior stroma. Myofibroblasts synthesized perlecan but were incompetent to incorporate the heparin sulfate proteoglycan into the nascent EBM. Corneal transparency was restored over several months even in fibrotic corneas, and this was associated with a return of EBM perlecan, myofibroblast disappearance and reabsorption of disordered extracellular matrix. Defective incorporation of perlecan into the regenerating EBM by subepithelial myofibroblasts, and likely their precursor cells, underlies the development and persistence of stromal fibrosis after PRK corneal injury.