Ultrastructural Aspects of Corneal Functional Recovery in Rats Following Intrastromal Keratocyte Injection.

IF 5 2区 医学 Q1 OPHTHALMOLOGY
Qian Ma, Andri K Riau, Robert D Young, James S Bell, Olga Shebanova, Nicholas J Terrill, Gary H F Yam, Evelina Han, Keith M Meek, Jodhbir S Mehta, Craig Boote
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Abstract

Purpose: Donor tissue shortfalls and postsurgical complications are driving novel corneal tissue regeneration approaches. Corneal stromal keratocytes (CSKs) have shown promise in promoting corneal repair and restoring transparency. We investigated the impact of intrastromal CSK injection on corneal ultrastructure and proteoglycan (PG) distribution in a rat injury model.

Methods: Rats were divided into four groups: normal (n = 12), injured (irregular phototherapeutic keratectomy centrally; n = 6), CSK (injured + human CSK intrastromal injection; n = 6), and PBS (injured + PBS injection; n = 6). Three weeks after treatment, corneas were examined by slit-lamp and optical coherence tomography. Corneal ultrastructure was analysed via small-angle x-ray scattering (collagen fibril diameter, interfibrillar spacing and matrix order), transmission electron microscopy with cuprolinic blue before and after chondroitinase digestion (CS/DS and KS PGs), and immunofluorescence staining (lumican and decorin).

Results: Irregular phototherapeutic keratectomy caused corneal opacity and significantly disrupted stromal ultrastructure, characterized by increased haze density (P < 0.0001), change in central corneal thickness (P = 0.0005), and interfibrillar spacing (P < 0.0001), along with decreased fibril diameter (P < 0.0001), matrix order (P < 0.0001), CS/DS (P < 0.0001) and KS (P < 0.0001) PGs, lumican, and decorin. CSK injection recovered corneal clarity and native stromal ultrastructure, with haze density (P = 0.8086), change in central corneal thickness (P = 0.9503), fibril diameter (P = 0.1139), interfibrillar spacing (P = 0.5879), matrix order (P = 0.9999), CS/DS (P = 0.9969) and KS (P = 0.2877) PGs, lumican, and decorin returning to normal. In contrast, the PBS group exhibited similar corneal injury responses to the injured group.

Conclusions: CSK injection resolved early stage corneal scarring by restoring stromal collagen arrangement and PG distribution, further endorsing its potential for treating corneal opacities.

角膜上皮内角质细胞注射后大鼠角膜功能恢复的超微结构研究。
目的:供体组织短缺和术后并发症正在推动新的角膜组织再生方法。角膜基质角质细胞(CSKs)在促进角膜修复和恢复透明度方面显示出前景。我们研究了角膜基质内注射CSK对大鼠角膜损伤模型超微结构和蛋白多糖(PG)分布的影响。方法:将大鼠分为4组:正常大鼠(n = 12)、损伤大鼠(中央不规则光疗性角膜切除术;n = 6)、CSK(受伤+人CSK基质内注射;n = 6), PBS(受伤+ PBS注射;n = 6)。治疗3周后,采用裂隙灯和光学相干断层扫描检查角膜。采用小角度x射线散射法(胶原原纤维直径、纤维间间距、基质有序)、软骨素酶解前后用铜丙蓝透射电镜(CS/DS和KS PGs)、免疫荧光染色法(lumican和decorin)分析角膜超微结构。结果:不规则光治疗性角膜切除术导致角膜混浊,间质超微结构明显破坏,表现为雾密度增加(P < 0.0001),角膜中央厚度改变(P = 0.0005),纤维间距改变(P < 0.0001),纤维直径(P < 0.0001),基质有序(P < 0.0001), CS/DS (P < 0.0001)和KS (P < 0.0001) PGs, lumican和decorin减少(P < 0.0001)。注射CSK恢复角膜清晰度和原基质超微结构,雾度密度(P = 0.8086)、角膜中央厚度(P = 0.9503)、原纤维直径(P = 0.1139)、原纤维间距(P = 0.5879)、基质有序(P = 0.9999)、CS/DS (P = 0.9969)、KS (P = 0.2877)、PGs、lumican、decorin恢复正常。相比之下,PBS组表现出与受伤组相似的角膜损伤反应。结论:CSK注射剂通过恢复间质胶原排列和PG分布来缓解早期角膜瘢痕形成,进一步证实了其治疗角膜混浊的潜力。
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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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