Md Yusof Alia, Abd Ghafar Norzana, Kamarudin Taty Anna, Chua Kien-Hui, Azmi Muhammad Fairuz, Ng Sook-Luan, Mohd Yusof Yasmin Anum
Department of Anatomy, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latiff, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia.
Department of Basic Science, Faculty of Health Sciences, Universiti Teknologi MARA Cawangan Selangor Puncak Alam, 42300, Puncak Alam, Selangor, Malaysia.
Cytotechnology. 2019 Dec;71(6):1121-1135. doi: 10.1007/s10616-019-00349-8. Epub 2019 Oct 12.
This study evaluated the effects of Gelam honey (GH) on ex vivo corneal fibroblast ulcer model via wound healing assay, gene expression and immunocytochemistry. Corneal fibroblasts from New Zealand white rabbits were culture expanded. The corneal fibroblast wound healing capacity was observed by creating a circular wound onto confluent monolayer cells cultured in basal medium (BM), BM with GH, serum-enriched basal medium (BMS) and BMS with GH respectively. Wound healing assay and phenotypic characterization of the corneal fibroblast were performed at different stages of wound closure. Expression of aldehyde dehydrogenase (ALDH), vimentin, α-smooth muscle actin (α-SMA), lumican, collagen I and matrix metalloproteinase 12 (MMP 12) were measured at day 1, day 3 and complete wound closure day. Corneal fibroblast cultured in BMS with GH demonstrated the fastest wound closure, at day 5 post wounding. The gene expressions of ALDH and vimentin were higher than control groups while α-SMA expression was lower, in GH enriched media. The expressions of lumican, collagen I and MMP 12 were also higher in cells cultured in GH enriched media compared to the control groups. GH was shown to promote in vitro corneal fibroblast wound healing and may be a potential natural adjunct in the treatment of corneal wound.
本研究通过伤口愈合试验、基因表达和免疫细胞化学方法，评估了吉拉蜂蜜（GH）对离体角膜成纤维细胞溃疡模型的影响。对新西兰白兔的角膜成纤维细胞进行培养扩增。分别在基础培养基（BM）、含GH的BM、富血清基础培养基（BMS）和含GH的BMS中，在汇合的单层细胞上制造圆形伤口，观察角膜成纤维细胞的伤口愈合能力。在伤口闭合的不同阶段进行角膜成纤维细胞的伤口愈合试验和表型特征分析。在第1天、第3天和伤口完全闭合日测量醛脱氢酶（ALDH）、波形蛋白、α平滑肌肌动蛋白（α-SMA）、核心蛋白聚糖、胶原蛋白I和基质金属蛋白酶12（MMP 12）的表达。在含GH的BMS中培养的角膜成纤维细胞在受伤后第5天显示出最快的伤口闭合。在富含GH的培养基中，ALDH和波形蛋白的基因表达高于对照组，而α-SMA表达较低。与对照组相比，在富含GH的培养基中培养的细胞中，核心蛋白聚糖、胶原蛋白I和MMP 12的表达也更高。结果表明，GH可促进体外角膜成纤维细胞伤口愈合，可能是治疗角膜伤口的一种潜在天然辅助剂。
