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The effects of acacia honey on in vitro corneal abrasion wound healing model.

Ker-Woon C, Abd Ghafar N, Hui CK, Mohd Yusof YA, Wan Ngah WZ - BMC Cell Biol. (2015)

Bottom Line: CEC cultured in AH-supplemented media showed higher percentage of wound closure compared to the controls.Gene expression of CK3 increased in AH-supplemented groups throughout the study.The protein expression of CEC cultured in all media was in accordance to their respective gene expressions.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Level 18, Pre-Clinical Block, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia. celine_kerwoon@yahoo.com.

ABSTRACT

Background: Acacia honey (AH) has been proven to improve skin wound healing, but its therapeutic effects on corneal epithelium has not been elucidated to date. This study aimed to investigate the effects of AH on cultured corneal epithelial cells (CEC) on in vitro corneal abrasion wound healing model. Six New Zealand white rabbits' CEC were isolated and cultured until passage 1. Circular wound area was created onto a confluent monolayer CEC using a corneal trephine which mimicked corneal abrasion and treated with 0.025% AH supplemented in basal medium (BM) and complete cornea medium (CCM). Wound healing was measured as the percentage of wound closure by the migration of CEC on day 0, day 3 and day 6, post wound creation. The morphological changes of CEC were assessed via phase contrast microscopy. Gene and protein expressions of cytokeratin (CK3), fibronectin and cluster of differentiation 44 (CD44) in AH treated groups and control groups were determined by real-time PCR and immunocytochemistry, respectively.

Results: Cultured CEC exhibited similar morphology of polygonal shaped cells in all culture media. CEC cultured in AH-supplemented media showed higher percentage of wound closure compared to the controls. Gene expression of CK3 increased in AH-supplemented groups throughout the study. Fibronectin expression was increased at the initial stage while CD44 expression was increased at day 3, post wound creation. The protein expression of CEC cultured in all media was in accordance to their respective gene expressions.

Conclusion: Supplementation of AH in BM and CCM media accelerates CEC wound closure of the in vitro corneal abrasion model by increasing the expression of genes and proteins associated with CEC wound healing.

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Related in: MedlinePlus

Immunocytochemistry for CK3 protein expression of CEC cultured in 4 different media; 1) BM medium (A, B, C), 2) BM supplemented with 0.025% AH (D, E, F), 3) CCM medium (G, H, I), and 4) CCM supplemented with 0.025% AH (J, K, L) at day 0, day 3 and day 6. Positive stained cells are marked with arrows. (Magnification × 50).
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Fig7: Immunocytochemistry for CK3 protein expression of CEC cultured in 4 different media; 1) BM medium (A, B, C), 2) BM supplemented with 0.025% AH (D, E, F), 3) CCM medium (G, H, I), and 4) CCM supplemented with 0.025% AH (J, K, L) at day 0, day 3 and day 6. Positive stained cells are marked with arrows. (Magnification × 50).

Mentions: Immunocytochemistry for CK3 protein was detected in CEC cultured in all media throughout the study period. CEC cultured in AH-enriched media showed increased immunodeposition of CK3 compared to controls which were in conformity with the gene expression in qRT-PCR analyses (Figure 7).Figure 7


The effects of acacia honey on in vitro corneal abrasion wound healing model.

Ker-Woon C, Abd Ghafar N, Hui CK, Mohd Yusof YA, Wan Ngah WZ - BMC Cell Biol. (2015)

Immunocytochemistry for CK3 protein expression of CEC cultured in 4 different media; 1) BM medium (A, B, C), 2) BM supplemented with 0.025% AH (D, E, F), 3) CCM medium (G, H, I), and 4) CCM supplemented with 0.025% AH (J, K, L) at day 0, day 3 and day 6. Positive stained cells are marked with arrows. (Magnification × 50).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4340287&req=5

Fig7: Immunocytochemistry for CK3 protein expression of CEC cultured in 4 different media; 1) BM medium (A, B, C), 2) BM supplemented with 0.025% AH (D, E, F), 3) CCM medium (G, H, I), and 4) CCM supplemented with 0.025% AH (J, K, L) at day 0, day 3 and day 6. Positive stained cells are marked with arrows. (Magnification × 50).
Mentions: Immunocytochemistry for CK3 protein was detected in CEC cultured in all media throughout the study period. CEC cultured in AH-enriched media showed increased immunodeposition of CK3 compared to controls which were in conformity with the gene expression in qRT-PCR analyses (Figure 7).Figure 7

Bottom Line: CEC cultured in AH-supplemented media showed higher percentage of wound closure compared to the controls.Gene expression of CK3 increased in AH-supplemented groups throughout the study.The protein expression of CEC cultured in all media was in accordance to their respective gene expressions.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, Level 18, Pre-Clinical Block, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000, Kuala Lumpur, Malaysia. celine_kerwoon@yahoo.com.

ABSTRACT

Background: Acacia honey (AH) has been proven to improve skin wound healing, but its therapeutic effects on corneal epithelium has not been elucidated to date. This study aimed to investigate the effects of AH on cultured corneal epithelial cells (CEC) on in vitro corneal abrasion wound healing model. Six New Zealand white rabbits' CEC were isolated and cultured until passage 1. Circular wound area was created onto a confluent monolayer CEC using a corneal trephine which mimicked corneal abrasion and treated with 0.025% AH supplemented in basal medium (BM) and complete cornea medium (CCM). Wound healing was measured as the percentage of wound closure by the migration of CEC on day 0, day 3 and day 6, post wound creation. The morphological changes of CEC were assessed via phase contrast microscopy. Gene and protein expressions of cytokeratin (CK3), fibronectin and cluster of differentiation 44 (CD44) in AH treated groups and control groups were determined by real-time PCR and immunocytochemistry, respectively.

Results: Cultured CEC exhibited similar morphology of polygonal shaped cells in all culture media. CEC cultured in AH-supplemented media showed higher percentage of wound closure compared to the controls. Gene expression of CK3 increased in AH-supplemented groups throughout the study. Fibronectin expression was increased at the initial stage while CD44 expression was increased at day 3, post wound creation. The protein expression of CEC cultured in all media was in accordance to their respective gene expressions.

Conclusion: Supplementation of AH in BM and CCM media accelerates CEC wound closure of the in vitro corneal abrasion model by increasing the expression of genes and proteins associated with CEC wound healing.

Show MeSH
Related in: MedlinePlus