Acacia honey accelerates in vitro corneal ulcer wound healing model

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Abstract

Background

The study aimed to evaluate the effects of Acacia honey (AH) on the migration, differentiation and healing properties of the cultured rabbit corneal fibroblasts.

Methods

Stromal derived corneal fibroblasts from New Zealand White rabbit ( n = 6) were isolated and cultured until passage 1. In vitro corneal ulcer was created using a 4 mm corneal trephine onto confluent cultures and treated with basal medium (FD), medium containing serum (FDS), with and without 0.025 % AH. Wound areas were recorded at day 0, 3 and 6 post wound creation. Genes and proteins associated with wound healing and differentiation such as aldehyde dehydrogenase (ALDH), vimentin, alpha-smooth muscle actin (α-SMA), collagen type I, lumican and matrix metalloproteinase 12 (MMP12) were evaluated using qRT-PCR and immunocytochemistry respectively.

Results

Cells cultured with AH-enriched FDS media achieved complete wound closure at day 6 post wound creation. The cells cultured in AH-enriched FDS media increased the expression of vimentin, collagen type I and lumican genes and decreased the ALDH, α-SMA and MMP12 gene expressions. Protein expression of ALDH, vimentin and α-SMA were in accordance with the gene expression analyses.

Conclusion

These results demonstrated AH accelerate corneal fibroblasts migration and differentiation of the in vitro corneal ulcer model while increasing the genes and proteins associated with stromal wound healing.

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Most cited references 38

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The molecular basis of corneal transparency.

John R. Hassell, David E Birk (2010)

The cornea consists primarily of three layers: an outer layer containing an epithelium, a middle stromal layer consisting of a collagen-rich extracellular matrix (ECM) interspersed with keratocytes and an inner layer of endothelial cells. The stroma consists of dense, regularly packed collagen fibrils arranged as orthogonal layers or lamellae. The corneal stroma is unique in having a homogeneous distribution of small diameter 25-30 nm fibrils that are regularly packed within lamellae and this arrangement minimizes light scattering permitting transparency. The ECM of the corneal stroma consists primarily of collagen type I with lesser amounts of collagen type V and four proteoglycans: three with keratan sulfate chains; lumican, keratocan, osteoglycin and one with a chondroitin sulfate chain; decorin. It is the core proteins of these proteoglycans and collagen type V that regulate the growth of collagen fibrils. The overall size of the proteoglycans are small enough to fit in the spaces between the collagen fibrils and regulate their spacing. The stroma is formed during development by neural crest cells that migrate into the space between the corneal epithelium and corneal endothelium and become keratoblasts. The keratoblasts proliferate and synthesize high levels of hyaluronan to form an embryonic corneal stroma ECM. The keratoblasts differentiate into keratocytes which synthesize high levels of collagens and keratan sulfate proteoglycans that replace the hyaluronan/water-rich ECM with the densely packed collagen fibril-type ECM seen in transparent adult corneas. When an incisional wound through the epithelium into stroma occurs the keratocytes become hypercellular myofibroblasts. These can later become wound fibroblasts, which provides continued transparency or become myofibroblasts that produce a disorganized ECM resulting in corneal opacity. The growth factors IGF-I/II are likely responsible for the formation of the well organized ECM associated with transparency produced by keratocytes during development and by the wound fibroblast during repair. In contrast, TGF-beta would cause the formation of the myofibroblast that produces corneal scaring. Thus, the growth factor mediated synthesis of several different collagen types and the core proteins of several different leucine-rich type proteoglycans as well as posttranslational modifications of the collagens and the proteoglycans are required to produce collagen fibrils with the size and spacing needed for corneal stromal transparency. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

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The keratocyte: corneal stromal cell with variable repair phenotypes.

Judith A. West-Mays, Dhruva Dwivedi (2006)

Keratocytes, also known as fibroblasts, are mesencyhmal-derived cells of the corneal stroma. These cells are normally quiescent, but they can readily respond and transition into repair phenotypes following injury. Cytokines and other growth factors that provide autocrine signals for stimulating wound responses in resident cells are typically presented by platelets at the site of an injury. However, due to the avascular nature of the cornea many of the environmental cues are derived from the overlying epithelium. Corneal epithelial-keratocyte cell interactions have thus been extensively studied in numerous in vivo corneal wound healing settings, as well as in in vitro culture models. Exposure to the different epithelial-derived factors, as well as the integrity of the epithelial substratum, are factors known to impact the keratocyte response and determine whether corneal repair will be regenerative or fibrotic in nature. Finally, the recent identification of bone-marrow derived stem cells in the corneal stroma suggests a further complexity in the regulation of the keratocyte phenotype following injury.

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Author and article information

Contributors

Norzana Abd Ghafar: norzanafi9506@yahoo.com , norzana@ukm.edu.my

Choy Ker-Woon: celine_kerwoon@yahoo.com

Chua Kien Hui: ckienhui@hotmail.com

Yasmin Anum Mohd Yusof: rahmatyasmin@yahoo.com

Wan Zurinah Wan Ngah: wanzurinah@yahoo.com

Journal

Journal ID (nlm-ta): BMC Complement Altern Med

Journal ID (iso-abbrev): BMC Complement Altern Med

Title: BMC Complementary and Alternative Medicine

Publisher: BioMed Central (London )

ISSN (Electronic): 1472-6882

Publication date (Electronic): 29 July 2016

Publication date PMC-release: 29 July 2016

Publication date Collection: 2016

Volume: 16

Electronic Location Identifier: 259

Affiliations

[1 ]Department of Anatomy, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur Malaysia

[2 ]Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur Malaysia

[3 ]Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur Malaysia

Article

Publisher ID: 1248

DOI: 10.1186/s12906-016-1248-0

PMC ID: 4966736

PubMed ID: 27473120

SO-VID: 85734a33-b106-4063-b6db-58996c4c42e1

License:

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

History

Date received : 22 April 2016

Date accepted : 23 July 2016

Funding

Funded by: FundRef http://dx.doi.org/10.13039/501100004515, Universiti Kebangsaan Malaysia;

Award ID: GGPM-2011-085

Award Recipient : Norzana Abd Ghafar

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ScienceOpen disciplines: Complementary & Alternative medicine

Keywords: acacia honey,corneal fibroblasts,in vitro corneal ulcer,wound healing

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ScienceOpen disciplines: Complementary & Alternative medicine

Keywords: acacia honey, corneal fibroblasts, in vitro corneal ulcer, wound healing

Acacia honey accelerates in vitro corneal ulcer wound healing model

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Abstract

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Conclusion

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The International Journal of Person Centered Medicine

Most cited references 38

The molecular basis of corneal transparency.

Superoxide and hydrogen peroxide in relation to mammalian cell proliferation.

The keratocyte: corneal stromal cell with variable repair phenotypes.

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