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

BMC Cell Biology

Wan Zurinah Wan Ngah²  Yasmin Anum Mohd Yusof²  Chua Kien Hui⁴  Norzana Abd Ghafar¹  Choy Ker-Woon³ 

[1] UKM Medical Molecular Biology Institute (UMBI), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, 56000, Malaysia;Department of Biochemistry, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, 56000, Malaysia;Department of Anatomy, Level 18, Pre-Clinical Block, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, 56000, Malaysia;Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre (UKMMC), Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, 56000, Malaysia

关键词: Corneal wound healing;    Corneal abrasion;    Migration;    Corneal epithelial cells;    Acacia honey;   

Others  :  1137100
DOI  :  10.1186/s12860-015-0053-9

【 摘 要 】

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.

【 授权许可】

   
2015 Ker-Woon et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Voon LV, See J, Wong TY: The epidemiology of ocular trauma in Singapore: perspective from the emergency service of a large tertiary hospital. Eye 2001, 15:75-81.
• [2]Hua L, Doll T: A series of 3 cases of corneal abrasion with multiple etiologies. Optometry 2010, 81(2):83-5.
• [3]Suzuki K, Saito J, Yanai R, Yamada N, Chikama T, Seki K, Nishida T: Cell-matrix and cell-cell interactions during corneal epithelial wound healing. Prog Retin Eye Res 2003, 22(2):113-33.
• [4]Lu L, Reinach PS, Kao WW: Corneal epithelial wound healing. Exp Biol Med 2001, 226:653-64.
• [5]Hattori A, Hozumi K, Ko JA, Chikama T, Oomikawa K, Kato J, Ishida K, Hoshi N, Katagiri F, Kikkawa Y, Nomizu M, Nishida T: Sequence specificity of the PHSRN peptide from fibronectin on corneal epithelial migration. Biochem Biophys Res Commun 2009, 379(2):346-50.
• [6]Germain L, Carrier P, Auger FA, Salesse C, Guérin SL: Can we produce a human corneal equivalent by tissue engineering? Prog Retin Eye Res 2000, 19(5):497-527.
• [7]Gomes JA, Amankwah R, Powell-Richards A, Dua HS: Sodium hyaluronate (hyaluronic acid) promotes migration of human corneal epithelial cells in vitro. Br J Ophthalmol 2004, 88(6):821-5.
• [8]Milder E, Vander J, Shah C, Garg S: Changes in antibiotic resistance patterns of conjunctival flora due to repeated use of topical antibiotics after intravitreal injection. Ophthalmology 2012, 119(7):1420-4.
• [9]Baudouin C, Labbe A, Liang H, Pauly A, Brignole-Baudouin F: Preservatives in eyedrops: the good, the bad and the ugly. Prog Retin Eye Res 2010, 29(4):312-34.
• [10]Aljadi AM, Kamaruddin MY: Evaluation of the phenolic contents and antioxidant capacities of two Malaysian floral honeys. Food Chem 2004, 85(4):513-8.
• [11]Hussein SZ, Yusoff KM, Makpol S, Yusof YA: Antioxidant capacities and total phenolic contents increase with gamma irradiation in two types of Malaysian honey. Molecules 2011, 16(9):6378-95.
• [12]Mandal S, DebMandal M, Pal NK, Saha K: Antibacterial activity of honey against clinical isolates of Escherichia coli, Pseudomonas aeruginosa and Salmonella enterica serovar Typhi. Asian Pac J Trop Med 2010, 3(12):961-4.
• [13]Pyrzynska K, Biesaga M: Analysis of phenolic acids and flavonoids in honey. TrAC Trends Analyt Chem 2009, 28(7):893-902.
• [14]Moniruzzaman M, Khalil MI, Sulaiman SA, Gan SH: Physicochemical and antioxidant properties of Malaysian honeys produced by Apis cerana, Apis dorsata and Apis mellifera. BMC Complement Altern Med 2013, 13(43):1-12.
• [15]Iftikhar F, Arshad M, Rasheed F, Amraiz D, Anwar P, Gulfraz M: Effects of acacia honey on wound healing in various rat models. Phytother Res 2010, 24(4):583-6.
• [16]Ker-Woon C, Abd Ghafar N, Hui CK, Mohd Yusof YA, Luan NS: Proliferative capacity of in vitro corneal epithelium: role of Acacia honey in the initial step of wound healing. J Med Bioeng 2014, 3(2):107-12.
• [17]Abd Ghafar N, Abd Rahman R, Che Hamzah J, Hui CK, Othman F, Aminuddin BS, Ruszymah BHI: Rabbit limbal epithelial cells maintains its stemness in serum-free and feeder layer-free culture system. Tissue Eng Regen Med 2007, 4(4):557-65.
• [18]Chang CY, Green CR, McGhee CN, Sherwin T: Acute wound healing in the human central corneal epithelium appears to be independent of limbal stem cell influence. Invest Ophthalmol Vis Sci 2008, 49(12):5279-86.
• [19]Dua HS, Gomes JA, Singh A: Corneal epithelial wound healing. Br J Ophthalmol 1994, 78:401-8.
• [20]Kadar T, Horwitz V, Sahar R, Cohen M, Cohen L, Gez R, Tveria L, Gutman H, Buch H, Fishbine E, Brandeis R, Dachir S, Amir A: Delayed loss of corneal epithelial stem cells in a chemical injury model associated with limbal stem cell deficiency in rabbits. Curr Eye Res 2011, 36(12):1098-107.
• [21]Takahashi H, Ohara K, Ohmura T, Takahashi R, Zieske JD: Glucose transporter 1 expression in corneal wound repair under high serum glucose level. Jpn J Ophthalmol 2000, 44:470-4.
• [22]Kumagai AK, Glasgow BJ, Pardridge WM: GLUT1 glucose transporter expression in the diabetic and nondiabetic human eye. Invest Ophthalmol Vis Sci 1994, 35(6):2887-94.
• [23]Cavia MM, Fernandez-Muino MA, Gomez-Alonso E, Montes-Perez MJ, Huidobro JF, Sancho MT: Evolution of fructose and glucose in honey over one year: influence of induced granulation. Food Chem 2002, 78:157-61.
• [24]Bang LM, Buntting C, Molan P: The effect of dilution on the rate of hydrogen peroxide production in honey and its implications for wound healing. J Altern Complement Med 2003, 9(2):267-73.
• [25]Mandal MD, Mandal S: Honey: its medicinal property and antibacterial activity. Asian Pac J Trop Biomed 2011, 1(2):154-60.
• [26]Burdon RH: Superoxide and hydrogen peroxide in relation to mammalian cell proliferation. Free Radic Biol Med 1995, 18(4):775-94.
• [27]Molan PC: The role of honey in the management of wounds. J Wound Care 1999, 8(8):415-8.
• [28]Chua LS, Abdul-Rahaman NL, Sarmidi MR, Aziz R: Multi-elemental composition and physical properties of honey samples from Malaysia. Food Chem 2012, 135(3):880-7.
• [29]Chen L, Meng Q, Kao W, Xia Y: IkB kinase beta regulates epithelium migration during corneal wound healing. PLoS One 2011, 6(1):1-11.
• [30]Boucher C, Ruiz JC, Thibault M, Buschmann MD, Wertheimer MR, Jolicoeur M, Durocher Y, De Crescenzo G: Human corneal epithelial cell response to epidermal growth factor tethered via coiled-coil interactions. Biomaterials 2010, 31(27):7021-31.
• [31]Steindl-Kuscher K, Boulton ME, Haas P, Dossenbach-Glaninger A, Feichtinger H, Binder S: Epidermal growth factor: the driving force in initiation of RPE cell proliferation. Graefes Arch Clin Exp Ophthalmol 2011, 249(8):1195-200.
• [32]Kumagai Y, Kurokawa MS, Ueno H, Kayama M, Tsubota K, Nakatsuji N, Kondo Y, Ueno S, Suzuki N: Induction of corneal epithelium-like cells from cynomolgus monkey embryonic stem cells and their experimental transplantation to damaged cornea. Cornea 2010, 29(4):432-8.
• [33]Notara M, Daniels JT: Characterisation and functional features of a spontaneously immortalised human corneal epithelial cell line with progenitor-like characteristics. Brain Res Bull 2010, 81(2–3):279-86.
• [34]Nakamura M, Sato N, Chikama T, Hasegawa Y, Nishida T: Fibronectin facilitates corneal epithelial wound healing in diabetic rats. Exp Eye Res 1997, 64:355-9.
• [35]Murakami J, Nishida T, Otori T: Coordinated appearance of beta 1 integrins and fibronectin during corneal wound healing. J Lab Clin Med 1992, 120(1):86-93.
• [36]Cai X, Foster CS, Liu JJ, Kupferman AE, Filipec M, Colvin RB, Lee SJ: Alternatively spliced fibronectin molecules in the wounded cornea: analysis by PCR. Invest Ophthalmol Vis Sci 1993, 34(13):3585-92.
• [37]Latvala T, Tervo K, Mustonen R, Tervo T: Expression of cellular fibronectin and tenascin in the rabbit cornea after excimer laser photorefractive keratectomy: a 12 month study. British J Ophthalmol 1995, 79:65-9.
• [38]Lai YH, Wang HZ, Lin CP, Hong SJ, Chang SJ: Endothelin-1 enhances corneal fibronectin deposition and promotes corneal epithelial wound healing after photorefractive keratectomy in rabbits. Kaohsiung J Med Sci 2008, 24(5):254-61.
• [39]Nickeleit V, Kaufman AH, Zagachin L, Dutt JE, Foster CS, Colvin RB: Healing corneas express embryonic fibronectin isoforms in the epithelium, subepithelial stroma and endothelium. Am J Pathol 1996, 49(2):549-58.
• [40]Saika S, Shiraishi A, Liu CY, Funderburgh JL, Kao CW, Converse RL, Kao WW: Role of Lumican in the corneal epithelium during wound healing. J Biol Chem 2000, 275(4):2607-12.
• [41]Crane IJ, Liversidge J: Mechanisms of leukocyte migration across the blood-retina barrier. Semin Immunopathol 2008, 30(2):165-77.
• [42]Desai VD, Wang Y, Simirskii VN, Duncan MK: CD44 expression is developmentally regulated in the mouse lens and increases in the lens epithelium after injury. Differentiation 2010, 79(2):111-9.
• [43]Lee JS, Hwang SH, Jeung G, Yoon S: The distribution of CD44 in wound healing of rabbit alkali burn. J Korean Ophthalmol Soc 1996, 37(10):1607-13.