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Fulltext Effect of acacia honey on cultured rabbit corneal keratocytes

Ker-Woon C, Abd Ghafar N, Hui CK, Mohd Yusof YA

BMC Cell Biol., 2014;15:19.
PMID: 24885607 DOI: 10.1186/1471-2121-15-19

Acacia honey is a natural product which has proven to have therapeutic effects on skin wound healing, but its potential healing effects in corneal wound healing have not been studied. This study aimed to explore the effects of Acacia honey (AH) on corneal keratocytes morphology, proliferative capacity, cell cycle, gene and protein analyses. Keratocytes from the corneal stroma of six New Zealand white rabbits were isolated and cultured until passage 1. The optimal dose of AH in the basal medium (FD) and medium containing serum (FDS) for keratocytes proliferation was identified using MTT assay. The morphological changes, gene and protein expressions of aldehyde dehydrogenase (ALDH), marker for quiescent keratocytes and vimentin, marker for fibroblasts were detected using q-RTPCR and immunocytochemistry respectively. Flowcytometry was performed to evaluate the cell cycle analysis of corneal keratocytes.

Matched MeSH terms: Corneal Keratocytes/cytology*; Corneal Keratocytes/drug effects*; Corneal Keratocytes/metabolism
Fulltext Effects of edible bird's nest (EBN) on cultured rabbit corneal keratocytes

Zainal Abidin F, Hui CK, Luan NS, Mohd Ramli ES, Hun LT, Abd Ghafar N

BMC Complement Altern Med, 2011;11:94.
PMID: 21992551 DOI: 10.1186/1472-6882-11-94

There has been no effective treatment or agent that is available for corneal injury in promoting corneal wound healing. Previous studies on edible bird's nest extract (EBN) had reported the presence of hormone-like substance; avian epidermal growth factor that could stimulate cell division and enhance regeneration. This study aimed to investigate the effects of EBN on corneal keratocytes proliferative capacity and phenotypical changes.

Matched MeSH terms: Corneal Keratocytes/cytology; Corneal Keratocytes/drug effects*
Phenotypic characterization of culture expanded rabbit limbal corneal keratocytes

Abd Ghafar N, Chua KH, Wan Ngah WZ, Che Hamzah J, Othman F, Abd Rahman R, et al.

Cell Tissue Bank, 2014 Mar;15(1):25-34.
PMID: 23292197 DOI: 10.1007/s10561-012-9360-y

The in vivo quiescent corneal stroma keratocytes need to be transformed to activated state in order to obtain sufficient number of cells either for monolayer evaluation or corneal stroma reconstruction. This study aimed to investigate the phenotypic characterization of corneal stromal cells during culture expansion from the limbal region of the cornea. Isolated corneal keratocytes from limbal tissue of New Zealand White Strain rabbits' corneas (n = 6) were culture expanded until three passages. Keratocytes morphology was examined daily with viability, growth rate, number of cell doubling and population doubling time were recorded at each passage. The expression of collagen type 1, aldehyde dehydrogenase (ALDH), lumican and alpha smooth muscle actin (α-SMA) were detected by RT-PCR. Immunocytochemistry was also used to detect ALDH, α-SMA, collagen type I and Cytokeratin-3 (CK3). Growth kinetic study revealed that the growth rate was low at the initial passage but increase to about two folds with concomitant reduction in population doubling time in later passages. Freshly isolated and cultured keratocytes expressed collagen type 1, ALDH and lumican but α-SMA expression was absent. However, α-SMA was expressed along with the other genes during culture expansion. Keratocytes at P1 expressed all the proteins except CK3. These results suggest that cultured keratocytes maintained most of the gene expression profile of native keratocytes while the emergence of α-SMA in serial passages showed a mix population of various phenotypes. The phenotypic characterization of monolayer keratocytes provides useful information before reconstruction of bioengineered tissue or in vitro pharmaceutical applications.

Matched MeSH terms: Corneal Keratocytes/cytology*; Corneal Keratocytes/transplantation
Fulltext In vivo slit scanning confocal microscopic observation in a patient with moderate and severe keratoconus: a case report

Mutalib HA, Ghosh S, Sharanjeet-Kaur, Ghoshal R

Clin Optom (Auckl), 2016;8:79-83.
PMID: 30214352 DOI: 10.2147/OPTO.S106421

A 22-year-old Indian female was referred to Sg Buloh hospital with the diagnosis of bilateral keratoconus. On examination, slit lamp biomicroscopy and corneal topography revealed stage 3 keratoconus in the right eye and stage 2 keratoconus in the left eye. Corneal cell morphology in both eyes was evaluated using confocal microscope. In qualitative observation, almost all corneal layers in right eye except endothelium were partially or completely obscured by haze. Additionally, morphological alterations, such as elongation of keratocyte nuclei and cluster of cells, and dark bands in the anterior stroma were observed in right eye. In the left eye, the amount of haze was less, allowing better visibility of the corneal layers compared with the right eye. The dark bands were evident in the posterior stroma. Quantitative analysis showed that anterior and posterior stromal keratocyte density and endothelium cell density were relatively low in the right eye (834.0, 700.5, and 2,133 cells/mm2, respectively) compared with the left eye (934.1, 750.6, and 2,361 cells/mm2, respectively). In this case, the right eye, exhibiting stage 3 keratoconus, showed more morphological alteration, particularly in the anterior stroma compared with the left eye with stage 2 keratoconus. Increased severity of the disease can explain these differences in corneal cell morphology.

Matched MeSH terms: Corneal Keratocytes
Fulltext Corneal Cell Morphology in Keratoconus: A Confocal Microscopic Observation

Ghosh S, Mutalib HA, Kaur S, Ghoshal R, Retnasabapathy S

Malays J Med Sci, 2017 Mar;24(2):44-54.
PMID: 28894403 MyJurnal DOI: 10.21315/mjms2017.24.2.6

PURPOSE: To evaluate corneal cell morphology in patients with keratoconus using an in vivo slit scanning confocal microscope.
METHODS: A cross-sectional study was conducted to evaluate the corneal cell morphology of 47 keratoconus patients and 32 healthy eyes without any ocular disease. New keratoconus patients with different disease severities and without any other ocular co-morbidity were recruited from the ophthalmology department of a public hospital in Malaysia from June 2013 to May 2014. Corneal cell morphology was evaluated using an in vivo slit-scanning confocal microscope. Qualitative and quantitative data were analysed using a grading scale and the Nidek Advanced Visual Information System software, respectively.
RESULTS: The corneal cell morphology of patients with keratoconus was significantly different from that of healthy eyes except in endothelial cell density (P = 0.072). In the keratoconus group, increased level of stromal haze, alterations such as the elongation of keratocyte nuclei and clustering of cells at the anterior stroma, and dark bands in the posterior stroma were observed with increased severity of the disease. The mean anterior and posterior stromal keratocyte densities and cell areas among the different stages of keratoconus were significantly different (P < 0.001 and P = 0.044, respectively). However, the changes observed in the endothelium were not significantly different (P > 0.05) among the three stages of keratoconus.
CONCLUSION: Confocal microscopy observation showed significant changes in corneal cell morphology in keratoconic cornea from normal healthy cornea. Analysis also showed significant changes in different severities of keratoconus. Understanding the corneal cell morphology changes in keratoconus may help in the long-term monitoring and management of keratoconus.

Matched MeSH terms: Corneal Keratocytes