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J. Funct. Biomater. 2012, 3(4), 760-775; doi:10.3390/jfb3040760
Article

Novel in Vitro Model for Keratoconus Disease

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Received: 29 August 2012; in revised form: 9 October 2012 / Accepted: 24 October 2012 / Published: 13 November 2012
(This article belongs to the Special Issue Corneal Scarring: Wound Healing and Biomaterials)
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Abstract: Keratoconus is a disease where the cornea becomes cone-like due to structural thinning and ultimately leads to compromised corneal integrity and loss of vision. Currently, the therapeutic options are corrective lenses for early stages and surgery for advanced cases with no in vitro model available. In this study, we used human corneal fibroblasts (HCFs) and compared them to human Keratoconus fibroblasts (HKCs) cultured in a 3-dimensional (3D) model, in order to compare the expression and secretion of specific extracellular matrix (ECM) components. For four weeks, the cells were stimulated with a stable Vitamin C (VitC) derivative ± TGF-β1 or TGF-β3 (T1 and T3, respectively). After four weeks, HKCs stimulated with T1 and T3 were significantly thicker compared with Control (VitC only); however, HCF constructs were significantly thicker than HKCs under all conditions. Both cell types secreted copious amounts of type I and V collagens in their assembled, aligned collagen fibrils, which increased in the degree of alignment upon T3 stimulation. In contrast, only HKCs expressed high levels of corneal scarring markers, such as type III collagen, which was dramatically reduced with T3. HKCs expressed α-smooth muscle actin (SMA) under all conditions in contrast to HCFs, where T3 minimized SMA expression. Fast Fourier transform (FFT) data indicated that HKCs were more aligned when compared to HCFs, independent of treatments; however, HKC’s ECM showed the least degree of rotation. HKCs also secreted the most aligned type I collagen under T3 treatment, when compared to any condition and cell type. Overall, our model for Keratoconus disease studies is the first 3D in vitro tissue engineered model that can mimic the Keratoconus disease in vivo and may be a breakthrough in efforts to understand the progression of this disease.
Keywords: Keratoconus disease; extracellular matrix; TGF-β3; fast fourier analysis; corneal fibroblasts Keratoconus disease; extracellular matrix; TGF-β3; fast fourier analysis; corneal fibroblasts
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Karamichos, D.; Zareian, R.; Guo, X.; Hutcheon, A.E.; Ruberti, J.W.; Zieske, J.D. Novel in Vitro Model for Keratoconus Disease. J. Funct. Biomater. 2012, 3, 760-775.

AMA Style

Karamichos D, Zareian R, Guo X, Hutcheon AE, Ruberti JW, Zieske JD. Novel in Vitro Model for Keratoconus Disease. Journal of Functional Biomaterials. 2012; 3(4):760-775.

Chicago/Turabian Style

Karamichos, Dimitrios; Zareian, Ramin; Guo, Xiaoqing; Hutcheon, Audrey E.; Ruberti, Jeffrey W.; Zieske, James D. 2012. "Novel in Vitro Model for Keratoconus Disease." J. Funct. Biomater. 3, no. 4: 760-775.


J. Funct. Biomater. EISSN 2079-4983 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert