TY - JOUR
T1 - Human corneal fibrosis
T2 - An in vitro model
AU - Karamichos, Dimitris
AU - Guo, Xiaoqing Q.
AU - Hutcheon, Audrey E.K.
AU - Zieske, James D.
PY - 2010/3
Y1 - 2010/3
N2 - Purpose. Corneal injury may ultimately lead to a scar by way of corneal fibrosis, which is characterized by the presence of myofibroblasts and improper deposition of extracellular matrix (ECM) components. TGF-β1 is known to stimulate overproduction and deposition of ECM components. Previously, an in vitro three-dimensional (3-D) model of a corneal stroma was developed by using primary human corneal fibroblasts (HCFs) stimulated with stable vitamin C (VitC). This model mimics corneal development. The authors postulate that with the addition of TGF-β1, a 3-D corneal scar model can be generated. Methods. HCFs were grown in four media conditions for 4 or 8 weeks: VitC only; VitC+TGF-β1 for the entire time; VitC+TGF-β1 for 1 week, then VitC only for 3 or 7 weeks; and VitC for 4 weeks, then VitC+TGF-β1 for 4 weeks. Cultures were analyzed with TEM and indirect immunofluorescence. Results. Compared with the control, addition of TGF-β1 increased construct thickness significantly, with maximum increase in constructs with TGF-β1 present for the entire time- 2.1- to 3.2-fold at 4 and 8 weeks, respectively. In all TGF-β- treated cultures, cells became long and flat, numerous filamentous cells were seen, collagen levels increased, and long collagen fibrils were visible. Smooth muscle actin, cellular fibronectin, and type III collagen expression all appeared to increase. Cultures between weeks 4 and 8 showed minimal differences. Conclusions. Human corneal fibroblasts stimulated by VitC and TGF-β1 appear to generate a model that resembles processes observed in human corneal fibrosis. This model should be useful in examining matrix deposition and assembly in a wound-healing situation.
AB - Purpose. Corneal injury may ultimately lead to a scar by way of corneal fibrosis, which is characterized by the presence of myofibroblasts and improper deposition of extracellular matrix (ECM) components. TGF-β1 is known to stimulate overproduction and deposition of ECM components. Previously, an in vitro three-dimensional (3-D) model of a corneal stroma was developed by using primary human corneal fibroblasts (HCFs) stimulated with stable vitamin C (VitC). This model mimics corneal development. The authors postulate that with the addition of TGF-β1, a 3-D corneal scar model can be generated. Methods. HCFs were grown in four media conditions for 4 or 8 weeks: VitC only; VitC+TGF-β1 for the entire time; VitC+TGF-β1 for 1 week, then VitC only for 3 or 7 weeks; and VitC for 4 weeks, then VitC+TGF-β1 for 4 weeks. Cultures were analyzed with TEM and indirect immunofluorescence. Results. Compared with the control, addition of TGF-β1 increased construct thickness significantly, with maximum increase in constructs with TGF-β1 present for the entire time- 2.1- to 3.2-fold at 4 and 8 weeks, respectively. In all TGF-β- treated cultures, cells became long and flat, numerous filamentous cells were seen, collagen levels increased, and long collagen fibrils were visible. Smooth muscle actin, cellular fibronectin, and type III collagen expression all appeared to increase. Cultures between weeks 4 and 8 showed minimal differences. Conclusions. Human corneal fibroblasts stimulated by VitC and TGF-β1 appear to generate a model that resembles processes observed in human corneal fibrosis. This model should be useful in examining matrix deposition and assembly in a wound-healing situation.
UR - http://www.scopus.com/inward/record.url?scp=77949879211&partnerID=8YFLogxK
U2 - 10.1167/iovs.09-3860
DO - 10.1167/iovs.09-3860
M3 - Article
C2 - 19875671
AN - SCOPUS:77949879211
SN - 0146-0404
VL - 51
SP - 1382
EP - 1388
JO - Investigative Ophthalmology and Visual Science
JF - Investigative Ophthalmology and Visual Science
IS - 3
ER -