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Corneal Tissue Engineering: An In Vitro Model of the Stromal-nerve Interactions of the Human Cornea
Published on: January 24, 2018
Human corneal fibrosis: an in vitro model
Dimitris Karamichos1, Xiaoqing Q Guo, Audrey E K Hutcheon
1Department of Ophthalmology, Harvard Medical School, Schepens Eye Research Institute, Boston, Massachusetts 02114, USA.
Investigative Ophthalmology & Visual Science
|October 31, 2009
Summary
Researchers developed a 3-D corneal fibrosis model using human corneal fibroblasts (HCFs) and TGF-beta1. This model mimics scar formation and aids in studying extracellular matrix deposition during wound healing.
Area of Science:
- Ophthalmology
- Tissue Engineering
- Cell Biology
Background:
- Corneal fibrosis, a result of injury, involves myofibroblasts and abnormal extracellular matrix (ECM) deposition.
- Transforming growth factor-beta1 (TGF-beta1) is known to promote ECM overproduction.
- A 3-D in vitro model using human corneal fibroblasts (HCFs) and vitamin C (VitC) mimics corneal development.
Purpose of the Study:
- To generate a 3-D corneal scar model by incorporating TGF-beta1 into an existing HCFs-VitC model.
- To investigate the effects of TGF-beta1 on ECM deposition and cellular morphology in a 3-D corneal stroma model.
Main Methods:
- HCFs were cultured in four conditions for 4 or 8 weeks: VitC only, VitC+TGF-beta1, sequential addition of TGF-beta1, or delayed TGF-beta1.
- Transmission electron microscopy (TEM) and indirect immunofluorescence were used for analysis.
Main Results:
- TGF-beta1 significantly increased construct thickness (2.1- to 3.2-fold at 4 and 8 weeks).
- TGF-beta1 induced elongated, flattened cells, increased collagen levels, and enhanced expression of smooth muscle actin, cellular fibronectin, and type III collagen.
- Minimal differences were observed between 4 and 8 weeks in cultures.
Conclusions:
- HCFs stimulated with VitC and TGF-beta1 effectively model human corneal fibrosis.
- This 3-D model is valuable for studying matrix deposition and assembly in wound healing contexts.
