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Engineering calcium deposits on polycaprolactone scaffolds for intravascular applications using primary human
Beili Zhu1, Steven R Bailey, C Mauli Agrawal
1Department of Biomedical Engineering, College of Engineering, University of Texas at San Antonio, TX 78249-1644, USA.
Journal of Tissue Engineering and Regenerative Medicine
|September 10, 2010
Summary
Researchers developed a new animal model for atherosclerotic occlusions with calcification using biodegradable scaffolds and human cells. The study identified optimal conditions for inducing calcification in vitro, paving the way for better disease modeling.
Area of Science:
- Biomaterials Science
- Vascular Biology
- Tissue Engineering
Background:
- Atherosclerotic occlusions frequently involve calcification, but current animal models inadequately replicate gradual artery occlusion and cell-mediated calcification.
- Developing a robust animal model is crucial for studying the mechanisms of chronic total occlusions and testing therapeutic strategies.
Purpose of the Study:
- To establish an animal model for chronic total occlusions with calcification using biodegradable scaffolds and primary human osteoblasts (HOBs).
- To determine the optimal dosage of transforming growth factor-beta 1 (TGF-β1) on polycaprolactone (PCL) scaffolds to induce in vitro calcification.
Main Methods:
- Primary human osteoblasts (HOBs) were cultured on PCL scaffolds with varying TGF-β1 loadings (0-100 ng) and dexamethasone (Dex).
- Calcification was assessed using alizarin red staining, DNA content, and alkaline phosphatase (ALP) activity over 28 days.
- Initial dose-finding experiments utilized TGF-β1 and Dex in well plates to identify effective calcification conditions.
Main Results:
- Optimal in vitro calcification was achieved with 0.02 ng/ml TGF-β1 and 10(-10) M Dex in initial well plate cultures.
- Without Dex, a TGF-β1 loading of 5 ng on PCL scaffolds promoted the most significant calcification, DNA synthesis, and ALP activity.
- TGF-β1 showed an inhibitory effect on scaffold calcification when cultured in Dex-supplemented medium.
Conclusions:
- A PCL-HOB construct shows potential for creating an in vivo animal model of atherosclerotic occlusion with calcification.
- The study identified specific TGF-β1 concentrations that influence HOB calcification on PCL scaffolds, with or without Dex.
- This research provides a foundation for developing improved animal models to study complex vascular diseases.
