Interaction of human valve interstitial cells with collagen matrices manufactured using rapid prototyping

Rapid prototyping is a novel process for the production of scaffolds of predetermined size and three-dimensional shape. The aim of the study was to determine the feasibility of this technology for producing scaffolds for tissue engineering an aortic valve and the optimal concentration of collagen processed in this manner that would maintain viability and promote proliferation of human valve interstitial cells. Scaffolds of 1%, 2% and 5% w/v bovine type-I collagen were manufactured using rapid prototyping. Valve interstitial cells isolated from three human aortic valves were seeded on the scaffolds and cultured for up to 4 weeks. Cell viability was assessed using the CellTiter 96 Aq(ueous) One Solution Cell Proliferation Assay and cell death by lactate dehydrogenase (LDH) measurement. Valve interstitial cells remained viable and proliferated within the collagen scaffolds. Cells consistently proliferated to a greater extent on 1% collagen scaffolds rather than either 2% or 5% collagen and after 4 weeks reached 212+/-33.1%, 139+/-25.9% and 129+/-38.3% (mean+/-SD) of their initial seeding density on 1%, 2% and 5% collagen scaffolds, respectively. LDH analysis demonstrated that there was minimal cell death indicating that the collagen scaffold was not toxic to human valve interstitial cells. Rapid prototyping provides a route to optimize biological scaffold designs for tissue engineering cardiac valves. This technology has the versatility to create scaffolds that are compatible with the specific needs of the valve interstitial cells and should enhance cell viability, proliferation and function.