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Preliminary Assessment Of De-novoCollagen Secretion By VIC-macrophage Co-culture On 3D Engineered Valve Tissues Versus A Porcine Bio-scaffold Valve.
Claudia Lorena Ponce Aportela, Oriana Marrone Mantovani, Sharan Ramaswamy.
Biomedical Engineering Department, Florida International University, Miami, FL, USA.

OBJECTIVE: To quantify collagen secreted by Rat Valvular Interstitial Cells (VICs) on co-cultures with Macrophages once seeded onto elastin-rich engineered ECM valves relative to raw porcine small intestinal submucosa (PSIS) bio-scaffold valves.
METHODS: In-house engineered ECM valves (n=4), as well as PSIS bio-scaffolds (n=4; Cormatrix, Roswell, GA), were seeded for 8 days in a rotisserie culture with Rat VICs. Subsequently, the valves were transferred to dynamic culture conditions simulating a heart rate of 70 bpm for 7 days. Next, bone marrow macrophages were seeded for 3 days of co-culture with the Rat VICs under static conditions. Sectioned constructs to a thickness of 16 Ám were then histologically assessed with Masson's Trichrome Stain (American Stat Lab, CA) to assess de-novo collagen formation. Samples were imaged at 20x magnification, and collagen quantified using a heat-mapping intensity code (MATLAB, Matworks, MA). The intensity average was reported as average ▒ SD. A T-test was done to analyze the statistical difference between the 2 groups.
RESULTS: Results of this preliminary study exhibited that the engineered ECM scaffolds had a higher de novo collagen content with an average of 0.776 ▒ 0.076 AU (Arbitrary Units) in contrast to Raw PSIS that averaged a total of 0.629▒ 0.037 AU. Additionally, a t-test between samples revealed a (p<0.05, p=0.02) representing a statistically significant increase in collagen in the Engineered ECM valves compared to Raw PSIS.

CONCLUSIONS: The preliminary study concludes that the engineered ECM scaffolds facilitate increased de novo collagen secretions by VICs-Macrophages co-culture compared to their identical co-culture on Raw PSIS scaffolds. Increased collagen in the engineered ECM represents a validation that engineered tissue valves could therefore facilitate regeneration compared to raw bio-scaffold valves-alone.This research is funded by the Florida Heart Research Foundation.


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