Human Endothelial Cell Response to Lipopolysaccharides from Periodontopathogens on Scaffold-collagen

Objectives: Compare the response to lipopolysaccharides (LPS) from Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans of human coronary endothelial cell (HCAEC) cultured in monolayer (2D) or on collagen scaffolds (3D)
Methods: Collagen type I dispersion (5 mg/mL) was poured into the transwell inserts, frozen (-80^oC) and lyophilized (24 h). Then, HCAEC were seeded on the apical surface of the obtained scaffolds and cultured (7 days, 37 ^oC, 5% CO₂). 3D cultures were incubated (24 h) with LPS-P.g or LPS-A.a (1 μg/mL). Transcription of 3 genes; secretion of 40 cytokines and 2 prostanoids and adhesion of monocytes to the endothelial monolayer were assessed with/without exposure to LPS. A principal component analysis (PCA) of secretion data was done, and the significance of the difference between the treatments was evaluated with ANOVA-Bonferroni or Kruskal-Wallis - U Mann Whitney tests (p <0.05).
Results: Transcription of COX-2, eNOS, and vWF decreased in LPS treated 3D cultures compared to their controls. Conversely, transcription of COX-2 increases in 2D compared to their controls, and no changes on eNOS and vWF transcription were seen. The concentration of 12 cytokines was found different in 3D compared to 2D cultures (p<0.05). Significant differences were found in the cytokine secretion profile of 3D and 2D treated and control cultures. In 3D cultures treated with LPS-Aa IL-8, RANTES, G-CSF, ICAM-1, IL-6, and TXA2 levels were significantly different from controls. The 2D cultures secreted G-CSF, IL-8, RANTES, ICAM-1, TNF-RI, PGI2, and TXA2. Monocyte adhesion to 2DCE cultures was increased after stimulation with LPS-A.a, however, in 3DCE cultures, the same was not observed. No differences were observed in cytokine and prostaglandin secretion in 3D or 2D cultures LPS-P.g treated, as well as in monocyte adhesion.
Conclusions: Gene transcription, protein secretion and monocyte adhesion of 3D and 2D cultures treated or not with LPSs were different.