Introduction: Fibrin is a natural polymer proposed as a cell scaffold for numerous tissue engineering purposes. Studies on fibrin optimisation for cell scaffolds have mainly focused on fibrinogen and thrombin content. However, other factors, such as factor XIII (FXIII), calcium, and chloride ions contained in fibrinogen and thrombin formulations, are also known to affect fibrin structure. In this study, such effects on cell viability during culturing were assessed. Materials and Methods: Different FXIII, calcium, and chloride concentration combinations were used to polymerize fibrin resulting in various amounts of cross-linking. The relative amounts of cross-links were estimated by gel transparency measurements (n¼3/conditions). Two compositions resulting in high and low amounts of cross-linking were then tested for cell viability. These carriers were seeded with C3H10T1/2 cell line (0.6 million cells/ ml) and cultured for 7 days (mediumþ5%FBS). Cell number, morphology and distribution on histological sections were assessed at day0 (D0) and day7 (D7), (n¼3/carrier-type/time/test). All fibrin carriers were synthesized as cylinder (8mm diameter, 4mm height) containing 20mg/ml fibrinogen and 13.5U/ml thrombin.
Results: Chloride increased gel cross-links, as well as FXIII (133þ/-1U, high transparency). Calcium decreased cross-links (60þ/-6U, low transparency) and countered the effect of FXIII.Cell proliferation at D7 was higher for lower (227þ/??26% of initial cell number) than for higher cross-linking (120þ/??1%).Cell spreading was observed in both types of carrier, but clusters were only seen for lower cross-linking. Conclusion: These results showed that FXIII, calcium and chloride affects fibrin structure and that less cross-linked fibrin
carriers allowed better proliferation of mesenchymal cells.
|Title of host publication||Conference Proceedings of TERMIS-EU 2008 meeting|
|Place of Publication||Portugal, Porto|
|Publication status||Published - 2008|
|Name||Tissue engineering. Part A|