(1) Background: To evaluate the effects of the direct and indirect contact of leukocyte and platelet-rich fibrin (L-PRF) on bone development, in an ex vivo embryonic chick femur model. (2) Methods: Both sections of L-PRF membranes (red and yellow portions) were evaluated with scanning electron microscopy and histochemical staining. The in vivo angiogenic activity was evaluated using a chorioallantoic membrane model. The osteogenic activity was assessed with an organotypic culture of embryonic chick femora through direct and indirect contact, and assessment was conducted by microtomographic and histological analysis. Descriptive statistics, One-Way ANOVA and Tukey's multiple comparisons tests were performed for datasets that presented a normal distribution, and Kruskal-Wallis tests were performed for non-parametric datasets. A significance level of 0.05 was considered. (3) Results: The L-PRF induced angiogenesis reflected by a higher number and a larger and more complex gauge in the vessels that invaded the membrane. The physical presence of the membrane over the bone (direct contact) unleashes the full potential of the L-PRF effects on bone growth enhancement. The greatest increase in mineral content was observed in the diaphysis region. (4) Conclusion: The L-PRF direct contact group presented higher values on mineral content for bone volume, bone surface and bone mineral density than the indirect contact and control groups.
Keywords: angiogenesis modulating agents; bone regeneration; bone transplantation; chorioallantoic membrane; platelet-rich fibrin.