Bioactive Fibrin Scaffolds for Use in Musculoskeletal Regenerative Medicine

Abstract Autologous fibrin matrices derived from the Leukocyte and Platelet Rich Plasma (L-PRP) and Leukocyte and Platelet Rich Fibrin (L-PRF) techniques present great potential to act as a bioactive scaffold in regenerative medicine, contributing to the maintenance of cell viability, proliferation stimulus and differentiation. In contrast, there are few studies that characterize the bioactive potential of these fibrin scaffolds by considering the process of production. The objective of this work was to characterize the intrinsic potential of maintaining cell viability of different fibrin scaffolds containing platelets and leukocytes. In order to achieve that, blood samples from a volunteer were collected and processed to obtain fibrin clots using the suggested techniques. To characterize the potential for in vitro viability, mesenchymal stem cells from human infrapatellar fat were used. The scaffolds were cellularized (1x105 cells/scaffolds) and maintained for 5 and 10 days under culture conditions with Dulbecco's Modified Eagle Medium, without addition of fetal bovine serum, and subsequently subjected to analyses by Fourrier transform infra-red spectroscopy, circular dichroism and fluorescence microscopy. The results demonstrated distinct intrinsic potential viability between the scaffolds, and L-PRP was responsible for promoting higher levels of viability in both periods of analysis. No viable cells were identified in the fibrin matrix used as controls. These results allow us to conclude that both fibrin substrates have presented intrinsic potential for maintaining cell viability, with superior potential exhibited by L-PRP scaffold, and represent promising alternatives for use as bioactive supports in musculoskeletal regenerative medicine.

Assessment of the metabolism of different strains of Bacillus megaterium

Bacillus megaterium is a promising host for expression of heterologous proteins. This paper reports the nutrient consumption patterns and production of metabolites for three different strains of B. megaterium, ATCC 14945, QMB 1551 and PV 361, which is QMB 1551 with seven constitutive plasmids deleted. 14 h cultivations in agitated flasks were run, for two different media: A (LB plus 10g/L glucose) and B (medium A, with the yeast extract replaced by tryptone). Strains PV361 and QMB 1551 showed higher maximum specific growth rates in medium B, reaching 0.42 h-1 and 0.48 h-1 respectively. The main by-products of the glucose overflow mechanism were acetate and lactate, for all three strains, which had preferential amino acids for substrate: Ala, Asp, Glu, Ser. No production of alcohols was detected.