Bone regeneration driven by a nano-hydroxyapatite/chitosan composite bioaerogel for periodontal regeneration.

The most recent progress in reconstructive therapy for the management of periodontitis and peri-implantitis bone defects has relied on the development of highly porous biodegradable bioaerogels for guided bone regeneration. The objective of this work was to evaluate in vitro the osteoinduction of periodontal-originating cells (human dental follicle mesenchymal cells, DFMSCs) promoted by a nano-hydroxyapatite/chitosan (nHAp/CS) bioaerogel, which was purified and sterilized by a sustainable technique (supercritical CO2). Moreover, the in vivo bone regeneration capacity of the nHAp/CS bioaerogel was preliminarily assessed as a proof-of-concept on a rat calvaria bone defect model. The quantification of DNA content of DFMSCs seeded upon nHAp/CS and CS scaffolds (control material) showed a significant increase from the 14th to the 21st day of culture. These results were corroborated through confocal laser scanning microscopy analysis (CLSM). Furthermore, the alkaline phosphatase (ALP) activity increased significantly on the 21st day, similarly for both materials. Moreover, the presence of nHAp promoted a significantly higher expression of osteogenic genes after 21 days when compared to CS scaffolds and control. CLSM images of 21 days of culture also showed an increased deposition of OPN over the nHAp/CS surface. The in vivo bone formation was assessed by microCT and histological analysis. The in vivo evaluation showed a significant increase in bone volume in the nHAp/CS test group when compared to CS and the empty control, as well as higher new bone formation and calcium deposition within the nHAp/CS structure. Overall, the present study showed that the nHAp/CS bioaerogel could offer a potential solution for periodontal and peri-implant bone regeneration treatments since the in vitro results demonstrated that it provided favorable conditions for DFMSC proliferation and osteogenic differentiation, while the in vivo outcomes confirmed that it promoted higher bone ingrowth.

Antimicrobial efficacy of sequentially applied eugenol against food spoilage micro-organisms.

AIMS: Compare the survival behaviour of food spoilage micro-organisms treated with sequential doses or all at once treatments of eugenol. METHODS AND RESULTS: Staphylococcus carnosus, Listeria innocua, Escherichia coli and Pseudomonas fluorescens were exposed to a minimal lethal concentration (MLC) initially, or to half doses over time, with first half dose applied immediately and a second half dose applied after 3, 4, 6 and 8 h, of eugenol. Direct plate counts were determined at regular time intervals. Population dynamics were analysed using a combined growth and mortality model. Effect of sequential dosing varied significantly between tested organisms. High antimicrobial efficacy on E. coli K12 was observed regardless of timing of the two doses. Reduced effectiveness was observed against Staph. carnosus and L. innocua the later the second half dose was applied. Complete cell reduction occurred after immediate exposure to full MLC dose, while sequential half doses were bacteriostatic regardless of application times. CONCLUSION: Time in between antimicrobial dose application had substantial impact on effectiveness, attributed to organisms becoming more tolerant. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to the evaluation of encapsulated antimicrobial systems, where antimicrobials are released over time and antimicrobial concentrations may only reach MLC levels after some time.