Balancing microbial and mammalian cell functions on calcium ion-modified implant surfaces.

Implant integration is a complex process mediated by the interaction of the implant surface with the surrounding ions, proteins, bacteria, and tissue cells. Although most implants achieve long-term bone-tissue integration, preventing pervasive implant-centered infections demands further advances, particularly in surfaces design. In this work, we analyzed classical microrough implant surfaces (only acid etched, AE; sandblasted then acid etching, SB + AE) and a new calcium-ion-modified implant surface (AE + Ca) in terms of soft- and hard-tissue integration, bacterial adhesion, and biofilm formation. We cultured on the surfaces primary oral cells from gingiva and alveolar bone, and three representative bacterial strains of the oral cavity, emulating oral conditions of natural saliva and blood plasma. With respect to gingiva and bone cells and in the presence of platelets and plasma proteins, AE + Ca surfaces yielded in average 86% higher adhesion, 44% more proliferation, and triggered 246% more synthesis of extracellular matrix biomolecules than AE-unmodified controls. Concomitantly, AE + Ca surfaces regardless of conditioning with saliva and/or blood plasma showed significantly less bacterial adhesion (67% reduction in average) and biofilm formation (40% reduction in average) than unmodified surfaces. These results highlight the importance of a calcium-rich hydrated interface to favor mammalian cell functions over microbial colonization at implant surfaces. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 421-432, 2018.

In vivo bactericidal efficacy of the Ti6Al4V surface after ultraviolet C treatment.

BACKGROUND: Biomaterial-associated infections are one of the most important complications in orthopedic surgery. The main goal of this study was to demonstrate the in vivo bactericidal effect of ultraviolet (UV) irradiation on Ti6Al4V surfaces. MATERIALS AND METHODS: An experimental model of device-related infections was developed by direct inoculation of Staphylococcus aureus into the canal of both femurs of 34 rats. A UV-irradiated Ti6Al4V pin was press-fit into the canal by retrograde insertion in one femur and the control pin was inserted into the contralateral femur. To assess the efficacy of UV radiation, the mean colony counts after inoculation in the experimental subjects and the control group were compared at different times of sacrifice and at different inoculum doses. RESULTS: At 72 h, the mean colony counts after inoculation in experimental femurs were significantly lower than those of the control group, with a reduction percentage of 76 % (p = 0.041). A similar difference between control and experimental pins was observed at 24 h using an inoculum dose <104 colony-forming units (CFU), for which the reduction percentage was 70.48 % (p = 0.017). CONCLUSION: The irradiated surface of Ti6Al4V is able to reduce early bacterial colonization of Ti6AlV pins located in the medullar channel and in the surrounding femur. The reductions depend on the initial inoculums used to cause infection in the animals and the greatest effects are detected for inoculums <104 CFU. LEVEL OF EVIDENCE: Not applicable.

Controlled RNA contamination and degradation and its impact on qPCR gene expression in S. epidermidis biofilms.

RNA quality is of utmost importance to perform gene expression quantification by qPCR. The classical methods used to determine RNA quality are based on electrophoresis and spectrophotometer assessment, namely A(260)/A(280) and A(260)/A(230) ratios. It was previously shown that due to the complex nature of Staphylococcus epidermidis biofilms, RNA extraction procedures could impact mRNA quality and thus accurate quantification. Herein, we contaminated and degraded RNA extracted from S. epidermidis biofilms, and assessed the effect on gene expression by qPCR. As expected, thermal degradation of RNA had a significant impact on gene expression on two out of the three tested genes. On the other hand, the contamination of the extracted RNA yielded an interesting result: while most contaminants did not changed the purity indicators or the integrity of RNA, significant changes on gene expression levels were found. This work confirms that poor RNA extraction has an important impact in qPCR quantification, emphasizing the consequences of carry-over contaminants on gene expression studies. Additionally, our results show that the parameters commonly used to assess the quality of extracted RNA from bacterial cultures seem to be insufficient to ensure reliable gene expression determination.