Comparison of the Therapeutic Efficacy of Antibiotic-Loaded Polymeric Tissue Scaffold and Bone Cement in the Regeneration of Infected Bone Tissue.

Background Local antibiotic applications have been used in chronic osteomyelitis and have been defined as an adjunctive treatment method. Biodegradable materials are also used for the same purpose by adding antibiotics. The fact that it does not require additional surgery to be removed is an important advantage. In this study, we intended to develop a new biodegradable drug-loaded polymeric scaffold with good antibiotic release and compare the microbiological results with antibiotic-impregnated bone cement. Methodology A tissue scaffold containing poly(2-hydroxyethyl methacrylate) (PHEMA) was prepared in our laboratory and loaded with ertapenem and daptomycin antibiotics. The surface morphology and pore geometries of drug-loaded and unloaded scaffolds were analyzed by a scanning electron microscope under vacuum. The dose-dependent antiproliferative effects of PHEMA scaffold, drug-loaded scaffold, cement, and drug-loaded cement on osteoblast cells were investigated. To evaluate drug release kinetics, the absorbance values of the scaffold loaded with ertapenem and daptomycin were measured with the spectrometer. For microbiological tests, ertapenem and daptomycin-impregnated cement and scaffold, as well as the control scaffold and cement samples, were investigated for their antibacterial activities on Staphylococcus aureus and Klebsiella pneumoniae strains using the disc diffusion method. These microorganisms are one of the most common microorganisms in osteomyelitis. Results The efficacy of antibiotic-impregnated scaffold and cement on both gram-negative and gram-positive microorganisms was investigated. The daptomycin zone diameter in S. aureus ATCC 29233 strain was 17 mm, whereas it was 24 mm for scaffold and 22 mm for cement. Scaffold was found to be more effective than cement against S. aureus strain. The K. pneumoniae ATCC BAA-2814 strain was found to be resistant to ertapenem, but the zone diameter was 21 mm for scaffold and 20 mm for cement. Ertapenem-loaded scaffold was found to be more effective than cement. It was found that the antimicrobial activity of the scaffold was higher than cement. When we evaluated the release profiles, for the daptomycin-loaded cement group, 98% of daptomycin was cumulatively released within 30 minutes, and for the daptomycin-loaded scaffold group, 100% of daptomycin was cumulatively released in six days. To compare ertapenem-loaded cement and scaffold, 98% of ertapenem was cumulatively released within 10 minutes in the cement group. For the scaffold group, 100% of ertapenem was cumulatively released in 17 days. We found that the scaffold released the antibiotic more slowly and for a longer duration. Therefore, it was thought that the scaffold would be more effective on biofilm and the treatment of osteomyelitis would be more successful. Conclusions The produced scaffold was compared with cement, and it was concluded that the scaffold had better release and antimicrobial efficacy. Scaffold is more advantageous than cement because it is bioeliminable. Thus, there is no need for a second surgical intervention with the likely prevention of mortality and morbidity. Because of all these features, the scaffold seems promising in the local treatment of osteomyelitis.

Proteomic Analysis of the Role of the Adenylyl Cyclase-cAMP Pathway in Red Blood Cell Mechanical Responses.

Red blood cell (RBC) deformability is modulated by the phosphorylation status of the cytoskeletal proteins that regulate the interactions of integral transmembrane complexes. Proteomic studies have revealed that receptor-related signaling molecules and regulatory proteins involved in signaling cascades are present in RBCs. In this study, we investigated the roles of the cAMP signaling mechanism in modulating shear-induced RBC deformability and examined changes in the phosphorylation of the RBC proteome. We implemented the inhibitors of adenylyl cyclase (SQ22536), protein kinase A (H89), and phosphodiesterase (PDE) (pentoxifylline) to whole blood samples, applied 5 Pa shear stress (SS) for 300 s with a capillary tubing system, and evaluated RBC deformability using a LORRCA MaxSis. The inhibition of signaling molecules significantly deteriorated shear-induced RBC deformability (p < 0.05). Capillary SS slightly increased the phosphorylation of RBC cytoskeletal proteins. Tyrosine phosphorylation was significantly elevated by the modulation of the cAMP/PKA pathway (p < 0.05), while serine phosphorylation significantly decreased as a result of the inhibition of PDE (p < 0.05). AC is the core element of this signaling pathway, and PDE works as a negative feedback mechanism that could have potential roles in SS-induced RBC deformability. The cAMP/PKA pathway could regulate RBC deformability during capillary transit by triggering significant alterations in the phosphorylation state of RBCs.

The role of heat stress on the age related protein carbonylation.

Since the proteins are involved in many physiological processes in the organisms, modifications of proteins have important outcomes. Protein modifications are classified in several ways and oxidative stress related ones take a wide place. Aging is characterized by the accumulation of oxidized proteins and decreased degradation of these proteins. On the other hand protein turnover is an important regulatory mechanism for the control of protein homeostasis. Heat shock proteins are a highly conserved family of proteins in the various cells and organisms whose expressions are highly inducible during stress conditions. These proteins participate in protein assembly, trafficking, degradation and therefore play important role in protein turnover. Although the entire functions of each heat shock protein are still not completely investigated, these proteins have been implicated in the processes of protection and repair of stress-induced protein damage. This study has focused on the heat stress related carbonylated proteins, as a marker of oxidative protein modification, in young and senescent fibroblasts. The results are discussed with reference to potential involvement of induced heat shock proteins. This article is part of a Special Issue entitled: Protein Modifications. BIOLOGICAL SIGNIFICANCE: Age-related protein modifications, especially protein carbonylation take a wide place in the literature. In this direction, to highlight the role of heat shock proteins in the oxidative modifications may bring a new aspect to the literature. On the other hand, identified carbonylated proteins in this study confirm the importance of folding process in the mitochondria which will be further analyzed in detail.