Environmentally persistent microbial contamination in agricultural soils: High risk of pathogenicity and antibiotic resistance.

Persistent microbial contamination commonly occurs in the environment. However, the characteristics and associated risks remain largely unknown. The coexistence of virulence factor genes (VFGs) and "last-resort" antibiotic resistance genes (LARGs) on human bacterial pathogens (HBPs) are notorious, creating ecological concerns and health risks. Herein, we explored the pathogenicity and antibiotic resistance levels of LARG-harboring HBPs in agricultural soils. Our findings revealed a high distribution level of VFGs and LARGs in soils (an absolute abundance up to 4.7 × 107 gene copies/g soil) by quantitative PCR (qPCR). Furthermore, most isolated LARG-harboring HBPs exhibited a 100 % lethality rate to Galleria mellonella. LARG-carrying plasmids had a low fitness cost to their host bacteria, implying the high adaptation of these plasmids within the HBPs. Most importantly, multiple LARG and VFG plasmid fusion and core genetic arrangements suggested that these LARG/VFG-linked plasmids endowed the stable and persistent horizontal spread of these genes in and/or cross the species and environments. This study not only unveiled high risk, multisource, compliance and stability aspects of environmentally persistent microbial contamination but also illuminated the importance of linking the phenotype-genotype-niche colonization of environmental microbial contamination within "One Health" framework.

Specific RNA m6A modification sites in bone marrow mesenchymal stem cells from the jawbone marrow of type 2 diabetes patients with dental implant failure.

The failure rate of dental implantation in patients with well-controlled type 2 diabetes mellitus (T2DM) is higher than that in non-diabetic patients. This due, in part, to the impaired function of bone marrow mesenchymal stem cells (BMSCs) from the jawbone marrow of T2DM patients (DM-BMSCs), limiting implant osseointegration. RNA N6-methyladenine (m6A) is important for BMSC function and diabetes regulation. However, it remains unclear how to best regulate m6A modifications in DM-BMSCs to enhance function. Based on the "m6A site methylation stoichiometry" of m6A single nucleotide arrays, we identified 834 differential m6A-methylated genes in DM-BMSCs compared with normal-BMSCs (N-BMSCs), including 43 and 790 m6A hypermethylated and hypomethylated genes, respectively, and 1 gene containing hyper- and hypomethylated m6A sites. Differential m6A hypermethylated sites were primarily distributed in the coding sequence, while hypomethylated sites were mainly in the 3'-untranslated region. The largest and smallest proportions of m6A-methylated genes were on chromosome 1 and 21, respectively. MazF-PCR and real-time RT-PCR results for the validation of erythrocyte membrane protein band 4.1 like 3, activity-dependent neuroprotector homeobox (ADNP), growth differentiation factor 11 (GDF11), and regulator of G protein signalling 2 agree with m6A single nucleotide array results; ADNP and GDF11 mRNA expression decreased in DM-BMSCs. Furthermore, gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that most of these genes were enriched in metabolic processes. This study reveals the differential m6A sites of DM-BMSCs compared with N-BMSCs and identifies candidate target genes to enhance BMSC function and improve implantation success in T2DM patients.

Shape-Memory ECM-Mimicking Heparin-Modified Nanofibrous Gelatin Scaffold for Enhanced Bone Regeneration in Sinus Augmentation.

Biomaterials with clinical maneuverability and predictable bone regeneration are needed in the field of maxillary sinus augmentation. Herein, gelatin was chemically modified with heparin that specifically interacted with and stabilized bone morphogenetic protein-2 (BMP-2). We then introduced thermally induced phase separation to form the injectable, shape-memory, highly porous scaffold for bone regeneration in sinus augmentation. The hydrated heparin-modified nanofibrous gelatin scaffolds (NH-GS) were demonstrated with high resilience and shape-memory property, both macroscopically and microscopically, making them injectable scaffolds and expected to be applied in sinus augmentation. This novel scaffold was verified to be biocompatible and an excellent matrix to support cell attachment, proliferation, and infiltration. Further, the growth factor-loaded NH-GS showed sustained release kinetics of BMP-2 through affinity-based scaffold-growth factor interaction, compared with BMP-2 loaded gelatin sponge (GS) and nanofibrous gelatin scaffold (NF). Both in vitro and in vivo experiments demonstrated that the BMP-2-loaded NH-GS exhibited the highest osteogenesis among the other groups. Taken together, this study introduces a new regenerative strategy in maxillary sinus augmentation, which is injectable with a predefined shape and structure and promotes bone regeneration through a more sustained BMP-2 release.

A biodegradable gelatin-based nanostructured sponge with space maintenance to enhance long-term osteogenesis in maxillary sinus augmentation.

The search for bone substitutes that are biodegradable, ensure space maintenance, and have osteogenic predictability, is ongoing in the field of sinus augmentation. We thus compared the bone regeneration potential of nanostructured sponges (NS-Sponge) with that of collagen-stabilized inorganic bovine bones (BO-Collagen), gelatin sponges (Gelatin), and blood clots (Cont) in sinus augmentation of rabbits. NS-Sponge was prepared by thermally induced phase separation with porogen leaching techniques. All the materials were non-hemolytic and cytocompatible. The porous and nanofibrous NS-Sponge showed better dimensional stability to support cell growth and osteogenic differentiation. In vivo, the sinus membrane collapsed in Cont and Gelatin, while BO-Collagen and NS-Sponge maintained the elevated height as assessed by come-beam computed tomography. Limited bone regeneration was observed in Cont and Gelatin. In the entire implanted area, histological analysis revealed a higher percentage of new bone area at 4 weeks of BO-Collagen treatment; however, a significantly greater increase in new bone area was observed after 12 weeks of NS-Sponge treatment. The 12-week remnant NS-Sponge material was significantly lower than the 4-week remnant material. Overall, NS-Sponge may be highly recommended for sinus augmentation, as it exhibits numerous advantages, including excellent operability, clear imaging characteristics, space maintenance, biodegradability, and superior osteogenic potential.