AbstractBackground: Ehrlichiaspp. are a group of intracellular parasitic bacteria primarily transmitted by ticks and exhibit a wide global distribution. Their ability to infect a diverse range of mammals, including humans, underscores their immense public health significance. Methods: We collected tick and dog blood samples in different areas of Hainan Province. The species of ticks were identified by standard PCR of the tick 16S rRNA gene. Detection of Ehrlichiainfection in ticks and dogs using nested PCR of the 16S rRNA gene of Ehrlichia spp.. The obtained sequences were subjected to comparison with other sequences cataloged in GenBank. Subsequently, risk factors affecting Ehrlichiainfection were statistically analysed. Results: All 631 ticks belonged to the Rhipicephaluslinnaei; 63 (9.98%) out of 631 ticks tested positive for Ehrlichia canis, while 140 (11.08%) out of 1264 dog blood samples were positive for E. canis. Notably, Ehrlichia ewingii and Ehrlichia chaffeensiswere not detected. The prevalence of Ehrlichia infection in dogs is correlated with factors such as age, breed, dewormer use, tick infestation, and living environment, while displaying no association with the gender of the dog. Conclusions: In Hainan Province, Rhipicephalus linnaei is the dominant tick species infecting dogs.Dogs are vulnerable to Ehrlichia infection, particularly rural and stray dogs.
Magnesium (Mg)-based implants were extensively developed and tested to improve the shortages of traditional hard metal implants. Unlike the nail, screw, and plate, pure Mg wire is rarely applied in the musculoskeletal system because of its poor mechanical properties. Therefore, we developed the magnesium–zinc–gadolinium (ZG21) alloy wire, which presented good mechanical properties. Before the in vivo study, the in vitro tests were carried out in this study. The ZG21 wire was scanned by SEM/EDS. The changing rate of weight and pH values were recorded during degradation. The corrosion interface was scanned by SEM/EDS. The cytotoxicity of metal extracts, Mg, Zn, and Gd ions was tested. The osteogenic and angiogenic potential was also evaluated. The ZG21 wire degraded at a stable speed in 14 days. The extracts were diluted ten times, and the correspondent ion concentration presented low cytotoxicity for cell lines of pre-osteoblasts, fibroblasts, and endothelial vessel cells. Pre-osteoblast cell lines cultured with 10% extracts presented significantly higher osteogenic potential. Endothelial vessel cell lines cultured with 2.5, 5, and 10 mM Mg2+ presented significantly higher angiogenic potential. The ZG21 wire could maintain an intact structure when making a surgical knot. Its degradation process and products presented low toxicity and potential for osteogenesis and angiogenesis. The ZG21 wire could be identified as a safe and bioactive material for further in vivo musculoskeletal regeneration.
After reconstructing the anterior cruciate ligament (ACL), unsatisfactory bone tendon interface healing may often induce tunnel enlargement at the early healing stage. With good biological features and high formability, Magnesium-Zinc-Gadolinium (ZG21) wires are developed to bunch the tendon graft for matching the bone tunnel during transplantation. Microstructure, tensile strength, degradation, and cytotoxicity of ZG21 wire are evaluated. The rabbit model is used for assessing the biological effects of ZG21 wire by Micro-CT, histology, and mechanical test. The SEM/EDS, immunochemistry, and in vitro assessments are performed to investigate the underlying mechanism. Material tests demonstrate the high formability of ZG21 wire as surgical suture. Micro-CT shows ZG21 wire degradation accelerates tunnel bone formation, and histologically with earlier and more fibrocartilage regeneration at the healing interface. The mechanical test shows higher ultimate load in the ZG21 group. The SEM/EDS presents ZG21 wire degradation triggered calcium phosphate (Ca-P) deposition. IHC results demonstrate upregulation of Wnt3a, BMP2, and VEGF at the early phase and TGFβ3 and Type II collagen at the late phase of healing. In vitro tests also confirmed the Ca-P in the metal extract could elevate the expression of Wnt3a, β catenin, ocn and opn to stimulate osteogenesis. Ex vivo tests of clinical samples indicated suturing with ZG21 wire did not weaken the ultimate loading of human tendon tissue. In conclusion, the ZG21 wire is feasible for tendon graft bunching. Its degradation products accelerated intra-tunnel endochondral ossification at the early healing stage and therefore enhanced bone-tendon interface healing in ACL reconstruction.
Biodegradable magnesium (Mg) materials are considered ideal as osteosynthesis implants. However, clinical application has proven complex. This is primarily associated with the issue of reducing the extent of implant degradation to a range acceptable for the human body, while simultaneously enhancing osteogenesis or osteoinduction. In the present study, a combination of Mg ions and low‑intensity pulsed ultrasound (LIPUS) treatment was applied in hFOB 1.19 human osteoblast cells as a potential strategy to resolve this issue. A total of 7,314 differentially expressed genes (DEGs) and 826 shared DEGs in hFOB1.19 osteoblast cells were identified by microarray analysis following treatment with Mg and/or LIPUS. Gene Ontology analysis demonstrated that among cells treated with a combination of Mg and LIPUS, DEGs were significantly enriched in various functional annotations, including 'wound healing', 'transforming growth factor beta receptor signaling pathway', 'transcription, DNA‑templated', 'receptor complex', 'nucleus', 'SMAD protein complex', 'DNA binding', 'metal ion binding' and 'GTPase activator activity'. Notably, the transforming growth factor (TGF)‑β, mitogen‑activated protein kinase (MAPK) and tumor necrosis factor (TNF) signaling pathways were preferentially overrepresented in the Mg and LIPUS combination group, which was subsequently confirmed by reverse transcription‑quantitative polymerase chain reaction. Furthermore, genes involved in osteoblast mineralization promotion, including bone morphogenetic protein 6, noggin, bone morphogenetic protein receptor (BMPR)1A, BMPR2 and SMAD 5/8, were significantly upregulated following combination treatment compared with the control group. Genes involved in the promotion of migration, including c‑Jun N‑terminal kinase, doublecortin, paxillin and Jun proto‑oncogene AP‑1 transcription factor subunit, were also upregulated in the combination treatment group compared with the control group. The DEG data were supported by morphological observations of the osteoblasts using alizarin red S staining and wound healing assays, which indicated that Mg and LIPUS combinative treatment had a synergistic effect on osteoblast mineralization and migration. Additionally, the combined treatment significantly upregulated metal transporter genes associated with Mg entry, including ATPase Na+/K+‑transporting subunit α1, cyclin and CBS domain divalent metal cation transport mediator 2, K+ voltage‑gated channel subfamily J member 14, transient receptor potential cation channel (TRP) subfamily M member 7 and TRP subfamily V member 2. In summary, the findings of the present study revealed that combined stimulation with Mg and LIPUS may exhibit a synergistic effect on human osteoblast bone formation through the TGF‑β, MAPK and TNF signaling pathways, while also facilitating Mg influx. The present study demonstrated the potential of combinative LIPUS and Mg treatment as a novel therapeutic strategy for enhancing the osteoinduction, biocompatibility and biosafety of biodegradable Mg implants.
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