One of the major challenges of implantology is to design nanoscale modifications of titanium implant surfaces inducing osseointegration. The aim of this study was to investigate the behavior of rat osteoblasts cultured on anodized TiO2 nanotubes of different crystallinity (amorphous and anatase phase) up to 24 days. TiO2 nanotubes were fabricated on VT1–0 titanium foil via a two-step anodization at 20 V using NH4F as an electrolyte. Anatase-phase samples were prepared by heat treatment at 500 °C for 1 h. VT1–0 samples with flat surfaces were used as controls. Primary rat osteoblasts were seeded over experimental surfaces for several incubation times. Scanning electron microscopy (SEM) was used to analyze tested surfaces and cell morphology. Cell adhesion and proliferation were investigated by cell counting. Osteogenic differentiation of cells was evaluated by qPCR of runt-related transcription factor 2 (RUNX2), osteopontin (OPN), integrin binding sialoprotein (IBSP), alkaline phosphatase (ALP) and osteocalcin (OCN). Cell adhesion and proliferation, cell morphology and the expression of osteogenic markers were affected by TiO2 nanotube layered substrates of amorphous and anatase crystallinity. In comparison with flat titanium, along with increased cell adhesion and cell growth a large portion of osteoblasts grown on the both nanostructured surfaces exhibited an osteocyte-like morphology as early as 48 h of culture. Moreover, the expression of all tested osteogenic markers in cells cultured on amorphous and anatase TiO2 nanotubes was upregulated at least at one of the analyzed time points. To summarize, we demonstrated that amorphous and anodized TiO2 layered substrates are highly biocompatible with rat osteoblasts and that the surface modification with about 1500 nm length nanotubes of 35 ± 4 (amorphous phase) and 41 ± 8 nm (anatase phase) in diameter is sufficient to induce their osteogenic differentiation. Such results are significant to the engineering of coating strategies for orthopedic implants aimed to establish a more efficient bone to implant contact and enhance bone repair.

Methods for obtaining osteoblast cultures from the calvaria of adult Wistar rats and 12-day-old embryos of these rats have been adapted for studying the biocompatibility and ossointegration of titanium-based implants. The osteoblast morphology and their differentiation into osteocytes on a titanium matrix with specially treated surface have been studied. It has been confirmed that two cultures of diploid rat cells obtained in the study can serve as efficient models for preclinical in vitro testing of nanostructured titanium implants for biocompatibility and osseointegration.

Relevance. The Sverdlovsk region is a highly endemic territory for infections transmitted by ixodic ticks. The possibilities of laboratory diagnostics of tick–borne infections in the routine practice of a clinician in the region today are limited mainly by testing blood serum for antibodies to the tick-borne encephalitis virus (TBE virus) and Lyme disease (LD) pathogens – Borrelia burgdorferi sensu lato complex, therefore, among tick-borne infections in the region currently mainly TBE and LD are registered. In case of negative results for antibodies to the pathogens of the above infections, the diagnosis may remain unknown. Aims. To study the etiological structure of infections transmitted by ixodic ticks in the Sverdlovsk region at the present stage, as well as to characterize the epidemiological and clinical features of the course of tick-borne infections detected in conditions of a combination of natural foci. Materials and methods. The study included 227 patients undergoing inpatient or outpatient treatment during the epidemic season of tick activity in 2021 (June-August) at LLS MO "New Hospital" (Urban Center of Natural Focal Infections). The case histories (epidemiological, clinical and laboratory indicators) were studied from each patient, as well as the material (blood) was examined prospectively and retrospectively using molecular biological (PCR) and serological (ELISA, planar protein biochip) methods. Results and discussion. During the study, six diseases were identified among the examined patients: TBE, LD (erythematous and nonerythematous forms), Borrelia miyamotoi disease (BMD), human granulocytic anaplasmosis (HGA), human monocytic ehrlichiosis (HME), as well as "Viral fever transmitted by arthropods" (VFTA). Among the examined patients, the proportion of patients with monoinfection was 49,5%, with mixed infection – 50,5%. Additional studies conducted by us using specific PCR for the presence DNA rickettsia and the causative agent of Q-fever in the blood of patients gave negative results. The general epidemiological characteristics for the above identified infections were the vector-borne mechanism, the prevalence of diseases in groups of middle-aged and elderly people, the largest number of cases of infection with pathogens of any tick-borne infection occurred in Yekaterinburg and its surroundings, for all infections, the duration of tick bite in most cases did not exceed one day; there were differences by sex, incubation period. Clinical symptoms were similar, except for the erythematous form of LD (the presence of erythema migrans at the byte site of tick), because the vast majority of patients had a general infectious syndrome; general laboratory indicators varied. Confirmation of the diagnosis was based on the results of the complex application of molecular biological and serological research methods. Conclusions. The modern etiological structure of infections transmitted by ixodic ticks in the Sverdlovsk region is represented not only by TBE and LD, but also by new disease to our country, such as BMD, HGA, HME, while a high proportion of mixed infections in various combinations has been revealed.

No abstract

No abstract