Vaccination intention among healthcare workers during the first wave of the coronavirus disease 2019 pandemic in relation to knowledge: a cross-sectional study in Croatia, Slovenia, Serbia, and Poland.

AIM: To analyze SARS-CoV-2 vaccination intention and acceptance in relation to the knowledge about coronavirus disease 2019 (COVID-19) among healthcare workers (HCWs) in Croatia, Slovenia, Serbia, and Poland. METHODS: In spring 2020, an online survey was distributed among HCWs by using snowball sampling. The questionnaire was fully completed by 623 respondents: 304 from Croatia, 86 from Slovenia, 90 from Serbia, and 143 from Poland. The survey collected data on demographic characteristics (age, gender, education), vaccination acceptance, and knowledge about COVID-19. RESULTS: A total of 31% of respondents declared their intention to be vaccinated when a vaccine against COVID-19 is available, and 45% were undecided. Vaccination intention was associated with age, educational level, and knowledge about the pandemic, and differed significantly among the countries. Younger HCWs (18-25 years) and those with higher education more frequently expressed vaccination acceptance. Vaccination acceptance score was not associated with gender. CONCLUSIONS: HCWs with higher knowledge were more likely to express vaccination intention. Improving the knowledge about COVID-19 and increasing HCWs' education might also increase vaccination acceptance among HCWs, and consequently in the general population.

Gene expression of cultured human chondrocytes as a model for assessing neutralization efficacy of soluble TNFα by TNFα antagonists.

Tumor necrosis factor-alpha (TNFα) antagonists are efficacious in the treatment of various immune-mediated inflammatory diseases. Because of rapidly growing demand for developing new or biosimilar versions of these biologicals, the need to create in vitro testing models that best represent physiological conditions is increasing. Primary human chondrocytes were used for potency evaluation and comparison between the molecular effects of anti-TNFα biologicals. Infliximab and etanercept were chosen to assess the suitability of chondrocyte cell culture for determination of anti-TNFα neutralization efficacy employing quantitative reverse transcription-polymerase chain reaction (qRT-PCR) technology. Use of both anti-TNFα biologics resulted in decrease of TNFα-stimulated expression of various matrix metalloproteinases, interleukins and other inflammation-related genes in our cell model. Significant differences in inhibition efficacy of etanercept and infliximab were observed, which were confirmed also on protein level. To evaluate the potency of anti-TNFα biologicals, a selection of TNFα-responsive target genes was made from the gene array data. The selected genes were employed in development of statistical model, which enables comparability of anti-TNFα biologicals. The presented analytical approach is suitable for assessment of the neutralization efficacy of various anti-TNFα biologicals. As such, it can be used for additional comprehensive characterization and comparability of TNF antagonists in preclinical drug testing.

Disc cell therapies: critical issues.

BACKGROUND: Disc cell therapies, in which cells are injected into the degenerate disc in order to regenerate the matrix and restore function, appear to be an attractive, minimally invasive method of treatment. Interest in this area has stimulated research into disc cell biology in particular. However, other important issues, some of which are discussed here, need to be considered if cell-based therapies are to be brought to the clinic. PURPOSE: Firstly, a question which is barely addressed in the literature, is how to identify patients with 'degenerative disc disease' who would benefit from cell therapy. Pain not disc degeneration is the symptom which drives patients to the clinic. Even though there are associations between back pain and disc degeneration, many people with even severely degenerate discs, with herniated discs or with spinal stenosis, are pain-free. It is not possible using currently available techniques to identify whether disc repair or regeneration would remove symptoms or prevent symptoms from occurring in future. Moreover, the repair process in human discs is very slow (years) because of the low cell density which can be supported nutritionally even in healthy human discs. If repair is necessary for relief of symptoms, questions regarding quality of life and rehabilitation during this long process need consideration. Also, some serious technical issues remain. Finding appropriate cell sources and scaffolds have received most attention, but these are not the only issues determining the feasibility of the procedure. There are questions regarding the safety of implanting cells by injection through the annulus whether the nutrient supply to the disc is sufficient to support implanted cells and whether, if cells are able to survive, conditions in a degenerate human disc will allow them to repair the damaged tissue. CONCLUSIONS: If cell therapy for treatment of disc-related disorders is to enter the clinic as a routine treatment, investigations must examine the questions related to patient selection and the feasibility of achieving the desired repair in an acceptable time frame. Few diagnostic tests that examine whether cell therapies are likely to succeed are available at present, but definite exclusion criteria would be evidence of major disc fissures, or disturbance of nutrient pathways as measured by post-contrast MRI.

Cell sources for nucleus pulposus regeneration.

PURPOSE: There is increasing interest in the development of cell therapy as a possible approach for the treatment of degenerative disc disease. To regenerate nucleus pulposus tissue, the cells must produce an appropriate proteoglycan-rich matrix, as this is essential for the functioning of the intervertebral disc. The natural environment within the disc is very challenging to implanted cells, particularly if they have been subcultured in normal laboratory conditions. The purpose of this work is to discuss parameters relevant to translating different proposed cell therapies of IVD into clinical use. RESULTS: Several sources of cells have been proposed, including nucleus pulposus cells, chondrocytes and mesenchymal stem cells derived from bone marrow or adipose tissue. There are some clinical trials and reports of attempts to regenerate nucleus pulposus utilising either autologous or allogenic cells. While the published results of clinical applications of these cell therapies do not indicate any safety issues, additional evidence will be needed to prove their long-term efficacy. CONCLUSION: This article discusses parameters relevant for successful translation of research on different cell sources into clinically applicable cell therapies: the influence of the intervertebral disc microenvironment on the cell phenotype, issues associated with cell culture and technical preparation of cell products, as well as discussing current regulatory requirements. There are advantages and disadvantages of each proposed cell type, but no strong evidence to favour any one particular cell source at the moment.

Barriers to Inter-Organisational Collaboration in the Preoperative Management of Patients with Osteoarthritis of the Hip or Knee.

Introduction: Integrated clinical pathways should provide the best and most efficient treatment. As no study on barriers to inter-organisational collaboration has investigated the barriers to unimplemented integrated clinical care in a country with less efficiently organised health system, the study aimed to identify these barriers in the preoperative management of patients with hip or knee osteoarthritis in Slovenia. Methods: A cross-sectional study was conducted using multiple methods, including a quantitative survey with participants involved in target patient groups, and in-depth interviews with involved key actors at micro, meso and macro levels in Slovenia. Results: Respondents predominantly expressed a lack of inter-organisational collaboration. The exposed barriers are individualistic culture, the level of development of the health system, financing, administration, and regulatory frame at the macro level, shortage of staff at the meso level, and the lack of technological standards, trust, communication, and perception of pressures at the micro level. Discussion and conclusion: In addition to the barriers identified in previous studies, our study shows that individualistic culture and the level of development of the health system at the macro level, manifested as a pressure on health professionals and other actors at the micro level, are important barriers to inter-organisational collaboration.

Influence of Psychological Factors on Vaccination Acceptance among Health Care Workers in Slovenia in Three Different Phases of the COVID-19 Pandemic.

COVID-19 vaccination acceptance among healthcare workers (HCWs) is very important to control the pandemic and to ensure the safety of HCWs and patients. As psychological factors may affect the decision to be vaccinated, the aim of this study was to investigate the influence of psychological factors on vaccination acceptance in different phases of the COVID-19 pandemic. A cross-sectional study using a web-based survey was conducted among HCWs in Slovenia at the beginning of the pandemic (N = 851), one month later (N = 86), and one year later (N = 145) when vaccines were already available. The results showed that the influence of psychological factors (anxiety, psychological burden, perceived infectability, and germ aversion) was specific for each survey period. At the beginning of the pandemic, vaccination intention was positively associated with anxiety. In the third survey period, anxiety was not exposed as a predictive factor for vaccination intention. However, comparison of vaccination status among groups with different levels of anxiety revealed an interesting distinction within those in favour of vaccination; in the group with minimal levels of anxiety, there was a relatively high share of respondents that were already vaccinated, whereas in the group with severe anxiety, most individuals intended to be vaccinated but hesitated to take action.

Attitudes of Nursing Students towards Vaccination and Other Preventive Measures for Limitation of COVID-19 Pandemic: Cross-Sectional Study in Three European Countries.

Several preventive measures have been applied to limit the COVID-19 pandemic, including successful the development and introduction of vaccines. The aim of this study was to investigate adherence to preventive measures and vaccination intentions among nursing students in three European countries and the factors associated with vaccination intention and advising vaccination. A cross-sectional study using convenience/snow-ball sampling strategy was performed in Slovenia, Poland, and Serbia between 12 February and 5 March 2021. Data from 872 eligible respondents were analyzed (mean age 23.5 ± 6.5 years, 89% female). Higher adherence to preventive behavior was declared by those working in healthcare (p < 0.001), engaged in COVID-19 departments (p < 0.001), had not had the disease yet (p < 0.001), and had children (p = 0.01). Those groups also expressed higher vaccination intention and advised vaccination to others. Higher vaccination intention and advising vaccination were mostly associated with belief in benefits of vaccine, trust in institutions, perceived effectiveness of vaccine, influence of social environment, protection of patients and perceived health care professionals' duty. Fear of side effects and general refusal of vaccines are the main reasons for vaccination hesitancy. The results of the study indicate how higher education institutions can support the development of appropriate professional attitudes and behaviors among nursing students.

Mental Health and Wellbeing at Schools: Health Promotion in Primary Schools with the Use of Digital Methods.

Mental health disorders among primary school children remain a crucial issue. Early health promotion interventions can positively affect and prevent the onset of mental disorders. Promising digital mental health methods have been implemented for adolescents and youths with scarce evidence among younger ages. Therefore, the aim of the current systematic review was to identify health promotion interventions on mental health and wellbeing, with the use of digital methods, delivered in primary school settings. Six digital interventions have been identified, three of which were targeting teachers and the others students. Regardless of the limited number of studies, the effectiveness of the web-based interventions upon teachers' knowledge and attitudes and the positive impact on children's behavioral improvements has been documented. The lack of adequate evidence highlights the need for further research in the field. The current review provides information for professionals working in primary schools useful for the design and implementation of effective mental health and wellbeing interventions.

Primary human alveolar bone cells isolated from tissue samples acquired at periodontal surgeries exhibit sustained proliferation and retain osteogenic phenotype during in vitro expansion.

OBJECTIVES: Bone tissue regeneration requires a source of viable, proliferative cells with osteogenic differentiation capacity. Periodontal surgeries represent an opportunity to procure small amounts of autologous tissues for primary cell isolation. Our objective was to assess the potential of human alveolar bone as a source of autologous osteogenic cells for tissue engineering and biomaterials and drug testing studies. MATERIALS AND METHODS: Alveolar bone tissue was obtained from 37 patients undergoing routine periodontal surgery. Tissue harvesting and cell isolation procedures were optimized to isolate viable cells. Primary cells were subcultured and characterized with respect to their growth characteristics, gene expression of osteogenic markers, alkaline phosphatase activity and matrix mineralization, under osteogenic stimulation. RESULTS: Alveolar bone cells were successfully isolated from 28 of the 30 samples harvested with bone forceps, and from 2 of the 5 samples obtained by bone drilling. The yield of cells in primary cultures was variable between the individual samples, but was not related to the site of tissue harvesting and the patient age. In 80% of samples (n = 5), the primary cells proliferated steadily for eight subsequent passages, reaching cumulative numbers over 10(10) cells. Analyses confirmed stable gene expression of alkaline phosphatase, osteopontin and osteocalcin in early and late cell passages. In osteogenic medium, the cells from late passages increased alkaline phosphatase activity and accumulated mineralized matrix, indicating a mature osteoblastic phenotype. CONCLUSIONS: Primary alveolar bone cells exhibited robust proliferation and retained osteogenic phenotype during in vitro expansion, suggesting that they can be used as an autologous cell source for bone regenerative therapies and various in vitro studies.

Bone grafts engineered from human adipose-derived stem cells in perfusion bioreactor culture.

We report engineering of half-centimeter-sized bone constructs created in vitro using human adipose-derived stem cells (hASCs), decellularized bone scaffolds, and perfusion bioreactors. The hASCs are easily accessible, can be used in an autologous fashion, are rapidly expanded in culture, and are capable of osteogenic differentiation. hASCs from four donors were characterized for their osteogenic capacity, and one representative cell population was used for tissue engineering experiments. Culture-expanded hASCs were seeded on fully decellularized native bone scaffolds (4 mm diameter x 4 mm thick), providing the necessary structural and mechanical environment for osteogenic differentiation, and cultured in bioreactors with medium perfusion. The interstitial flow velocity was set to a level necessary to maintain cell viability and function throughout the construct volume (400 microm/s), via enhanced mass transport. After 5 weeks of cultivation, the addition of osteogenic supplements (dexamethasone, sodium-beta-glycerophosphate, and ascorbic acid-2-phosphate) to culture medium significantly increased the construct cellularity and the amounts of bone matrix components (collagen, bone sialoprotein, and bone osteopontin). Medium perfusion markedly improved the distribution of cells and bone matrix in engineered constructs. In summary, a combination of hASCs, decellularized bone scaffold, perfusion culture, and osteogenic supplements resulted in the formation of compact and viable bone tissue constructs.

Comparison of articular and auricular cartilage as a cell source for the autologous chondrocyte implantation.

Articular (medial femoral condyle) and auricular cartilage (anithelix) was compared as a cell source for the autologous joint repair. Cells isolated from five human cadaveric donors were cultured parallel in the monolayer cultures and in the 3D alginate hydrogel constructs for 1 week. Cell morphology was controlled by the fluorescent microscopy and gene expressions of type I collagen (COL1), type II collagen (COL2), aggrecan (AGR), versican (VER), and elastin (ELS) were analyzed by the real-time polymerase chain reaction. COL1 and ELS, predominant in the phenotype of auricular biopsy, were statistically lower in the articular biopsies. Even though COL2 and AGR decreased in monolayers of both cell sources, the dedifferentiation process affected auricular cells intensely. Cells embedded in the alginate hydrogel directly after the isolation did not exhibit the dedifferentiated phenotype. Additionally, COL1, COL2, AGR, and VER were comparable between the two sources. ELS however, remained higher in the auricular cells regardless of the culture type. The study indicates that auricular chondrocytes cultured in a 3D environment immediately after the isolation have a neo-cartilage potential for the articular surface reconstruction.

Quantitative analysis of gene expression in human articular chondrocytes assigned for autologous implantation.

Autologous chondrocyte implantation (ACI) relies on the implantation of in vitro expanded cells. The aim was to study the dedifferentiation of human articular chondrocytes under different cultivating conditions [days 0-10 in the primary culture (P0); passages in a monolayer from P0 to P3; monolayer vs. alginate and monolayer vs. alginate/agarose hydrogels] using real-time PCR analysis. The relative gene expressions for collagen type I and II, aggrecan and versican were quantified and the corresponding differentiation indexes (Col2/Col1, Agr/Ver) were calculated. The values of both differentiation indexes decreased exponentially with time in the P0 monolayer culture, and continued with a significant decrease over the subsequent monolayer passages. On the contrary, the chondrocytes seeded in either of the hydrogels significantly increased the indexes compared to their parallel monolayer cultures. These results indicate that alginate and alginate/agarose hydrogels offer an appropriate environment for human articular chondrocytes to redifferentiate after being expanded in vitro. Therefore the three-dimensional (3D) hydrogel chondrocyte cultures present not only surgical, but also biological advantage over the classic suspension-periosteum chondrocyte implantation.

Nucleus pulposus repair with cultured autologous elastic cartilage derived chondrocytes.

Low back pain is one of the most common medical conditions in the Western world. Disc degeneration, an inevitable process of ageing, is one of the major causes of low back pain. Autologous chondrocyte transplantation (ACT) is an increasingly popular method of addressing pathological disorders of cartilage. The purpose of our study was to determine whether autologous chondrocytes from elastic cartilage could survive and synthesise a cartilage specific matrix in the intervertebral disc of rabbits. Sixteen lumbar intervertebral discs (IVD) of New Zealand White rabbits were analysed. In 6 IVD, the nucleus pulposus was evacuated and replaced with tissue engineered autologous chondrocytes from auricular cartilage. In the second group, only the nucleus pulposus was evacuated from 6 IVD, with no chondrocytes implantation. Four non-operated IVD were used as a control. Six months after the operation, the animals were euthanized and the IVD were analysed histologically. Autologous cartilage implants were well tolerated by the host for up to six months in vivo. There was only hyaline-like cartilage in the place of the nucleus pulposus. We could not detect any elastic fibres in the new cartilage matrix. In IVD from which only the nucleus pulposus was evacuated and no chondrocytes were implanted, just fibrous tissue was found instead of nucleus pulposus. The overall histological analysis of new cartilage produced after implantation in our study confirmed the hypothesis that ACT from auricular cartilage can be implanted into the IVD instead of the nucleus pulposus and that a significant percentage of implanted chondrocytes survive and produce hyaline-like cartilage.

The outcome of autologous chondrocyte transplantation treatment of cartilage lesions in the knee.

Recent results of the clinical outcome of autologous chondrocyte transplantation (ACT) treatment in a group of 28 patients with focal femoral condyle cartilage lesions revealed a correlation trend with the quality of the in vitro cell culture matrix-protein synthesis. No impact of the patients' age and chondrocyte cryopreservation prior to implantation was observed. Further studies are needed to confirm the preliminary results.