Cumulative loading increases and loading asymmetries persist during walking for people with a transfemoral bone-anchored limb.

BACKGROUND: A bone-anchored limb (BAL) is an alternative to a traditional socket-type prosthesis for people with transfemoral amputation. Early laboratory-based evidence suggests improvement in joint and limb loading mechanics during walking with a BAL compared to socket prosthesis use. However, changes in cumulative joint and limb loading measures, which may be predictive of degenerative joint disease progression, remain unknown. RESEARCH QUESTION: Do cumulative total limb and hip joint loading during walking change using a BAL for people with unilateral transfemoral amputation, compared to prior socket prosthesis use? METHODS: A case-series cohort of eight participants with prior unilateral transfemoral amputation who underwent BAL hardware implantation surgery were retrospectively analyzed (4 M/4 F; BMI: 27.7 ± 3.1 kg/m2; age: 50.4 ± 10.2 years). Daily step count and whole-body motion capture data were collected before (using socket prosthesis) and one-year after BAL hardware implantation. Cumulative total limb and hip joint loading and between-limb loading symmetry metrics were calculated during overground walking at both time points and compared using Cohen's d effect sizes. RESULTS: One year after BAL hardware implantation, participants demonstrated bilateral increases in cumulative total limb loading (amputated: d = -0.65; intact: d = -0.72) and frontal-plane hip moment (amputated: d = -1.29; intact: d = -1.68). Total limb loading and hip joint loading in all planes remained asymmetric over time, with relative overloading of the intact limb in all variables of interest at the one-year point. SIGNIFICANCE: Despite increases in cumulative total limb and hip joint loading, between-limb loading asymmetries persist. Habitual loading asymmetry has been implicated in contributing to negative long-term joint health and onset or progression of degenerative joint diseases. Improved understanding of methods to address habitual loading asymmetries is needed to optimize rehabilitation and long-term joint health as people with transfemoral amputation increase physical activity when using a BAL.

Toward a disruptive, minimally invasive small finger joint implant concept: Cellular and molecular interactions with materials in vivo.

Osteoarthritis (OA) poses significant therapeutic challenges, particularly OA that affects the hand. Currently available treatment strategies are often limited in terms of their efficacy in managing pain, regulating invasiveness, and restoring joint function. The APRICOT® implant system developed by Aurora Medical Ltd (Chichester, UK) introduces a minimally invasive, bone-conserving approach for treating hand OA (https://apricot-project.eu/). By utilizing polycarbonate urethane (PCU), this implant incorporates a caterpillar track-inspired design to promote the restoration of natural movement to the joint. Surface modifications of PCU have been proposed for the biological fixation of the implant. This study investigated the biocompatibility of PCU alone or in combination with two surface modifications, namely dopamine-carboxymethylcellulose (dCMC) and calcium-phosphate (CaP) coatings. In a rat soft tissue model, native and CaP-coated PCU foils did not increase cellular migration or cytotoxicity at the implant-soft tissue interface after 3 d, showing gene expression of proinflammatory cytokines similar to that in non-implanted sham sites. However, dCMC induced an amplified initial inflammatory response that was characterized by increased chemotaxis and cytotoxicity, as well as pronounced gene activation of proinflammatory macrophages and neoangiogenesis. By 21 d, inflammation subsided in all the groups, allowing for implant encapsulation. In a rat bone model, 6 d and 28 d after release of the periosteum, all implant types were adapted to the bone surface with a surrounding fibrous capsule and no protracted inflammatory response was observed. These findings demonstrated the biocompatibility of native and CaP-coated PCU foils as components of APRICOT® implants. STATEMENT OF SIGNIFICANCE: Hand osteoarthritis treatments require materials that minimize irritation of the delicate finger joints. Differing from existing treatments, the APRICOT® implant leverages polycarbonate urethane (PCU) for minimally invasive joint replacement. This interdisciplinary, preclinical study investigated the biocompatibility of thin polycarbonate urethane (PCU) foils and their surface modifications with calcium-phosphate (CaP) or dopamine-carboxymethylcellulose (dCMC). Cellular and morphological analyses revealed that both native and Ca-P coated PCU elicit transient inflammation, similar to sham sites, and a thin fibrous encapsulation in soft tissues and on bone surfaces. However, dCMC surface modification amplified initial chemotaxis and cytotoxicity, with pronounced activation of proinflammatory and neoangiogenesis genes. Therefore, native and CaP-coated PCU possess sought-for biocompatible properties, crucial for patient safety and performance of APRICOT® implant.

Molecular mechanisms of poor osseointegration in irradiated bone: In vivo study in a rat tibia model.

AIM: Radiotherapy is associated with cell depletion and loss of blood supply, which are linked to compromised bone healing. However, the molecular events underlying these effects at the tissue-implant interface have not been fully elucidated. This study aimed to determine the major molecular mediators associated with compromised osseointegration due to previous exposure to radiation. MATERIALS AND METHODS: Titanium implants were placed in rat tibiae with or without pre-exposure to 20 Gy irradiation. Histomorphometric, biomechanical, quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay analyses were performed at 1 and 4 weeks after implantation. RESULTS: The detrimental effects of irradiation were characterized by reduced bone-implant contact and removal torque. Furthermore, pre-exposure to radiation induced different molecular dysfunctions such as (i) increased expression of pro-inflammatory (Tnf) and osteoclastic (Ctsk) genes and decreased expression of the bone formation (Alpl) gene in implant-adherent cells; (ii) increased expression of bone formation (Alpl and Bglap) genes in peri-implant bone; and (iii) increased expression of pro-inflammatory (Tnf) and pro-fibrotic (Tgfb1) genes in peri-implant soft tissue. The serum levels of pro-inflammatory, bone formation and bone resorption proteins were greater in the irradiated rats. CONCLUSIONS: Irradiation causes the dysregulation of multiple biological activities, among which perturbed inflammation seems to play a common role in hindering osseointegration.

Lumbopelvic movement coordination during walking improves with transfemoral bone anchored limbs: Implications for low back pain.

BACKGROUND: Low back pain (LBP) is more prevalent in patients with transfemoral amputation using socket prostheses than able-bodied individuals, in part due to altered spinal loading caused by aberrant lumbopelvic movement patterns. Early evidence surrounding bone-anchored limb functional outcomes is promising, yet it remains unknown if this novel prosthesis influences LBP or movement patterns known to increase its risk. RESEARCH QUESTION: How are self-reported measures of LBP and lumbopelvic movement coordination patterns altered when using a unilateral transfemoral bone-anchored limb compared to a socket prosthesis? METHODS: Fourteen patients with unilateral transfemoral amputation scheduled to undergo intramedullary hardware implantation for bone-anchored limbs due to failed socket use were enrolled in this longitudinal observational cohort study (7 F/7 M, Age: 50.2±12.0 years). The modified Oswestry Disability Index (mODI) (self-reported questionnaire) and whole-body motion capture during overground walking were collected before (with socket prosthesis) and 12-months following bone-anchored limb implantation. Lumbopelvic total range of motion (ROM) and continuous relative phase (CRP) segment angles were calculated during 10 bilateral gait cycles. mODI, total ROM, CRP and CRP variabilities were compared between time points. RESULTS: mODI scores were significantly reduced 12-months after intramedullary hardware implantation for the bone-anchored limb (P = 0.013). Sagittal plane trunk and pelvis total ROM during gait were reduced after implantation (P = 0.001 and P < 0.001, respectively). CRP values were increased (more anti-phase) in the sagittal plane during single limb stance and reduced (more in-phase) in the transverse plane during pre-swing of the amputated limb gait cycle (P << 0.001 and P = 0.029, respectively). No differences in CRP values were found in the frontal plane. SIGNIFICANCE: Decreases in mODI scores and lumbopelvic ROM, paired with the changes in lumbopelvic coordination, indicate that bone-anchored limbs may reduce LBP symptoms and reduce compensatory movement patterns for people with unilateral transfemoral amputation.

Modulation of gene expression and bone formation by expanded and dense polytetrafluoroethylene membranes during guided bone regeneration: An experimental study.

BACKGROUND: Nonresorbable membranes promote bone formation during guided bone regeneration (GBR), yet the relationships between membrane properties and molecular changes in the surrounding tissue are largely unknown. AIM: To compare the molecular events in the overlying soft tissue, the membrane, and the underlying bone defect during GBR using dual-layered expanded membranes versus dense polytetrafluoroethylene (PTFE) membranes. MATERIALS AND METHODS: Rat femur defects were treated with either dense PTFE (d-PTFE) or dual-layered expanded PTFE (dual e-PTFE) or left untreated as a sham. Samples were collected after 6 and 28 days for gene expression, histology, and histomorphometry analyses. RESULTS: The two membranes promoted the overall bone formation compared to sham. Defects treated with dual e-PTFE exhibited a significantly higher proportion of new bone in the top central region after 28 days. Compared to that in the sham, the soft tissue in the dual e-PTFE group showed 2-fold higher expression of genes related to regeneration (FGF-2 and FOXO1) and vascularization (VEGF). Furthermore, compared to cells in the d-PTFE group, cells in the dual e-PTFE showed 2.5-fold higher expression of genes related to osteogenic differentiation (BMP-2), regeneration (FGF-2 and COL1A1), and vascularization (VEGF), in parallel with lower expression of proinflammatory cytokines (IL-6 and TNF-α). Multiple correlations were found between the molecular activities in membrane-adherent cells and those in the soft tissue. CONCLUSION: Selective surface modification of the two sides of the e-PTFE membrane constitutes a novel means of modulating the tissue response and promoting bone regeneration.

The effect of lacosamide in peripheral neuropathic pain: A randomized, double-blind, placebo-controlled, phenotype-stratified trial.

BACKGROUND: Neuropathic pain is common and difficult to treat. The sodium channel blocker lacosamide is efficacious in animal models of pain, but its effect on neuropathic pain in humans is inconclusive. METHODS: In a multicentre, randomized, double-blinded placebo-controlled phenotype stratified trial, we examined if lacosamide produced better pain relief in patients with the irritable nociceptor phenotype compared to those without. The primary outcome was the change in daily average pain from baseline to last week of 12 weeks of treatment. Secondary and tertiary outcomes included pain relief, patient global impression of change and presence of 30% and 50% pain reduction. RESULTS: The study was prematurely closed with 93 patients included and 63 randomized to lacosamide or placebo in a 2:1 ratio, of which 49 fulfilled the per protocol criteria and was used for the primary objective. We did not find a better effect of lacosamide in patients with the irritable nociceptor phenotype, the 95% CI for the primary objective was 0.41 (-1.2 to 2.0). For all patients randomized, lacosamide had no effect on the primary outcome, but significantly more patients were responders to lacosamide than during placebo, with an NNT of 4.0 (95% CI 2.3-16.1) and 5.0 (95% CI 2.8-24.5) for 30% and 50% pain reduction respectively. We did not identify any predictors for response. Lacosamide was generally well tolerated. CONCLUSION: We could not confirm that lacosamide was more efficacious in patients with the irritable nociceptor type, but the study was prematurely closed, so we cannot exclude a small difference. SIGNIFICANCE: Treatment of neuropathic pain is often a trial and error process. Little is known about which patient benefit from which kind of medication. The sodium channel blocker lacosamide shows variable effect on neuropathic pain. Pain sensory phenotype, as defined by quantitative sensory testing, did not predict response to treatment with lacosamide.

Leptin receptor gene deficiency minimally affects osseointegration in rats.

Metabolic syndrome represents a cluster of conditions such as obesity, hyperglycaemia, dyslipidaemia, and hypertension that can lead to type 2 diabetes mellitus and/or cardiovascular disease. Here, we investigated the influence of obesity and hyperglycaemia on osseointegration using a novel, leptin receptor-deficient animal model, the Lund MetS rat. Machined titanium implants were installed in the tibias of animals with normal leptin receptor (LepR+/+) and those harbouring congenic leptin receptor deficiency (LepR-/-) and were left to heal for 28 days. Extensive evaluation of osseointegration was performed using removal torque measurements, X-ray micro-computed tomography, quantitative backscattered electron imaging, Raman spectroscopy, gene expression analysis, qualitative histology, and histomorphometry. Here, we found comparable osseointegration potential at 28 days following implant placement in LepR-/- and LepR+/+ rats. However, the low bone volume within the implant threads, higher bone-to-implant contact, and comparable biomechanical stability of the implants point towards changed bone formation and/or remodelling in LepR-/- rats. These findings are corroborated by differences in the carbonate-to-phosphate ratio of native bone measured using Raman spectroscopy. Observations of hypermineralised cartilage islands and increased mineralisation heterogeneity in native bone confirm the delayed skeletal development of LepR-/- rats. Gene expression analyses reveal comparable patterns between LepR-/- and LepR+/+ animals, suggesting that peri-implant bone has reached equilibrium in healing and/or remodelling between the animal groups.

Targeting Pseudomonas aeruginosa quorum sensing with sodium salicylate modulates immune responses in vitro and in vivo.

Introduction: Chronic infections are a major clinical challenge in hard-to-heal wounds and implanted devices. Pseudomonas aeruginosa is a common causative pathogen that produces numerous virulence factors. Due to the increasing problem of antibiotic resistance, new alternative treatment strategies are needed. Quorum sensing (QS) is a bacterial communication system that regulates virulence and dampens inflammation, promoting bacterial survival. QS inhibition is a potent strategy to reduce bacterial virulence and alleviate the negative impact on host immune response. Aim: This study investigates how secreted factors from P. aeruginosa PAO1, cultured in the presence or absence of the QS inhibitor sodium salicylate (NaSa), influence host immune response. Material and methods: In vitro, THP-1 macrophages and neutrophil-like HL-60 cells were used. In vivo, discs of titanium were implanted in a subcutaneous rat model with local administration of P. aeruginosa culture supernatants. The host immune response to virulence factors contained in culture supernatants (+/-NaSa) was characterized through cell viability, migration, phagocytosis, gene expression, cytokine secretion, and histology. Results: In vitro, P. aeruginosa supernatants from NaSa-containing cultures significantly increased THP-1 phagocytosis and HL-60 cell migration compared with untreated supernatants (-NaSa). Stimulation with NaSa-treated supernatants in vivo resulted in: (i) significantly increased immune cell infiltration and cell attachment to titanium discs; (ii) increased gene expression of IL-8, IL-10, ARG1, and iNOS, and (iii) increased GRO-α protein secretion and decreased IL-1ß, IL-6, and IL-1α secretion, as compared with untreated supernatants. Conclusion: In conclusion, treating P. aeruginosa with NaSa reduces the production of virulence factors and modulates major immune events, such as promoting phagocytosis and cell migration, and decreasing the secretion of several pro-inflammatory cytokines.

High risk of dairy cow mortality in early lactation.

BACKGROUND: On-farm mortality among dairy cows constitutes a major problem in modern dairy production. Detailed knowledge about when cows die may help guide the prevention of future deaths. METHODS: Data from 46,927 on-farm dairy cow deaths reported to the Danish Cattle Database were analysed to evaluate the distribution of deaths in relation to time after calving and the effect of parity and breed. RESULTS: Of all on-farm deaths, 15.2% occurred within 7 days after calving, 32.2% within 30 days and 49.4% within 90 days. Older cows generally died sooner after calving compared to younger cows, and Jersey cows died later than other breeds. LIMITATIONS: The results are based on a large and reliable dataset but include only data from Danish dairy cows. CONCLUSION: The distribution of on-farm deaths was very uneven, with a large proportion of deaths occurring during the first few weeks after calving, especially in older cows.

Magnesium implant degradation provides immunomodulatory and proangiogenic effects and attenuates peri-implant fibrosis in soft tissues.

Implants made of magnesium (Mg) are increasingly employed in patients to achieve osteosynthesis while degrading in situ. Since Mg implants and Mg2+ have been suggested to possess anti-inflammatory properties, the clinically observed soft tissue inflammation around Mg implants is enigmatic. Here, using a rat soft tissue model and a 1-28 d observation period, we determined the temporo-spatial cell distribution and behavior in relation to sequential changes of pure Mg implant surface properties and Mg2+ release. Compared to nondegradable titanium (Ti) implants, Mg degradation exacerbated initial inflammation. Release of Mg degradation products at the tissue-implant interface, culminating at 3 d, actively initiated chemotaxis and upregulated mRNA and protein immunomodulatory markers, particularly inducible nitric oxide synthase and toll-like receptor-4 up to 6 d, yet without a cytotoxic effect. Increased vascularization was demonstrated morphologically, preceded by high expression of vascular endothelial growth factor. The transition to appropriate tissue repair coincided with implant surface enrichment of Ca and P and reduced peri-implant Mg2+ concentration. Mg implants revealed a thinner fibrous encapsulation compared with Ti. The detailed understanding of the relationship between Mg material properties and the spatial and time-resolved cellular processes provides a basis for the interpretation of clinical observations and future tailoring of Mg implants.

Prevalence of lameness in dairy cows: A literature review.

Lameness in dairy cows has major negative impacts on animal welfare and production economy. While previous studies have evaluated the prevalence of lameness in single countries, the present literature review is the first overview of the prevalence of lameness in dairy cows globally. This literature review identified 53 studies reporting prevalence of lameness among representative samples of dairy cows and fulfilling a number of specified inclusion criteria (e.g., at least 10 herds and 200 cows, and locomotion scoring by trained observers). A total of 414,950 cows from 3945 herds were included in these 53 studies, which spanned a 30-year period (1989-2020) and included herds from six continents, with the majority from Europe and North America. Across the studies, the mean prevalence of lameness (typically defined as score 3-5 on a 1-5 scale) was 22.8% with a median of 22.0% and a range between studies from 5.1% to 45%, and a within herd range from 0% to 88%. The mean prevalence of severely lame cows (typically defined as score 4-5 on a 1-5 scale) was 7.0% with a median of 6.5% and a range between studies from 1.8% to 21.2%, and a within herd range from 0% to 65%. Over time, it appears that the prevalence of lameness has changed very little. Several different locomotion scoring systems and definitions of (severe) lameness were used across the 53 studies, and this may have affected the reported lameness prevalence. Sampling of herds and cows, inclusion criteria and representativeness also differed between studies. This review offers recommendations for the future capture of information on lameness in dairy cows and identifies potential knowledge gaps.

Bone without borders - Monetite-based calcium phosphate guides bone formation beyond the skeletal envelope.

Calcium phosphates (CaP) represent an important class of osteoconductive and osteoinductive biomaterials. As proof-of-concept, we show how a multi-component CaP formulation (monetite, beta-tricalcium phosphate, and calcium pyrophosphate) guides osteogenesis beyond the physiological envelope. In a sheep model, hollow dome-shaped constructs were placed directly over the occipital bone. At 12 months, large amounts of bone (∼75%) occupy the hollow space with strong evidence of ongoing remodelling. Features of both compact bone (osteonal/osteon-like arrangements) and spongy bone (trabeculae separated by marrow cavities) reveal insights into function/need-driven microstructural adaptation. Pores within the CaP also contain both woven bone and vascularised lamellar bone. Osteoclasts actively contribute to CaP degradation/removal. Of the constituent phases, only calcium pyrophosphate persists within osseous (cutting cones) and non-osseous (macrophages) sites. From a translational perspective, this multi-component CaP opens up exciting new avenues for osteotomy-free and minimally-invasive repair of large bone defects and augmentation of the dental alveolar ridge.

Osseointegrated prostheses for the rehabilitation of patients with transfemoral amputations: A prospective ten-year cohort study of patient-reported outcomes and complications.

Background: Osseointegrated implants for patients with transfemoral amputations (TFAs) are a novel treatment under development, and prospective long-term evidence is lacking. The objectives were to determine patient-reported outcomes (PROs) and complications after ten years compared to before treatment and to compare the first five-year period with the later five-year period with regard to the outcomes. Methods: In a nonrandomized, prospective cohort study, patients with TFAs treated between 1999 and 2007 with the Osseointegrated Prosthesis for the Rehabilitation of Amputees (OPRA) system (n â€‹= â€‹51) (28 men/23 women; mean age at amputation: 32 years old; mean age at treatment: 44 years old in a single university hospital were followed for ten years. PROs included the Questionnaire for Persons with a Transfemoral Amputation (Q-TFA, four scores 0-100) and the Short Form 36 Health Survey (SF-36, ten scores 0-100) and were answered before treatment and until the ten-year follow-up after treatment. Analyses of differences in PRO scores were conducted using Wilcoxon's signed rank test. The implant survival and revision-free rates with respect to adverse events (implant revision, mechanical complications, and deep infections) were presented as Kaplan-Meier graphs with 95% confidence intervals (CIs). The incidences of events per ten and five person-years were calculated. Spearman's correlation analysis was used for analyses of associations between adverse events. Results: PROs showed statistically significant mean improvements between baseline and the ten-year follow-up with regard to all Q-TFA scores: the prosthetic use score (+36), prosthetic mobility score (+18), problem score (-28) and global score (+38) (all p â€‹< â€‹0.001), and the SF-36 physical functioning score (+26, p â€‹< â€‹0.001) and physical component score (+6, p â€‹< â€‹0.01). No PROs showed a statistically significant deterioration. Over the ten years, 12 patients were lost (one lost to follow-up, one dropped out of the study, two died, and eight had implants removed (four before five years and four between five and ten years). At ten years, the revision-free survival rates were 83% (CI: 69%-91%), 65% (CI: 49%-77%) and 17% (CI: 7%-29%) for implant revision, deep infection and mechanical complications, respectively. Mechanical complications, 3.9 per 10 person-years (CI: 2.2-5.1) constituted the most common serious adverse event and were more common during the last five years than during the first five years (p â€‹< â€‹0.001). No significant difference in the incidence of deep infections was observed between the earlier and the later five-year periods: 0.3 per 5 person-years (CI: 0.1-0.5) vs. 0.3 per person-years (CI: 0.1-0.5) (p â€‹= â€‹0.740). Correlation analyses between the earlier and later five years revealed a positive association between deep infections and implant removal (0.57, p â€‹< â€‹0.001) and between mechanical complications and adverse events (0.65, p â€‹< â€‹0.001). Conclusion: Improved PROs were demonstrated ten years after the introduction of a novel principle for bone anchorage of amputation prostheses. Nevertheless, an increasing rate of mechanical complications is of concern.

Non-invasive sampling procedure revealing the molecular events at different abutments of bone-anchored hearing systems-A prospective clinical pilot study.

Purpose: To investigate the molecular activities in different compartments around the bone-anchored hearing system (BAHS) with either electropolished or machined abutments and to correlate these activities with clinical and microbiological findings. Materials and methods: Twelve patients received machined or electropolished abutments after implant installation of BAHS. Peri-abutment fluid and tissue were collected from baseline to 12 months. Gene expression of cytokines and factors related to tissue healing and inflammation, regeneration and remodelling, as well as bacterial recognition were determined using quantitative-polymerase chain reaction (qPCR). The clinical status was evaluated using the Holgers scoring system, and bacterial colonisation was investigated by culturing. Results: The gene expression of inflammatory cytokines (IL-8, IL-1ß, and IL-10) and bacteria-related Toll-like receptors (2 and 4) was higher in the peri-abutment fluid than at baseline and in the peri-abutment tissue at 3 and 12 months. Conversely, the expression of genes related to tissue regeneration (Coll1a1 and FOXO1) was higher in the tissue samples than in the peri-abutment fluid at 3 and 12 months. Electropolished abutments triggered higher expression of inflammatory cytokines (IL-8 and IL-1ß) (in peri-abutment fluid) and regeneration factor FOXO1 (in peri-abutment tissue) than machined abutments. Several cytokine genes in the peri-abutment fluid correlated positively with the detection of aerobes, anaerobes and Staphylococcus species, as well as with high Holger scores. Conclusion: This study provides unprecedented molecular information on the biological processes of BAHS. Despite being apparently healed, the peri-abutment fluid harbours prolonged inflammatory activity in conjunction with the presence of different bacterial species. An electropolished abutment surface appears to be associated with stronger proinflammatory activity than that with a machined surface. The analysis of the peri-abutment fluid deserves further verification as a non-invasive sampling and diagnostic procedure of BAHS.

Effect of lameness on feeding behavior of zero grazed Jersey dairy cows.

The dairy industry faces major challenges with high levels of lameness, in parallel to an increased consumer focus on animal welfare. This encourages farmers to consider more robust breeds, such as Jersey cows. As little is known about the behavior of this breed under loose housing conditions, the present study sought to describe the feeding behavior of lame and non-lame Jersey cows in different parities. Such breed-specific information of behavioral changes is needed for breed-specific herd management decisions and may contribute to identifying animals that are susceptible to developing lameness in the future, thus reducing impacts on the welfare and production of cows. Feeding data from 116 Danish Jersey cows were collected using automatic feeders, and lameness status was assessed by technicians every second week. The cows were kept in a loose housing system, with cubicles, a slatted concrete floor, and automatic milking robots. Eating time per visit and per day, the number of visits per day, and intervals between meals were analyzed using generalized linear mixed effects models. The effect of lameness was not significant for any variable. Primiparous Jersey cows had significantly longer eating times per day, shorter meal intervals, and a lower number of visits per day than older Jersey cows. Week in lactation affected the eating time per visit and per day, the number of visits, and between-meal intervals. In conclusion, we found no differences between lame and non-lame Jersey cows but between parities, which disagree with previous research on other breeds, suggesting that Jersey cows not just differ in size and looks but also in their behavioral reaction when lame. Although data from only one herd of a research center were used, this study has demonstrated the need for further research about breed-specific differences and their implications for the health and welfare of the animals.

The Role of Titanium Particles and Ions in the Pathogenesis of Peri-Implantitis.

Titanium (Ti) particles and ions have been investigated in recent years as important factors in the pathogenesis of peri-implantitis. However, their role in the pathogenesis is yet to be fully understood. A review of pertinent literature was performed in various databases to determine the current position of Ti particles and ions role in the pathogenesis of peri-implantitis. There are several in vitro, preclinical and clinical published studies that have addressed the role of Ti particles and ions in the pathogenesis of peri-implantitis. These studies explored the effect of Ti particles and ions in the pathogenesis of peri-implantitis with respect to foreign body reaction, cellular response, epigenetic mechanisms, namely DNA methylation, and the oral microbiome. Studies have shown that the release of Ti particles/ions during implant insertion, early healing stages, late healing stages, and treatments during peri-implantitis might contribute to peri-implantitis through different mechanisms, such as foreign body reaction, cellular response, DNA methylation, and shaping the oral microbiome by increasing dysbiosis. However, further studies are needed to elucidate the complex interactions between all these mechanisms and Ti particles/ions in the pathogenesis and progression of peri-implantitis.

Welfare of equidae during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of animal welfare legislation. This opinion deals with the protection of horses and donkeys during transport. While the opinion focuses primarily on road transport of horses, there are specific sections dealing with the transport of horses on roll-on-roll-off ferries, horses transported by air and the transport of donkeys. In addition, the opinion covers welfare concerns in relation to a specific scenario identified by the European Commission related to the transport of horses on long journeys to slaughterhouses. Current practices related to transport of horses during the different stages (preparation, loading and unloading, transit and the journey breaks) are described. Overall, 13 welfare consequences were identified as being highly relevant for the welfare of horses during transport based on severity, duration and frequency of occurrence: gastro-enteric disorders, handling stress, heat stress, injuries, isolation stress, motion stress, prolonged hunger, prolonged thirst, respiratory disorders, resting problems, restriction of movement, sensory overstimulation and separation stress. These welfare consequences and their animal-based measures are described. A variety of hazards were identified related to factors such as inexperienced/untrained handlers, lack of horse training, structural deficiencies of vehicles/facilities, poor driving skills/conditions, horse separation/regrouping, unfavourable microclimatic and environmental conditions and poor husbandry practices. The opinion contains general and specific conclusions in relation to the different stages of transport. Recommendations to prevent hazards and correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and for space allowance. The development of welfare consequences over time was assessed in relation to maximum journey time.

Welfare of pigs during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of the animal welfare legislation. The present Opinion deals with protection of pigs during transport. The welfare of pigs during transport by road is the main focus, but other means of transport are also covered. Current practices related to transport of pigs during the different stages (preparation, loading/unloading, transit and journey breaks) are described. Overall, 10 welfare consequences were identified as highly relevant for the welfare of pigs during transport based on the severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, prolonged hunger, prolonged thirst, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A variety of hazards were identified, mainly relating to factors such as mixing of unfamiliar pigs, inappropriate handling methods and devices, the use of pick-up pens, inexperienced/untrained handlers, structural deficiencies of vehicles and facilities, poor driving conditions, unfavourable microclimatic and environmental conditions and poor husbandry practices leading to these welfare consequences. The Opinion contains general and specific conclusions relating to the different stages of transport of pigs. Recommendations to prevent hazards and to correct or mitigate welfare consequences are made. Recommendations were also developed to define quantitative thresholds for microclimatic conditions and minimum space allowance within means of transport. The development of the welfare consequences over time was assessed in relation to maximum journey duration. The Opinion covers specific animal transport scenarios identified by the European Commission relating to transport of cull sows and 'special health status animals', and lists welfare concerns associated with these.

Welfare of small ruminants during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of animal welfare legislation. The present Opinion deals with the protection of small ruminants (sheep and goats) during transport. The main focus is on welfare of sheep during transport by road but other means of transport and concerns for welfare of goats during transport are also covered. Current practices related to transport of sheep during the different stages (preparation, loading and unloading, transit and journey breaks) are described. Overall, 11 welfare consequences were identified as being highly relevant for the welfare of sheep during transport based on severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, predation stress, prolonged hunger, prolonged thirst, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A wide variety of hazards, mainly relating to inappropriate or aggressive handling of animals, structural deficiencies of vehicles and facilities, unfavourable microclimatic and environmental conditions and poor husbandry practices, leading to these welfare consequences were identified. The Opinion contains general and specific conclusions in relation to the different stages of transport. Recommendations to prevent hazards and to correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and spatial thresholds (minimum space allowance). The development of welfare consequences over time were assessed in relation to maximum journey time. The Opinion covers specific animal transport scenarios identified by the European Commission relating to the export of sheep by livestock vessels, export of sheep by road, roll-on-roll-off vessels and 'special health status animals', and lists welfare concerns associated with these.

Welfare of cattle during transport.

In the framework of its Farm to Fork Strategy, the Commission is undertaking a comprehensive evaluation of the animal welfare legislation. The present Opinion deals with protection of cattle (including calves) during transport. Welfare of cattle during transport by road is the main focus, but other means of transport are also covered. Current practices related to transport of cattle during the different stages (preparation, loading/unloading, transit and journey breaks) are described. Overall, 11 welfare consequences were identified as being highly relevant for the welfare of cattle during transport based on severity, duration and frequency of occurrence: group stress, handling stress, heat stress, injuries, motion stress, prolonged hunger, prolonged thirst, respiratory disorders, restriction of movement, resting problems and sensory overstimulation. These welfare consequences and their animal-based measures are described. A variety of hazards, mainly relating to inexperienced/untrained handlers, inappropriate handling, structural deficiencies of vehicles and facilities, poor driving conditions, unfavourable microclimatic and environmental conditions, and poor husbandry practices leading to these welfare consequences were identified. The Opinion contains general and specific conclusions relating to the different stages of transport for cattle. Recommendations to prevent hazards and to correct or mitigate welfare consequences have been developed. Recommendations were also developed to define quantitative thresholds for microclimatic conditions within the means of transport and spatial thresholds (minimum space allowance). The development of welfare consequences over time was assessed in relation to maximum journey duration. The Opinion covers specific animal transport scenarios identified by the European Commission relating to transport of unweaned calves, cull cows, the export of cattle by livestock vessels, the export of cattle by road, roll-on-roll-off ferries and 'special health status animals', and lists welfare concerns associated with these.