Search Results (429)

Background: The role of collagen membrane fixation during guided bone regeneration (GBR) remains debatable, particularly in post-extraction sockets with buccal defects and concomitant immediate implant placement. This study evaluated whether or not fixation with titanium pins improved regenerative outcomes. Methods: Six adult Beagle dogs received bilateral extractions of the fourth mandibular premolars. An implant was immediately placed in both the distal alveoli, and standardized buccal bone defects (5 mm height, 3–2 mm width) were prepared. All defects were filled with a slowly resorbing equine xenograft and covered by a resorbable pericardium membrane. At the test sites, the membrane was apically fixed with pins, while no fixation was applied to the control sites. After 3 months of healing, histomorphometric analyses were performed. Results: The vertical bone gain of the buccal crest was 3.2 mm in the test sites (pin group) and 2.9 mm in the control sites (no-pin) (p > 0.754). No significant difference was found in terms of bone-to-implant contact (BIC). However, residual graft particles were located significantly more coronally in the pin group compared to the no-pin group (p = 0.021). Morphometric analyses revealed similar new bone formation within the groups, but with higher amounts of residual xenograft and soft tissue in the pin group. Conclusions: Membrane fixation did not significantly enhance vertical bone gain, and although the slightly higher regeneration in the pin group (3.2 mm vs. 2.9 mm) may hold clinical relevance in esthetically sensitive areas and osseointegration, it appeared to limit coronal migration of the grafting material. Full article

Soil salinization poses a significant constraint to agricultural productivity. However, certain plant growth-promoting bacteria (PGPB) can mitigate salinity stress and enhance crop performance. In this study, a bacterial isolate, R-18, isolated from saline-alkali soil in Ningxia, China, was identified as Enterobacter bugandensis based on 16S rRNA gene sequencing. The isolate was characterized for its morphological, biochemical, and plant growth-promoting traits and was evaluated for its potential to alleviate NaCl-induced stress in maize (Zea mays L.) under hydroponic conditions. Isolate R-18 exhibited halotolerance, surviving at NaCl concentrations ranging from 2.0% to 10.0%, and alkaliphilic adaptation, growing at pH 8.0–11.0. Biochemical assays confirmed it as a Gram-negative bacterium, displaying positive reactions in the Voges–Proskauer (V–P) tests, catalase activity, citrate utilization, fluorescent pigment production, starch hydrolysis, gelatin liquefaction, and ammonia production, while testing negative for the methyl red and cellulose hydrolysis. Notably, isolate R-18 demonstrated multiple plant growth-promoting attributes, including nitrogen fixation, phosphate and potassium solubilization, ACC deaminase activity, and indole-3-acetic acid (IAA) biosynthesis. Under 100 mM NaCl stress, inoculation with isolate R-18 significantly enhanced maize growth, increasing plant height, stem dry weight, root fresh weight, and root dry weight by 20.64%, 47.06%, 34.52%, and 31.25%, respectively. Furthermore, isolate R-18 improved ion homeostasis by elevating the K⁺/Na⁺ ratio in maize tissues. Physiological analyses revealed increased chlorophyll and proline content, alongside reduced malondialdehyde (MDA) levels, indicating mitigated oxidative damage. Antioxidant enzyme activity was modulated, with decreased superoxide dismutase (SOD) and peroxidase (POD) activities but increased catalase (CAT) activity. These findings demonstrated that Enterobacter bugandensis R-18 effectively alleviated NaCl-induced growth inhibition in maize by enhancing osmotic adjustment, reducing oxidative stress, and improving ion balance. Full article

Cranial cruciate ligament (CrCL) rupture is a common orthopedic disorder in dogs, leading to stifle joint instability and progressive osteoarthritis. This study aimed to develop and biomechanically evaluate a novel intra-articular reconstruction system designed to mimic the natural ligament and restore joint stability following CrCL excision. The system consisted of a 3D-printed thermoplastic polyurethane (TPU) graft, cerclage wire, and H-button fixation. Fourteen pelvic limbs from mature mixed-breed cadaveric dogs were used. The inclination angle, dimensions, volume, tensile strength, and elongation of the native CrCL were measured. Seven CrCL-deficient stifles were reconstructed using the proposed system and tested biomechanically. The native CrCL showed a significantly higher tensile strength than the TPU graft; however, the TPU demonstrated a greater flexibility. The reconstruction system successfully stabilized the joint and provided repeatable fixation. Significant correlations were found between CrCL volume and both age and body weight. These findings support the mechanical suitability of the proposed system for ex vivo stifle stabilization and highlight the potential of 3D-printed TPU in ligament reconstruction. Further in vivo studies are recommended to assess long-term performance, including implant integration, tissue remodeling, and clinical outcomes. Full article

Snails at hydrothermal vents rely on symbiotic bacteria for nutrition; however, the specifics of these associations in adapting to such extreme environments remain underexplored. This study investigated the community structure and metabolic potential of bacteria associated with two Indian Ocean vent snails, Chrysomallon squamiferum and Gigantopelta aegis. Using microscopic, phylogenetic, and metagenomic analyses, this study examines bacterial communities inhabiting the foot and gland tissues of these snails. G. aegis exhibited exceptionally low bacterial diversity (Shannon index 0.14–0.18), primarily Gammaproteobacteria (99.9%), including chemosynthetic sulfur-oxidizing Chromatiales using Calvin–Benson–Bassham cycle and methane-oxidizing Methylococcales in the glands. C. squamiferum hosted significantly more diverse symbionts (Shannon indices 1.32–4.60). Its black variety scales were dominated by Campylobacterota (67.01–80.98%), such as Sulfurovum, which perform sulfur/hydrogen oxidation via the reductive tricarboxylic acid cycle, with both Campylobacterota and Gammaproteobacteria prevalent in the glands. The white-scaled variety of C. squamiferum had less Campylobacterota but a higher diversity of heterotrophic bacteria, including Delta-/Alpha-Proteobacteria, Bacteroidetes, and Firmicutes (classified as Desulfobacterota, Pseudomomonadota, Bacteroidota, and Bacillota in GTDB taxonomy). In C. squamiferum, Gammaproteobacteria, including Chromatiales, Thiotrichales, and a novel order “Endothiobacterales,” were chemosynthetic, capable of oxidizing sulfur, hydrogen, or iron, and utilizing the Calvin–Benson–Bassham cycle for carbon fixation. Heterotrophic Delta- and Alpha-Proteobacteria, Bacteroidetes, and Firmicutes potentially utilize organic matter from protein, starch, collagen, amino acids, thereby contributing to the holobiont community and host nutrition accessibility. The results indicate that host species and intra-species variation, rather than the immediate habitat, might shape the symbiotic microbial communities, crucial for the snails’ adaptation to vent ecosystems. Full article

Apomixis-mediated fixation of heterosis could transform hybrid breeding in alfalfa (Medicago sativa), a globally important forage crop. The parthenogenesis-inducing morphogenetic regulator BABY BOOM (BBM) represents a promising candidate for enabling this advancement. Here, we identified BBM homologs from three alfalfa genomes, characterized their promoter regions, and cloned a 2082 bp MsBBM gene encoding a 694-amino acid nuclear-localized protein. Three alfalfa BBM gene promoters primarily contained light- and hormone-responsive elements. Phylogenetic and conserved domain analyses of the MsBBM protein revealed a high sequence similarity with M. truncatula BBM. Expression profiling demonstrated tissue-specific accumulation of MsBBM transcripts, with the highest expression in the roots and developing pods. Hormonal treatments differentially regulated MsBBM. Expression was upregulated by GA₃ (except at 4 h) and SA, downregulated by NAA, MeJA (both except at 8 h), and ABA (except at 4 h), while ETH treatment induced a transient expression peak at 2 h. As an AP2/ERF family transcription factor showing preferential expression in young embryos, MsBBM likely participates in reproductive development and may facilitate apomixis. These findings establish a molecular framework for exploiting MsBBM to enhance alfalfa breeding efficiency through heterosis fixation. Full article

Background and Objective: Intervertebral disc degeneration (IVDD) is a leading cause of lower back pain with limited regenerative treatments. Among emerging regenerative approaches, growth factor-based therapies, such as recombinant human transforming growth factor-beta (Rh-TGF-β), have shown potential for disc regeneration but are hindered by rapid degradation and uncontrolled release by direct administration. Additionally, mechanical stress elevates heat shock protein 90 (HSP-90), impairing cell function and extracellular matrix (ECM) production. This study aimed to investigate a dual self-cross-linked alginate di-aldehyde (ADA) hydrogel system for the sustained delivery of Rh-TGF-β and epigallocatechin gallate (EGCG) to enhance protein stability, regulate release, and promote disc regeneration by targeting both regenerative and stress-response pathways. Methods: ELISA and UV-Vis spectrophotometry assessed Rh-TGF-β and EGCG release profiles. A rat tail IVDD model was established with an Ilizarov-type external fixator for loading, followed by hydrogel treatment with or without bioactive agents. Disc height, tissue structure, and protein expression were evaluated via radiography, histological staining, immunohistochemistry, and Western blotting. Results: The hydrogel demonstrated a biphasic release profile with 100% Rh-TGF-β released over 60 days and complete EGCG release achieved within 15 days. Treated groups showed improved disc height, structural integrity, and proteoglycan retention revealed by histological analysis and elevated HSP-90 expression by immunohistochemistry. In contrast, Western blot analysis confirmed that EGCG effectively downregulated HSP-90 expression, suggesting a reduction in mechanical stress-induced degeneration. Conclusions: ADA hydrogel effectively delivers therapeutic agents, offering a promising strategy for IVDD treatment. Full article

Synthetic biology has fundamentally advanced cell engineering and helped to develop effective therapeutics such as chimeric antigen receptor (CAR)-T cells. For these applications, the detection, localization, and quantification of heterologous fusion proteins assembled from interchangeable building blocks is of high importance. The V5 tag, a 14-residue epitope tag, offers promising characteristics for these applications but has only rarely been used in this context. Thus, we have systematically evaluated the murine anti-V5 tag antibody mu_SV5-Pk1 as well as its humanized version, hu_SV5-Pk1, to analyze cells expressing V5-tagged receptors in samples from various in vitro and in vivo experiments. We found that the V5 tag signal on cells is affected by certain fixation and detachment reagents. Immunohistochemistry (IHC) on formalin-fixed paraffin-embedded (FFPE) mouse tissue samples was performed to sensitively detect cells in tissue. We improved IHC by applying the hu_SV5-Pk1 monoclonal antibody (mAb) to avoid cross-reactivity within and unspecific background signals arising on fixed mouse tissue. Conversely, the absence of unspecific binding by the mu_SV5-Pk1 mAb was evaluated on 46 human normal or cancer tissues. Our findings present a robust toolbox for utilizing the V5 tag and cognate antibodies in synthetic biology applications. Full article

Orthopedic implant technology has historically seen difficulties in attaining long-term stability and biological integration, leading to complications such as implant loosening, wear debris production, and heightened infection risk. Nanotechnology provides a revolutionary method for addressing these constraints through the introduction of materials characterized by exceptional biocompatibility, durability, and integration potential. Nanomaterials (NMs), characterized by distinctive surface topographies and elevated surface area-to-volume ratios, facilitate improved osseointegration and provide regulated medication release, thereby creating a localized therapeutic milieu surrounding the implant site. To overcome the long-standing constraints of conventional implants, such as poor osseointegration, low mechanical fixation, immunological rejection, and implant-related infections, nanotechnology is causing a revolution in the field of orthopedic research. NMs are ideally suited for orthopedic applications due to their exceptional features, including increased tribology, wear resistance, prolonged drug administration, and excellent tissue regeneration. Because of their nanoscale size, they can imitate the hierarchical structure of real bone, which in turn encourages the proliferation of cells, lowers the risk of infection, and helps with the mending of bone fractures. This article will investigate the wide-ranging possibilities of nanostructured ceramics, polymers, metals, and carbon materials in bone tissue engineering, diagnostics, and the treatment of implant-related infections, bone malignancies, and bone healing. In addition, this paper will provide a basic overview of the most recent discoveries in nanotechnology driving the future of translational orthopedic research. It will also highlight safety evaluations and regulatory requirements for orthopedic devices. Full article

Background/Objectives: Advanced destruction of the ankle and subtalar joints due to neuropathy, chronic infection, or inflammatory conditions presents a major surgical challenge, often resulting in limb amputation. This descriptive retrospective study aims to evaluate outcomes of reconstructive surgery in patients, in whom limb preservation was prioritized over amputation despite significant soft tissue and osseous involvement. Methods: Between January 2013 and December 2022, 31 reconstructive procedures were performed on 29 patients (16 women and 13 men) with severe hindfoot deformities. Etiologies included Charcot arthropathy (55%), osteomyelitis (25%), combined pathology (10%), and rheumatoid deformity with skin defect (10%). Surgical procedures included tibiotalocalcaneal arthrodesis (39%), astragalectomy with tibiocalcaneal arthrodesis (32%), tibiotalar arthrodesis (23%), and multistage procedures (6%). Fixation methods varied based on the extent of deformity and infection. The union was assessed via radiographs and CT imaging, and outcomes were statistically analyzed using Fisher’s exact test. Results: Successful arthrodesis was achieved in 74% of cases (23/31). The union rate was significantly influenced by the type and level of fixation (p = 0.0199), with the lowest rate observed in tibiotalocalcaneal arthrodesis using external fixation (17%). Complications included surgical site infection or abscess in 42% of cases, requiring reoperation in 35%. Limb amputation was ultimately necessary in five patients (16%). Conclusions: Despite high complication rates, limb-preserving reconstructive surgery remains a viable alternative to amputation in selected high-risk patients with severe hindfoot pathology. Appropriate preoperative planning, tailored surgical strategy, and patient compliance are essential to achieving functional limb salvage and restoring weight-bearing capacity. Full article

Background and Objectives: Adequate soft tissue thickness and keratinized mucosa are essential for the long-term health and esthetics of the peri-implant area. A porcine acellular dermal matrix (PADM) has shown promise in augmenting soft tissue, but reliable fixation remains a challenge. Materials and Methods: This case series describes the use of a PADM fixed with resorbable magnesium screws (NOVAMag^®) in three patients requiring peri-implant soft tissue augmentation. The grafts were stabilized with magnesium screws on the buccal side. The clinical outcomes were evaluated over a period of 3–6 months using STL imaging and direct measurements. Results: All patients showed an improvement in their mucosal volume and keratinization. The mean vertical increase in soft tissue was 0.87 ± 0.16 mm and the mean horizontal increase was 1.00 ± 0.13 mm. The mucosal thickness increased from a baseline value of 1.0–1.2 mm to 1.9–2.1 mm, and the width of the keratinized mucosa improved by an average of 1.0 mm. No complications were observed, and in all cases there was tension-free healing and esthetic results. Conclusions: A PADM in combination with resorbable magnesium fixation screws offers a predictable and minimally invasive solution to improve peri-implant soft tissue with favourable short-term volumetric and esthetic results. Full article

Low phosphorus (P) stress impacts nitrogen (N) metabolism in soybeans. This study investigated the effects of exogenous cytokinin (Trans-Zeatin) on soybean N metabolism under low P stress by treating seeds with Trans-Zeatin and analyzing N accumulation, ¹⁵N abundance, nodule N fixation accumulation, nodule N fixation rate, nodule nitrogenase activity, soluble protein content, and free amino acid profiles. The results showed that exogenous cytokinin enhanced N accumulation in aboveground tissues, roots, and nodules, as well as nodule N fixation accumulation and fixation rate (from day 35 onward) under low P stress. Additionally, it promoted both acetylene reduction activity (ARA) and specific nitrogenase activity (SNA) in soybean nodules. By increasing the absorption of fertilizer-derived N, exogenous cytokinin alleviated the inhibitory effects of low P stress on the early growth and development of soybeans. Notably, under low P conditions, exogenous cytokinin significantly elevated the soluble protein content in nodules. However, the underlying mechanisms governing changes in free amino acid profiles require further investigation. This study provides a theoretical foundation for developing strategies to regulate soybean N metabolism under low P stress. Full article

Histological techniques are essential for studying small crustaceans’, such as Neocaridina shrimp, anatomy and physiology. However, their small size and rapid tissue autolysis present challenges for fixation and processing. This study aimed to optimize histological methods for Neocaridina shrimp by evaluating different protocols for fixation, decalcification, and enzymatic digestion. Shrimp were fixed using 10% neutral-buffered formalin (NBF) and Bouin’s or Davidson’s fluid with or without modifications such as trypsin digestion, decalcification, or abdomen removal. Tissue preservation, section quality, and staining properties were assessed. Davidson’s fluid consistently gave generally acceptable fixation results, with minimal autolysis and good tissue preservation. Trypsin digestion increased tissue damage and autolysis, particularly in the liver and pancreas. Decalcification improved the quality of the sections; however, it increased autolysis and resulted in less specific staining. The optimal protocol involved the removal of the abdomen, followed by fixation in Davidson’s fluid and decalcification, which resulted in rapid penetration of the fixative, minimal autolysis, and a beneficial effect on staining. This study highlights the importance of adapting histological methods to the specific characteristics of small crustaceans and provides a basis for future research on Neocaridina shrimp. Implementing these optimized techniques will improve the quality and reliability of histological analyses in crustacean research, deepening the understanding of their biology and facilitating their use as model organisms in various scientific fields. Full article

Lentil is a nutritionally valuable legume crop, rich in protein, carbohydrates, amino acids, and vitamins, and is also used as green manure. Symbiotic nitrogen fixation (SNF) plays a crucial role in lentil growth and development, yet there is limited research on isolating and identifying lentil rhizobia related to nodulation and nitrogen fixation. This study employed tissue block isolation, line purification, and molecular biology to isolate, purify, and identify rhizobial strains from lentils, analyzing their physiological characteristics, including bromothymol blue (BTB) acid and alkali production capacity, antibiotic resistance, salt tolerance, acid and alkali tolerance, growth temperature range, and drought tolerance simulated by PEG6000. Additionally, the nodulation capacity of these rhizobia was assessed through inoculation experiments using the identified candidate strains. The results showed that all isolated rhizobial strains were resistant to Congo red, and nifH gene amplification confirmed their potential as nitrogen fixers. Most strains were positive for H₂O₂ and BTB acid and base production, with a preference for alkaline environments. In terms of salt tolerance, the strains grew normally at 0.5–2% NaCl, and six strains were identified as salt stress resistant at 4% NaCl. The temperature range for growth was between 4 °C and 49 °C. Antibiotic assays revealed resistance to ampicillin and low concentrations of streptomycin, while kanamycin significantly inhibited growth. Two drought-tolerant strains, TG25 and TG55, were identified using PEG6000-simulated drought conditions. Inoculation with candidate rhizobial strains significantly increased lentil biomass, highlighting their potential for enhancing crop productivity. Full article

External fixators are frequently used to treat complex fractures with multiple bone fragments and soft tissue injuries. Inaccurate assessment of bone union and premature removal of the fixator may necessitate revisions and prolong treatment. The decision to remove an external fixator typically depends on an orthopedist’s experience. The accuracy of diagnosis can be improved by using a force sensor integrated into the modified external fixator according to Mitkovic. A sensor measuring axial compressive force is mounted between two vertical rods. Experiments with the modified external fixator were carried out using three different axial loads delivered by the universal electromechanical testing machine (UETM) and a sensor that detected the corresponding axial force. Springs with variable rigidity were used to simulate bone callus stiffness. Low rigidity springs represented a high elasticity callus, whereas high rigidity springs represented a callus with lower elasticity. The results show that the force detected by the sensor was nearly identical to the force delivered by the UETM while the callus did not form, decreased as spring rigidity increased, and eventually zeroed out as the leg healed. The findings indicated that using modified external fixator according to Mitkovic can help orthopedists assess bone healing more accurately. Full article

Background: Tibial plateau fractures (TPFs) are complex injuries often leading to long-term complications such as knee instability, limited range of motion, and osteoarthritis. Accurate diagnostic evaluations combining subjective and objective assessments are essential for identifying functional limitations, guiding rehabilitation, and improving recovery outcomes. This study examines the role of diagnostic predictors in differentiating recovery trajectories in two groups of patients treated for closed TPFs by open reduction and internal fixation (ORIF), comparing patients with less severe fractures and patients with more severe fractures (BCFs). Methods: A consecutive series of patients with a diagnosis of TPFs treated by ORIF at our institution between 2009 and 2016 were analyzed in this retrospective study. All injured patients were divided according to the Schatzker classification into two groups: mono-condylar (MCF) and bi-condylar (BCF) fracture patient groups. Diagnostic evaluations included patient-reported outcome measures (PROMs) such as KOOS, IKDC, and AKSS, alongside objective assessments of functional recovery using dynamometers, force platform tests (single-leg stance and squat jump variations), and measurements of active range of motion (AROM). Results: A total of 28 patients were included: 17 in the MCF patient group (Schatzker: 12 II; 5 III; 0 IV) and 11 in the BCF patient group (Schatzker: 6 V; 5 VI). Patients with less severe MCFs exhibited significantly better recovery outcomes, including higher KOOS (86.0 vs. 64.6, p = 0.04), IKDC (80.3 vs. 64.6, p = 0.04), and AKSS (95.3 vs. 70.5, p = 0.02) scores. They also demonstrated greater knee flexion (122.8° vs. 105.5°, p = 0.04) and faster neuromuscular recovery, as evidenced by higher rates of force development (RFD) during dynamic performance tests. Conversely, patients with more severe BCFs showed lower RFD values, indicating slower recovery and greater rehabilitation challenges. Conclusions: Integrating diagnostic tools like PROMs, AROM, and neuromuscular performance tests provides valuable insights into recovery after ORIF for TPFs. Fracture severity significantly impacts functional recovery patients with MCFs showing better outcomes and faster neuromuscular recovery, while subjects with BCFs require a longer rehabilitation treatment focusing on neuromuscular re-education and soft tissue recovery. Full article