Bone Formation Research Articles

Effects of vitamin K2 administration on guided bone regeneration in diabetic rats.

The present study aimed to investigate the histomorphometric and immunohistochemical impacts of vitamin K2 on guided bone regeneration (GBR) in calvarial critical-size defects (CSDs) in diabetic rats. A total of 30 rats were used in this study, comprising 12 non-diabetic (control) rats and 18 with streptozotocin-nicotinamide-induced experimental Diabetes mellitus (DM). In all rats, two calvarial CSDs were created: one defect was left empty (E), the other was treated with bovine-derived bone graft and collagen-based resorbable membrane (GM). Study groups were as follows: control rats administered saline (n = 6, C-E and C-GM groups) or vitamin K2 (n = 6, CK-E and CK-GM groups) and diabetic rats administered saline (n = 6, DM-E and DM-GM groups) or vitamin K2 (n = 6, DMK-E and DMK-GM groups). After 4 weeks of saline or vitamin K2 administration, the rats were euthanized. Bone defect healing and new bone formation were assessed histomorphometrically, and osteocalcin and osteopontin levels were examined immunohistochemically. Percentage of new bone formation was greater in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 3.86 (95% CI = 16.38-28.61), d = 1.86, (95% CI = 10.74-38.58), respectively, p < .05]. Bone defect healing scores were higher in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 2.69 (95% CI = -2.12 to -0.87), d = 3.28 (95% CI = 0.98-1.91), respectively, p < .05]. Osteocalcin expression levels were elevated in CK-GM vs. CK-E, in DMK-GM vs. DMK-E [d = 1.19 (95% CI = 0.08-1.41), d = 1.10 (95% CI = 0.02-1.22), respectively p < .05]. Vitamin K2 enhanced osteocalcin expression levels in DMK-E vs. DM-E [d = 2.78, (95% CI = 0.56-1.53), p < .05] and in DMK-GM vs. DM-GM [d = 2.43, (95% CI = 0.65-2.10), p < .05]. Osteopontin expression was enhanced in defects treated with GM vs. E defects [C-GM vs. C-E, d = 1.56 (95% CI = 0.38-2.01); CK-GM vs. CK-E, d = 1.91 (95% CI = 0.49-1.72); DM-GM vs. DM-E, d = 2.34 (95% CI = -1.12 to -0.50); DMK-GM vs. DMK-E, d = 2.00 (95% CI = 0.58-1.91), p < .05]. The research findings suggest that administering vitamin K2 in GBR for rats with DM favorably impacts bone healing in CSDs, presenting an adjunctive strategy for bone regeneration.

Evaluation of Potential Roles of Zinc Finger Homeobox 3 (Zfhx3) Expressed in Chondrocytes and Osteoblasts on Skeletal Growth in Mice.

Bone formation is tightly modulated by genetically encoded molecular proteins that interact to regulate cellular differentiation and secretion of bony matrix. Many transcription factors are known to coordinate these events by controlling gene transcription within networks. However, not all factors involved are known. Here, we identified a novel function forZinc Finger Homeobox 3(Zfhx3), a gene encoding a transcription factor, as a regulator of bone metabolism. We knocked outZfhx3conditionally in mice in either chondrocytes or osteoblasts and characterized their bones by micro-CT in 12-week-old mice. We observed a negative effect in linear bone growth in both knockout mice but reduced bone mass only in mice withZfhx3deleted in osteoblasts. Loss of Zfhx3 expression in osteoblasts affected trabecular bone mass in femurs and vertebrae in both sexes but influenced cortical bone volume fraction only in females. Moreover, transcriptional analysis of femoral bones in osteoblastZfhx3conditional knockout mice revealed a reduced expression of osteoblast genes, and histological evaluation of trabecular bones suggests that Zfhx3 causes changes in bone formation and not resorption. The loss of Zfhx3 causes reductions in trabecular bone area and osteoid volume, but no changes in the expression of osteoclast differentiation markers or number of TRAP stained osteoclasts. These studies introduce Zfhx3 as a relevant factor toward understanding gene regulatory networks that control bone formation and development of peak bone mass.

Loss of Hdac4 in Osteoprogenitors Impairs Postnatal Trabecular and Cortical Bone Formation, Resulting in a Dwarfism and Osteopenia Phenotype in Mice.

HDAC4 is a class II histone deacetylation protein with a well-characterized role in chondrocyte differentiation and skeletal development, and dysregulated expression or haploinsufficiency of Hdac4 leads to skeletal formation and malformation disorders. The early lethality of hdac4 ablation mice hindered further investigation of its role in postnatal bone growth and development. Therefore, this study aims to investigate the significant role of Hdac4 in postnatal endochondral bone development using two mouse models with conditional deletion of Hdac4 in Sp7-expressing osteoprogenitors or chondrocytes and monitored postnatal bone development. The phenotype of Acan-CreERT2; Hdac4fl/fl mice largely resembled that of conventional Hdac4-/- mice. But phenotypic characterizations of mice with Hdac4 inactivation in Sp7-expressing osteoprogenitors (Sp7-Cre; Hdac4fl/fl) showed dwarfism with body and limb shortening and remarkable skeletal defects. Micro-computed tomography analysis of tibias further demonstrated that loss of Hdac4 expression impaired bone formation and microarchitecture, mainly characterized by dysplasia of trabecular and cortical bone in young mice. Our in vivo and in vitro data support a crucial role for Hdac4 in regulating osteoblast proliferation and differentiation, bone matrix protein production, angiogenesis, and ultimately trabecular and cortical bone formation. Moreover, RNA-seq analysis implicated Hdac4 in the regulation of key genes and pathways necessary to affect the accumulation of extracellular matrix, biological processes related to signal transduction, and skeletal growth. Collectively, our data show that postnatal expression of Hdac4 in Sp7-expressing osteoprogenitors provides essential regulatory oversight of endochondral bone formation, bone morphology, and homeostasis.

Ghrelin alleviated TiO2 NPs-induced inhibition of endochondral osteogenesis and promoted longitudinal growth of long bones in juvenile rats via Wnt/β-catenin signaling pathway

Titanium dioxide nanoparticles (TiO2 NPs) are widely used in children’s daily necessities and foods, and their health hazards to children have attracted particular attention. Childhood is a critical time for accelerated bone growth and development. Current studies revealed that TiO2 NPs exposure causes bone damage in juvenile rats; however, the underlying mechanism is unknown. Ghrelin is a polypeptide hormone that is considered to be a candidate factor for regulating bone growth and development. In this research, 3-week-old juvenile male rats were administered 0, 100 or 200 mg/kg TiO2 NPs and 50 μg/kg ghrelin for 4 weeks to explore the underlying mechanism of TiO2 NPs-induced bone damage, and the protective effect of ghrelin. Our results revealed that TiO2 NPs resulted in decreased synthesis of bone growth-related hormones, disturbed bone metabolism, and destruction of bone structure. Further mechanism studies showed that TiO2 NPs inhibited Wnt/β-catenin pathway, reduced collagen synthesis, inhibited chondrocyte proliferation and differentiation, promoted chondrocyte apoptosis, and inhibited endochondral osteogenesis, ultimately leading to long bone longitudinal growth retardation and osteoporosis. Ghrelin alleviated the negative effects of TiO2 NPs-induced bone growth in juvenile rats by upregulating the Wnt/β-catenin signaling pathway. This study provided a reference for the clinical treatment of growth retardation and idiopathic short stature in juvenile children caused by environmental pollutants.

Comparison of Denosumab with Romosozumab in the treatment of male osteoporosis: a retrospective cohort study

We aimed to investigate the efficacy of romosozumab treatment compared with that of denosumab in especially male osteoporosis patients. This retrospective cohort study included 174 Japanese male patients receiving either denosumab or romosozumab for 12 months. Propensity score matching extracted 50 patients per treatment group for standardization of group characteristics. The endpoints include the rate of change in the bone mineral density (BMD) of the lumbar spine, total hip, and femoral neck after 12 months of treatment as well as the changes in serum bone metabolism markers. The mean 12-month percentage increase in the lumbar spine BMD from baseline was significantly greater with romosozumab (13.0% ±1.7%) than with denosumab (4.5%±0.6%) (P < 0.01). The total hip and femoral neck BMD exhibited a similar trend at 12 months; however, no significant between-group differences were observed. With denosumab, bone formation, and resorption marker levels significantly decreased at 6 and 12 months. Conversely, with romosozumab, the levels of bone formation markers increased transiently at 6 months before returning to baseline, whereas bone resorption markers significantly decreased at both time points. Romosozumab demonstrated significantly superior effects over denosumab in improving BMD, especially of the lumbar spine, suggesting that romosozumab can be used for treating male osteoporosis.

Tannic acid-modified magnesium oxychloride bone cement with high water resistance and osteogenic properties

Magnesium oxychloride bone cement (MOC) is a bone replacement material that exhibits excellent mechanical properties and is non-toxic. However, the material exhibits poor water resistance, as the 5-phase composition of MOC decomposes readily in the presence of water. Tannic acid (TA) is an organic acid with favorable biocompatibility, containing a large amount of -OH that can form chelates with metal ions. Therefore, we plan to control spill Mg2+ in MOC using TA to improve its water resistance. The results demonstrate that the addition of TA resulted in a denser structure of MOC, an improved stability of the 5-phase in solution, a reduced degradation rate and pH value of MOC in body fluids, and the generation of hydroxyapatite in vitro. Furthermore, the MOC modified with TA demonstrated enhanced cell proliferation and increased new bone formation in cellular and animal experiments. This high-strength, highly bioactive material system is adaptable to patient-specific requirements and has significant potential for clinical translation.

Regulatory role of Heparan sulfate in leptin signaling

Leptin, a hormone mainly secreted by adipocytes, has attracted significant attention since its discovery in 1994. Initially known for its role in appetite suppression and energy regulation, leptin is now recognized for its influence on various physiological processes, including immune response, bone formation, and reproduction. It exerts its effects by binding to receptors and initiating an intracellular signaling cascade. Heparan sulfate (HS) is known to regulate the intracellular signaling of various ligands. HS is present as the glycan portion of HSPGs on cell surfaces and in intercellular spaces, with diverse structures due to extensive sulfation and epimerization. Although HS chains on HSPGs are involved in many physiological processes, the detailed effects of HS chains on leptin signaling are not well understood.This study examined the role of HS chains on HSPGs in leptin signaling using Neuro2A cells expressing the full-length leptin receptor (LepR). We showed that cell surface HS was essential for efficient leptin signaling. Enzymatic degradation of HS significantly reduced leptin-induced phosphorylation of downstream molecules, such as signal transducer and activator of transcription 3 and p44/p42 Mitogen-activated protein kinase. In addition, HS regulated LepR expression and internalization, as treatment with HS-degrading enzymes decreased cell surface LepR. HS was also found to exhibit a weak interaction with LepR. Enzymatic removal of HS enhanced the interaction between LepR and low-density lipoprotein receptor-related protein 1, suggesting that HS negatively regulates this interaction. In conclusion, HS plays a significant role in modulating LepR availability on the cell surface, thereby influencing leptin signaling. These findings provide new insights into the complex regulation of leptin signaling and highlight potential therapeutic targets for metabolic disorders and obesity.

Association between musculoskeletal sonographic features and response to treatment in patients with psoriatic arthritis.

To investigate the association between musculoskeletal sonographic features and clinical features, as well as treatment outcomes, in patients with active psoriatic arthritis (PsA). A prospective cohort study was conducted involving patients with active PsA. Disease activity was assessed clinically at baseline and 3-6 months after initiating therapy, with a Disease Activity Index for PsA (DAPSA) score calculated. A baseline ultrasound examination of 64 joints, 28 tendons and 16 entheses evaluated the following lesions: synovitis, peritenonitis, enthesitis, tenosynovitis, new bone formation and erosions. Total scores for each lesion and total inflammatory and structural scores were calculated. The association between baseline sonographic scores and treatment outcomes was assessed using Cox proportional hazards models (for drug persistence) and generalised estimating equation models for DAPSA change. A total of 135 treatment periods (107 patients) were analysed. Multivariable analysis showed that a greater reduction in DAPSA score at follow-up was associated with higher baseline synovitis (β -3.89), peritenonitis (β -3.93) and enthesitis structural scores (β -2.91). Additionally, the total inflammatory score independently predicted DAPSA change (β -5.23) regardless of the total structural damage score. Drug persistence was analysed in 105 treatment periods, revealing that a higher sonographic erosion score was associated with earlier drug discontinuation (adjusted HR 1.28, 95% CI 1.03 to 1.61). The study results provide preliminary evidence supporting the utility of musculoskeletal ultrasound in predicting treatment response and drug persistence in PsA.

Elucidating the Mechanism of Large-Diameter Titanium Dioxide Nanotubes in Protecting Osteoblasts Under Oxidative Stress Environment: The Role of Fibronectin and Albumin Adsorption.

Large-diameter titanium dioxide nanotubes (TNTs) have shown promise in preserving osteoblast function under oxidative stress (OS) in vitro. However, their ability to enhance osteogenesis in vivo under OS conditions and the underlying mechanisms remain unclear. This study aimed to evaluate the osteogenic potential of 110 nm TNTs (TNT110) compared to 30 nm TNTs (TNT30) in an aging rat model exhibiting OS, and to investigate the mechanisms involved. Surface properties of TNTs were characterized, and in vitro and in vivo experiments were conducted to assess their osteoinductive effects under OS. Transcriptomic, proteomic analyses, and Western blotting were performed to investigate the protective mechanisms of TNT110 on osteoblasts. Protein adsorption studies focused on the roles of fibronectin (FN) and albumin (BSA) in modulating osteoblast behavior on TNT110. In both in vitro and in vivo experiments, TNT110 significantly improved new bone formation and supported osteoblast survival under OS conditions. Subsequent ribonucleic acid sequencing results indicated that TNT110 tended to attenuate inflammatory responses and reactive oxygen species (ROS) expression while promoting endoplasmic reticulum (ER) stress and extracellular matrix receptor interactions, all of which are crucial for osteoblast survival and functionality. Further confirmation indicated that the cellular behavior changes of osteoblasts in the TNT110 group could only occur in the presence of serum. Moreover, proteomic analysis under OS conditions revealed the pivotal roles of FN and BSA in augmenting TNT110's resistance to OS. Surface pretreatment of TNT110 with FN/BSA alone could beneficially influence the early adhesion, spreading, ER activity, and ROS expression of osteoblasts, a trend not observed with TNT30. TNT110 effectively protects osteoblast function in the OS microenvironment by modulating protein adsorption, with FN and BSA synergistically enhancing osteogenesis. These findings suggest TNT110's potential for use in implants for elderly patients.

MicroRNA-324 mediates bone homeostasis and the regulation of osteoblast and osteoclast differentiation and activity

MicroRNAs (miRNAs) modulate the expression of other RNA molecules. One miRNA can target many transcripts, allowing each miRNA to play key roles in many biological pathways. Defects in bone homeostasis result in common age-related diseases including osteoporosis. Serum levels of miR-324-3p positively correlate with several features of bone maintenance. In contrast here, using in vivo micro-computed tomography and histology, global miR-324-null mice demonstrated increased bone mineral density and both trabecular and cortical thickness, with effect magnitudes increasing with age. The bone marrow of miR-324-null mice had reduced lipid content while TRAP staining revealed a decrease in osteoclasts, with histomorphometry demonstrating an increased rate of bone formation. Ex vivo assays showed that the high bone mass phenotype of miR-324-null mice resulted from both increased osteoblast activity and decreased osteoclastogenesis. RNA-seq analysis of osteoblasts, osteoclasts and bone marrow macrophages and target validation assays identified that the osteoclast fusion regulator Pin1 and the master osteogenic regulator Runx2 were targets of miR-324-5p in osteoclast lineage cells and osteoblasts, respectively. Indeed, in vitro Runx2 overexpression recapitulated the increased osteogenesis and decreased adipogenesis phenotype observed in vivo by the loss of miR-324. Overall, these data demonstrate the importance of miR-324 in bone homeostasis by regulating aspects of both bone formation and remodelling. Elucidation of pathways regulated by miR-324 offer promise for the treatment of bone diseases such as osteoporosis.

Remineralizing capacity of zinc oxide eugenol sealer following the addition of nanohydroxyapatite-tyrosine amino acid: An in vivo animal study

ObjectiveAlthough several sealers have been developed, the zinc oxide eugenol (ZOE) sealer is still used in many private dental clinics. This study aimed to compare the biocompatibility and remineralizing capacity οf ZOE sealer with the addition of nanohydroxyapatite-tyrosine amino acid. MethodsThis study was conducted on Twenty rabbits. The rabbits were divided into four groups based on the test observation period (3, 7, 21, and 28 days) following surgical implantation. General anesthesia was administered to each rabbit, and a subcutaneous incision of approximately 1±0.5 cm was made along the symphyseal area of the mandible of each rabbit. Four bone cavities were generated in the interdental space of the lower jaw between the central and molar teeth of each rabbit, with one longitudinal subcutaneous incision. The ZOE sealers were mixed according to the manufacturer's guidelines and directly inserted within the cavities generated at the end of each test period. The animals were sacrificed, and bone biopsy was carried out at the site of testing. The biopsy samples were obtained and subjected to histological analysis using a low-power light microscope (Olympus C ⅹ 21, Japan) and immunohistochemistry using Ki67 antibody. ResultsThe collected information was examined by parametric statistical tests using SPSS software version ‘‘22.” One-way ANOVA and post-hoc Duncan’s tests were used to measure the significance among various groups, with statistical significance set, when “P≤0.01”. ConclusionThe 20% mixed endodontic sealer displayed excellent outcomes compared to other experimental groups, as identified by higher new bone formation at every evaluation period.

OMIBONE: Omics-driven computer model of bone regeneration for personalized treatment

Treatment of bone fractures are standardized according to the AO classification, which mainly refers to the mechanical stabilization required in a given situation but neglect individual differences due to patient's healing potential or accompanying diseases. Specially in elderly or immune-compromised patients, the complexity of individual constrains on a biological as well as mechanical level are hard to account for. Here, we introduce a novel framework that allows to predict bone regeneration outcome using combined proteomic and mechanical analyses in a computer model. The framework uses Ingenuity Pathway Analysis (IPA) software to link protein changes to alterations in biological processes and integrates these in an Agent-Based Model (ABM) of bone regeneration. This combined framework allows to predict bone formation and the potential of an individual to heal a given fracture setting. The performance of the framework was evaluated by replicating the experimental setup of a mouse femur fracture stabilized with an intramedullary pin. The model was informed by serum derived proteomics data. The tissue formation patterns were compared against experimental data based on x-ray and histology images. The results indicate the framework potential in predicting an individual's bone formation potential and hold promise as a concept to enable personalized bone healing predictions for a chosen fracture fixation.

The Role of Fabricated Coral Shell Powder in the Healing of Mandibular Bone Gap in Dogs

Background: The reconstruction of mandibular bone defects poses a real challenge and difficulty for surgeons; biomaterial bone substitutes are the most used material for reconstructing mandibular bone defects. Objectives: This study explored the role of fabricated hydroxyapatite (HAp) powder from the coral shell in healing critical size mandible gaps in dogs. Methods: HAp was prepared using the hydrothermal method from coral shells. Characterization of the fabricated coral shell was done by x-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), and energy dispersive x-ray spectroscopy (EDX). The designed research was performed on 18 dogs of both sexes (mean weight: 20±0.5 kg, mean age: 2±0.6 years). The samples were divided into two equal groups. Animals underwent experimental defects at the ventral surface of the lower mandible about 14.5 mm. Results: The results of XRD represented high crystallinity, the EDX results indicated the surface morphology of distributed particles of calcium, phosphorous, carbon, and oxygen, respectively, and the FESEM results suggested that the surface morphology of HAp appears as a spherical particle that regularly distributed within the sample. In the HAp group, at 30 days, the FESEM images show that the defective gap completely closed, and the center of the defect was filled with a thick layer of osteoid matrix. Radiographically, the HAp group at 30 days post-surgery indicated a well-defined circular radiolucent bone gap at the caudal portion of the mandible, with a partially sclerosed margin. Macroscopically, at 30 days, the gap appears very small and is invaded by new bone formation. Conclusion: In conclusion, recycling HAp from coral shells has practical value in the reconstitution of the mandibular gap, and the radiological and critical properties of prepared HAp emphasize this outcome.

Laser powder bed fusion printed poly-ether-ether-ketone/bioactive glass composite scaffolds with dual-scale pores for enhanced osseointegration and bone ingrowth

Although poly-ether-ether-ketone (PEEK) implants hold significant medical promise, their bioinert nature presents challenges in osseointegration and bone ingrowth within clinical contexts. To mitigate these challenges, the present study introduces Diamond PEEK/bioactive glass (BG) composite scaffolds, characterized by macro/micro dual-porous structures, precisely fabricated via laser powder bed fusion (LPBF) technology. The findings indicate that an increase in BG content within these scaffolds significantly augments their hydrophilicity and hydroxyapatite formation capacities. Stress-strain curve analysis demonstrates reliable load-bearing stability across all scaffold types. In vitro assessments confirmed the non-cytotoxicity of PEEK/BG samples and demonstrated improved osteogenic differentiation and mineralization with increased BG incorporation. Further, in vivo experiments illustrated that the Diamond porous structure of these scaffolds facilitated bone growth, an effect notably amplified with higher BG content. Particularly in groups with 15 wt% and 25 wt% BG scaffolds, new bone formation was observed not only within the macropores of the Diamond structure but also within the micropores inside the scaffold rod, suggesting an almost seamless fusion with the new bone. This demonstrates the scaffolds’ effective osteointegration and bone ingrowth properties. This study conclusively established the effectiveness of Diamond-structured PEEK/BG composite scaffolds, fabricated via LPBF, in bone repair. It highlights the crucial role of BG in enhancing osteogenic potential through interaction with the macro/micro pores of the scaffold. Statement of significanceThis study addresses the bioinert nature of PEEK implants by developing Diamond-structured PEEK/bioactive glass (BG) composite scaffolds by laser powder bed fusion. The dual-porous macro/microstructure enhances hydrophilicity and hydroxyapatite formation, vital for bone regeneration. By adjusting the BG content, we controlled the melt viscosity and sintering rate, leading to the formation of beneficial microscale pores. These pores resolve the issue of ineffective bioactive fillers in previous LPBF-fabricated scaffolds, enhancing the osteogenic potential of BG and inducing superior bone ingrowth and osseointegration. In vitro and in vivo analyses show enhanced osteogenic differentiation, mineralization, and bone growth, underscoring the clinical potential of these scaffolds for bone repair.

Physical and histological comparison of three-dimensional interconnected porous carbonate apatite bone graft fabricated with cylindrical and spherical granules

Three-dimensional interconnected porous carbonate apatite (3D-CO3Ap) bone grafts facilitate rapid bone replacement because of their structure, which supports cellular infiltration. Existing 3D-CO3Ap grafts have demonstrated limited mechanical strength, with a maximum compressive strength of 3 MPa. By fusing polymer-bound CaCO3 granules, followed by the debinding and compositional transformation of the granules via a dissolution–precipitation reaction, we successfully fabricated 3D-CO3Ap grafts that exhibited remarkably enhanced compressive strengths, which were recorded at 14.9 ± 3.7 MPa. The use of spherical granules, which provide more uniform and effective intergranular contacts than cylindrical granules, was key to achieving higher mechanical strength and superior bone formation. 3D-CO3Ap developed with spherical granules not only exhibited higher compressive strength, but also enhanced new bone formation compared with those fabricated with cylindrical granules. This study underscores the significance of porous microstructure design in 3D-CO3Ap, highlighting its critical role in optimizing both mechanical properties and bone regeneration capabilities.

Investigation of the Effects of Heavy Metals (Copper, Cobalt, Manganese, Selenium, and Zinc) on Fish Immune Systems – An Overview

Abstract Aquaculture, as a strategic and developmental industry, plays an important role in ensuring food security and economic stability within countries. This crucial sector faces various influences, including infectious and nutritional diseases. Certain minerals function as essential nutrients, playing a main role in processes across all aquatic animals when present in appropriate dietary proportions. Studies showed that some heavy metals serve as a catalyst for various biological functions, including the maintenance of colloidal systems, acid-base balance, bone formation, and the regulation of fundamental natural elements such as vitamins, hormones, and minerals. Fish acquire essential elements crucial for their well-being through both diet and water sources. Adequate levels of essential elements, such as iron, zinc, copper, cobalt, manganese, and selenium, in the diet contribute to enhanced growth, increased survival rates, improved disease resistance, and heightened specific immunity in fish. When incorporated in appropriate quantities, these elements play a pivotal role in disease prevention, fostering the production of high-quality fish, minimizing economic losses, and solidifying aquaculture as a robust and profitable industry. This article aims to delve into the immunological effects induced by metal elements, specifically copper (Cu), cobalt (Co), manganese (Mn), zinc (Zn), and selenium (Se), in diverse fish species. The objective is to underscore the significance of this exploration, paving the way for the development of more potent immune supplements in the future. These supplements have the potential to improve growth, fortify immune responses, and enhance resilience against diseases in various fish species.

Filamin B knockdown impairs differentiation and function in mouse pre-osteoblasts via aberrant transcription and alternative splicing

ObjectiveFilamin B (FLNB) encodes an actin-binding protein that is known to function as a novel RNA-binding protein involved in cell movement and signal transduction and plays a pivotal role in bone growth. This study aimed to investigate possible FLNB function in the skeletal system by characterizing the effecs of FLNB knockdown in mouse preosteoblast cells. MethodsStable FLNB MC3T3-E1 knockdown cells were constructed for RNA-seq and alternative splicing event (ASE) analysis of genes involved in osteoblast differentiation and function that may be regulated by FLNB. Standard transwell, MTT, ALP, qPCR, Western blot, and alizarin red staining assays were used to assess functional changes of FLNB-knockdown MC3T3-E1 cells. ResultsAnalysis of differentially expressed genes (DEGs) in FLNB knockdown cells revealed enrichment for genes related to osteoblast proliferation, differentiation and migration, such as ITGA10, Cebpβ, Grem1, etc. Alternative splicing (AS) analysis showed changes in the predominant mRNA isoforms of skeletal development-related genes, especially Tpx2 and Evc. Functional asslysis indicated that proliferation, migration, and differentiation were all inhibited upon FLNB knockdown in MC3T3-E1 cells compared to that in vector control cells. ConclusionsFLNB participates in regulating the transcription and AS of genes required for osteoblast development and function, consequently affecting growth and development in MC3T3-E1 cells.

Alpha-calcium sulfate hemihydrate incorporated with tri-calcium phosphate and hydroxyapatite bioceramics as potential scaffold for bone regeneration

The development of artificial grafts, including biodegradable materials and non-biodegradable ones, for bone regeneration is a significant advance in the field of biomaterial science. Notably, bioceramics such as calcium sulfate hemihydrate and calcium phosphates have attracted extensive attention from researchers due to their natural bone-like composition. However, some requirements related to mechanical properties, resorption ability, osteoinduction and osteoconduction of each bioceramics have not been effectively controlled to make them ideal materials for bone regeneration applications. In this study, composites of β-tricalcium phosphate, hydroxyapatite, and α-calcium sulfate hemihydrate (β-TCP/HA/α-CSH) were synthesized using wet mixing method in order to create a template that degrades in sync with bone growth, offer sufficient mechanical strength during healing process, and has appropriate setting time for precise surgical application. The β-TCP/HA compositional phase with the ratio of 80/20 (% in weight) would be used for the entire study meanwhile the influence of α-CSH on the behaviour of β-TCP/HA/α-CSH composites would be investigated. The successful preparation of β-TCP/HA/α-CSH composite was evaluated by FTIR, XRD, and SEM. In vitro testing has further demonstrated the commendable biodegradability of β-TCP/HA/α-CSH composites and its ability for cell adhesion and migration, suggesting that β-TCP/HA/α-CSH composites would be a promising biomaterial for bone engineering.

Mesenchymal stem cell-derived protein extract induces periodontal regeneration: MSC-protein induces periodontal regeneration

BackgroundPeriodontal disease is characterized by chronic inflammation and destruction of supporting periodontal tissues, ultimately leading to tooth loss. In recent years, “cell-free treatment” without stem cell transplantation has attracted considerable attention for tissue regeneration. This study investigated the effects of extracts of mesenchymal stem cells (MSC-extract) and their protein components (MSC-protein) on the proliferation and migration of periodontal ligament (PDL) cells and whether MSC-protein can induce periodontal regeneration. MethodsMSC-extract and MSC-protein were obtained by subjecting mesenchymal stem cells (MSCs) to freeze–thaw cycles and acetone precipitation. Cell proliferation was examined using a WST-8 assay and Ki67 immunostaining, and cell migration was examined using Boyden chambers. The MSC-protein content was analyzed using liquid chromatography-mass spectrometry, protein arrays, and enzyme-linked immunosorbent assays (ELISAs). Gene expression in MSC-protein-treated PDL cells was examined using RNA-sequencing and Gene Ontology analyses. The regenerative potential of MSC-protein was examined using micro-computer tomography (CT) and histological analyses after transplantation into a rat periodontal defect model. ResultsMSC-extract and MSC-protein promoted the proliferation and migration of PDL cells. Protein array and ELISA revealed that MSC-protein contained high concentrations of basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF). Exogenous bFGF promoted the proliferation and migration of PDL cells. Furthermore, the transplantation of MSC-protein enhanced periodontal tissue regeneration with the formation of new alveolar bone and PDLs. ConclusionsThese results indicate that the MSC-protein promotes the proliferation and migration of PDL cells and induces significant periodontal tissue regeneration, suggesting that the MSC-protein could be used as a new cell-free treatment for periodontal disease.

Proboscis Lateralis of the Nose

Abstract Proboscis lateralis (PL) of the nose is a very uncommon craniofacial anomaly characterized by a rudimentary tubular nose-like structure seen a little away from the midline of the face. It may be associated with other anomalies such as heminasal aplasia, mental retardation, callosal agenesis, microphthalmia, heminasal hypoplasia, and atypical clefting syndrome. Patients with this congenital craniofacial malformation need proper evaluation with the help of a computed tomography scan to assess the growth of facial and skull bones. The esthetic aspects and psychological problems are often a concern for patient’s families. The cartilaginous part or bony support of the nose can be reconstructed following proper growth of the face. Here, we presented a male neonate of 15 days with PL of the nose. There was no evidence of cleft lip or cleft palate. Parents were counseled for staged repair at an early age to avoid anxiety and to allow proportionate growth of the nose and face.