Rat Femur Research Articles

Interaction between ferroptosis and TNF-α: Impact in obesity-related osteoporosis.

The relationship of obesity and osteoporosis has been widely studied over the past years. However, the implications of obesity for bone health remain controversial, and the underlying molecular mechanism is not yet fully understood. This study demonstrated that high-fat diet-induced obesity leads to significantly decreased bone volume/tissue volume (BV/TV), trabecular number (Tb.N), and cortical thickness (Ct.Th) of male rat femur after mechanical loading effects of body weight were controlled. HFD-induced obese rats exhibited attenuated expression of ferroptosis inhibitory protein SLC7A11 and GPX4 in bone tissues, which was correlated with elevated serum TNF-α concentration. Ferroptosis inhibitor administration could effectively rescue decreased osteogenesis-associated type H vessels and osteoprogenitors, and downregulate serum levels of TNF-α to ameliorate bone loss in obese rats. Since ferroptosis and TNF-α both affect bone and vessel formation, we further investigated the interaction between ferroptosis and TNF-α, and its impact in osteogenesis and angiogenesis in vitro. In human osteoblast-like MG63 and umbilical vein endothelial cells (HUVEC), TNF-α/TNFR2 signaling promoted cystine uptake and GSH biosynthesis to provide protection against low-dose ferroptosis inducer erastin. While, TNF-α/TNFR1 facilitated ferroptosis in the presence of high-dose erastin through ROS accumulation. Moreover, TNF-α regulated ferroptosis-induced osteogenic and angiogenic dysfunctions based on its ferroptosis regulatory role. Meanwhile, ferroptosis inhibitors could reduce intracellular ROS overproduction and enhance osteogenesis and angiogenesis in TNF-α-treated MG63 and HUVECs. This study revealed the interaction between ferroptosis and TNF-α and its impact in osteogenesis and angiogenesis, which provides new insights into the pathogenesis and regenerative therapy of obesity-related osteoporosis.

Three-dimensional adipose-derived stem cell spheroids exert potent therapeutic effects on rat femur osteochondral defects

BackgroundSpheroids can allow three-dimensional (3D) cell culture without scaffolds, potentially promoting the production of growth factors from adipose-derived stem cells (ADSCs). We hypothesized that ADSC spheroids exert more favourable effects on osteochondral defects than ADSCs in two-dimensional (2D) cultures. The purpose of this study was to compare the therapeutic effects of 2D and 3D cultures of ADSCs on osteochondral defects using animal models. MethodsRat femoral osteochondral defects were created. When creating osteochondral defects, phosphate‐buffered saline, 2D ADSCs, or ADSC spheroids as a 3D culture were administered on to the lesion. At 2, 4, 6, 8, 10 and 12 weeks post-surgery, knee tissues were harvested and evaluated via histological examination. The expression of genes related to growth factors and apoptosis were compared between 2D and 3D ADSCs. ResultsHistologically, the repair of osteochondral defects was significantly enhanced in 3D ADSCs than in 2D ADSCs in terms of the Wakitani score and cartilage repair rate. In 3D ADSCs, TGF-β1, VEGF, HGF and BMP-2 were significantly upregulated, while apoptosis was suppressed in the early phase. ConclusionThe therapeutic effects of 3D ADSC spheroids on osteochondral defects were more potent than those of 2D ADSCs. The upregulated expression of growth factors and suppression of apoptosis could contribute to promoting these therapeutic effects. Overall, ADSC spheroids can help treat osteochondral defects.

Stromal Lineage Precursors from Rodent Femur and Tibia Bone Marrows after Hindlimb Unloading: Functional Ex Vivo Analysis.

Rodent hindlimb unloading (HU) model was developed to elucidate responses/mechanisms of adverse consequences of space weightlessness. Multipotent mesenchymal stromal cells (MMSCs) were isolated from rat femur and tibia bone marrows and examined ex vivo after 2 weeks of HU and subsequent 2 weeks of restoration of load (HU + RL). In both bones, decrease of fibroblast colony forming units (CFU-f) after HU with restoration after HU + RL detected. In CFU-f and MMSCs, levels of spontaneous/induced osteocommitment were similar. MMSCs from tibia initially had greater spontaneous mineralization of extracellular matrix but were less sensitive to osteoinduction. There was no recovery of initial levels of mineralization in MMSCs from both bones during HU + RL. After HU, most bone-related genes were downregulated in tibia or femur MMSCs. After HU + RL, the initial level of transcription was restored in femur, while downregulation persisted in tibia MMSCs. Therefore, HU provoked a decrease of osteogenic activity of BM stromal precursors at transcriptomic and functional levels. Despite unidirectionality of changes, the negative effects of HU were more pronounced in stromal precursors from distal limb-tibia. These observations appear to be on demand for elucidation of mechanisms of skeletal disorders in astronauts in prospect of long-term space missions.

Chronic intermittent hypobaric hypoxia ameliorates osteoporosis after spinal cord injury through balancing osteoblast and osteoclast activities in rats.

As a common complication of spinal cord injury (SCI), most SCI patients suffer from osteoporosis. In our previous study, chronic intermittent hypobaric hypoxia (CIHH) could promote bone fracture healing. We speculated that it may act a role in the progression of osteoporosis. The current study purposed to explore the role of CIHH in the osteoporosis triggered by SCI in rats. A SCI-induced SCI model was established by completed transection at T9-T10 spinal cord of Wistar rats. One week after SCI, the rats were conducted toCIHH treatment (PB = 404 mmHg, Po2 = 84 mmHg) 6 hours a day for continuously 7 weeks. The results of X-radiography and Micro-CT assessment demonstrated that compared with sham rats, the areal bone mineral density (BMD), bone volume to tissue volume, volumetric BMD, trabecular thickness, trabecular number, and trabecular connectivity were decreased. Trabecular bone pattern factor, trabecular separation, as well as structure model index were increased at the distal femur and proximal tibia of SCI rats, which were effectively reversed by CIHH treatment. Histomorphometry showed that CIHH treatment increased bone formation of SCI rats, as evidenced by the increased osteoid formation, the decreased number and surface of TRAP-positive osteoclasts. Furthermore, ELISA and real time PCR results showed that the osteoblastogenesis-related biomarkers, such as procollagen type 1 N-terminal propeptide, osteocalcin in serum, as well as ALP and OPG mRNAs in bone tissue were decreased, while the osteoclastogenesis-related biomarkers, including scleorostin in serum and RANKL and TRAP mRNAs in bone tissue were increased in SCI rats. Importantly, the deviations of aforementioned biomarkers were improved by CIHH treatment. Mechanically, the protective effects of CIHH might be at least partly mediated by hypoxia-inducible factor-1 alpha (HIF-1α) signaling pathway. The present study testified that CIHH treatment ameliorates osteoporosis after SCI by balancing osteoblast and osteoclast activities in rats.

The effects of cephalexin on fracture healing in a rat femur fracture model.

The aim of this study was to examine the effects of cephalexin on the fracture union histomorphometrically, radiologically, biomechanically, immunohistochemically, and histopathologically in a rat femur fracture model and to evaluate the effects of the antibiotics to be used in the prophylaxis of fracture infection on the union of the fracture. A total of 48 male Wistar rats were divided into four groups as two-week control (C2) and cephalexin (CEP2) and four-week control (C4) and cephalexin (CEP4). After establishment of standard fracture model on right femurs, 60 mg/kg/day of cephalexin was applied to CEP2 and CEP4 by oral gavage. Radiological, biomechanical, histopathological, immunohistochemical, and histomorphometric examinations were performed on amputated femurs. Callus volume of CEP4 group significantly increased compared to CEP2 group (p=0.005), while no significant difference was found in the bone mineral density and callus/bone volume among the groups (p>0.05). There was no significant difference in flexural strength between the C4 and CEP4 groups (p=0.093). Histological healing scores increased from Week 2 to Week 4 (p=0.002) and inflammation scores decreased in both control and cephalexin groups (p=0.010 and p=0.008); however, no significant difference was found in healing and inflammation scores (p>0.05). The CD34+ immunoreactivity in the CEP2 group was significantly higher than the C2 group (p=0.029). Collagen type III level was significantly lower in the CEP2 and CEP4 groups compared to the corresponding control groups (p=0.008 and p=0.016, respectively). Cephalexin did not exert any radiological, histopathological, histomorphometric, biomechanical, and immunohistochemical adverse effects on the femoral fracture healing model in rats; however, it showed positive effects on CD34 and Collagen type III levels. Based on these findings, antibiotherapy with cephalexin may be considered as a safe treatment for fracture union.

The mechanism of palmatine-mediated intestinal flora and host metabolism intervention in OA-OP comorbidity rats.

ErXian decoction is a Chinese herbal compound that can prevent and control the course of osteoarthritis (OA) and osteoporosis (OP). OP and OA are two age-related diseases that often coexist in elderly individuals, and both are associated with dysregulation of the gut microbiome. In the initial study, Palmatine (PAL) was obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and network pharmacological screening techniques, followed by 16S rRNA sequencing and serum metabolomics of intestinal contents, to explore the mechanism of PAL in the treatment of OA and OP. The rats selected for this study were randomly divided into three groups: a sham group, an OA-OP group and a PAL group. The sham group was intragastrically administered normal saline solution, and the PLA group was treated with PAL for 56 days. Through microcomputed tomography (micro-CT), ELISA, 16S rRNA gene sequencing and non-targeted metabonomics research, we explored the potential mechanism of intestinal microbiota and serum metabolites in PAL treatment of OA-OP rats. Palmatine significantly repair bone microarchitecture of rat femur in OA-OP rats and improved cartilage damage. The analysis of intestinal microflora showed that PAL could also improve the intestinal microflora disorder of OA-OP rats. For example, the abundance of Firmicutes, Bacteroidota, Actinobacteria, Lactobacillus, unclassified_f_Lachnospiraceae, norank_f_Muribaculaceae, Lactobacillaceae, Lachnospiraceae and Muribaculaceae increased after PAL intervention. In addition, the results of metabolomics data analysis showed that PAL also change the metabolic status of OA-OP rats. After PAL intervention, metabolites such as 5-methoxytryptophol, 2-methoxy acetaminophen sulfate, beta-tyrosine, indole-3-carboxylic acid-O-sulfate and cyclodopa glucoside increased. Association analysis of metabolomics and gut microbiota (GM) showed that the communication of multiple flora and different metabolites played an important role in OP and OA. Palmatine can improve cartilage degeneration and bone loss in OA-OP rats. The evidence we provided supports the idea that PAL improves OA-OP by altering GM and serum metabolites. In addition, the application of GM and serum metabolomics correlation analysis provides a new strategy for uncovering the mechanism of herbal treatment for bone diseases.

A kaolin/calcium incorporated shape memory and antimicrobial chitosan-dextran based cryogel as an efficient haemostatic dressing for uncontrolled hemorrhagic wounds

Increased mortalities associated with uncontrolled and excessive bleeding is still of paramount concern in the clinics, caregivers and military medics. Herein, we designed a shape memory cryogel based on chitosan (C) and functionalized-dextran (D), incorporated with Kaolin (K) and calcium (Ca2+) as haemostatic agents. The developed cryogel (CDKCa) exhibits a uniform interconnected porous architecture with profound fluid absorption ability, rapid blood clotting, stable clot formation and good antibacterial activity. The CDKCa elucidates significantly less clotting time (~30 s; in-vitro) and increased aggregation and activation of platelets/red blood cells in comparison to the control groups and commercial dressings (Axiostat and QuikClot). The developed CDKCa also significantly reduced the aPTT and PT values by ~58 % and 31 % respectively, leading to the activation of intrinsic and extrinsic coagulation cascades. The CDKCa cryogel displays enhanced mechanical stability, flexibility and a good shape memory, a property quintessential to cease uncontrolled bleeding in irregular and non-compressible wounds. Further, the Kaolin and Ca2+ incorporated shape memory CDKCa cryogel demonstrates a rapid blood coagulation and stable clot formation in different compressible and non-compressible rat liver and femur hemorrhagic models. In summary, the endorsed results of CDKCa suggest that the design, fabrication and excellent clotting ability may attribute to high haemostatic efficiency of CDKCa dressing and have a great potential to prevent uncontrollable hemorrhages.

Regulation of Wnt Signaling Pathway by Costic Acid Derivative, An Efficient Strategy for Treatment of Glucocorticoid‐Induced Osteoporosis in Rat Model

AbstractOsteoporosis leads to a decrease in bone mass, loss of mechanical strength, increase in fracture susceptibility and is a serious medical problem worldwide. The present study investigated the effect of costic acid derivative (CA) on dexamethasone (DXM)‐induced osteoporosis in vitro in osteoblasts and in vivo in rat a model. The results demonstrated that DXM‐mediated reduction in osteoblast viability was effectively prevented by CA treatment in dose‐dependent manner. The DXM‐induced loss of osteoblast viability was completely reversed by CA treatment at 4 μM dose. Treatment of the primary osteoblasts with 0. 5 and 4 μM CD for 72 h reversed DXM‐mediated reduction in ALP activity. The CA treatment significantly (P>0.05) reversed DXM‐mediated lowering of osteoclacin, collagen type‐1 and osteonectin mRNA expression. The CA treatment at 1.5, 3 and 6 mg/kg doses significantly (P>0.05) reversed DXM‐mediated decrease in BMD in rat femurs. Treatment of the rats with CA reversed DXM‐mediated changes in osteocalcin and CTX levels in serum samples. Administration of CA to the rats reversed DXM‐mediated decrease in Wnt, β‐catenin expression and increase in SOST and Dickkopf‐1 level. Docking studies showed that CA interacts with mammalian Wnt‐frizzled complex (6AHY) through conventional H‐bonding. Thus, CA inhibits DXM‐induced dysfunction of osteoblast differentiation in vitro and inhibits osteoporosis development in vivo in rat model. Therefore, CA may be developed as a potential therapeutic agent for treatment of bone damage induced by glucocorticoid (GC)‐mediated osteoporosis.

Surface modification of titanium implant for repairing/improving microenvironment of bone injury and promoting osseointegration

• A coating consist of MnO 2 nanoenzyme and Sr ions was fabricated on Ti implant. • MnO 2 nanoenzyme alleviated OA and O 2 deficiency at the injury site. • Sr ions released from the surface of implant promoted the growth of MSCs. • The modified Ti implant significantly accelerate bone regeneration in vivo . Bone injury and implantation operation are often accompanied by microenvironment damage of bone tissue, which seriously affects the process of osseointegration of implants, especially for titanium (Ti)-based bioinert materials. Thus, repairing or improving the microenvironment of damaged bone tissue is of great significance for bone rescue, reconstruction, and regeneration still a major medical challenge. Oxidative stress (OS) and oxygen (O 2 ) deficiency are considered to be specific physiological signals of the bone-injury microenvironment. From the above background, a coating consisting of manganese dioxide (MnO 2 ) nanoenzyme and strontium (Sr) ions was fabricated on the surface of the Ti implant via a one-step hydrothermal treatment. MnO 2 nanoenzyme presented in the coating alleviated OS and O 2 deficiency at the injury site by catalyzing the decomposition of abundant endogenous H 2 O 2 around the modified Ti implants into O 2 . In addition, Sr ions were released from the surface of the implant at a certain rate in a body-fluid environment, further promoting the adhesion, growth, and osteogenic differentiation of mesenchymal stem cells. More importantly, a Sprague Dawley rat femur model demonstrated that the modified Ti implant showed significant potential to accelerate bone tissue reconstruction in vivo . In summary, the present system provides a new idea for the treatment of bone injury and the development of new orthopedic implants.

Effects of Different Numerical Methods on the Fracture Prediction Accuracy for Cortical Bone Structure under Bending Load

Three numerical methods, including element instantaneous failure, continuum damage mechanics, and extended finite element methods, are mainly used to simulate the fracture in cortical bone structure. Although many simulations focus on the cortical bone fracture, few have investigated the differences in prediction accuracy among the three numerical methods. The purpose of this study was to evaluate the prediction accuracy and applicability of the three numerical methods in simulating cortical bone fracture under bending load. The rat femur samples were first used to perform the three-point bending experiment. Then, the three numerical methods were respectively used to conduct fracture simulation on the femoral finite element models. Each result was compared with the experimental data to determine the prediction accuracy. The results showed that fracture simulation based on the continuum damage mechanics method was in better agreement with the experimental results, and observable differences in the failure processes could be seen in the same model under the three simulations due to various element failure strategies. The numerical method that was suitable for simulating cortical bone fracture under bending load was determined; meanwhile, the variations in the failure simulations were observed, and the cause of the variations in the predicted results using different numerical methods was also discussed, which may have potential to improve the prediction accuracy of cortical bone fracture.

Properties of Calcium Phosphate/Hydrogel Bone Grafting Composite on the Model of Diaphyseal Rat Femur’s Defect: Experimental Study

Background. The problem of bone defects replacement is relevant nowadays, that is why many scientists create new synthetic bone substitutes, but the ideal material has not been found so far.
 The aims of the study: 1) to determine the suitability of the monocortical defect model in the rat femur diaphysis with additional prophylactic reinforcement with a bone plate for assessing the biological properties of implanted materials using the commercially available ChronOS material as an example; 2) to assess of the osteoconductive properties of composite materials based on poly(ethylene glycol)diacrylate and octacalcium phosphate with architecture Kelvin and gyroid types on the developed model.
 Methods. A prospective study, level of evidence II. A monocortical defect of the rat femoral diaphysis (length 7 mm) was produced under anaesthesia in aseptic conditions and fixed with a polyetheretherketone plate and six titanium screws. In the control group, the defect was left empty. In other groups, blocks of one of three materials were implanted сhronOS and composites of poly(ethylene glycol)diacrylate and octacalcium phosphate with 3D-printed Kelvin and gyroid architectures. After 3 and 6 weeks, the rats were sacrificed, and histological examination of the defect zone was performed. The amount of newly formed bone tissue was histometricly assessed, followed by statistical processing of the results.
 Results. All rats have reached the planned endpoint, and there were no infectious complications or loss of fixation. Histological examination of the defect zone revealed minimal bone growth in the Control group, rather slow bone formation in the Gyroid group, and statistically significantly more pronounced bone formation in the pores of the materials in the Kelvin and Chronos groups.
 Conclusions. Bone defect in this model was not spontaneously filled with bone tissue and allowed us to study the biological properties of bone substitutes (the ability to biodegrade and osteoconductive properties). The osteoconductive properties of a composite material based on poly(ethylene glycol)diacrylate and octacalcium phosphate with a Kelvin architecture are higher than with a gyroid architecture and are comparable to that of the сhronOS.

Effects of Eleutherine bulbosa (mill.) urb. bulb extract on mice glucocorticoid-induced osteoporosis models.

Low bone mass accompanied by microarchitectural alterations in the bone that cause fragility fractures is known as secondary osteoporosis and occurs when there is an underlying condition or medication present. Eleutherine bulbosa bulb extract has been shown to affect bone because of its content, which can help osteoblast differentiation and inhibit osteoclast differentiation. This study aimed to assess the effects of 70% ethanol extract of E. bulbosa Bulbs (EBE) from Pasuruan-East Java on blood calcium levels, osteoblast cell count, and bone density of trabecular femur in osteoporosis rats. Six groups of 30 female Wistar rats were created. There were no test materials offered to the healthy group; the negative group received 0.5% CMC; the positive group received alendronate 0.9 mg/kg BW; and the dose group received 30, 60, and 120 mg/kg BW. Glucocorticoid (Dexamethasone) 0.1015 mg/kg BW/day induction was given to all groups except the healthy group to create osteoporosis rats for approximately four weeks. Then they were given oral therapy for approximately 28 days. Followed by the determination of blood calcium levels, the number of osteoblast cells, and bone density of the rat femur trabecular. The result showed that E. bulbosa bulbs extract could raise blood calcium levels and bone density percentage at doses of 60 and 120 mg/kg BW, as well as raise osteoblast cell levels at doses of 120 mg/kg BW. The findings indicate that E.bulbosa bulb extract is a potential complementary medicine for osteoporosis.

In vivo degradation and bone reaction of long-term fixation with a magnesium alloy made by twin-roll casting in a rat femur model.

The effect of casting parameters on the microstructure and corrosion resistance of Mg alloys is still limited, especially in clinical animal experiments. We prepared a new magnesium rare earth alloy (Mg-Re, where Re is Ce or La) by vertical two-roll casting and Mg-A by further rolling. The microstructure characteristics, degradation behavior, and bone reaction of the two alloys were studied. Ti, Mg-Re, and Mg-A alloy plates were implanted in a rat femur model, and their degradation behavior was observed 48 weeks later. In vivo experiments showed no significant changes around the femur in the Ti group, excluding external factors that may cause bone remodeling and lead to new bone formation. Mg-A induces more new bone formation than Mg-Re, which meets the necessary conditions to prevent pathological fracture. The specimen staining and sectioning showed that the liver and heart of rats implanted with magnesium alloys had no pathological changes and the cell structure was normal, similar to that of rats without a magnesium alloy. Mg-A alloy has good healing potential as a biodegradable implant material.

Development of a Metaphyseal Non-Union Model in the Osteoporotic Rat Femur.

The aim of this current study was to establish a metaphyseal femoral non-union model in osteoporotic rats by comparing a power tool versus a manual tool for fracture creation. Twelve adult female Sprague Dawley rats were ovariectomized (OVX) and received a special diet for 6 weeks. Biweekly pQCT measurements confirmed a significant reduction in the cancellous and total bone mineral density in OVX rats compared to control (CTRL) animals. After 6 weeks, OVX rats underwent surgery creating a distal metaphyseal osteotomy, either using a piezoelectric- (n = 6) or a manual Gigli wire (n = 6) saw. Fractures were stabilized with a Y-shaped mini-locking plate. Within each group, three rats received Alginate directly into the fracture gap. OVX animals gained more weight over 8 weeks compared to CTRL animals. pQCT analysis showed a significant difference in the volumetric cancellous bone mineral density between OVX and CTRL rats. A histological examination of the osteoporotic phenotype was completed. Radiographic evaluation and Masson-Goldner trichrome staining with the piezoelectric saw failed to demonstrate bony bridging or a callus formation. New bone formation and complete healing were seen after 6 weeks in the Gigli group. For the creation of a metaphyseal atrophic non-union in the osteoporotic bone, a piezoelectric saw should be used.

Segmental defect healing in the presence or absence of recombinant human BMP2: Novel insights from a rat model.

This study defined and compared the course of native, impaired and growth factor-stimulated bone regeneration in a rat femoral defect model. A mid-diaphyseal defect with rigid internal fixation was surgically created in the right femur of male Fischer rats and serially analyzed over 36 weeks. Native bone regeneration was modeled using a sub-critical, 1 mm size defect, which healed uneventfully. Critical size defects of 5 mm were used to analyze impaired bone regeneration. In a third group, the 5 mm defects were filled with 11 µg of recombinant human bone morphogenetic protein 2 (rhBMP2) impregnated onto an absorbable collagen sponge, modeling its clinical use. Native bone regeneration was characterized by endochondral ossification with progressive remodeling to ultimately resemble intact femora. An endochondral response was also observed under conditions of impaired bone regeneration, but by week 8 medullary capping occurred with fibrofatty consolidation of the tissue within the defect, resembling an atrophic non-union. rhBMP2 treatment was associated with prolonged inflammatory cytokine expression and rapid intramembranous bone formation occurring with reduced expression of cartilage-associated collagens. Between weeks 4 and 36, rhBMP2-treated bones demonstrated decreased trabecular number and increased trabecular separation, which resulted in inferior mechanical properties compared with bones that healed naturally. Clinical Significance: Recombinant human bone morphogenetic protein 2 (rhBMP2) is used clinically to promote healing of long bones. Our data suggest that it drives intramembraneous ossification producing an inferior regenerate that deteriorates with time. Clinical outcomes would be improved by technologies favoring endochondral regenerative ossification.

Biological properties of hydroxyapatite coatings on titanium dioxide nanotube surfaces using negative pressure method.

Titanium (Ti) exhibits superior biocompatibility and mechanical properties but is bioinert, while hydroxyapatite (HA) possesses excellent osteogenesis and is widely used for the modification of Ti surface coatings. However, the synthesis of homogeneous and stable HA on metallic materials is still a major challenge. In this study, porous titanium dioxide nanotube arrays were prepared on Ti surface by anodic oxidation, loaded with calcium and phosphorus precursors by negative pressure immersion, and HA coating was formed by in situ crystallization of calcium and phosphorus on the surface by hydrothermal heating. Scanning electron microscopy (SEM), X-ray diffraction (XRD), and bonding strength were conducted to confirm the surface characteristics of each group. The cell proliferation, mineralization degree, and alkaline phosphatase (ALP) activity of MC3T3-E1 cells on samples were calculated and compared in vitro experiments. Cylindrical samples were implanted into rat femurs to evaluate biocompatibility and osteogenesis in vivo. The results showed that HA crystals successfully synthesized in TiO2 nanotubes, enhancing the bonding strength of HA coating and Ti substrate under negative pressure. Moreover, HA coating on Ti substrate remarkably enhanced cell proliferation and osteogenic differentiation activity in vitro, and improved new bone formation as well as osseointegration in vivo.

Bone Zinc Is Poorly Released in Young, Growing Rats Fed Marginally Zinc-Restricted Diet

The effect of marginal zinc restriction on bone zinc release was studied. Male weanling rats were fed zinc-adequate diet (12 mg zinc/kg diet) for 1 wk, intubated with 65Zn, refed the zinc-adequate diet for an additional week, and divided into one baseline group and five marginally zinc-restricted groups (6 mg/kg) and five corresponding zinc-adequate pair-fed groups. Significantly lower (P < 0.05) 65Zn radioactivity (10-20%) in the first week was observed in femurs of rats fed the marginally zinc-restricted diet without further change in the following weeks. Pair-fed rats showed higher 65Zn radioactivity in femurs in the second week, and the level was maintained. Similar results were found in tibia 65Zn radioactivity. Muscle zinc concentration was higher in the first week (P < 0.05) but subsequently returned to baseline levels in the second week through the fifth week. The results support the existence of two zinc pools in the skeleton, one a small, rapidly turning-over pool (10-20% in size), and the other a slowly turning-over pool. When rats suffer marginal zinc restriction, zinc from the readily turning-over bone pool is utilized for soft tissue needs, followed by decreased growth rate to maintain zinc homeostasis.

Micron/Submicron Scaled Hierarchical Ti Phosphate/Ti Oxide Hybrid Coating on 3D Printed Scaffolds for Improved Osteointegration.

Three-dimensional (3D) printed implants have attracted substantial attention in the field of personalized medicine, but negative impacts on mechanical properties or initial osteointegration have limited their application. To address these problems, we prepared hierarchical Ti phosphate/Ti oxide (TiP-Ti) hybrid coatings on 3D printed Ti scaffolds. The surface morphology, chemical composition, and bonding strength of the scaffolds were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), contact angle measurement, X-ray diffraction (XRD), and scratch test. In vitro performance was analyzed by colonization and proliferation of rat bone marrow mesenchymal stem cells (BMSCs). In vivo osteointegration of the scaffolds in rat femurs was assessed by micro-CT and histological analyses. The results demonstrated improved cell colonization and proliferation as well as excellent osteointegration obtained by incorporation of our scaffolds with the novel TiP-Ti coating. In conclusion, micron/submicron scaled Ti phosphate/Ti oxide hybrid coatings on 3D printed scaffolds have promising potential in future biomedical applications.

MiR-218-5p-dependent SOCS3 downregulation increases osteoblast differentiation inpostmenopausal osteoporosis

BackgroundPostmenopausal osteoporosis (POP) is a prevalent skeletal disease among elderly women. Previous study indicated that suppressor of cytokine signaling 3 (SOCS3) participates in the regulation of bone marrow stromal cell (BMSC) osteogenesis. Here, we further investigated the exact function and mechanism of SOCS3 in POP progression.MethodsBMSCs were isolated from Sprague–Dawley rats and treated with Dexamethasone (Dex). Alizarin Red staining and ALP activity assays were applied to assess the osteogenic differentiation of rat BMSCs under the indicated conditions. Osteogenic genes (ALP, OPN, OCN, COL1) mRNA levels were determined using quantitative RT-PCR. Luciferase reporter assay verified the interaction between SOCS3 and miR-218-5p. Rat models of POP were established in ovariectomized (OVX) rats to detect the in vivo effects of SOCS3 and miR-218-5p.ResultsWe found that silencing SOCS3 antagonized the suppressive effects of Dex on the osteogenic differentiation of BMSCs. SOCS3 was found to be targeted by miR-218-5p in BMSCs. The SOCS3 levels were negatively modulated by miR-218-5p in femurs of POP rats. MiR-218-5p upregulation promoted the BMSC osteogenic differentiation, while SOCS3 overexpression reversed the effects of miR-218-5p. Moreover, SOCS3 was highly expressed and miR-218-5p was downregulated in the OVX rat models, and silencing SOCS3 or overexpressing miR-218-5p alleviated POP in OVX rats by promoting osteogenesis.ConclusionSOCS3 downregulation mediated by miR-218-5p increases osteoblast differentiation to alleviate POP.

The effect of alginate scaffolds on bone healing in defects formed with drilling model in rat femur diaphysis.

Alginate (ALG) is a biocompatible and biodegradable polymer. Mechanical weakness is one of the main problems for the alginate-based scaffolds. Various plasticizer additives or modifications tested to improve the mechanical properties. In the presented study, ALG plasticized with triacetin (TA), and tributyl citrate (TBC) than tested on bone healing. In the presented study, the alginate modified with triacetin or tributyl citrate. In-vitro, and in-vivo efficiency of the scaffolds tested on bone tissue regeneration. Scaffolds fabricated by solvent casting, and physicochemical characterizations performed. Monocytes (THP-1) cultured with scaffolds, and macrophage-released cytokines was determined. In-vivo efficacy of the scaffolds was tested in the rat drill hole model. Alginate and tributyl citrate-modified scaffolds have no cytotoxic effect on osteoblastic cells (MC-3T3). Tributyl citrate modification increased tumor necrosis factor-alpha (TNF-alpha) level but did not increase interleukin -1 beta (IL-1 beta) level. In vivo studies showed that osteoblastic growth was significant in alginate and triacetin-modified scaffolds. However, the best values for osteoclastic activity and osteoid tissue formation seen in the triacetin modification. The results demonstrated that the modified alginate scaffolds were more successful than non-modified alginate scaffolds and can used as long-term bone repairing treatments.