Rat Femoral Model Research Articles

Effects of Vancomycin/Tobramycin-Doped Ceramic Composite (Polyvinyl Alcohol Composite–Vancomycin/Tobramycin–Polymeric Dicalcium Phosphate Dihydrate) in a Rat Femur Model Implanted With Contaminated Porous Titanium Cylinders

BackgroundPeriprosthetic joint infection (PJI) remains common and problematic. We hypothesized that using a bioceramic that provided rapid release of the antibiotics (vancomycin [VAN] or VAN and tobramycin [VAN and TOB]) from a polyvinyl-alcohol-composite (PVA) combined with a delayed and sustained antibiotic release from polymeric-dicalcium-phosphate-dihydrate (PDCPD) ceramic would inhibit S. aureus-associated implant infections. MethodsA total of 50 male Sprague Dawley rats were randomly divided into 5 groups—I: negative control; II: bacteria only; III: bacteria + saline wash; IV: bacteria + PVA-VAN-PDCPD, and V: bacteria + PVA-VAN-TOB-PDCPD. A porous titanium (Ti) implant was press-fit into the rat knee. S. aureus-containing broth was added into the joint space creating a PJI. After 1 week, the joints from groups III to V were washed with saline and the fluid collected for bacterial quantification. This was followed by saline irrigation treatment (groups III to V) and application of the antibiotic-loaded PVA-PDCPD bioceramic (groups IV and V). On day 21, joint fluid was collected, and the implants harvested for bacterial quantification. ResultsNo bacteria were isolated from the negative control (group I). The positive control (group II) was positive on both days 7 and 21. Bacteria were still present on day 21 in the fluid and implant in group III. Groups (IV and V) showed a decrease in the bacterial burden in the fluid and implant on day 21. There were significant differences in bacteria levels in the collected wash fluid and on the implant at day 21 between the saline wash (group III) and treatment groups (IV and V). ConclusionsIn this animal model of acute periprosthetic infection, treatment with PVA-VAN-PDCPD and PVA-VAN/TOB-PDCPD reduced bacterial load in the infected joint and the infected Ti implant. Application of PVA-VAN-PDCPD and/or PVA-VAN/TOB-PDCPD after saline irrigation could be used as an addition to the treatment of PJI.

The osteoinductive potential of different root-end filling materials in a rat femur model

In pediatric dentistry, the enduring success of root-end filling materials employed for the establishment of apical plugs in immature teeth undergoing endodontic intervention is contingent upon their possessing a robust osteoinductive capacity. Hence, the primary objective of this study was to histologically assess the osteoinductive potential of four distinct dental materials, specifically recommended for retrograde filling applications, utilizing an animal hard tissue model. Within the designed experimental model, two bone defects measuring 2 mm in diameter and 2 mm in depth were created in both femurs of a cohort comprising 21 male Wistar albino rats. The first defect in the right femur was left blank as the control group, and Neo MTA Plus was placed in the second defect. The EndoSequence BCRRM Fast Set Putty was placed in the first of the defects on the left femur, and Endo Repair was placed in the second defect. Subjects were sacrificed after 7, 14, and 28 days of follow-up, and sections were examined to assess the degree of inflammation, connective tissue formation, and new bone formation. The data were statistically evaluated with Kruskal‒Wallis and post hoc Dunn's tests using SPSS 12 software. The bone healing levels of the Neo MTA Plus group were significantly higher than those of the other groups in all periods (p < 0.05). Bone organization in all groups decreased over time, and fibrous tissue was enriched. The results of this study demonstrate that NeoMta Plus has superior osteoinductive properties compared to other materials but that EndoSequence and Endo Repair have the potential to be developed.

Comparative study in vivo of the osseointegration of 3D-printed and plasma-coated titanium implants.

Total hip arthroplasty is a common surgical treatment for elderly patients with osteoporosis, particularly in postmenopausal women. In such cases, highly porous acetabular components are a favorable option in achieving osseointegration. However, further discussion is needed if use of such acetabular components is justified under the condition of normal bone mass. To determine the features of osseointegration of two different types of titanium implants [3-dimensional (3D)-printed and plasma-coated titanium implants] in bone tissue of a distal metaphysis in a rat femur model. This study was performed on 20 white male laboratory rats weighing 300-350 g aged 6 mo. Rats were divided into two groups of 10 animals, which had two different types of implants were inserted into a hole defect (2 × 3 mm) in the distal metaphysis of the femur: Group I: 3D-printed titanium implant (highly porous); Group II: Plasma-coated titanium implant. After 45 and 90 d following surgery, the rats were sacrificed, and their implanted femurs were extracted for histological examination. The relative perimeter (%) of bone trabeculae [bone-implant contact (BIC%)] and bone marrow surrounding the titanium implants was measured. Trabecular bone tissue was formed on the 45th day after implantation around the implants regardless of their type. 45 d after surgery, group I (3D-printed titanium implant) and group II (plasma-coated titanium implant) did not differ in BIC% (83.51 ± 8.5 vs 84.12 ± 1 .73; P = 0.838). After 90 d, the BIC% was higher in group I (87.04 ± 6.99 vs 81.24 ± 7.62; P = 0.049), compared to group II. The relative perimeter of the bone marrow after 45 d did not differ between groups and was 16.49% ± 8.58% for group I, and 15.88% ± 1.73% for group II. Futhermore, after 90 d, in group I the relative perimeter of bone marrow was 1.4 times smaller (12.96 ± 6.99 vs 18.76 ± 7.62; P = 0.049) compared to the relative perimeter of bone marrow in group II. The use of a highly porous titanium implant, manufactured with 3D printing, for acetabular components provides increased osseointegration compared to a plasma-coated titanium implant.

The effect of medical biodegradable magnesium alloy in vivo degradation and bone response in a rat femur model with long-term fixation.

It is of great significance to understand the effect of the different corrosion behaviors of magnesium (Mg) alloys manufactured using different casting methods and implanted with different methods on the long-term implantation to expand the application of Mg-based biomedical implants. The effects of four different casting and rolling speeds on the microstructure of an Mg-rare earth (Mg-Re) alloy were analyzed using electron backscatter diffraction (EBSD). Four Mg alloys were obtained using vertical two-roll casting (TRC) at 10m/min, 16m/min, 24m/min, and 30m/min, and their microstructure, corrosion behavior and bone reaction in vivo were studied. The corrosion resistance of the alloy increases with an increase in casting speed and finer grain size of the cast-rolled parts. The Mg-Re alloys with TRC-10m/min and TRC-30m/min were selected for animal experiments. The two Mg alloys were made into metal rods and inserted into the rat femur to simulate the effect of Mg-Re on femoral healing under an injury condition. The rods were implanted for a long time to judge the effects of the Mg-Re alloy on the body. The TRC-30m/min implants obtained highly mature new bone tissue in the case of bone injury. The in vivo experiments showed that the corrosion resistance of the TRC-30m/min implant was better than that of the TRC-10m/min implant. After 32 weeks of implantation, there were no pathological changes in the liver, heart, or kidney of rats in the TRC-30m/min group, and the cell structure was normal.

Impacts of residual 3D printing metal powders on immunological response and bone regeneration: an in vivo study

Residual powder is a defect in powder bed fusion-based additive manufacturing (3D printing), and it is difficult to completely remove it from as-printed materials. In addition, it is not necessary to apply 3D printed implants with residual powder in the clinic. The immunological response triggered by the residual powder is an important area of study in medical research. To further understand the possible immunological reactions and hidden dangers caused by residual powders in vivo, this study compared the immunological reactions and osteolysis caused by typical powders for four implant materials: 316 L stainless steel, CoCrMo, CP-Ti, and Ti-6Al-4V (particle size range of 15–45 μm), in a mouse skull model. Furthermore, the possible immunological responses and bone regeneration induced by the four 3D printed implants with residual powder in a rat femur model were compared. In the mouse skull model, it was found that the 316L-S, CoCrMo-S, and especially the 316L-M powders, upregulated the expression of pro-inflammatory factors, increased the ratio of RANKL/OPG, and activated more functional osteoclasts, resulting in more severe bone resorption compared with those in other groups. In the rat femur model, which is more suitable for clinical practice, there is no bone resorption in implants with residual powders, but they show good bone regeneration and integration ability because of their original roughness. The results indicate that the expressions of inflammatory cytokines in all experimental groups were the same as those in the control group, showing good biological safety. The results answered some critical questions related to additively manufactured medical materials in vivo and indicated that as-printed implants may have great potential in future clinical applications.Graphical

The Use of Retraction Sutures in End-To-Side Microvascular Anastomosis—A Novel Technical Innovation in Experimental Rat Femoral Model with Successful Use in STA-MCA Bypass in Moyamoya Disease

A narrow working space, small diameters, and the tendency to collapse with clamps make cerebral microvascular anastomosis challenging. A retraction suture (RS) is a novel technique to keep the recipient vessel lumen open during the bypass. To provide a step-by-step overview of RS for end-to-side (ES) microvascular anastomosis on rat femoral vessels and successful use for STA-MCA bypass in Moya Moya disease (MMD) patients. A prospective experimental study with approval from the Institutional Animal Ethics Committee. Femoral vessels ES anastomoses were performed on Sprague Daley rats. The rat model used three types of RS (Adventitial, Luminal, and Flap-retraction sutures). An ES-interrupted anastomosis was done. The rats were observed for an average period of 16.18±5.65 days; the patency was assessed by re-exploration. The immediate patency on the STA-MCA bypasses was confirmed with intra-operative indocyanine green (ICG) angiography and micro-doppler; delayed patency with MRI and DSA after 3-6 months. In the rat model, 45 anastomoses were performed, 15 each using the three subtypes. The immediate patency was 100%. Delayed patency was 42/43 (97.67%), and two rats died during observation. In the clinical series, 59 STA-MCA bypasses were done in 44 patients (average age -18.14±11.09 years) using RS. The follow-up imaging was available for 41/59 patients. Both immediate and delayed patency (41/41 at six months) was 100%. The RS allows continuous visualization of the vessel lumen, reduces the handling of intimal edges, and avoids incorporating the back wall in sutures; thus improving anastomosis patency.

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.

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.

Sustained Release of Antifungal and Antibacterial Agents from Novel Hybrid Degradable Nanofibers for the Treatment of Polymicrobial Osteomyelitis

This study aimed to develop a drug delivery system with hybrid biodegradable antifungal and antibacterial agents incorporated into poly lactic-co-glycolic acid (PLGA) nanofibers, facilitating an extended release of fluconazole, vancomycin, and ceftazidime to treat polymicrobial osteomyelitis. The nanofibers were assessed using scanning electron microscopy, tensile testing, water contact angle analysis, differential scanning calorimetry, and Fourier-transform infrared spectroscopy. The in vitro release of the antimicrobial agents was assessed using an elution method and a high-performance liquid chromatography assay. The in vivo elution pattern of nanofibrous mats was assessed using a rat femoral model. The experimental results demonstrated that the antimicrobial agent-loaded nanofibers released high levels of fluconazole, vancomycin, and ceftazidime for 30 and 56 days in vitro and in vivo, respectively. Histological assays revealed no notable tissue inflammation. Therefore, hybrid biodegradable PLGA nanofibers with a sustainable release of antifungal and antibacterial agents may be employed for the treatment of polymicrobial osteomyelitis.

High-strength biodegradable zinc alloy implants with antibacterial and osteogenic properties for the treatment of MRSA-induced rat osteomyelitis

Implant-related infections caused by drug-resistant bacteria remain a major challenge faced by orthopedic surgeons. Furthermore, ideal prevention and treatment methods are lacking in clinical practice. Here, based on the antibacterial and osteogenic properties of Zn alloys, Ag and Li were selected as alloying elements to prepare biodegradable Zn-Li-Ag ternary alloys. Li and Ag addition improved the mechanical properties of Zn-Li-Ag alloys. The Zn-0.8Li-0.5Ag alloy exhibited the highest ultimate tensile strength (>530 MPa). Zn-Li-Ag alloys showed strong bactericidal effects on methicillin-resistant Staphylococcus aureus (MRSA) in vitro. RNA sequencing revealed two MRSA-killing mechanisms exhibited by the Zn-0.8Li-0.5Ag alloy: cellular metabolism disturbance and induction of reactive oxygen species production. To verify that the therapeutic potential of the Zn-0.8Li-0.5Ag alloy is greater than that of Ti intramedullary nails, X-ray, micro-computed tomography, microbiological, and histological analyses were conducted in a rat femoral model of MRSA-induced osteomyelitis. Treatment with Zn-0.8Li-0.5Ag alloy implants resulted in remarkable infection control and favorable bone retention. The in vivo safety of this ternary alloy was confirmed by evaluating vital organ functions and pathological morphologies. We suggest that, with its good antibacterial and osteogenic properties, Zn-0.8Li-0.5Ag alloy can serve as an orthopedic implant material to prevent and treat orthopedic implant-related infections.

Octacalcium Phosphate/Gelatin Composite (OCP/Gel) Enhances Bone Repair in a Critical-sized Transcortical Femoral Defect Rat Model.

Bone grafting is widely used to treat large bone defects. A porous composite of a bioactive octacalcium phosphate material with gelatin sponge (OCP/Gel) has been shown to biodegrade promptly and be replaced with new bone both in animal models of a membranous bone defect and a long bone defect. However, it is unclear whether OCP/Gel can regenerate bone in more severe bone defects, such as a critical-size transcortical defect. Using an in vivo rat femur model of a standardized, transcortical, critical-size bone defect, we asked: Compared with a Gel control, does OCP/Gel result in more newly formed bone as determined by (1) micro-CT evaluation, (2) histologic and histomorphometric measures, and (3) osteocalcin staining and tartrate-resistant acid phosphatase staining? Thirty-four 12-week-old male Sprague-Dawley rats (weight 356 ± 25.6 g) were used. Gel and OCP/Gel composites were prepared in our laboratory. Porous cylinders 3 mm in diameter and 4 mm in height were manufactured from both materials. The OCP/Gel and Gel cylinders were implanted into a 3-mm-diameter transcortical critical-size bone defect model in the left rat femur. The OCP/Gel and Gel were randomly assigned, and the cylinders were implanted. The biological responses of the defect regions were evaluated radiologically and histologically. At 4 and 8 weeks after implantation, CT evaluation, histological examination of decalcified samples, and immunostaining were quantitatively performed to evaluate new bone formation and remaining bone graft substitutes and activity of osteoblasts and osteoclast-like cells (n = 24). Qualitative histological evaluation was performed on undecalcified samples at 3 weeks postimplantation (n = 10). CT and decalcified tissue analysis was not performed blinded, but an analysis of undecalcified specimens was performed under blinded conditions. Radiologic analysis revealed that the OCP/Gel group showed radiopaque regions around the OCP granules and at the edge of the defect margin 4 weeks after implantation, suggesting that new bone formation occurred in two ways. In contrast, the rat femurs in the Gel group had a limited radiopaque zone at the edge of the defect region. The amount of new bone volume analyzed by micro-CT was higher in the OCP/Gel group than in the Gel group at 4 and 8 weeks after implantation (​​4 weeks after implantation: OCP/Gel versus Gel: 6.1 ± 1.6 mm 3 versus 3.4 ± 0.7 mm 3 , mean difference 2.7 [95% confidence interval (CI) 0.9 to 4.5]; p = 0.002; intraclass correlation coefficient [ICC] 0.72 [95% CI 0.29 to 0.91]; 8 weeks after implantation: OCP/Gel versus Gel: 3.9 ± 0.7 mm 3 versus 1.4 ± 1.1 mm 3 , mean difference 2.5 [95% CI 0.8 to 4.3]; p = 0.004; ICC 0.81 [95% CI 0.47 to 0.94]). Histologic evaluation also showed there was a higher percentage of new bone formation in the OCP/Gel group at 4 and 8 weeks after implantation (​​4 weeks after implantation: OCP/Gel versus Gel: 31.2% ± 5.3% versus 13.6% ± 4.0%, mean difference 17.6% [95% CI 14.2% to 29.2%]; p < 0.001; ICC 0.83 [95% CI 0.53 to 0.95]; 8 weeks after implantation: OCP/Gel versus Gel: 28.3% ± 6.2% versus 9.5% ± 1.9%, mean difference 18.8% [95% CI 11.3% to 26.3%]; p < 0.001; ICC 0.90 [95% CI 0.69 to 0.97]). Bridging of the defect area started earlier in the OCP/Gel group than in the Gel group at 4 weeks after implantation. Osteocalcin immunostaining showed that the number of mature osteoblasts was higher in the OCP/Gel group than in the Gel group at 4 weeks (OCP/Gel versus Gel: 42.1 ± 6.5/mm 2 versus 17.4 ± 5.4/mm 2 , mean difference 24.7 [95% CI 16.2 to 33.2]; p < 0.001; ICC 0.99 [95% CI 0.97 to 0.99]). At 4 weeks, the number of osteoclast-like cells was higher in the OCP/Gel composite group than in the Gel group (OCP/Gel versus Gel: 3.2 ± 0.6/mm 2 versus 0.9 ± 0.4/mm 2 , mean difference 2.3 [95% CI 1.3 to 3.5]; p < 0.001; ICC 0.79 [95% CI 0.35 to 0.94]). OCP/Gel composites induced early bone remodeling and cortical bone repair in less time than did the Gel control in a rat critical-size, transcortical femoral defect, suggesting that OCP/Gel could be used as a bone replacement material to treat severe bone defects. In a transcortical bone defect model of critical size in the rat femur, the OCP/Gel composite demonstrated successful bone regeneration. Several future studies are needed to evaluate the clinical application of this interesting bone graft substitute, including bone formation capacity in refractory fracture and spinal fusion models and the comparison of bone strength after repair with OCP/Gel composite to that of autologous bone.

3D printing of poly(butylene adipate-co-terephthalate) (PBAT)/niobium containing bioactive glasses (BAGNb) scaffolds: Characterization of composites, in vitro bioactivity, and in vivo bone repair.

This study aimed to produce poly(butylene adipate-co-terephthalate) (PBAT)/niobium containing bioactive glasses (BAGNb) composites scaffolds produced by fused deposition modeling (FDM) printing and evaluate their physicochemical and biological properties in vitro and in vivo. The composite filaments were produced by melt-extrusion with the addition of 10 wt% of BAGNb (PBAT/BAGNb). Filaments without BAGNb were produced as the control group (PBAT). The filaments were characterized and were used to produce 3D-printed scaffolds using FDM. The scaffolds' structure and surface properties were assessed. In vitro cell, proliferation, and cell mineralization analysis were performed. In vivo data was obtained in the rat femur model (n=10), and the bone repair was assessed after 15, 30, and 60 postoperative days. The printed structures presented 69.81% porosity for the PBAT/BAGNb group and 74.54% for the PBAT group. Higher cell mineralization was observed for the PBAT/BAGNb group. The in vivo data showed that the PBAT/BAGNb presented new bone formation comparable to positive controls. The combination of PBAT and BAGNb in 3D-printed scaffolds may be an alternative to produce bioactive materials with controllable shapes and properties for bone regeneration treatments.

UV photocatalytic activity of titanium dioxide (TiO2) surface contaminated with bacterial biofilm: Implications for photo-restoration of osteoconductivity

Ultraviolet (UV) light-mediated activation of titanium dioxide (TiO2) cleans the surface microenvironment through photocatalysis, but it is unknown whether this occurs when TiO2 surfaces is contaminated with bacterial biofilms. We therefore formed bacterial biofilms on TiO2 surfaces through culture with oral microorganisms from rats, which were subsequently exposed to high-intensity broadband UV light for 12 min. Osteoblast attachment, proliferation, and phenotypes were significantly compromised on biofilm-contaminated TiO2 surfaces, but UV treatment restored these biological activities to native baselines of TiO2 surfaces. The strength of bone-implant integration was 18.3 N for original implants, 1.5 N for biofilm-contaminated implants, and 30.5 N for biofilm-contaminated/UV-treated implants in a rat femur model after two weeks of healing. Histologically, there was limited, fragmented bone formation around biofilm-contaminated implants separated by thick fibrous tissue, while biofilm-contaminated/UV-treated implants induced robust bone formation with extensive direct bone-implant contact. Lipopolysaccharide (LPS) deposited on biofilm-contaminated TiO2 surfaces which was decomposed and removed by UV treatment. Notably, biofilm-contaminated TiO2 surfaces became superhydrophilic after UV treatment despite the persistence of carbon and nitrogen compounds, and UV treatment significantly restored the surface morphology of the innate titanium on the biofilm-contaminated TiO2 surfaces. In summary, bacterial biofilm severely compromised titanium osteoconductivity, but treatment of contaminated titanium with UV light significantly restored osteoconductivity through substantially decreased accumulation of carbon, nitrogen, and LPS; the re-emergence of micro-topography; and the induction of superhydrophilicity, paving the way for photoenergy-mediated debridement of TiO2 surface for clinical benefit.

Probing the role of methyl methacrylate release from spacer materials in induced membrane bone healing.

In the induced membrane (IM) technique for bone reconstruction, a poly(methyl methacrylate) (PMMA) spacer is implanted to induce formation of a foreign body membrane around the defect site. Membrane development is essential for later bone grafting success, yet the mechanism by which the IM promotes bone regeneration remains unknown, as are the ways that spacer composition plays a role in the membrane's healing potential. This study investigated the impact of leached methyl methacrylate (MMA)-the major monomeric component of PMMA-on IM development. In vitro cell culture found that MMA elution did not impact endothelial cell or mesenchymal stem cell proliferation. For in vivo analysis, we advanced a streamlined rat femoral model to efficiently study the influence of spacer properties on IM characteristics. Comparison of membrane formation around polycaprolactone (PCL), MMA-eluting PCL (high-dose PCL-MMA and low-dose PCL-MMA), and surgical PMMA revealed robust membranes enveloped all groups after 4 weeks in vivo, with elevated expression of osteogenic bone morphogenetic protein-2 and angiogenic vascular endothelial growth factor compared with the surrounding muscle and bone tissues. Growth factor quantitation in IM tissue found no statistically significant difference between groups. New bone growth, vascularization, and CD163+ macrophage populations surrounding the polymer implants were also quantified; and blood vessel formation around high-dose PCL-MMA was found to be significantly decreased compared with PCL alone. To the best of our knowledge, these findings represent the first time that results have been obtained about the characteristics of membranes formed around PCL in the IM setting.

Zn-0.4Li alloy shows great potential for the fixation and healing of bone fractures at load-bearing sites

Biodegradable orthopedic implants, designed to promote vigorous bone healing and be safely absorbed by the body, are developing rapidly to complement the deficiency of conventional implants. Limited strength, however, has blocked their path to high load-bearing applications. In this study, we report a pioneering application of implants made of high-strength biodegradable Zn-Li alloy in a high load-bearing rabbit shift fracture model. The Zn-0.4Li based bone plates and screws showed comparable performance in bone fracture fixation compared to the Ti-6Al-4 V counterpart. The fracture was healed completely after 6 months. The osteogenic activity of Zn-0.4Li alloy, as a result of biodegradation, was verified in a rat femur model. Furthermore, the underlying mechanism could include activation of the PI3K-AKT pathway and stimulation of metallothionein proteins. The empirical findings here reveal the great potential of Zn-Li based alloys for degradable biomaterials in high load-bearing applications.

Improved osseointegration using porcine xenograft compared to demineralized bone matrix for the treatment of critical defects in a small animal model.

Autograft (AG) is the gold standard bone graft due to biocompatibility, osteoconductivity, osteogenicity, and osteoinductivity. Alternatives include allografts and xenografts (XG). We investigated the osseointegration and biocompatibility of a decellularized porcine XG within a critical defect animal model. We hypothesized that the XG will result in superior osseointegration compared to demineralized bone matrix (DBM) and equivalent immune response to AG. Critical defects were created in rat femurs and treated with XG, XG plus bone morphogenetic protein (BMP)-2, DBM, or AG. Interleukin (IL)-2 and IFN-gamma levels (inflammatory markers) were measured from animal blood draws at 1week and 1month post-operatively. At 1month, samples underwent micro-positron-emission tomography (microPET) scans following 18-NaF injection. At 16weeks, femurs were retrieved and sent for micro-computerized tomography (microCT) scans for blinded grading of osseointegration or were processed for histologic analysis with tartrate resistant acid phosphatase (TRAP) and pentachrome. Enzyme linked immunosorbent assay testing demonstrated greater IL-2 levels in the XG vs. AG 1week post-op; which normalized by 28days post-op. MicroPET scans showed increased uptake within the AG compared to all groups. XG and XG+BMP-2 showed a trend toward increased uptake compared with DBM. MicroCT scans demonstrated increased osseointegration in XG and XG+BMP groups compared to DBM. Pentachrome staining demonstrated angiogenesis and endochondral bone formation. Furthermore, positive TRAP staining in samples from all groups indicatedbone remodeling. These data suggest that decellularized and oxidized porcine XG is biocompatible and at least equivalent to DBM in the treatment of a critical defect in a rat femur model.

The Double Stitch Everting Technique in the End-to-Side Microvascular Anastomosis: Validation of the Technique Using a Randomized N-of-1 Trial.

The double stitch everting (DSE) technique, in which time is won by leaving the needle inside the vessel wall in-between stitching, is a modification of the end-to-side (ETS) anastomosis in the interest of reducing anastomosis time. This ensures proper wall eversion, intima-to-intima contact, and improved suture symmetry. We designed an N-of-1 randomized trial with each microsurgeon as their own control. We included 10 microsurgeons of different levels of experience who were then asked to perform classic and DSE ETS anastomoses on the chicken leg and rat femoral models. Every anastomosis was cut and evaluated using blinded assessment. Two-way analysis of variance (ANOVA) and multivariable logistic regression were used to analyze the results and for confounder adjustment. A total of 210 anastomoses were performed, of which 177 on the chicken leg and 43 on the rat femoral artery and vein. From the 210 anastomoses, 111 were performed using the classic technique and 99 using the DSE technique. The mean anastomosis time was 28.8 ± 11.3 minutes in the classic group and 24.6 ± 12 minutes in the DSE group (p < 0.001, t-test). There was a significant reduction (p < 0.001, two-way ANOVA) in the number of mistakes when using the DSE technique (mean 5.5 ± 2.6) compared with those using the classic technique (mean 7.7 ± 3.4). The DSE technique for ETS anastomoses improves anastomoses times in experienced and moderately experienced microsurgeons while also improving or maintaining suture symmetry and lowering the number of mistakes.

In vivo degradation behaviour and bone response of a new Mg-rare earth alloy immobilized in a rat femoral model

A new type of Mg-rare earth (Ce, La) and the AZ31 alloy sheets were prepared by vertical twin-roll casting (TRC) technology under identical casting conditions, and their microstructural features, degradation behaviours and bone responses were investigated. The microstructural characterization showed that the Mg-RE (rare earth) exhibited a higher amorphous forming ability than the AZ31. Moreover, the results of electrochemical impedance spectroscopy (EIS) and potentiodynamic polarization indicated that the Mg-RE sheets displayed a higher corrosion resistance compared with the AZ31 sheets. Additionally, the Ti, Mg-RE and AZ31 sheet implants were immobilized and implanted in a rat femur model to observe degradation behavior during 16 weeks. in vivo tests showed that no significant change in the femur surrounding the Ti group, which excluded the external factor that the new bone formation resulting from bone remodeling. Furthermore, the Mg-RE group induced more newly formed bones, which met the necessary conditions for the prevention of pathological fractures. Therefore, the novel Mg-RE alloy appear to hold a healing candidate as the biodegradable implant material.

Puerarin inhibits titanium particle-induced osteolysis and RANKL-induced osteoclastogenesis via suppression of the NF-κB signaling pathway.

Osteolysis around the prosthesis and subsequent aseptic loosening are the main causes of prosthesis failure. Inflammation due to wear particles and osteoclast activation are the key factors in osteolysis and are also potential targets for the treatment of osteolysis. However, it is not clear whether puerarin can inhibit chronic inflammation and alleviate osteolysis. In this study, we investigated the effect of puerarin on Ti particle‐induced inflammatory osteolysis in vivo in rat femoral models and in vitro in receptor activator of nuclear factor kappa‐B ligand (RANKL)‐induced osteoclast activation models. Our in vivo results showed that puerarin significantly inhibited Ti particle‐induced osteolysis and the expression of matrix metallopeptidase 9 (MMP‐9), nuclear factor of activated T cells 1 (NFATc1), tumour necrosis factor (TNF)‐α and interleukin (IL)‐6. In vitro, puerarin prevented RANKL‐induced osteoclast differentiation, bone resorption and F‐actin ring formation in a concentration‐dependent manner. Furthermore, puerarin decreased the phosphorylation of p65 and prevented p65 moving from the cytoplasm to the nucleus. Puerarin also reduced the expression of osteoclast‐specific factors and inhibited the inflammatory response. In conclusion, our study proves that puerarin can block the NF‐κB signalling pathway to inhibit osteoclast activation and inflammatory processes, which provides a new direction for the treatment of osteolysis‐related diseases.

Influence of collagen membrane on bone quality in titanium mesh reconstructions-Study in rats.

New bone formation and tissue remodeling are the major challenges in implantology today. Titanium meshes have demonstrated reconstructive potential for vertical bone gain. However, the soft tissue healing is technically sensitive to the surgical procedure. The combined usage of collagen membrane and specification of the meshes may ensure greater predictability. Therefore, this study aims to evaluate the influence of collagen membrane on the quality of the new bone formation in guided bone regeneration (GBR) procedures with different titanium meshes. Twenty-eight Wistar rats were randomly allocated into four main experimental groups, according to mesh pore size inμm: Group P300 (titanium meshes, with 0.3-mm thickness and 3-mm pore size; n=7); Group P175 (titanium meshes, with 0.3-mm thickness and 1.75-mm pore size; n=7); Group P85: (titanium meshes, with 0.04-mm thickness and 0.85-mm pore size; n=7); Group P15: (titanium meshes. with 0.04-mm thickness and 0.15-mm pore size; n=7). The femurs of each animal were subdivided into test and control groups: Test: bovine bone graft associated with porcine collagen and collagen membrane was used; control: bovine bone graft associated with porcine collagen was used without association with collagen membrane. Bone quality evaluation by in vivo microtomography and histologic analysis were performed. Bone volume formation was similar between groups (P>0.05). However, the titanium meshes with pore size>1mm demonstrated higher mineral bone density in comparison with meshes with pore size<1mm (P<0.05), regardless of the combined usage of collagen membrane. All groups showed a spongy bone formation after 30days. Combined usage of collagen membrane in GBR procedures with titanium mesh did not show improvements in new bone quality in rat femur model. However, titanium mesh pore size specifications may influence bone quality.