Rejuvenation of tendon stem/progenitor cells for functional tendon regeneration through platelet-derived exosomes loaded with recombinant Yap1.

The regenerative capabilities including self-renewal, migration and differentiation potentials shift from the embryonic phase to the mature period of endogenous tendon stem/progenitor cells (TSPCs) characterize restricted functions and disabilities following tendon injuries. Recent studies have shown that tendon regeneration and repair rely on multiple specific transcription factors to maintain TSPCs characteristics and functions. Here, we demonstrate Yap, a Hippo pathway downstream effector, is associated with TSPCs phenotype and regenerative potentials through gene expression analysis of tendon development and repair process. Exosomes have been proven an efficient transport platform for drug delivery. In this study, purified exosomes derived from donor platelets are loaded with recombinant Yap1 protein (PLT-Exo-Yap1) via electroporation to promote the stemness and differentiation potentials of TSPCs in vitro. Programmed TSPCs with Yap1 import maintain stemness and functions after long-term passage in vitro. The increased oxidative stress levels of TSPCs are related to the phenotype changes in duplicative senescent processes. The results show that treatment with PLT-Exo-Yap1 significantly protects TSPCs against oxidative stressor-induced stemness loss and senescence-associated secretory phenotype (SASP) through the NF-κB signaling pathway. In addition, we fabricate an Exos-Yap1-functioned GelMA hydrogel with a parallel-aligned substrate structure to enhance TSPCs adhesion, promote cell stemness and force regenerative cells toward the tendon lineage for in vitro and in vivo tendon regeneration. The application of Exos-Yap1 functioned implant assists new tendon-like tissue formation with good mechanical properties and locomotor functions in a full-cut Achilles tendon defect model. Thus, PLT-Exo-Yap1-functionalized GelMA promotes the rejuvenation of TSPCs to facilitate functional tendon regeneration. STATEMENT OF SIGNIFICANCE: This is the first study to explore that the hippo pathway downstream effector Yap is involved in tendon aging and repair processes, and is associated with the regenerative capabilities of TSPCs. In this syudy, Platelet-derived exosomes (PLT-Exos) act as an appropriate carrier platform for the delivery of recombinant Yap1 into TSPCs to regulate Yap activity. Effective Yap1 delivery inhibit oxidative stress-induced senescence associated phenotype of TSPCs by blocking ROS-mediated NF-κb signaling pathway activation. This study emphasizes that combined application of biomimetic scaffolds and Yap1 loaded PLT-Exos can provide structural support and promote rejuvenation of resident cells to assist functional regeneration for Achilles tendon defect, and has the prospect of clinical setting.

Sandwich Biomimetic Scaffold Based Tendon Stem/Progenitor Cell Alignment in a 3D Microenvironment for Functional Tendon Regeneration.

Tendon injuries are some of the most commonly diagnosed musculoskeletal diseases. Tendon regeneration is sensitive to the topology of the substitute as it affects the cellular microenvironment and homeostasis. To bionic in vivo three-dimensional (3D) aligned microenvironment, an ordered 3D sandwich model was used to investigate the cell response in the tendon. First, high-resolution 3D printing provided parallel-grooved topographical cues on the hydrogel surface. Then the cells were seeded on its surface to acquire a 2D model. Afterward, an additional hydrogel coating layer was applied to the cells to create the 3D model. The interaction between cells and order structures in three-dimensions is yet to be explored. The study found that the tendon stem/progenitor cells (TSPCs) still maintain their ordering growth in the 3D model as in the 2D model. The study also found that the 3D-aligned TSPCs exhibited enhanced tenogenic differentiation through the PI3K-AKT signaling pathway and presented a less inflammatory phenotype than those in the 2D model. The in vivo implantation of such a 3D-aligned TSPC composite promoted tendon regeneration and mitigated heterotopic ossification in an Achilles defect model. These findings demonstrated that 3D-aligned TSPCs within a biomimetic topology environment are promising for functional tendon regeneration.

A Novel Suture-Preset Spring Plate System (SSPS) for Comminuted Coronoid Process Fracture in the Elbow.

OBJECTIVE: This study is aimed to investigate the clinical outcomes of a novel SSPS for fixation of the comminuted coronoid fracture. METHODS: A retrospective study was carried out in the patients with comminuted fractures of the coronoid treated by SPSS fixation between January 2014 and December 2018. A total of 17 patients (17 sides) was included in our study, including 11 male and six female, with a mean age range from 18 to 60. All cases started to functional rehabilitation immediately after the operation. Clinical outcomes were evaluated both radiographically and functionally at the follow-up visit, including the elbow instability, range of motion and Mayo elbow performance score (MEPS). RESULTS: According to the O'Driscoll classification system, there was two side of type 1.2, two of type 2.1, four of type 2.2, three of type 2.3, two of 3.1 and four of type 3.2. The surgery was carried out by Kocher and anteromedial approach in 12 patients, posterior and anteromedial approach in four, anterior approach in one. The average operation time and intraoperative blood loss was 129.41±43.87 min and 115.29±104.65 ml. The median follow-up time was 9 months (range, 6 to 15 months). The mean flexion, extension, pronation and supination motion was 138.76±8.67 degrees, 20.00±13.58, 82.94±5.32and 74.12±14.39 respectively at final follow up. The mean MEPS score was 89.76±8.46, including 11 excellent, 3 good and 3 fair result. The mean VAS score was 1.94±0.97. The mean union time of coronoid fractures was 2.77±0.31 months according to the established standard of healing. There were no significant differences in clinical outcomes among groups according to the O'Driscoll classification (P > .05) and ligament repair strategy (P > .05). No patient underwent instability or dislocation of the elbow during follow up. There were two cases with mild ulnar nerve symptoms which recovered totally at follow up. Meanwhile, there were three cases with heterotopic ossification of the elbow. CONCLUSION: Our findings demonstrated that the SSPS can provide a reliable fixation for the comminuted coronoid fracture with satisfactory clinical outcomes.

Non-Invasive Muscular Atrophy Causes Evaluation for Limb Fracture Based on Flexible Surface Electromyography System.

Muscular atrophy after limb fracture is a frequently occurring complication with multiple causes. Different treatments and targeted rehabilitation procedures should be carried out based on the causes. However, bedside evaluation methods are invasive in clinical practice nowadays, lacking reliable non-invasive methods. In this study, we propose a non-invasive flexible surface electromyography system with machine learning algorithms to distinguish nerve-injury and limb immobilization-related atrophy. First, a flexible surface electromyography sensor was designed and verified by in vitro tests for its robustness and flexibility. Then, in vivo tests on rats proved the reliability compared with the traditional invasive diagnosis method. Finally, this system was applied for the diagnosis of muscular atrophy in 10 patients. The flexible surface electromyography sensor can achieve a max strain of 12.0%, which ensures close contact with the skin. The in vivo tests on rats show great comparability with the traditional invasive diagnosis method. It can achieve a high specificity of 95.28% and sensitivity of 98.98%. Application on patients reaches a relatively high specificity of 89.44% and sensitivity of 91.94%. The proposed painless surface electromyography system can be an easy and accurate supplementary for bedside muscular atrophy causes evaluation, holding excellent contact with the body.

One-Stage Closed Intramedullary Nailing for Delayed Femoral Fracture in Multiple Injured Patients.

OBJECTIVE: To tackle the challengeable dilemma of delayed femoral fracture, a technique of fixator-assisted closed reduction and intramedullary nailing at one stage was introduced herein and its clinical results were investigated. METHODS: A retrospective study was conducted on delayed femoral shaft fracture between February 2008 and January 2017. The multiple injured patients aged from 18 to 60 years with delayed femoral fracture was included. All patients were treated by one-stage internal fixation technique and followed up for more than 1 year. Outcome measures including the operation time, intraoperative blood loss and limb alignment, healing time of fracture, visual analog scale (VAS), and range of knee motion were recorded and evaluated. RESULTS: A total of 13 patients (16 sides) with a mean injury severity score (ISS) of 32.77 ± 9.98 (range, 19 to 52) participated in the investigation. The median length of time-after-fracture was 38 days (range, 21 to 110 days). The average shortening distance of the fracture ends was 35.48 ± 19.24 mm (range, 10.00 to 79.00 mm). The average surgery time was 192.19 ± 29.38 min for unilateral femoral fracture, with blood loss of 587.50 ± 232.02 ml. The postoperative discrepancy of lower limb was 3.87 ± 2.52 mm. No patient had vascular and neurologic complications due to the lengthening. All fractures healed successfully with a mean time of 2.98 ± 0.57 months. The mean VAS and maximal knee flexion were 1.63 ± 1.09 and 131.25° ± 5.32° at final follow-up, respectively. CONCLUSIONS: Our findings indicated that fixator-assisted closed reduction and intramedullary nailing at one stage is an effective treatment for delayed femoral fracture with satisfactory functional recovery.

Prediction of Bedridden Duration of Hospitalized Patients by Machine Learning Based on EMRs at Admission.

Being bedridden is a frequent comorbid condition that leads to a series of complications in clinical practice. The present study aimed to predict bedridden duration of hospitalized patients based on EMR at admission by machine learning. The medical data of 4345 hospitalized patients who were bedridden for at least 24 hours after admission were retrospectively collected. After preprocessing of the data, features for modeling were selected by support vector machine recursive feature elimination. Thereafter, logistic regression, support vector machine, and extreme gradient boosting algorithms were adopted to predict the bedridden duration. The feasibility and efficacy of above models were evaluated by performance indicators. Our results demonstrated that the most important features related to bedridden duration were Charlson Comorbidity Index, age, bedridden duration before admission, mobility capability, and perceptual ability. The extreme gradient boosting algorithm showed the best performance (accuracy, 0.797; area under the curve, 0.841) when compared with support vector machine (accuracy, 0.771; area under the curve, 0.803) and logistic regression (accuracy, 0.765; area under the curve, 0.809) algorithms. Meanwhile, the extreme gradient boosting algorithm had a higher sensitivity (0.856), specificity (0.650), and F1 score (0.858) than that of support vector machine algorithm (0.843, 0.589, and 0.841) and logistic regression (0.852, 0.545, and 0.839), respectively. These findings indicate that machine learning based on EMRs at admission is a feasible avenue to predict the bedridden duration. The extreme gradient boosting algorithm shows great potential for further clinical application.

A 3D-printed PRP-GelMA hydrogel promotes osteochondral regeneration through M2 macrophage polarization in a rabbit model.

Osteochondral regeneration is an orchestrated process of inflammatory immunity, host cell response, and implant degradation in tissue engineering. Here, the effects of a platelet-rich plasma (PRP)-gelatin methacryloyl (GelMA) hydrogel scaffold fabricated using the digital micro-mirror device (DMD) technique for osteochondral repair were investigated in a rabbit model. GelMA hydrogels with different PRP concentrations were fabricated, and their roles in bone marrow mesenchymal stem cells (BMSCs) and macrophage polarization in vitro were investigated. The incorporation of 20% PRP into the hydrogel showed optimal effects on the proliferation, migration, and osteogenic and chondrogenic differentiation of BMSCs. The 20% PRP-GelMA (v/v) hydrogel also promoted M2 polarization with high expression of Arg1 and CD206. Compared to the 20% PRP group, the 50% PRP group showed similar biological roles in BMSCs but less extent of osteogenesis. In the vivo study, the 20% PRP-GelMA composite was used for osteochondral reconstruction and showed more cartilage and subchondral bone regeneration than that observed using the pure GelMA hydrogel. The PRP-GelMA group exhibited more M2 macrophage infiltration and less M1 macrophage presentation at three time points as compared to the nontreatment group. The expression of Arg1 in the PRP-GelMA group increased significantly at 6 weeks but decreased to a lower level at 12 weeks, while CD163 showed sustained high expression until 18 weeks. Our findings demonstrated that the 3D-printed PRP-GelMA composite could promote osteochondral repair through immune regulation by M2 polarization and could be a potential candidate for osteochondral tissue engineering. STATEMENT OF SIGNIFICANCE: PRP-GelMA hydrogels promoted the migration and osteogenic and chondrogenic differentiation of BMSCs. PRP-GelMA hydrogels participated in immune regulation and M1-to-M2 transition of macrophages. PRP-GelMA hydrogels coordinated and promoted several overlapping osteochondral repair events, including dynamic immune regulation, chemotaxis of MSCs, and osteochondral differentiation. PRP-GelMA hydrogels showed superior cartilage and subchondral bone repair properties.

In vitro and in vivo evaluation of antibacterial activity of polyhexamethylene guanidine (PHMG)-loaded TiO2 nanotubes.

Artificial joint replacement is an effective surgical method for treating end-stage degenerative joint diseases, but peripheral bacterial infection of prosthesis can compromise the effect of the surgery. Herein, antibacterial effects of titanium dioxide nanotubes (TNTs) coated with polyhexamethylene guanidine (PHMG) were examined via in vitro and in vivo experiments. TNTs with a pore diameter 46.4 ± 5.9 nm and length of 300-500 nm for the slice and 650-800 nm for the rod were fabricated by anodization. Then, 3.46 ± 0.40 mg and 1.27 ± 0.28 mg of PHMG were coated onto the TNT slice and rod, respectively. In vitro studies of the release of PHMG showed that the antibacterial agent was released in two stages: initial burst release and relatively slow release. In vitro and in vivo antibacterial studies showed that the PHMG-loaded TNTs (PHMG-TNTs) had excellent antibacterial abilities to prevent bacterial infections. Clinical pathological analysis of rabbit femurs indicated that the implanted PHMG-TNTs had no apparent pathological changes. Real-time quantitative reverse transcription polymerase chain reaction analysis of the femur tissues around the implants showed that the expression of osteogenic-related genes, including runt-related transcription factor 2, osteocalcin, alkaline phosphatase, bone sialoprotein, bone morphogenetic protein 2 and vascular endothelial growth factor A, was significantly upregulated in the PHMG-TNT implanted group as compared to the other groups. Overall, these findings provide a promising approach for the fabrication of antibacterial and bone biocompatible titanium-based implants in orthopedics.

Comparison of Leukocyte-Rich Platelet-Rich Plasma and Leukocyte-Poor Platelet-Rich Plasma on Achilles Tendinopathy at an Early Stage in a Rabbit Model.

BACKGROUND: Tendinopathy is still a great challenge in clinical practice, and the role of platelet-rich plasma (PRP) is controversial. The influence of leukocytes on tendinopathy at an early stage has not been defined so far. PURPOSE: To compare the effects of leukocyte-rich PRP (Lr-PRP) and leukocyte-poor PRP (Lp-PRP) on Achilles tendinopathy when applied at an early stage. STUDY DESIGN: Controlled laboratory study. METHODS: A rabbit Achilles tendinopathy model was induced by a collagenase injection. A week later, treatments were applied randomly on local Achilles tendon lesions: (1) 200 µL of Lr-PRP (16 legs), (2) 200 µL of Lp-PRP (16 legs), and (3) 200 µL of saline (16 legs). At 3 and 6 weeks after the collagenase injection, outcomes were evaluated by histology, magnetic resonance imaging (MRI), real-time polymerase chain reaction analysis, immunohistochemistry, and transmission electron microscopy (TEM). RESULTS: The Lr-PRP group had a lower T2 signal intensity (P = .0377) and smaller diameter (P = .0193) and cross-sectional area (P = .0194) than the Lp-PRP group on MRI. Histologically, the Lr-PRP group had better scores than the Lp-PRP group (P = .0284 and P = .0188, respectively). Compared with the Lp-PRP group, higher gene expression and more protein synthesis of collagen I (P = .0160 and P = .0309, respectively) and CD163 (P < .0001 and P = .0411, respectively) were found in the Lr-PRP group. Considering TEM and biomechanical testing, the Lr-PRP group demonstrated more mature collagen fibers (P < .0001), a larger fiber diameter (P = .0005), a higher failure load (P = .00417), and higher tensile stress (P < .0001) than the Lp-PRP group. CONCLUSION: Lr-PRP had more beneficial effects than Lp-PRP when delivered at an early stage during tendon repair. CLINICAL RELEVANCE: Here, we showed that tendinopathy influenced the curative effects of PRP in vivo. An early-stage application of Lr-PRP had more benefits for the repair of tendinopathy than Lp-PRP in a rabbit model, which will supplement guidelines of PRP treatment on tendinopathy clinically.

Detection of extremity chronic traumatic osteomyelitis by machine learning based on computed-tomography images: A retrospective study.

Despite the availability of a series of tests, detection of chronic traumatic osteomyelitis is still exhausting in clinical practice. We hypothesized that machine learning based on computed-tomography (CT) images would provide better diagnostic performance for extremity traumatic chronic osteomyelitis than the serological biomarker alone. A retrospective study was carried out to collect medical data from patients with extremity traumatic osteomyelitis according to the criteria of musculoskeletal infection society. In each patient, serum levels of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), and D-dimer were measured and CT scan of the extremity was conducted 7 days after admission preoperatively. A deep residual network (ResNet) machine learning model was established for recognition of bone lesion on the CT image. A total of 28,718 CT images from 163 adult patients were included. Then, we randomly extracted 80% of all CT images from each patient for training, 10% for validation, and 10% for testing. Our results showed that machine learning (83.4%) outperformed CRP (53.2%), ESR (68.8%), and D-dimer (68.1%) separately in accuracy. Meanwhile, machine learning (88.0%) demonstrated highest sensitivity when compared with CRP (50.6%), ESR (73.0%), and D-dimer (51.7%). Considering the specificity, machine learning (77.0%) is better than CRP (59.4%) and ESR (62.2%), but not D-dimer (83.8%). Our findings indicated that machine learning based on CT images is an effective and promising avenue for detection of chronic traumatic osteomyelitis in the extremity.

Intratendon delivery of leukocyte-rich platelet-rich plasma at early stage promotes tendon repair in a rabbit Achilles tendinopathy model.

Tendinopathy is a great obstacle in clinical practice due to its poor regenerative capacity. The influence of different stages of tendinopathy on effects of leukocyte-rich platelet-rich plasma (Lr-PRP) has not been elucidated. The aim of this study is to investigate the optimal time point for delivery of Lr-PRP on tendinopathy. A tendinopathy model was established by local collagenase injection on the rabbit Achilles tendon. Then after collagenase induction, following treatments were applied randomly on the lesion: (a) 200 µl of Lr-PRP at 1 week (PRP-1 group), (b) 200 µl of saline at 1 week (Saline-1 group), (c) 200 µl of Lr-PRP at 4 weeks (PRP-2 group), and (d) 200 µl of saline at 4 weeks (Saline-2 group). Six weeks after collagenase induction, outcomes were assessed by magnetic resonance imaging, cytokine quantification, gene expression, histology, and transmission electron microscopy. Our results demonstrated that PRP-1 group had the least cross-sectional area and lesion percent of the involved tendon, as well as the lowest signal intensity in magnetic resonance imaging among all groups. However, the PRP-2 group showed larger cross-sectional area than saline groups. Enzyme-linked immunosorbent assay indicated that PRP-1 group had a higher level of interleukin-10 but lower level of interleukin-6 when compared with PRP-2 and saline groups. Meanwhile, the highest expression of collagen (Col) 1 in PRP-1 and Col 3, matrix metalloproteinase (MMP)-1, and MMP-3 in PRP-2 was found. Histologically, the PRP-1 showed better general scores than PRP-2, and no significant difference was found between the PRP-2 and saline groups. For transmission electron microscopy, PRP-1 had the largest mean collagen fibril diameter, and the PRP-2 group showed even smaller mean collagen fibril diameter than saline groups. In conclusion, intratendon delivery of Lr-PRP at early stage showed beneficial effect for repair of tendinopathy but not at late stage. For translation of our results to clinical circumstances, further studies are still needed.

Platelet-rich gel-incorporated silk scaffold promotes meniscus regeneration in a rabbit total meniscectomy model.

Aim: To investigate whether platelet-rich gel (PRG) incorporation could promote meniscal regeneration of the silk scaffold. Materials & methods: A PRG-incorporated silk sponge was fabricated for reconstruction of the meniscus in a rabbit meniscectomy model. Subsequently, characterization of the scaffold, as well as the in vitro cytocompatibility and in vivo function was evaluated. Results: Our results showed that the PRG-incorporated silk scaffold provided a sustained release of TGF-ß1 over 1 week. The PRG enhanced the cytocompatibility in vitro and cell infiltration in vivo of the silk sponge. Meanwhile, the implantation of the composite in situ ameliorated the cartilage degeneration in knee at 3 months. Conclusion: These findings indicated that PRG-incorporated silk scaffold could promote functional regeneration of the meniscus and effectively prevented subsequent osteoarthritis after meniscectomy.

A collagen-coated sponge silk scaffold for functional meniscus regeneration.

Tissue engineering is a promising solution for meniscal regeneration after meniscectomy. However, in situ reconstruction still poses a formidable challenge due to multifunctional roles of the meniscus in the knee. In this study, we fabricate a silk sponge from 9% (w/v) silk fibroin solution through freeze drying and then coat its internal space and external surface with collagen sponge. Subsequently, various characteristics of the silk-collagen scaffold are evaluated, and cytocompatibility of the construct is assessed in vitro and subcutaneously. The efficacy of this composite scaffold for meniscal regeneration is evaluated through meniscus reconstruction in a rabbit meniscectomy model. It is found that the internally coated collagen sponge enhances the cytocompatibility of the silk sponge, and the external layer of collagen sponge significantly improves the initial frictional property. Additionally, the silk-collagen composite group shows more tissue ingrowth and less cartilage wear than the pure silk sponge group at 3 months postimplantation in situ. These findings thus demonstrate that the composite scaffold had less damage to the joint surface than the silk alone through promoting functional meniscal regeneration after meniscectomy, which indicates its clinical potential in meniscus reconstruction.

Exogenous stromal derived factor-1 releasing silk scaffold combined with intra-articular injection of progenitor cells promotes bone-ligament-bone regeneration.

Anterior cruciate ligament (ACL) is one of the most difficult tissues to heal once injured. Ligament regeneration and tendon-bone junction healing are two major goals of ACL reconstruction. This study aimed to investigate the synergistic therapeutic effects of Stromal cell-derived factor 1 (SDF-1)-releasing collagen-silk (CSF) scaffold combined with intra-articular injection of ligament-derived stem/progenitor cells (LSPCs) for ACL regeneration and the amelioration in the long-term complication of osteoarthritis (OA). The stem cell recruitment ability of CSF scaffold and the multipotency, particularly the tendon forming ability of LSPCs from rabbits were characterized in vitro, while the synergistic effect of the CSF scaffold and LSPCs for ACL regeneration and OA amelioration were investigated in vivo at 1, 3, and 6 months with a rabbit ACL reconstruction model. The CSF scaffold was used as a substitute for the ACL, and LSPCs were injected into the joint cavity after 7 days of the ACL reconstruction. CSF scaffold displayed a controlled release pattern for the encapsulated protein for up to 7 days with an increased stiffness in the mechanical property. LSPCs, which exhibited highly I Collagen and CXCR4 expression, were attracted by SDF-1 and successfully relocated into the CSF scaffold at 1 month in vivo. At 3 and 6 months post-treatment, the CSF scaffold combined with LSPCs (CSFL group) enhanced the regeneration of ACL tissue, and promoted bone tunnel healing. Furthermore, the OA progression was impeded efficiently. Our findings here provided a new strategy that using stem cell recruiting CSF scaffold with tissue-specific stem cells, could be a promising solution for ACL regeneration. STATEMENT OF SIGNIFICANCE: In this study, we developed a silk scaffold with increased stiffness and SDF-1 controlled release capacity for ligament repair. This advanced scaffold transplantation combined with intra-articular injection of LSPCs (which was isolated from rabbit ligament for the first time in this study) promoted the regeneration of both the tendinous and bone tunnel portion of ACL. This therapeutic strategy also ameliorated cartilage degeneration and reduced the severity of arthrofibrosis. Hence, combining LSPCs injection with SDF-1-releasing silk scaffold is demonstrated as a therapeutic strategy for ACL regeneration and OA treatment in the clinic.

The effect of pathological fractures on the prognosis of patients with osteosarcoma: a meta-analysis of 14 studies.

Osteosarcoma is a leading cause of malignant tumor related death. We conducted a meta-analysis to evaluate the association between pathological fractures and prognosis in patients with osteosarcoma. We searched PubMed, Web of Science, and Embase for studies published until May 15, 2017. Crude and adjusted relative risk (RR) with 95% confidence intervals were used to compare data between the case and control groups. Fourteen studies and 3910 patients were included in the final meta-analysis. No statistically significant difference was detected between the pathological fracture and non-pathological fracture groups in local recurrences analysis (RR = 1.102, 95% CI: 0.813-1.495, P = 0.531); however, a statistically significant difference was found between group in distant metastasis (RR = 1.424, 95% CI: 1.089-1.862, P = 0.01). For survival rates, the following RRs were calculated: 3-year overall survival (OS) (RR = 0.736, 95% CI: 0.593-0.912, P = 0.005); 5-year OS (RR = 0.889, 95% CI: 0.791-0.999, P = 0.049); 3-year event-free survival (EFS) (RR = 0.812, 95% CI: 0.682-0.966, P = 0.018); and 5-year EFS (RR = 0.876, 95% CI: 0.785-0.978, P = 0.019). The pooled estimate of RR was 0.673 (95% CI: 0.364-1.244, P = 0.206) for local recurrence in the amputation and limb salvage groups. In conclusion, our analysis indicated that there were no differences in local recurrence and local recurrence after limb salvage between patients with or without a fracture. Additionally, the patients with pathological fracture had a higher risk of distant metastasis and lower 3-year OS, 5-year OS, 3-year EFS, and 5-year EFS. Considering the limitations of this study, we believe that future large-scale studies should be performed to confirm our conclusions.

Intratendon Delivery of Leukocyte-Poor Platelet-Rich Plasma Improves Healing Compared With Leukocyte-Rich Platelet-Rich Plasma in a Rabbit Achilles Tendinopathy Model.

BACKGROUND: Chronic tendinopathy is a commonly occurring clinical problem that affects both athletes and inactive middle-aged patients. Although some studies have shown that different platelet-rich plasma (PRP) preparations could exert various therapeutic effects in vitro, the role of leukocytes in PRP has not yet been defined under tendinopathy conditions in vivo. PURPOSE: This study compared the effects of the intratendon delivery of leukocyte-poor PRP (Lp-PRP) versus leukocyte-rich PRP (Lr-PRP) in a rabbit chronic tendinopathy model in vivo. STUDY DESIGN: Controlled laboratory study. METHODS: Four weeks after a local injection of collagenase in the Achilles tendon, the following treatments were randomly administered on the lesions: injections of (1) 200 µL of Lp-PRP (n = 8), (2) 200 µL of Lr-PRP (n = 8), or (3) 200 µL of saline (n = 8). Healing outcomes were assessed at 4 weeks after therapy with magnetic resonance imaging (MRI), cytokine quantification, real-time polymerase chain reaction analysis of gene expression, histology, and transmission electron microscopy (TEM). RESULTS: MRI revealed that the Lr-PRP and saline groups displayed higher signal intensities compared with the Lp-PRP group with T2 mapping. Histologically, the Lp-PRP group displayed significantly better general scores compared with the Lr-PRP ( P = .001) and saline ( P < .001) groups. Additionally, TEM showed that the Lp-PRP group had larger collagen fibril diameters than the Lr-PRP group ( P < .001). Enzyme-linked immunosorbent assay showed a significantly lower level of catabolic cytokine IL-6 in the Lp-PRP group compared with the Lr-PRP ( P = .001) and saline ( P = .021) groups. The Lp-PRP group displayed significantly increased expression of collagen I compared with the saline group ( P = .004) but not the Lr-PRP group. Both the Lp-PRP and Lr-PRP groups exhibited significantly lower matrix metalloproteinase (MMP)-1 and MMP-3 expression levels compared with the saline group. However, only the Lp-PRP group displayed significantly higher expression of TIMP-1 than the saline group ( P = .024). CONCLUSION: Compared with Lr-PRP, Lp-PRP improves tendon healing and is a preferable option for the clinical treatment of tendinopathy. CLINICAL RELEVANCE: PRP is widely used in the clinical management of chronic tendinopathy. However, the clinical results are ambiguous. It is imperative to understand the influence of leukocytes on PRP-mediated tissue healing in vivo, which could facilitate the better clinical management of chronic tendinopathy. Further studies are needed to translate our findings to the clinical setting.

Single nucleotide polymorphisms of ABCB1 gene and response to etanercept treatment in patients with ankylosing spondylitis in a Chinese Han population.

BACKGROUND: Etanercept was highly recommended for patients with ankylosing spondylitis (AS), as its efficacy has been confirmed in AS, while genetic polymorphisms, by affecting drug metabolism or drug receptor, lead to interindividual variability in drug disposition and efficacy. Therefore, this study aims to investigate whether ABCB1 gene polymorphisms can predict therapeutic response to etanercept in patients with AS. METHODS: A total of 185 patients with AS in our hospital were recruited into our study from December 2012 to May 2015. The frequency distributions of genotype and allele of rs2032582, rs1128503, and rs1045642 were detected by polymerase chain reaction (PCR) and electrophoresis verification enzyme products method. AS patients received etanercept treatment for 12 weeks, followed by this would be evaluated by the bath AS disease activity index (BASDAI) score improvement and the assessment of spondyloArthritis international society 20/50/70 (ASAS20/50/70) score improvements to explore the relationship between genotype of ABCB1 gene polymorphisms and therapeutic response to etanercept in patients with AS. RESULTS: After 12 weeks, the BASDAI score mean improvement value of rs2032582 A/A genotype was 2.87 ±â€Š0.52. The ratios of patients with rs2032582 A/A genotype reaching the BASDAI50 and ASAS20 evaluation criteria were 64.29% and 92.86%, respectively. The results indicated that efficacy of etanercept was promoted in rs2032582 A/A genotype. The BASDAI score mean improvement value of rs1128503 C/C genotype was 2.79 ±â€Š0.54 after 12 weeks. The ratios of patients with rs1128503 C/C genotype reaching the BASDAI50 and ASAS20 evaluation criteria were 66.67% and 93.94%, respectively. The results indicated that efficacy of etanercept was promoted in rs1128503 C/C genotype. However, no significant associations were observed between rs1045642 and therapeutic response to etanercept in AS patients. CONCLUSION: ABCB1 gene rs2032582 and rs1128503 polymorphisms may be associated with the efficacy of etanercept in AS patients. ABCB1 gene polymorphisms can act as biological indicators of etanercept efficacy.

Ectopic tissue engineered ligament with silk collagen scaffold for ACL regeneration: A preliminary study.

Anterior cruciate ligament (ACL) reconstruction remains a formidable clinical challenge because of the lack of vascularization and adequate cell numbers in the joint cavity. In this study, we developed a novel strategy to mimic the early stage of repair in vivo, which recapitulated extra-articular inflammatory response to facilitate the early ingrowth of blood vessels and cells. A vascularized ectopic tissue engineered ligament (ETEL) with silk collagen scaffold was developed and then transferred to reconstruct the ACL in rabbits without interruption of perfusion. At 2weeks after ACL reconstruction, more well-perfused cells and vessels were found in the regenerated ACL with ETEL, which decreased dramatically at the 4 and 12week time points with collagen deposition and maturation. ACL treated with ETEL exhibited more mature ligament structure and enhanced ligament-bone healing post-reconstructive surgery at 4 and 12weeks, as compared with the control group. In addition, the ETEL group was demonstrated to have higher modulus and stiffness than the control group significantly at 12weeks post-reconstructive surgery. In conclusion, our results demonstrated that the ETEL can provide sufficient vascularity and cellularity during the early stages of healing, and subsequently promote ACL regeneration and ligament-bone healing, suggesting its clinic use as a promising therapeutic modality. STATEMENT OF SIGNIFICANCE: Early inflammatory cell infiltration, tissue and vessels ingrowth were significantly higher in the extra-articular implanted scaffolds than theses in the joint cavity. By mimicking the early stages of wound repair, which provided extra-articular inflammatory stimulation to facilitate the early ingrowth of blood vessels and cells, a vascularized ectopic tissue engineered ligament (ETEL) with silk collagen scaffold was constructed by subcutaneous implantation for 2weeks. The fully vascularized TE ligament was then transferred to rebuild ACL without blood perfusion interruption, and was demonstrated to exhibit improved ACL regeneration, bone tunnel healing and mechanical properties.

[Analysis on the long-term effects of modified double endobutton technique in the treatment of Tossy type III acromioclavicular joint dislocations].

OBJECTIVE: To investigate the long-term clinical effects of modified double Endobutton technique for the treatment of acromioclavicular joint dislocations of Tossy type III. METHODS: A retrospective study was done in 42 patients with acromioclavicular joint dislocations of Tossy type III treated with modified double Endobutton technique from December 2008 to December 2010. There were 24 males and 18 females, ranging in age from 21 to 56 years old (averaged, 32.5 years old). All the patients were treated with open reduction, coracoclavicular ligament reconstruction using double Endobutton technique, and repair of acromioclavicular ligament. The Karlsson system was used to evaluate therapeutic effects. The distance from coracoid to clavicle was measured to evaluate reduction loss. RESULTS: All the patients were followed up, and the duration ranged from 2.0 to 3.2 years (averaged,2.4 years). According to Karlsson system, 32 patients got an A degree and 10 patients got a B degree at three months post-operatively; 26 patients got an A degree and 16 patients got a B degree at the latest follow-up; 6 patients got an A degree at 3 months after operation lowered to B degree at the latest follow-up. The coracoid-clavicle distance increased from (26.91 +/- 0.91) mm at 3 months after operation to (27.41 +/- 1.10) mm at the latest follow-up. Te patients treated with over-reduction during operation or with heavy physical labour work after operation had obvious widened coracoid-clavicle distance. Bone absorption was found around the plate in most cases, mainly in the clavicular side. CONCLUSION: Treatment for acromioclavicular joint dislocations of Tossy type III with modified double Endobutton technique has satisfactory early clinical results. But with time passing, loss of reduction and bone absorption around the plate could be observed, and clinical outcomes of some cases downgrade during the long-term follow-up.

[One-stage compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone for the treatment of chronic calcaneal osteomyelitis].

OBJECTIVE: To explore the treatment of chronic calcaneal osteomyelitis with bone defect after debridement and evaluate its clinical outcomes. METHODS: From June 2009 to June 2011, 52 patients with chronic calcaneal osteomyelitis were treated with stage-one compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone,including 12 females and 40 males with an average age of 43 years old ranging from 18 to 67. According to Cierny-Mader classification, there were 34 cases with stage III and 18 with stage IV. There were 32 cases on right side and 20 on left,with a course of 6 months to 3 years. The area of soft tissue wound ranged from 3.0 cm x l.5 cm to 23.0 cm x l2.0 cm. The clinical effects were evaluated according to infection controlling, calcium sulfate absorption,bone defect repair and heel functional recovery. RESULTS: All patients were followed up for 2 to 3.8 years (averaged 2.8 years). Primary healing was achieved in 52 patients. Two cases of recurrence were found post-operatively, 1 case in 3 months and another in 5 months,which were cured after a second operation. Bone repair healing was gained in 1.5 to 3.5 months (averaged 2.5 months). Complete radiological absorption of calcium sulfate was found in 1.2 to 3 months(averaged 2.2 months). Local exudation after removal of drainage tube had been persisting in 10 patients for 2 to 3 months, which was consistent with the time when cacium sulfate were totally absorbed. Flap had partial necrosis in 4 cases,and the wounds were closed after appropriate treatment finally. The mean Maryland score was 88.15±7.70. There were excellent results in 32 cases, good in 14, fair in 6. CONCLUSION: A satisfactory short-term clinical results can be gained by one-stage compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone in chronic calcaneal osteomyelitis, but the long-term results need further follow-up. And much more study is also demanded to reduce the exudation of calcium sulfate.