Mid-term Clinical Outcomes of "Light Bulb" Core Decompression with Arthroscopic Assistance in Peri-collapse Osteonecrosis of the Femoral Head: A Retrospective Comparative Study.

OBJECTIVE: Nontraumatic osteonecrosis of the femoral head (ONFH) is commonly encountered in orthopedics. Without early clinical intervention, most patients with peri-collapse of the ONFH will develop femoral head necrosis and eventually require hip replacement surgery. The aim of this study is to evaluate clinical outcomes in patients with ONFH who underwent "light bulb" core decompression (CD) with arthroscopic assistance and to compare them with the outcomes of those treated with traditional procedures. METHODS: A retrospective review of patients with Stage II and IIIA (Peri-collapse) radiographic findings based on the Association Research Circulation Osseous (ARCO) stage for ONFH who underwent "light bulb" CD with or without arthroscopic assistance by a single-surgeon team between March 2014 and December 2018 was performed. All patients were followed up for a minimum of 2 years. The visual analogue scale (VAS) pain score, Harris hip score (HHS), and radiological imaging were evaluated. The categorical parameters were analyzed by chi-square test and the continuous variables conforming to a normal distribution were analyzed by Student's t-test. RESULTS: The study included a total of 39 patients (18 and 21 patients in the with and without arthroscopic assistance groups, respectively), with a mean age of 40.3 years and a mean follow-up of 22.2 months. Overall, there was a better VAS score in the arthroscopic assistance group than in the control group (p < 0.05), There was a significant difference in HHS (80.1 ± 9.2 vs 75.1 ± 12.7) at the last follow-up (p < 0.05). The rate of good and excellent outcomes was 94%. Similarly, there was no significant difference in the total rate of complications or conversion to THA. CONCLUSION: With arthroscopic assistance, "light bulb" CD could be achieved via hip arthroscopy with less trauma, and it offered the opportunity for more precise evaluation and monitoring for therapy and yielded better VAS scores after surgery and better hip function outcomes at the last follow-up.

A non-negative feedback self-distillation method for salient object detection.

Self-distillation methods utilize Kullback-Leibler divergence (KL) loss to transfer the knowledge from the network itself, which can improve the model performance without increasing computational resources and complexity. However, when applied to salient object detection (SOD), it is difficult to effectively transfer knowledge using KL. In order to improve SOD model performance without increasing computational resources, a non-negative feedback self-distillation method is proposed. Firstly, a virtual teacher self-distillation method is proposed to enhance the model generalization, which achieves good results in pixel-wise classification task but has less improvement in SOD. Secondly, to understand the behavior of the self-distillation loss, the gradient directions of KL and Cross Entropy (CE) loss are analyzed. It is found that KL can create inconsistent gradients with the opposite direction to CE in SOD. Finally, a non-negative feedback loss is proposed for SOD, which uses different ways to calculate the distillation loss of the foreground and background respectively, to ensure that the teacher network transfers only positive knowledge to the student. The experiments on five datasets show that the proposed self-distillation methods can effectively improve the performance of SOD models, and the average Fß is increased by about 2.7% compared with the baseline network.

Soluble thrombomodulin alleviates Diquat-induced acute kidney injury by inhibiting the HMGB1/IκBα/NF-κB signalling pathway.

Our research aimed to investigate whether soluble thrombomodulin (sTM) relieved Diquat (DQ)-induced acute kidney injury (AKI) via HMGB1/IκBα/NF-κB signaling pathways. An AKI rat model was constructed using DQ. Pathological changes in renal tissue were detected by HE and Masson staining. Gene expression was determined using qRT-PCR, IHC, and western blotting. Cell activity and apoptosis were analysed using CCK-8 and Flow cytometry, respectively. An abnormal kidney structure was observed in DQ rats. The levels of blood urea nitrogen (BUN), creatinine (CRE), uric acid (UA), oxidative stress, and inflammatory responses in the DQ group increased on the 7th day but decreased on the 14th day, compared with the control group. Additionally, HMGB1, sTM, and NF-kappaB (NF-κB) expression had increased in the DQ group compared with the control group, while the IκKα and IκB-α levels had decreased. In addition, sTM relieved the damaging effects of diquat on renal tubular epithelial cell viability, apoptosis, and the inflammatory response. The levels of HMGB1, TM, and NF-κB mRNA and protein were significantly decreased in the DQ + sTM group compared with the DQ group. These findings indicated that sTM could relieve Diquat-induced AKI through HMGB1/IκBα/NF-κB signaling pathways, which provides a treatment strategy for Diquat-induced AKI.

An immune genes signature for predicting mortality in sepsis patients.

A growing body of evidence indicates that the immune system plays a central role in sepsis. By analyzing immune genes, we sought to establish a robust gene signature and develop a nomogram that could predict mortality in patients with sepsis. Herein, data were extracted from the Gene Expression Omnibus and Biological Information Database of Sepsis (BIDOS) databases. We enrolled 479 participants with complete survival data using the GSE65682 dataset, and grouped them randomly into training (n = 240) and internal validation (n = 239) sets based on a 1:1 proportion. GSE95233 was set as the external validation dataset (n=51). We validated the expression and prognostic value of the immune genes using the BIDOS database. We established a prognostic immune genes signature (including ADRB2, CTSG, CX3CR1, CXCR6, IL4R, LTB, and TMSB10) via LASSO and Cox regression analyses in the training set. Based on the training and validation sets, the Receiver Operating Characteristic curves and Kaplan-Meier analysis revealed that the immune risk signature has good predictive power in predicting sepsis mortality risk. The external validation cases also showed that mortality rates in the high-risk group were higher than those in the low-risk group. Subsequently, a nomogram integrating the combined immune risk score and other clinical features was developed. Finally, a web-based calculator was built to facilitate a convenient clinical application of the nomogram. In summary, the signature based on the immune gene holds potential as a novel prognostic predictor for sepsis.

[Short-term effectiveness of high tibial osteotomy combined with arthroscopic surgery for knee varus arthritis and the results of secondary arthroscopic exploration].

Objective: To investigate the effectiveness of high tibial osteotomy (HTO) combined with arthroscopic surgery to treat medial compartment knee osteoarthritis (KOA) and secondary arthroscopic exploration to evaluate the outcome of cartilage and meniscus. Methods: A clinical data of 57 patients with medial compartment KOA with varus deformity of lower extremities admitted between August 2014 and October 2018 were retrospectively analyzed. There were 23 males and 34 females with an average age of 51.2 years (range, 41-63 years). The disease duration ranged from 2 to 8 years, with an average of 4.7 years. The preoperative femorotibial angle was (179.86±4.69)°, the relative position of the lower limb mechanical axis passing through the tibial plateau was 24.21%±6.98%, and the posterior slope of the tibial plateau was (5.23±1.45)°. The Kellgren-Lawrence grade of knee joint was grade Ⅱ in 22 cases and grade Ⅲ in 35 cases. The preoperative Hospital for Special Surgery (HSS) score, Lysholm score, and visual analogue scale (VAS) score were 59.1±7.3, 48.8±7.6, and 6.2±1.1, respectively. Arthroscopic exploration was performed during the operation to record the articular cartilage degeneration in the weight-bearing area of the medial compartment (Outerbridge grade Ⅰ in 18 cases, grade Ⅱ in 30 cases, and grade Ⅲ in 9 cases) and the condition of the medial meniscus injury, and the corresponding treatment was performed. The coronal force line was adjusted according to the preoperative Kellgren-Lawrence grade of the knee joint during the operation. After operation, the relative position of the lower limb mechanical axis passing through the tibial plateau, the femorotibial angle, and the posterior slope of the tibial plateau were measured; the Kellgren-Lawrence grade of the knee joint was recorded; the Outerbridge grade of articular cartilage degeneration and the meniscus outcome were evaluated by combining with the MRI of the knee joint at 1 year after operation and the second arthroscopic exploration when the internal fixator was removed. The function and pain of the knee were evaluated by Lysholm score, HSS score, and VAS score. Results: All the 57 patients were followed up 36-58 months with an average of 42.1 months. Incisions healed by first intention, and no neurovascular injury, intraarticular or hinge fractures occurred during operation, and no postoperative complications such as deep vein thrombosis of lower limbs and internal fixation failure occurred. All the osteotomy sites healed at 3 months after operation. At 1 year after operation, the internal fixator was removed, and the second arthroscopic exploration showed that there were 15 cases of Outerbridge grade Ⅰ, 31 cases of grade Ⅱ, and 11 cases of grade Ⅲ in the weight-bearing area of the medial compartment, and there was no significant difference when compared with preoperative grade ( Z=31.992, P=0.997); there was no cartilage degeneration in other compartments. Meniscus healing was seen in the injured meniscus, and no injury was seen in the normal meniscus. At last follow-up, there were 19 cases of Kellgren-Lawrence grade Ⅱ and 38 cases of grade Ⅲ, and there was no significant difference when compared with preoperative grade ( Z=49.049, P=0.764). The relative position of the lower limb mechanical axis passing through the tibial plateau was 59.16%±2.87%, and the femorotibial angle was (171.54±3.39)°, which significantly improved when compared with those before operation ( P<0.001). The posterior slope of the tibial plateau was (5.65±1.22)°, which was not significantly different from that before operation ( t=-1.673, P=0.096). The HSS score, Lysholm score, and VAS score were 82.3±7.7, 83.4±6.4, and 1.6±1.1 respectively, which were significantly different from those before operation ( P<0.001). Conclusion: HTO combined with arthroscopic surgery for medial compartment KOA with varus deformity of lower extremities can effectively improve the force line of lower extremities, relieve pain symptoms, and improve joint function, with satisfactory short-term effectiveness, and without significant progress in articular cartilage or meniscus injury after operation.

Dual-path transformer-based network with equalization-generation components prediction for flexible vibrational sensor speech enhancement in the time domain.

The flexible vibrational sensor (FVS) has the potential to become a popular wearable communication device because of its natural noise shielding characteristics and soft materials. However, FVS speech faces a severe loss of frequency components. To improve speech quality, a time-domain neural network model based on the dual-path transformer combined with equalization-generation components prediction (DPT-EGNet) is proposed. More specifically, the DPT-EGNet consists of five modules, namely the pre-processing module, dual-path transformer module, equalization module, generation module, and post-processing module. The dual-path transformer module is leveraged to extract the local and global contextual relationship of long-term speech sequences, which is extremely beneficial for inferring the missing components. The equalization and generation modules are designed according to the characteristics of FVS speech, which further improve the speech quality by simulating the inversion process of the speech distortion. The experimental results demonstrate that the proposed model effectively improves the quality of FVS speech; the average perceptual evaluation of speech quality (PESQ), short-time objective intelligibility (STOI), and composite measure for overall speech quality (COVL) scores of three males and three females are relatively increased by 64.19%, 29.63%, and 101.37%, which is superior to other baseline models developed in different domains. The proposed model also has significantly lower complexity than the others.

[Research progress of tissue engineering technology in promoting revascularization of necrotic femoral bone tissue].

OBJECTIVE: To summarize the research progress of tissue engineering technology to promote bone tissue revascularization in osteonecrosis of the femoral head (ONFH). METHODS: The relevant domestic and foreign literature in recent years was extensively reviewed. The mechanism of femoral head vascularization and the application progress of tissue engineering technology in the promotion of ONFH bone tissue revascularization were summarized. RESULTS: Rebuilding or improving the blood supply of the femoral head is the key to the treatment of ONFH. Tissue engineering is a hot spot in current research. It mainly focuses on the three elements of seed cells, scaffold materials, and angiogenic growth factors, combined with three-dimensional printing technology and drug delivery systems to promote the revascularization of the femoral bone tissue. CONCLUSION: The strategy of revascularization of the femoral head can improve the local blood supply and delay or even reverse the progression of ONFH disease.

Carbon Nanodots Inhibit Oxidized Low Density Lipoprotein-Induced Injury and Monocyte Adhesion to Endothelial Cells Through Scavenging Reactive Oxygen Species.

Oxidized low density lipoprotein (Ox-LDL) is a known biomarker of inflammation and atherosclerosis, a leading cause of death worldwide. As a new class of nanomaterials, carbon nanodots (CNDs) are widely used in bioimaging, diagnostics, and drug delivery. However, there is no current report on how these CNDs affect the cardiovascular system, particularly their potential in mediating endothelial inflammatory dysfunction. This study examined effects of CNDs on Ox-LDL-mediated endothelial dysfunction. CNDs significantly inhibited Ox-LDL-mediated adhesion of monocytes to human microvascular endothelial cells (HMEC-1), in human microvascular endothelial cells (HMEC-1). CNDs significantly inhibited Ox-LDL-mediated adhesion of monocytes to endothelial cells, which is an essential step in the development of atherosclerosis. Further, CNDs significantly inhibited OxLDL-induced expression of interleukin-8 (IL-8), a vital cytokine on monocyte adhesion to the endothelial cells. These results demonstrate CNDs possess anti-inflammatory properties. CNDs also protect cells against Ox-LDL-induced cytotoxicity. Electron paramagnetic resonance (EPR) spectroscopy studies demonstrated direct reactive oxygen species-scavenging by CNDs. This result indicates that the anti-inflammatory properties of CNDs are most likely due to their direct scavenging of reactive oxygen species. Animal studies involving mice did not show any morphological or physical changes between the CNDs and control groups. Our study provides evidence of potential of CNDs in reducing Ox-LDL-mediated inflammation and cytotoxicity in HMEC-1.

Quadriceps strength and knee joint function in patients with severe knee extension contracture following arthroscopic-assisted mini-incision quadricepsplasty.

PURPOSE: To evaluate reserve quadriceps function and improve knee activity in patients with severe knee extension contracture following arthroscopic-assisted mini-incision quadricepsplasty as well as post-operative complications. METHODS: From 2012 to 2019, 32 patients with severe knee extension contractures (less than 45° range of flexion) were treated with an all-arthroscopic release technique. The clinical results, including range of motion (ROM), quadriceps function (quadriceps index, QI), and knee function, were evaluated, and MRI of the healed tendon after partial quadricepsplasty was performed. The patellar track and length during knee flexion were measured on three normal knees under fluoroscopy. Three formalin-fixed lower limbs were used to mimic severely contracted quadriceps to evaluate the extension of the patellar track. RESULTS: The median follow-up time was 2.1 years (1-5 years). The average QI was 92.0 ± 6.2, and the quadriceps muscle strength was increased from 3.28 to 4.72. At the final follow-up, 90% of the patients had no difficulty going upstairs, going downstairs, or rising from a chair. The ROM improved by 25.69 ± 3.6 preoperatively to 105.88 ± 6.6 at the final follow-up (P < 0.001). The open surgery showed that a 2-cm extension could be achieved by partly cutting the quadriceps tendon, and two cuts achieved a total extension of 5.2 ± 0.52 cm. The patellar tracking distance was 7.7 ± 0.43 cm, and the gap between the patella and femur was also reduced. CONCLUSION: Partial quadricepsplasty of the rectus femoris extended the contracted quadriceps and maintained quadriceps strength, allowing for full knee flexion and satisfactory clinical outcomes of knee function with few complications.

FGF2: a key regulator augmenting tendon-to-bone healing and cartilage repair.

Ligament/tendon and cartilage injuries are clinically common diseases that perplex most clinicians. Because of the lack of blood vessels and nerves, their self-repairing abilities are rather poor. Therefore, surgeries are necessary and also widely used to treat ligament/tendon or cartilage injuries. However, after surgery, there are still many problems that affect healing. In recent years, it has been found that exogenous FGF2 plays an important role in the repair of ligament/tendon and cartilage injuries and exerts a synergistic effect with endogenous FGF2. Therefore, FGF2 can be used as a new type of biomolecule to accelerate tendon-to-bone healing and cartilage repair after injury.

Fibroblast growth factor 2-induced human amniotic mesenchymal stem cells combined with autologous platelet rich plasma augmented tendon-to-bone healing.

OBJECTIVE: The purpose of this study was to explore the effect of fibroblast growth factor 2 (FGF-2) on collagenous fibre formation and the osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs) in vitro, as well as the effect of FGF-2-induced hAMSCs combined with autologous platelet-rich plasma (PRP) on tendon-to-bone healing in vivo. METHODS: In vitro, hAMSCs were induced by various concentrations of FGF-2 (0, 10, 20, and 40 â€‹ng/ml) for 14 days, and the outcomes of ligamentous differentiation and osteogenic differentiation were detected by quantitative real-time reverse transcription PCR, Western blot, immunofluorescence, and picrosirius red staining. In addition, a lentivirus carrying the FGF-2 gene was used to transfect hAMSCs, and transfection efficiency was detected by quantitative real time reverse transcription PCR (qRT-PCR) and Western blot. In vivo, the effect of hAMSCs transfected with the FGF-2 gene combined with autologous PRP on tendon-to-bone healing was detected via histological examination, as well as biomechanical analysis and radiographic analysis. RESULTS: In vitro, different concentrations of FGF-2 (10, 20, and 40 â€‹ng/ml) all promoted the ligamentous differentiation and osteogenic differentiation of hAMSCs, and the low concentration of FGF-2 (10 â€‹ng/ml) had a good effect on differentiation. In addition, the lentivirus carrying the FGF-2 gene was successfully transfected into hAMSCs with an optimal multiplicity of infection (MOI) (50), and autologous PRP was prepared successfully. In vivo, the hAMSCs transfected with the FGF-2 gene combined with autologous PRP had a better effect on tendon-to-bone healing than the other groups (p â€‹< â€‹0.05), as evidenced by histological examination, biomechanical analysis, and radiographic analysis. CONCLUSION: hAMSCs transfected with the FGF-2 gene combined with autologous PRP could augment tendon-to-bone healing in a rabbit extra-articular model. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: hAMSCs transfected with the FGF-2 gene combined with autologous PRP may be a good clinical treatment for tendon-to-bone healing, especially for acute sports-related tendon-ligament injuries.

A Nomogram-Based Prediction for Severe Pneumonia in Patients with Coronavirus Disease 2019 (COVID-19).

BACKGROUND: The outbreak of a novel coronavirus disease 2019 (COVID-19) is currently ongoing worldwide. A proportion of COVID-19 patients progress rapidly to acute respiratory failure. OBJECTIVE: We aimed to build a model to predict the risk of developing severe pneumonia in patients with COVID-19 in the early stage. METHODS: Data from patients who were confirmed to have COVID-19 and were admitted within 7 days from the onset of respiratory symptoms were retrospectively collected. The patients were classified into severe and non-severe groups according to the presence or absence of severe pneumonia during 1-2 weeks of follow-up. The clinical characteristics and laboratory indicators were screened by cross-validation based on LASSO regression to build a prediction model presented by a nomogram. The discrimination and stability, as well as the prediction performance of the model, were analysed. RESULTS: The neutrophil-lymphocyte ratio, monocyte counts, eosinophil percentage, serum lactate dehydrogenase level and history of diabetes mellitus were collected for the model. Bootstrap resampling showed the apparent C-statistics, and the brier scores were 0.929 and 0.098. The optimism of the C-statistics and brier score was 0.0172 and -0.019, respectively. The adjusted C-statistics and brier score were 0.9108 and 0.1169, respectively. The optimal cut-off value of the total nomogram score was determined to be 119 according to the maximal Youden index. The sensitivity, specificity, positive predictive value, and negative predictive value for differentiating the presence and absence of severe pneumonia were 83%, 89%, 74%, and 94%, respectively. CONCLUSION: In our study, the neutrophil-lymphocyte ratio, monocyte counts, eosinophil percentage, serum lactate dehydrogenase level and history of diabetes mellitus showed great discrimination and stability for the prediction of the presence of severe pneumonia and were selected for the model.

Immune characteristics distinguish patients with severe disease associated with SARS-CoV-2.

This single-center, retrospective study aimed to explore the immune characteristics of COVID-19 and biomarkers to predict the severity of this disease. Patients infected with SARS-CoV-2 (n = 215) treated at the First Affiliated Hospital of Nanchang University from January 24 to March 12, 2020, were included in the study and classified into severe and non-severe groups. Peripheral immunocyte count and cytokine statuses were compared. The correlation between immune status, cytokine levels, and disease severity was analyzed. Leukocyte numbers were normal in both groups; however, they were relatively high (7.19 × 109/L) in patients of the severe group. Leukocyte distributions differed between the two groups; the severe group had a higher percentage of neutrophils and lower percentage of lymphocytes compared with the non-severe group, and absolute lymphocyte numbers were below normal in both groups, and particularly deficient in patients in the severe group. Lymphocyte counts have negative correlation with duration of hospital period whereas neutrophil count has no significant correlation with it. Of tested cytokines, IL-6 levels were significantly higher in the severe group (P = 0.0418). Low level of lymphocyte predicts severity of COVID-19. IL-6 levels were significantly higher in the severe group, especially in some extremely severe patients. But we did not detect the significant correlation between severity of COVID-19 with IL-6 level which may be due to limited case numbers. Our observations encourage future research to understand the underlying molecular mechanisms and to improve treatment outcome of COVID-19.

miR-134-5p inhibition reduces infarct-induced cardiomyocyte apoptosis via Creb1 upregulation.

BACKGROUND: Following the recent discovery that microRNA-134-5p (miR-134-5p) is elevated in the early stages of acute myocardial infarction (AMI), we examined the specific role of miR-134-5p in cardiomyocytes during AMI. METHODS: To study miR-134-5p's role in the context of AMI, we used a combination of in vitro experiments in H2O2-treated or hypoxic cardiomyocyte cell cultures as well as in vivo experiments in a murine model of AMI. RESULTS: H2O2- and hypoxia-induced cardiomyocyte injury upregulated miR-134-5p expression. miR-134-5p overexpression increased cardiomyocyte apoptosis, whereas miR-134-5p inhibition reduced cardiomyocyte apoptosis. We discovered that the transcription factor cAMP-responsive element binding protein 1 (Creb1) is a functional target of miR-134-5p responsible for regulating cardiomyocyte apoptosis. In vivo AMI resulted in the upregulation and downregulation of miR-134-5p and Creb1 in the infarct area, respectively. Circulating miR-134-5p levels were also increased at days 1 and 2 post-AMI. Modulation of myocardial miR-124-5p expression by intramyocardial injection of antagomiR-134-5p or agomiR-134-5p significantly affected cardiomyocyte apoptosis, infarct size, and cardiac function in vivo. CONCLUSIONS: miR-134-5p/Creb1 axis dysregulation plays a role in hypoxia- or oxidative stress-induced cardiomyocyte apoptosis as well as AMI. Circulating miR-134-5p may show promise as a biomarker for AMI or post-AMI cardiac dysfunction. Manipulating the miR-134-5p/Creb1 axis through either inhibition of miR-134-5p or overexpression of Creb1 may show promise as a novel therapeutic strategy to attenuate cardiac dysfunction following AMI.

FGF-2-Induced Human Amniotic Mesenchymal Stem Cells Seeded on a Human Acellular Amniotic Membrane Scaffold Accelerated Tendon-to-Bone Healing in a Rabbit Extra-Articular Model.

BACKGROUND: FGF-2 (basic fibroblast growth factor) has a positive effect on the proliferation and differentiation of many kinds of MSCs. Therefore, it represents an ideal molecule to facilitate tendon-to-bone healing. Nonetheless, no studies have investigated the application of FGF-2-induced human amniotic mesenchymal stem cells (hAMSCs) to accelerate tendon-to-bone healing in vivo. OBJECTIVE: The purpose of this study was to explore the effect of FGF-2 on chondrogenic differentiation of hAMSCs in vitro and the effect of FGF-2-induced hAMSCs combined with a human acellular amniotic membrane (HAAM) scaffold on tendon-to-bone healing in vivo. METHODS: In vitro, hAMSCs were transfected with a lentivirus carrying the FGF-2 gene, and the potential for chondrogenic differentiation of hAMSCs induced by the FGF-2 gene was assessed using immunofluorescence and toluidine blue (TB) staining. HAAM scaffold was prepared, and hematoxylin and eosin (HE) staining and scanning electron microscopy (SEM) were used to observe the microstructure of the HAAM scaffold. hAMSCs transfected with and without FGF-2 were seeded on the HAAM scaffold at a density of 3 × 105 cells/well. Immunofluorescence staining of vimentin and phalloidin staining were used to confirm cell adherence and growth on the HAAM scaffold. In vivo, the rabbit extra-articular tendon-to-bone healing model was created using the right hind limb of 40 New Zealand White rabbits. Grafts mimicking tendon-to-bone interface (TBI) injury were created and subjected to treatment with the HAAM scaffold loaded with FGF-2-induced hAMSCs, HAAM scaffold loaded with hAMSCs only, HAAM scaffold, and no special treatment. Macroscopic observation, imageological analysis, histological assessment, and biomechanical analysis were conducted to evaluate tendon-to-bone healing after 3 months. RESULTS: In vitro, cartilage-specific marker staining was positive for the FGF-2 overexpression group. The HAAM scaffold displayed a netted structure and mass extracellular matrix structure. hAMSCs or hAMSCs transfected with FGF-2 survived on the HAAM scaffold and grew well. In vivo, the group treated with HAAM scaffold loaded with FGF-2-induced hAMSCs had the narrowest bone tunnel after three months as compared with other groups. In addition, macroscopic and histological scores were higher for this group than for the other groups, along with the best mechanical strength. CONCLUSION: hAMSCs transfected with FGF-2 combined with the HAAM scaffold could accelerate tendon-to-bone healing in a rabbit extra-articular model.

Enhanced tenogenic differentiation and tendon-like tissue formation by Scleraxis overexpression in human amniotic mesenchymal stem cells.

Tendon and ligament injuries are not uncommon in clinics and have poor self-healing capacity due to their bloodless and slow-proliferative nature. Promoting the repair or reconstruction of an injured structure is an urgent problem. While Scleraxis (Scx) is a highly specific tendon cell marker, its function has not been explored to a large extent. Hence, Recombinant adenovirus was used to study the influence of Scx overexpression on directional differentiation of human amniotic mesenchymal stem cells (hMSCAs). hAMSCs modified with Scx could dramatically enhance the gene expression of tendon-related molecules, containing Scx, collagens I and III, Tenascin-C, fibronectin, matrix metalloproteinase-2 (MMP-2), lysyl oxidase-1 (LOX-1) and Tenomodulin at all-time points (P < 0.05), and the secretion of collagen I and III, fibronectin and Tenascin-C over time (P < 0.05) but did not impact the cell proliferation capacity (P > 0.05). Immunofluorescence staining showed the cobweb-like fusion of collagen I and fibronectin in the AdScx group on day 7, with higher average fluorescence intensity than the control (P < 0.05). After mixing with Matrigel, transplants were subcutaneously implanted in nude mice, obvious inflammation and rejection of immune response were not observed and HE staining showed a histological feature of swirl of fibers is closely linked in parallel in hAMSCs modified with Scx. On the contrary, in the control group, an unorganized connective structure with cell distributed randomly was spotted. The results of promoted directional differentiation of stem cells and the spatial structure of the normal tendon tissue in three-dimensional space manifested that Scx can be used as a specific marker for tendon cells, and as a positive regulator for directional differentiation of hAMSCs, which is possible to be applied to novel therapeutics for clinical tendon and ligament injury by hAMSCs modified with Scx.

Enhancement of in vitro proliferation and bioactivity of human anterior cruciate ligament fibroblasts using an in situ tissue isolation method and basic fibroblast growth factor culture conditions: A pilot analysis.

BACKGROUND: Previous studies have reported poor proliferation and bioactivity of human anterior cruciate ligament fibroblasts (hACLFs) after injury. As hACLFs are one of the most significant and indispensable source of seed cells in constructing tissue-engineered ligament, enhancing hACLF proliferation would offer favorable cellular-biological ability and induce the extracellular matrix secretion of hACLFs after loading on multiple types of scaffolds. Enhancing the bioactivity of hACLFs would improve tissue repair and functional recovery after tissue-engineered ligament transplantation. This study compared cells prepared by collagenase digestion and the in situ culture of tissue pieces and investigated the effect of basic fibroblast growth factor (bFGF) on hACLFs. METHODS: Six adult patients participated in this study. Of these patients, tissues from three were compared after culture establishment through collagenase digestion or in situ tissue isolation. hACLF phenotypic characteristics were assessed, and the effect of bFGF on hACLF cultures was observed. hACLFs cultured with and without bFGF served as the experimental and control groups, respectively. Cell Counting Kit-8 was used to detect proliferation. The expression of ligament-related genes and proteins was evaluated by immunofluorescence staining, real-time polymerase chain reaction (PCR) assays, and Western blot assays. RESULTS: The morphology of hACLFs isolated using the two methods differed after the 2nd passage. The proliferation of cells obtained by in situ culture was higher than that of cells obtained by collagenase digestion. hACLFs cultured with bFGF after the 3rd passage exhibited a higher proliferation rate than the controls. Immunofluorescence staining, real-time PCR, and Western blot analysis showed a significant increase in ligament-related gene and protein expression in the hACLFs cultured with bFGF. CONCLUSIONS: The in situ isolation of tissue pieces enhanced hACLF proliferation in vitro, and the hACLFs exhibited phenotypic characteristics of fibroblasts. hACLFs cultured with bFGF exhibited increased hACLF bioactivity.

All-arthroscopic release for treating severe knee extension contractures could improve the knee range of motion and the mid-term functional outcomes.

PURPOSE: To evaluate the safety, feasibility, and effectiveness of an all-arthroscopic technique for the intra- and extraarticular release of severe knee extension contractures. METHODS: From 2012 to 2016, 25 patients with severe knee extension contractures (less than 45° range of flexion) were treated with an all-arthroscopic release technique. The patients underwent intra- and extraarticular arthroscopic release and arthroscopic-assisted mini-incision quadriceps plasty. The post-operative rehabilitation was initiated the first day after the procedures. Comprehensive clinical follow-up evaluations including the range-of-motion (ROM) assessment, the Lysholm score, and the International Knee Documentation Committee (IKDC) score were performed on all patients. RESULTS: The median follow-up time was 28 months (range 12-65 months). The ROM improved from 23.9° ± 7.5° pre-operatively to 105.9° ± 6.5° at the final follow-up (P < 0.001). In addition, the Lysholm score increased from 59.9 ± 5.2 pre-operatively to 89.7 ± 3.3 (P < 0.001). The IKDC score increased from 47.6 ± 3.4 pre-operatively to 91.7 ± 2.4 (P < 0.001). All patients were satisfied with their final ROM and functional outcomes. CONCLUSION: The all-arthroscopic release technique was a safe, feasible and effective method for treating severe knee extension contractures. The severe knee extension contractures may be successfully addressed by the all-arthroscopic release technique during our clinical practice. LEVEL OF EVIDENCE: IV.

Differentiation of Human Amniotic Mesenchymal Stem Cells into Human Anterior Cruciate Ligament Fibroblast Cells by In Vitro Coculture.

Anterior cruciate ligament injuries are common in humans, though cellular components of the knee have little regenerative or proliferation potential. This study investigated the differentiation of human amnion-derived mesenchymal stem cells (hAMSCs) into human anterior cruciate ligament fibroblasts (hACLFs) in vitro through induction with bFGF and TGF-ß1 with coculture systems. Groups A and B comprised hAMSCs at the 3rd passage cultured with and without bFGF and TGF-ß1, respectively; Groups C and D consisted of hAMSCs and hACLFs in monolayer coculture with and without bFGF and TGF-ß1, respectively; Groups E and F were composed of hAMSCs and hACLFs in Transwell coculture with and without bFGF and TGF-ß1, respectively. Cell morphology and proliferation were recorded. Protein expression and relative mRNA expression were evaluated in each group. Cell proliferation was significantly higher in the induced groups than in the noninduced groups. Protein expression increased over time with the highest expression observed in Group E. mRNA levels were significantly higher in Group E than in other groups. This study is the first to demonstrate the use of the Transwell coculture system for this purpose, and hAMSCs were successfully differentiated into hACLFs. Thus, hAMSCs may be a superior choice for hACLF differentiation via Transwell coculture.

[In vitrodifferentiation of human amniotic mesenchymal stem cells into ligament fibroblasts after induced by transforming growth factor ß 1 and vascular endothelial growth factor].

Objective: To investigate whether human amniotic mesenchymal stem cells (hAMSCs) have the characteristics of mesenchymal stem cells (MSCs) and the differentiation capacity into ligament fibroblasts in vitro. Methods: The hAMSCs were separated through trypsin and collagenase digestion from placenta, the phenotypic characteristics of hAMSCs were detected by flow cytometry, the cytokeratin-19 (CK-19) and vimentin expression of hAMSCs were tested through immunofluorescence staining. The hAMSCs at the 3rd passage were cultured with L-DMEM/F12 medium containing transforming growth factor ß 1 (TGF-ß 1) and vascular endothelial growth factor (VEGF) as the experimental group and with single L-DMEM/F12 medium as the control group. The morphology of hAMSCs was observed by inverted phase contrast microscope; the cellular activities and ability of proliferation were examined by cell counting kit-8 (CCK-8) method; the ligament fibroblasts related protein expressions including collagen type I, collagen type III, Fibronectin, and Tenascin-C were detected by immunofluorescence staining; specific mRNA expressions of ligament fibroblasts and angiogenesis including collagen type I, collagen type III, Fibronectin, α-smooth muscle actin (α-SMA), and VEGF were measured by real-time fluorescence quantitative PCR. Results: The hAMSCs presented monolayer and adherent growth under inverted phase contrast microscope; the flow cytometry results demonstrated that hAMSCs expressed the MSCs phenotypes; the immunofluorescence staining results indicated the hAMSCs had high expression of the vimentin and low expression of CK-19; the hAMSCs possessed the differentiation ability into the osteoblasts, chondroblasts, and lipoblasts. The CCK-8 results displayed that cells reached the peak of growth curve at 7 days in each group, and the proliferation ability in the experimental group was significantly higher than that in the control group at 7 days ( P<0.05). The immunofluorescence staining results showed that the expressions of collagen type I, collagen type III, Fibronectin, and Tenascin-C in the experimental group were significantly higher than those in the control group at 5, 10, and15 days after culture ( P<0.05). The real-time fluorescence quantitative PCR results revealed that the mRNA relative expressions had an increasing tendency at varying degrees with time in the experimental group ( P<0.05). The relative mRNA expressions of collagen type I, collagen type III, Fibronectin, α-SMA, and VEGF in the experimental group were significantly higher than those in the control group at the other time points ( P<0.05), but no significant difference was found in the relative mRNA expressions of collagen type I, collagen type III, and VEGF between 2 groups at 5 days ( P>0.05). Conclusion: The hAMSCs possesses the characteristics of MSCs and good proliferation ability which could be chosen as seed cell source in tissue engineering. The expressions of ligament fibroblasts and angiogenesis related genes could be up-regulated, after induction in vitro, and the synthesis of ligament fibroblasts related proteins could be strengthened. In addition, the application of TGF-ß 1 and VEGF could be used as growth factors sources in constructing tissue engineered ligament.