LncRNA 4930581F22Rik promotes myogenic differentiation by regulating the ERK/MAPK signaling pathway.

The skeletal muscle is the largest organ in mammals and is the primary motor function organ of the body. Our previous research has shown that long non-coding RNAs (lncRNAs) are significant in the epigenetic control of skeletal muscle development. Here, we observed progressive upregulation of lncRNA 4930581F22Rik expression during skeletal muscle differentiation. Knockdown of lncRNA 4930581F22Rik hindered skeletal muscle differentiation and resulted in the inhibition of the myogenic markers MyHC and MEF2C. Furthermore, we found that lncRNA 4930581F22Rik regulates myogenesis via the ERK/MAPK signaling pathway, and this effect could be attenuated by the ERK-specific inhibitor PD0325901. Additionally, in vivo mice injury model results revealed that lncRNA 4930581F22Rik is involved in skeletal muscle regeneration. These results establish a theoretical basis for understanding the contribution of lncRNAs in skeletal muscle development and regeneration.

The role of macrophage polarization in tendon healing and therapeutic strategies: Insights from animal models.

Tendon injuries, a common musculoskeletal issue, usually result in adhesions to the surrounding tissue, that will impact functional recovery. Macrophages, particularly through their M1 and M2 polarizations, play a pivotal role in the inflammatory and healing phases of tendon repair. In this review, we explore the role of macrophage polarization in tendon healing, focusing on insights from animal models. The review delves into the complex interplay of macrophages in tendon pathology, detailing how various macrophage phenotypes contribute to both healing and adhesion formation. It also explores the potential of modulating macrophage activity to enhance tendon repair and minimize adhesions. With advancements in understanding macrophage behavior and the development of innovative biomaterials, this review highlights promising therapeutic strategies for tendon injuries.

Multi-metal electrocatalyst with crystalline/amorphous structure for enhanced alkaline water/seawater hydrogen evolution.

The development of well-defined nanomaterials as non-noble metal electrocatalysts has broad application prospect for hydrogen generation technology. Recently, multi-metal electrocatalysts for hydrogen evolution reaction (HER) have attracted extensive attention due to their high catalytic performance arising from the synergistic effect of multi-metal interaction. However, most multi-metal catalysts suffer from the limited synergistic effect because of poor interfacial compatibility between different components. Here, a novel multi-metal catalyst (Ni/MoO2@CoFeOx) nanosheet with a crystalline/amorphous structure is demonstrated, which shows high HER activity. Ni/MoO2@CoFeOx exhibits an ultra-low overpotential of 18, 39, and 93 mV at 10 mA cm-2 in alkaline water, alkaline seawater and natural seawater, respectively, which outperformances most of the state-of-the-art non-noble metal compounds. In addition, the catalyst shows exceptional stability under 500 mA cm-2 in alkaline solution. In-situ Raman and other advanced structural characterization confirms the excellent catalytic activity is mainly contributed by: (1) the strong synergistic effect of multi-metal components provides multiple active sites in the catalytic process; (2) the crystalline/amorphous interface in Ni/MoO2@CoFeOx boosts the catalytically active sites and structure stability; (3) the crystalline phase enhances the intrinsic conductivity greatly; and (4) the amorphous phase provides abundant unsaturated sites for improved intrinsic catalytic activity. This work provides a feasible way to design electrocatalyst with high activity and stability for practical applications.

Reconstructing avascular necrotic femoral head through a bioactive ß-TCP system: From design to application.

A variety of techniques have been used for treating avascular necrosis of the femoral head (ANFH), but have frequently failed. In this study, we proposed a ß-TCP system for the treatment of ANFH by boosting revascularization and bone regeneration. The angio-conductive properties and concurrent osteogenesis of the highly interconnected porous ß-TCP scaffold were revealed and quantified through an in vivo model that simulated the ischemic environment of ANFH. Mechanical test and finite element analysis showed that the mechanical loss caused by tissue necrosis and surgery was immediately partially compensated after implantation, and the strength of the operated femoral head was adaptively increased and eventually returned to normal bone, along with continuous material degradation and bone regeneration. For translational application, we further conducted a multi-center open-label clinical trial to assess the efficacy of the ß-TCP system in treating ANFH. Two hundred fourteen patients with 246 hips were enrolled for evaluation, and 82.1% of the operated hips survived at a 42.79-month median follow-up. The imaging results, hip function, and pain scores were dramatically improved compared to preoperative levels. ARCO stage Ⅱ disease outperformed stage Ⅲ in terms of clinical effectiveness. Thus, bio-adaptive reconstruction using the ß-TCP system is a promising hip-preserving strategy for the treatment of ANFH.

Short-Term Clinical Response and Changes in the Fecal Microbiota and Metabolite Levels in Patients with Crohn's Disease After Stem Cell Infusions.

Recent studies have shown a close relationship between the gut microbiota and Crohn's disease (CD). This study aimed to determine whether mesenchymal stem cell (MSC) treatment alters the gut microbiota and fecal metabolite pathways and to establish the relationship between the gut microbiota and fecal metabolites. Patients with refractory CD were enrolled and received 8 intravenous infusions of MSCs at a dose of 1.0 × 106 cells/kg. The MSC efficacy and safety were evaluated. Fecal samples were collected, and their microbiomes were analyzed by 16S rDNA sequencing. The fecal metabolites at baseline and after 4 and 8 MSC infusions were identified by liquid chromatography-mass spectrometry (LC--MS). A bioinformatics analysis was conducted using the sequencing data. No serious adverse effects were observed. The clinical symptoms and signs of patients with CD were substantially relieved after 8 MSC infusions, as revealed by changes in weight, the CD activity index (CDAI) score, C-reactive protein (CRP) level, and erythrocyte sedimentation rate (ESR). Endoscopic improvement was observed in 2 patients. A comparison of the gut microbiome after 8 MSC treatments with that at baseline showed that the genus Cetobacterium was significantly enriched. Linoleic acid was depleted after 8 MSC treatments. A possible link between the altered Cetobacterium abundance and linoleic acid metabolite levels was observed in patients with CD who received MSCs. This study enabled an understanding of both the gut microbiota response and bacterial metabolites to obtain more information about host-gut microbiota metabolic interactions in the short-term response to MSC treatment.

Emerging role of the RNA-editing enzyme ADAR1 in stem cell fate and function.

Stem cells are critical for organism development and the maintenance of tissue homeostasis. Recent studies focusing on RNA editing have indicated how this mark controls stem cell fate and function in both normal and malignant states. RNA editing is mainly mediated by adenosine deaminase acting on RNA 1 (ADAR1). The RNA editing enzyme ADAR1 converts adenosine in a double-stranded RNA (dsRNA) substrate into inosine. ADAR1 is a multifunctional protein that regulate physiological processes including embryonic development, cell differentiation, and immune regulation, and even apply to the development of gene editing technologies. In this review, we summarize the structure and function of ADAR1 with a focus on how it can mediate distinct functions in stem cell self-renewal and differentiation. Targeting ADAR1 has emerged as a potential novel therapeutic strategy in both normal and dysregulated stem cell contexts.

Messenger RNA electroporated hepatitis B virus (HBV) antigen-specific T cell receptor (TCR) redirected T cell therapy is well-tolerated in patients with recurrent HBV-related hepatocellular carcinoma post-liver transplantation: results from a phase I trial.

BACKGROUND AND AIMS: Liver transplantation (LT) is the primary curative option for cirrhotic patients with early-stage hepatocellular carcinoma (HCC). However, tumor recurrence occurs in 15-20% of cases with unfavorable prognosis. We have developed a library of T cell receptors (TCRs) specific for different hepatitis B virus (HBV) antigens, restricted by different molecules of human leucocyte antigen (HLA)-class I, to redirect T cells against HBV antigens (Banu in Sci Rep 4:4166, 2014). We further demonstrated that these transiently functional T cells specific for HBV obtained through messenger RNA (mRNA) electroporation can eliminate HCC cells expressing HBV antigens in vitro and in vivo (Kah in J Clin Invest 127:3177-3188, 2017). A phase I clinical trial for patients with HCC recurrence post-liver transplant was conducted to assess the safety, tolerability, and anti-tumor efficacy of transiently functional HBV-TCR T cells. Here, we report the clinical findings with regard to the safety and anti-tumor efficacy of mRNA electroporated HBV-specific TCR-T cells. (ClinicalTrials.gov identifier: NCT02719782). PATIENTS AND METHODS: A total of six patients with HBV-positive recurrent HCC post-liver transplant and HLA-matched to TCR targeting hepatitis B surface antigen (HBsAg) or hepatitis B core antigen (HBcAg) (HLA-A*02:01/HBsAg, HLA-A*11:01/HBcAg, HLA-B*58:01/HBsAg or HLA-C*08:01/HBsAg) were enrolled in this study. The primary objective was to assess the safety of short-lived mRNA electroporated HBV-TCR T cells based on the incidence and severity of the adverse event (AE) graded per National Cancer Institute Common Terminology Criteria for Adverse Events (NCI CTCAE), Version 4.0. The secondary objective was to determine the effectiveness of HBV-TCR T cells as per RECIST 1.1 criteria. Patients were followed up for survival for 2 years post-end of treatment. RESULTS: The median age of the six patients was 35.5 years (range: 28-47). The median number of HBV-TCR T cell infusions administered was 6.5 (range: 4-12). The treatment-related AE included grade 1 pyrexia. This study reported no cytokine release syndrome nor neurotoxicity. One patient remained alive and five were deceased at the time of the data cutoff (30 April 2020). CONCLUSION: This study has demonstrated that multiple infusions of mRNA electroporated HBV-specific TCR T cells were well-tolerated in patients with HBV-positive recurrent HCC post-liver transplant.

Role of lncRNA Has2os in Skeletal Muscle Differentiation and Regeneration.

Long non-coding RNAs (lncRNAs) regulate a series of physiological processes and play an important role in development, metabolism and disease. Our previous studies showed that lncRNAs involved in skeletal muscle differentiation. Here, we demonstrated that lncRNA Has2os is highly expressed in skeletal muscle and significantly elevated during skeletal cell differentiation. The knockdown of Has2os inhibited myocyte fusion and impeded the expression of the myogenic factors MyHC and Mef2C. Mechanically, Has2os regulates skeletal muscle differentiation by inhibiting the JNK/MAPK signaling pathway. Furthermore, we also revealed that Has2os is involved in the early stage of regeneration after muscle injury, and the JNK/MAPK signaling pathway is activated at both protein and mRNA levels during early repair. Our results demonstrate the new function of lncRNA Has2os, which plays crucial roles during skeletal muscle differentiation and muscle regeneration, providing a basis for the therapy of lncRNA-related muscle diseases.

Human umbilical cord-derived mesenchymal stem cells ameliorate experimental colitis by normalizing the gut microbiota.

BACKGROUND: Crohn's disease (CD) is a chronic non-specific inflammatory bowel disease. Current CD therapeutics cannot fundamentally change the natural course of CD. Therefore, it is of great significance to find new treatment strategies for CD. Preclinical and clinical studies have shown that mesenchymal stromal cells (MSCs) are a promising therapeutic approach. However, the mechanism by which MSCs alleviate CD and how MSCs affect gut microbes are still unclear and need further elucidation. METHODS: We used 2,4,6-trinitrobenzenesulfonic acid (TNBS) to induce experimental colitis in mice and analysed the microbiota in faecal samples from the control group, the TNBS group and the TNBS + MSC group with faecal 16S rDNA sequencing. Subsequent analyses of alpha and beta diversity were all performed based on the rarified data. PICRUStII analysis was performed on the 16S rRNA gene sequences to infer the gut microbiome functions. RESULTS: MSC Treatment improved TNBS-induced colitis by increasing survival rates and relieving symptoms. A distinct bacterial signature was found in the TNBS group that differed from the TNBS + MSC group and controls. MSCs prevented gut microbiota dysbiosis, including increasing α-diversity and the amount of Bacteroidetes Firmicutes and Tenericutes at the phylum level and decreasing the amount of Proteobacteria at the phylum level. MSCs alleviated the increased activities of sulphur and riboflavin metabolism. Meanwhile some metabolic pathways such as biosynthesis of amino acids lysine biosynthesis sphingolipid metabolism and secondary bile acid biosynthesis were decreased in the TNBS group compared with the control group and the TNBS + MSC group CONCLUSIONS: Overall, our findings preliminarily confirmed that colitis in mice is closely related to microbial and metabolic dysbiosis. MSC treatment could modulate the dysregulated metabolism pathways in mice with colitis, restoring the abnormal microbiota function to that of the normal control group. This study provides insight into specific intestinal microbiota and metabolism pathways linked with MSC treatment, suggesting a new approach to the treatment of CD.

Nanosized-Ag-doped porous ß-tricalcium phosphate for biological applications.

The treatment of infectious or potentially infective bone defects remains a major problem in clinical practice. Silver has the ability to potentiate antibiotics against resistant bacterial strains. In order to reduce the risk of long-term infections, it is necessary for the biomaterial scaffold to release Ag+ in a controlled manner during the entire healing process. In this study, given the antimicrobial characteristics of nanosized Ag (NSAg), we synthesized ß-tricalcium phosphate (ß-TCP) doped with 5 and 10 wt% NSAg (5 wt% NSAgTCP and 10 wt% NSAgTCP, respectively). The NSAgTCP composites exhibited similar macroporous structures to pure ß-TCP. The NSAgTCP samples were examined by scanning electron microscopy at 10,000-times magnification, which revealed that silver was still present at the nanometer scale. X-ray diffraction revealed that silver does not change the crystalline properties of ß-TCP. In addition, we observed that the mechanical strength of NSAgTCP increased with increasing amounts of added Ag. The antibacterial, physical, and chemical properties of NSAgTCP were investigated in vitro. We found that NSAgTCP is effective at inhibiting the growth of Staphylococcus aureus and Escherichia coli and is not cytotoxic to human bone marrow mesenchymal stem cells. Moreover, it does not hinder liver or kidney function when tested in vivo. As the bioceramic degrades, Ag ions are slowly released and new bone is formed. No significant cytotoxic effects were observed even when 10 wt% NSAgTCP was used. NSAgTCP has the ability to simultaneously repair bone defects and act as an anti-infective agent; hence, we expect that this material, with its good bone-repairing and anti-infective properties, will find wide spread use as a novel bone substitute.

[Minimally invasive treatment for osteonecrosis of the femoral head in ARCO stage â ¡ and â ¢ with bioceramic system].

OBJECTIVE: To perfect the theory system of minimally invasive treatment for osteonecrosis of the femoral head (ONFH) with ß tricalcium phosphate (ß-TCP) bioceramic system and evaluate the effectiveness. METHODS: Eighteen New Zealand white rabbits aged 7-8 months were used to establish an animal model to verify the vascularization of porous ß-TCP bioceramic rods. Micro-CT based three-dimensional reconstruction and fluorescence imaging were used to display the new blood vessels at 4, 8, and 12 weeks after operation. The inserting depth, number and diameter of vessels in the encapsulated area were analyzed. Nine pig femoral specimens were randomly divided into 3 groups ( n=3): group A was normal femur; group B had cavity (core decompression channel+spherical bone defect in femoral head); in group C, mixed bioceramic granules were implanted to fill the defect in femoral head, and porous ß-TCP bioceramic rod was implanted into decompression channel. The stiffness and yield load of specimens were analyzed by biomechanical test. A multicenter retrospective study was conducted to analyze 200 patients (232 hips) with femoral head necrosis treated with bioceramic system in 7 hospitals in China between January 2012 and July 2018. There were 145 males and 55 females, with an average age of 42 years (range, 17-76 years). According to the Association Research Circulation Osseous (ARCO) stage, 150 hips were in stage Ⅱ and 82 hips in stage Ⅲ. Postoperative imaging assessment was carried out regularly, and hip function was evaluated by Harris score. The effectiveness of ARCO stage Ⅱ and Ⅲ was also compared. RESULTS: Animal experiments showed that blood vessels could grow into the encapsulated area and penetrate it at 12 weeks. The inserting depth, number and diameter of blood vessels in the encapsulated area gradually increased, and there was significant difference between different time points ( P<0.05). Biomechanical tests showed that the stiffness and yield load of specimens in groups B and C were significantly lower than those in group A, while the yield load in group B were significantly lower than that in group C ( P<0.05). The stiffness in group C was restored to 41.52%±3.96% in group A, and the yield load was restored to 46.14%±7.85%. Clinical study showed that 200 patients were followed up 6-73 months, with an average of 22.7 months. At last follow-up, 12 patients (16 hips) underwent total hip arthroplasty, and the hip survival rate was 93.10%. According to the imaging evaluation, 184 hips (79.31%) were stable and 48 (20.69%) were worse. Harris score (79.3±17.3) was significantly higher than that before operation (57.3±12.0) ( t=18.600, P=0.000). The excellent rate of hip function was 64.22% (149/232). The survival rate of hip joint, imaging score and Harris score of patients in ARCO stage Ⅱ were better than those in ARCO stage Ⅲ ( P<0.05). CONCLUSION: ß-TCP bioceramic system can guide the abundant blood supply of greater trochanter and femoral neck to the femoral head to promote repair; it can partly restore the mechanical properties of the femoral head and neck in the early stage, providing a new minimally invasive hip-preserving method for patients with ONFH, especially for those in early stage.

Novel 3D-printed prosthetic composite for reconstruction of massive bone defects in lower extremities after malignant tumor resection.

OBJECTIVE: To introduce a novel 3D-printed prosthetic composite for reconstruction of massive bone defects after resection for bone malignancy of lower extremities. The design concept, surgical technique, and the preliminary outcomes were elaborated. METHODS: Patients with primary malignant tumors of lower extremities requiring tumor resection and reconstruction were recruited between Jun 2015 and Nov 2018. Patient-specific 3D-printed prostheses were designed according to preoperative imaging data. After tumor resection, reconstruction was performed with composites consisting of 3D- printed prosthesis, beta-tricalcium phosphate (ß-TCP) bioceramics and/or vascularized fibula. All patients underwent regular follow-up postoperatively. The functional outcomes were assessed by the Musculoskeletal Tumor Society score (MSTS). Oncological outcomes, imaging results, and complications were recorded and analyzed. RESULTS: Ten cases averaging 12.90 years of age participated in this study. There were five femur and five tibia reconstructions. The mean follow-up period was 16.90 months. At last follow-up, all patients were alive without tumor recurrence. Average MSTS functional score was 80.33 ± 11.05%. All prostheses were intact and stable without failure or systemic breakage. No serious complications occurred after the operation. Postoperative X-ray, computed tomography (CT) and single-photon emission computed tomography (SPECT) showed an ideal integration between the bone and the prosthetic composite. Moreover, vascularized fibula and implanted ß-TCP bioceramics indicated relatively high metabolic activity in vivo. CONCLUSIONS: Patient-specific 3D-printed prostheses combined with ß-TCP bioceramics and/or vascularized fibula provide an excellent option for reconstruction of massive bone defects after lower extremity malignant tumor extirpation. Short-term follow up showed promising clinical results in recovering lower limb function, promoting osseointegration and reducing complications.

Collagen/ß-TCP composite as a bone-graft substitute for posterior spinal fusion in rabbit model: a comparison study.

ß-TCP bioceramic, as a kind of biocompatible and biodegradable artificial bone scaffolds, is increasingly used to supplement lamina autografts when performing instrumented or non-instrumented spinal fusion, clinically, although solid fusion is not always achieved. The addition of collagen to ß-TCP appears to be a potential strategy to improve bone regeneration, thereby enhancing the rate of spinal fusion. This study aimed to compare the fusion in collagen/ß-TCP composite, ß-TCP and autologous bone in a posterior spinal fusion model. The fusion grade evaluated radiography was greater in the collagen/ß-TCP group than in the ß-TCP group (p < 0.05). Stiffness and yield strength of the fused segments in collagen/ß-TCP group were comparable to that in autogenous bone group. Histological analysis revealed that the proportion of new bone formation in collagen/ß-TCP group were significantly greater than in ß-TCP group (p < 0.05). In addition, bone deposition rate in the collagen/ß-TCP group was greater than in the ß-TCP group (p < 0.05) and comparable to that in the autogenous bone group. We therefore concluded that collagen/ß-TCP is superior to ß-TCP alone in facilitating posterior spinal fusion. The addition of collagen to ß-TCP represents a simple strategy that couples the osteogenic effect, providing a promising alternative to autologous bone in the clinical treatment of spinal disorders.

Minimally invasive treatment for osteonecrosis of the femoral head with angioconductive bioceramic rod.

PURPOSE: To describe the rationale, the surgical technique, and the short-term follow-up results of a new minimally invasive treatment of osteonecrosis of the femoral head (ONFH) with an angioconductive bioceramic rod (ABR) implant. METHODS: Sixty-two patients (72 hips) with ARCO stage IIA-IIIC ONFH treated with the minimally invasive ABR from January 2012 to December 2016 were reviewed (17 females, 45 males, mean age 44.49). This technique used the angioconductive properties of the porous implant to repair the necrosis by driving vascularization from the trochanter to the necrotic area. Patients had a mean follow-up period of 26.74 months. The outcomes were evaluated by hip joint survival, radiograph, and the Harris Hip Score (HHS). The complications occurred during the treatment period were recorded. RESULTS: No serious post-operative complications occurred during the treatment. The overall joint survival rate was 90.27%, with seven conversions to THA. Improvements were observed in 23 (31.95%) hips, 24 (33.33%) hips remained stable, and 25 (34.72%) hips had worse results according to the radiographic evaluation. The mean HHS at the end follow-up significantly improved compared to the pre-operative mean HHS (82.27 vs 58.14, p < 0.001). In both radiographic evaluation and HHS, the treatment was more effective on patients beneath 44 years old (p < 0.05); ARCO stage II compared to stage III (p < 0.05); and China-Japan Friendship Hospital (CJFH) type C compared to CJFH type L (p < 0.05). CONCLUSIONS: The minimally invasive treatment of ONFH with ABR showed promising results in delaying or even terminating the progression of the necrosis and improving hip function, especially in younger patients and in the early stages of the disease.

IL-32γ promotes integrin αvß6 expression through the activation of NF-κB in HSCs.

Hepatic stellate cell (HSC) activation is important in the pathogenesis of liver fibrosis. However, the molecular mechanism of HSC activation is not completely understood. In the present study, it was demonstrated that interleukin-32γ (IL-32γ) is capable of enhancing intefgrin αvß6 expression by inducing integrin αvß6 promoter activity in a dose-dependent manner in HSCs. Furthermore, it was determined that nuclear factor κB (NF-κB) activation is required for IL-32γ-induced integrin αvß6 expression. Increased integrin αvß6 expression is then able to activate HSCs. These results indicate that NF-κB activation is required for IL-32γ to induce integrin αvß6 expression and consequently promote HSC activation. Therefore, IL-32γ activates HSCs and therefore may be associated with hepatic fibrogenesis. These results may enable the development of novel effective strategies to treat hepatic fibrosis.

Influence of biological scaffold regulation on the proliferation of chondrocytes and the repair of articular cartilage.

PURPOSE: To investigate the effects of hard tissue engineering scaffold (the material is ß-TCP) with different micro-structures on the proliferation of chondrocytes, and the influence of its composite erythrocytes on the repair of articular cartilage defects. METHODS: Rabbit cartilage cells were on ß-TCP bioceramic scaffold with different micro-structures in vitro, the proliferation growth trend of chondrocytes within the scaffold was calculated, and a optimal micro-structure suitable for cartilage cell growth was determined. Composite chondrocytes were implanted into rabbit models of articular cartilage defects, and the repair situation was observed. RESULTS: the bioceramic scaffold with an inner diameter of 120 µm and an aperture of 500-630 µm was suitable for the growth of cartilage cells. Scaffold loaded with second generation of cartilage cell suspension got a top histological score of 20.76±2.13, which was closely similar to that of normal cartilage. CONCLUSION: When loaded with the second generation of cartilage cells, the ß-TCP biological ceramic scaffold with a pore size of 500-630 µm, and an inner diameter of 120 µm, shows a best repairing effect on animal articular cartilage defects.

Beta-tricalcium phosphate granules improve osteogenesis in vitro and establish innovative osteo-regenerators for bone tissue engineering in vivo.

The drawbacks of traditional bone-defect treatments have prompted the exploration of bone tissue engineering. This study aimed to explore suitable ß-tricalcium phosphate (ß-TCP) granules for bone regeneration and identify an efficient method to establish ß-TCP-based osteo-regenerators. ß-TCP granules with diameters of 1 mm and 1-2.5 mm were evaluated in vitro. The ß-TCP granules with superior osteogenic properties were used to establish in vivo bioreactors, referred to as osteo-regenerators, which were fabricated using two different methods. Improved proliferation of bone mesenchymal stem cells (BMSCs), glucose consumption and ALP activity were observed for 1-2.5 mm ß-TCP compared with 1-mm granules (P < 0.05). In addition, BMSCs incubated with 1-2.5 mm ß-TCP expressed significantly higher levels of the genes for runt-related transcription factor-2, alkaline phosphatase, osteocalcin, osteopontin, and collagen type-1 and the osteogenesis-related proteins alkaline phosphatase, collagen type-1 and runt-related transcription factor-2 compared with BMSCs incubated with 1 mm ß-TCP (P < 0.05). Fluorochrome labelling, micro-computed tomography and histological staining analyses indicated that the osteo-regenerator with two holes perforating the femur promoted significantly greater bone regeneration compared with the osteo-regenerator with a periosteum incision (P < 0.05). This study provides an alternative to biofunctionalized bioreactors that exhibits improved osteogenesis.

A Novel Adeno-Associated Virus-Based Genetic Vaccine Encoding the Hepatitis C Virus NS3/4 Protein Exhibits Immunogenic Properties in Mice Superior to Those of an NS3-Protein-Based Vaccine.

More than 170 million individuals worldwide are infected with hepatitis C virus (HCV), and up to an estimated 30% of chronically infected individuals will go on to develop progressive liver disease. Despite the recent advances in antiviral treatment of HCV infection, it remains a major public health problem. Thus, development of an effective vaccine is urgently required. In this study, we constructed novel adeno-associated virus (AAV) vectors expressing the full-length NS3 or NS3/4 protein of HCV genotype 1b. The expression of the NS3 or NS3/4 protein in HepG2 cells was confirmed by western blotting. C57BL/6 mice were intramuscularly immunised with a single injection of AAV vectors, and the resultant immune response was investigated. The AAV2/rh32.33.NS3/4 vaccine induced stronger humoral and cellular responses than did the AAV2/rh32.33.NS3 vaccine. Our results demonstrate that AAV-based vaccines exhibit considerable potential for the development of an effective anti-HCV vaccine.

[Progress on strategies to promote vascularization in bone tissue engineering].

With the continuous development of bone tissue engineering, a variety of emerging bone graft materials provided various methods for repairing bone defects. Early and rapid accomplishment of revascularization of materials interior after implantation of bone transplantation materials is a difficulty faced to bone tissue engineering. Blood vessels ingrowth provides the requisite netritional support for the regeneration reconstruction of bone tissue, for this reason, vascularization plays a significant role in bone tissue engineering. However,there is not a golden standard strategy of vascularization at present. Scaffold materials, cells and growth factors still are three indispensable elements in tissue engineering, and are cardinal points of the promoting vascularization strategies. Multiple growth factors or multiple cells combined with scaffolds, which are hot spots, have obtained excellent vascularization. This review focused on the comprehensive strategies for promoting the successful vascularization of tissue engineered scaffolds.

In vitro comparison of three rifampicin loading methods in a reinforced porous ß-tricalcium phosphate scaffold.

The antibiotic compound, rifampicin (RFP), was loaded into porous reinforced ß-tricalcium phosphate (ß-TCP) scaffolds using three different solution adsorption methods. This resulted in drug delivery systems (DDS) generated by vacuum adsorption (VA), dynamic adsorption (DA), and static adsorption (SA). In vitro examination of the drug loading and release profiles of the DDS indicated that the unit mass of RFP loaded into the scaffold by the VA method (0.44 mg/g) was higher than that achieved by SA (0.42 mg/g) or DA (0.38 mg/g) (P < 0.05). The mechanical strength had no significant change after RFP-loading (P > 0.05). Moreover, there were no significant differences among the mechanical strength of three ß-TCP DDS generated by loading RFP using SA, DA, and VA (P > 0.05). In vitro release testing showed an initial burst release of RFP from the three different DDS within the first 3 h and in the first 51 h, the cumulative release of RFP from VA-DDS, DA-DDS, and SA-DDS had reached 56.2, 83.6, and 88.6 %, respectively. Complete RFP release had occurred from VA-DDS, DA-DDS, and SA-DDS after 23, 17, and 15 days, respectively. As the VA-DDS method showed improved RFP loading and a more sustained drug release, this method is recommended for solution adsorption drug loading into porous ß-TCP scaffolds to form a DDS.