The Protective Effect of Cordycepin On Alcohol-Induced Osteonecrosis of the Femoral Head.

BACKGROUND: Alcohol abuse is known to be a leading risk factor for atraumatic osteonecrosis of the femoral head (ONFH), in which the suppression of osteogenesis plays a critical role. Cordycepin benefits bone metabolism; however, there has been no study to determine its effect on osteonecrosis. METHODS: Human bone mesenchymal stem cells (hBMSCs) were identified by multi-lineage differentiation. Alkaline phosphatase (ALP) activity, RT-PCR, western blots, immunofluorescent assay and Alizarin red staining of BMSCs were evaluated. A rat model of alcohol-induced ONFH was established to investigate the protective role of cordycepin against ethanol. Hematoxylin & eosin (H&E) staining and micro-computerized tomography (micro-CT) were performed to observe ONFH. Apoptosis was assessed by TdT-mediated dUTP nick end labeling (TUNEL). Immunohistochemical staining was carried out to detect OCN and COL1. RESULTS: Ethanol significantly suppressed ALP activity, decreased gene expression of OCN and BMP2, lowered levels of RUNX2 protein, and reduced immunofluorescence staining of OCN and COL1 and calcium formation of hBMSCs. However, these inhibitory effects were attenuated by cordycepin co-treatment at concentrations of 1 and 10 µg/mL Moreover, it was revealed that the osteo-protective effect of cordycepin was associated with modulation of the Wnt/ß-catenin pathway. In vivo, by micro-CT, TUNEL and immunohistochemical staining of OCN and COL1, we found that cordycepin administration prevented alcohol-induced ONFH. CONCLUSION: Cordycepin treatment to enhance osteogenesis may be considered a potential therapeutic approach to prevent the development of alcohol-induced ONFH.

The synthesis of cycloalka[b]furans via an Au(i)-catalyzed tandem reaction of 3-yne-1,2-diols.

An Au(i)-catalyzed cyclization/1,2-rearrangement/aromatization cascade of 3-yne-1,2-diols has been successfully realized. This reaction not only provides a new and efficient strategy for the synthesis of substituted cycloalka[b]furan compounds as well as their derivatives, but might also facilitate related biological studies.

Advantages of pure platelet-rich plasma compared with leukocyte- and platelet-rich plasma in promoting repair of bone defects.

BACKGROUND: High levels of pro-inflammatory cytokines in leukocyte- and platelet-rich plasma (L-PRP) may activate the nuclear factor κB (NF-κB) pathway to counter the beneficial effect of the growth factors on bone regeneration. However, to date, no relevant studies have substantiated this. METHODS: L-PRP and pure platelet-rich plasma (P-PRP) were isolated. The in vitro effects of L-PRP and P-PRP on the proliferation, viability and migration of human bone marrow-derived mesenchymal stem cells (HBMSCs) and EaHy926, tube formation of EaHy926, and osteogenic differentiation of HBMSCs were assessed by cell counting, flow cytometry, scratch assay, tube formation assay, and real-time quantitative polymerase chain reaction (RT-PCR), western blotting and Alizarin red staining, respectively. The in vitro effects of L-PRP and P-PRP on the nuclear translocation of NF-κB p65, mRNA expression of inducible nitric oxide synthase and cyclooxygenase-2, and production of prostaglandin E2 and nitric oxid were assessed by western blotting, RT-PCR, enzyme-linked immunosorbent assay and Griess reaction, respectively. The in vivo effects of L-PRP or P-PRP preprocessed ß-tricalcium phosphate (ß-TCP) on the calvarial defects in rats were assessed by histological and immunofluorescence examinations. RESULTS: P-PRP, which had similar platelet and growth factors concentrations but significantly lower concentrations of leukocytes and pro-inflammatory cytokines compared with L-PRP, promoted the proliferation, viability and migration of HBMSCs and EaHy926, tube formation of EaHy926 and osteogenic differentiation of HBMSCs in vitro, compared with L-PRP. The implantation of P-PRP preprocessed ß-TCP also yielded better histological results than the implantation of L-PRP preprocessed ß-TCP in vivo. Moreover, L-PRP treatment resulted in the activation of the NF-κB pathway in HBMSCs and EaHy926 in vitro while the postoperative delivery of caffeic acid phenethyl ester, an inhibitor of NF-κB activation, enhanced the histological results of the implantation of L-PRP preprocessed ß-TCP in vivo. CONCLUSIONS: Leukocytes in L-PRP may activate the NF-κB pathway via the increased pro-inflammatory cytokines to induce the inferior effects on bone regeneration of L-PRP compared with P-PRP. Hence, P-PRP may be more suitable for bone regeneration compared with L-PRP, and the combined use of P-PRP and ß-TCP represents a safe, simple, and effective alternative option for autogenous bone graft in the treatment of bone defects.

Breviscapine attenuates lead­induced myocardial injury by activating the Nrf2 signaling pathway.

The present study investigated the therapeutic potential of breviscapine (Bre) in mitigating lead (Pb)-induced myocardial injury through activation of the nuclear factor erythroid-2 related factor 2 (Nrf2) pathway. Rat cardiomyocytes (H9C2 cells) were exposed to Pb to model Pb poisoning, and various parameters, including cell viability, apoptosis and reactive oxygen species (ROS) production, were assessed using Cell Counting Kit-8, flow cytometry and 2',7'-dichlorfluoresceindiacetate assays, respectively. Additionally, a rat model of Pb poisoning was established in which blood Pb levels were measured using a graphite furnace atomic absorption spectrophotometer, and alterations in myocardial tissue, oxidative stress markers, inflammatory indicators, protein expression related to apoptosis and the Nrf2 pathway were evaluated via histopathology, ELISA and western blotting. The results showed that Bre treatment enhanced cell viability, decreased apoptosis, and reduced ROS production in Pb-exposed H9C2 cells. Moreover, Bre modulated oxidative stress markers and inflammatory factors while enhancing the expression of proteins in the Nrf2 pathway. In a rat model, Bre mitigated the lead-induced increase in blood Pb levels and myocardial injury biomarkers, and reversed the downregulation of Nrf2 pathway proteins. In conclusion, the current findings suggested that Bre mitigates Pb-induced myocardial injury by activating the Nrf2 signaling pathway, highlighting its potential as a therapeutic agent for protecting the heart from the harmful effects of Pb exposure. Further research is required to elucidate the exact mechanisms and explore the clinical applicability of Bre in mitigating Pb-induced myocardial damage.

Real-time elastography for the differentiation of benign and malignant breast lesions: a meta-analysis.

The prognostic significance of ultrasound real-time elastography (RTE) in patients with breast lesions is controversial. There are two different diagnostic methods: the elasticity score (ES) and the strain ratio (SR). A meta-analysis was performed using a random-effect model to assess the overall sensitivity and specificity of RTE in the differentiation of breast lesions. MEDLINE, EMBASE, PubMed, and the Cochrane Library before February 2011 were searched. A total of 22 studies, which included 4,713 breast nodules in 4,266 patients were analyzed. The overall mean sensitivity and specificity for the diagnosis of malignant breast lesions by RTE were 0.834 [95% confidence interval (CI) 0.814-0.853] and 0.842 (95% CI, 0.829-0.854) for ES, and 0.883 (95% CI, 0.844-0.916) and 0.814 (95% CI, 0.786-0.839) for SR, respectively. RTE has a high sensitivity and specificity in the evaluation of breast lesions and can potentially reduce unnecessary breast biopsies.

Alleviation of synovitis caused by joint instability with application of platelet-rich plasma.

INTRODUCTION: Synovitis is an early set and persistent change during the progression of osteoarthritis, which causes symptoms such as pain and swelling of the joints. Platelet-rich plasma (PRP) can release quantities of growth factors and cytokines, and is proved effected in promoting restoration of multiple soft tissues. MATERIALS AND METHODS: In this study, twenty rabbits were used to establish animal model of synovitis, with the method of severing the anterior and posterior cruciate ligaments and removing the medial meniscus. Then the rabbits were evenly divided into the PRP group and the control group, and each group received injections of PRP and saline respectively once a week for 3 weeks consecutively, with the first injection administered 3 weeks postoperatively. RESULTS: The platelet count, the concentrations of growth factors in PRP and whole blood were investigated, and the IL-1ß concentration in the joint fluid was detected via ELISA. The synovium as well as the adjacent articular cartilage were collected for histological assessment. The platelet concentration in PRP is 6.8 fold of that in the whole blood. The IL-1ß level in the PRP group was lower than that in the control group 2 and 3 weeks after the administration of PRP. Histological investigation showed that the inflammatory reaction of the synovium and impact on the cartilage was much abated in the PRP group. CONCLUSIONS: PRP can effectively alleviate the synovitis caused by osteoarthritis following loss of joint stability. Given the autologous origin, its low cost and low risk features of PRP, it's a promising choice for OA patients to control the symptoms caused by synovitis.

Diminished membrane recruitment of Akt is instrumental in alcohol-associated osteopenia via the PTEN/Akt/GSK-3ß/ß-catenin axis.

Alcohol is considered a leading risk factor for osteopenia. Our previous research indicated that the Akt/GSK-3ß/ß-catenin pathway plays a critical role in the ethanol-induced antiosteogenic effect in bone mesenchymal stem cells (BMSCs). PI3K/Akt is negatively regulated by the phosphatase and tensin homolog (PTEN) phosphatase. In this study, we found that ethanol increased PTEN expression in the BMSCs and bone tissue of ethanol-treated Sprague-Dawley rats. PTEN upregulation impaired Akt recruitment to the plasma membrane and suppressed Akt phosphorylation at Ser473, thereby inhibiting Akt/GSK3ß/ß-catenin signaling and the expression of COL1 and OCN in BMSCs in vitro and in vivo. The results of in vivo assays indicated that PTEN inhibition protected bone tissue against ethanol. Interestingly, our data revealed that following ethanol stimulation, PTEN and PTEN pseudogene 1 (PTENP1) mRNA expression was increased in a time-dependent manner, resulting in an increased PTEN protein level. In addition, ethanol upregulated PTEN expression and decreased PTEN phosphorylation (p-PTEN), indicating an increase in functional PTEN levels. In summary, the ethanol-mediated transcriptional and post-transcriptional regulation of PTEN impaired downstream Akt/GSK3ß/ß-catenin signaling and BMSC osteogenic differentiation. Therefore, we propose that Akt/GSK3ß/ß-catenin activation via PTEN inhibition may be a potential therapeutic approach for preventing the development of alcohol-induced osteopenia.

Ag-loaded MgSrFe-layered double hydroxide/chitosan composite scaffold with enhanced osteogenic and antibacterial property for bone engineering tissue.

Bone tissue engineering scaffolds for the reconstruction of large bone defects should simultaneously promote osteogenic differentiation and avoid postoperative infection. Herein, we develop, for the first time, Ag-loaded MgSrFe-layered double hydroxide/chitosan (Ag-MgSrFe/CS) composite scaffold. This scaffold exhibits three-dimensional interconnected macroporous structure with a pore size of 100-300 µm. The layered double hydroxide nanoplates in the Ag-MgSrFe/CS show lateral sizes of 200-400 nm and thicknesses of ∼50 nm, and the Ag nanoparticles with particle sizes of ∼20 nm are uniformly dispersed on the scaffold surfaces. Human bone marrow-derived mesenchymal stem cells (hBMSCs) present good adhesion, spreading, and proliferation on the Ag-MgSrFe/CS composite scaffold, suggesting that the Ag and Sr elements in the composite scaffold have no toxicity to hBMSCs. When compared with MgFe/CS composite scaffold, the Ag-MgSrFe/CS composite scaffold has better osteogenic property. The released Sr2+ ions from the composite scaffold enhance the alkaline phosphatase activity of hBMSCs, promote the extracellular matrix mineralization, and increase the expression levels of osteogenic-related RUNX2 and BMP-2. Moreover, the Ag-MgSrFe/CS composite scaffold possesses good antibacterial property because the Ag nanoparticles in the composite scaffold effectively prevent biofilm formation against S. aureus. Hence, the Ag-MgSrFe/CS composite scaffold with excellent osteoinductivity and antibacterial property has a great potential for bone tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 863-873, 2018.

Comparative evaluation of the effects of platelet­rich plasma formulations on extracellular matrix formation and the NF­κB signaling pathway in human articular chondrocytes.

Concentrated leukocytes in leukocyte and platelet­rich plasma (L­PRP) may deliver increased levels of pro­inflammatory cytokines to activate the nuclear factor (NF)­κB signaling pathway, to counter or overwhelm the beneficial effects of growth factors on cartilage regeneration. However, to date, no relevant studies have substantiated this. In the present study, L­PRP and pure platelet­rich plasma (P­PRP) were prepared, and leukocytes, platelets, pro­inflammatory cytokines and growth factor concentrations were quantified; they were then used to treat human articular chondrocytes (HACs). Pyrrolidine dithiocarbamate (PDTC; 50 µM) was used to inhibit the activation of NF­κB. The nuclear translocation of NF­κB p65 and the protein expression of cartilaginous markers (collagen II, aggrecan and sex­determining region Y­box 9) were determined using western blot analysis. The mRNA expression of NF­κB­dependent inflammatory mediators, including inducible nitric oxide synthase and cyclooxygenase­2, and cartilaginous markers were determined using reverse transcription­quantitative polymerase chain reaction analysis. The production of prostaglandin E2, nitric oxide and glycosaminoglycan (GAG) were quantified using enzyme­linked immunosorbent assays, the Griess reaction and a 1,9­dimethylmethylene blue assay, respectively. The results demonstrated that L­PRP induced the nuclear translocation of NF­κB p65, upregulated the mRNA expression of NF­κB­dependent inflammatory mediators and upregulated the production of their products, whereas P­PRP, which had similar growth factor concentrations but significantly lower pro­inflammatory cytokine concentrations than L­PRP, did not. P­PRP promoted the mRNA and protein expression levels of cartilaginous markers and the production of GAG more effectively, compared with L­PRP. Furthermore, inhibition of the activation of NF­κB by PDTC enhanced the effects of L­PRP on extracellular matrix formation in the HACs to a level similar to that of P­PRP. These findings suggested that leukocytes in L­PRP activated the NF­κB signaling pathway via the delivery of interleukin­1ß and tumor necrosis factor­α to counter the beneficial effects of growth factors on extracellular matrix formation in HACs. Therefore, P­PRP may be more suitable for the treatment of osteoarthritis.

Erythrocyte sedimentation rate and fibrinogen concentration of whole blood influences the cellular composition of platelet-rich plasma obtained from centrifugation methods.

Erythrocyte sedimentation rate (ESR), which reflects the sedimentation rate of platelets, leukocytes and erythrocytes in response to centrifugal force, may influence the cellular composition of platelet-rich plasma (PRP) obtained via centrifugation methods. However, no relevant studies have substantiated this. In the present study, blood was collected from 40 healthy volunteers and used to prepare PRP with two plasma-based preparation systems [YinPRP and Plasma Rich in Growth Factor (PRGF) systems] and two buffy coat-based systems (RegenPRP and WEGOPRP systems) in a single-donor model. Volumes of PRP and platelet-poor plasma (PPP) that were removed in the preparation process were recorded. Analyses of ESR, haematocrit, C-reaction protein, coagulation, serum glucose and serum lipid of the whole blood used for PRP preparation were performed to evaluate the levels of ESR and the factors known to influence it. Whole blood analysis was performed to evaluate the cellular composition of PRP. Results demonstrated that there were marked positive correlations between the ESR of the whole blood used for PRP preparation and PPP removal efficiencies, platelet concentrations, platelet capture efficiencies and platelet enrichment factors of PRP formulations obtained from plasma-based systems, and PRP yield efficiency of RegenPRP and PPP removal efficiency of WEGOPRP. Furthermore, there were marked negative correlations between ESR and concentrations and enrichment factors of platelets, leukocytes and erythrocytes of RegenPRP. Fibrinogen concentration of the whole blood, which had a marked positive correlation with ESR, also influenced the cellular composition of PRP. These findings may increase the understanding of PRP preparation and provide substantial evidence for the individualised optimisation of PRP preparation systems used in clinical practice.

Comparative evaluation of leukocyte- and platelet-rich plasma and pure platelet-rich plasma for cartilage regeneration.

Platelet-rich plasma (PRP) has gained growing popularity in the treatment of articular cartilage lesions in the last decade. However, the potential harmful effects of leukocytes in PRP on cartilage regeneration have seldom been studied in vitro, and not at all in vivo yet. The objective of the present study is to compare the effects of leukocyte- and platelet-rich plasma (L-PRP) and pure platelet-rich plasma (P-PRP) on cartilage repair and NF-κB pathway, in order to explore the mechanism underlying the function of leukocytes in PRP in cartilage regeneration. The constituent analysis showed that P-PRP had significantly lower concentrations of leukocytes and pro-inflammatory cytokines compared with L-PRP. In addition, cell proliferation and differentiation assays indicated P-PRP promoted growth and chondrogenesis of rabbit bone marrow mesenchymal stem cells (rBMSC) significantly compared with L-PRP. Despite similarity in macroscopic appearance, the implantation of P-PRP combining rBMSC in vivo yielded better cartilage repair results than the L-PRP group based on histological examination. Importantly, the therapeutic effects of PRP on cartilage regeneration could be enhanced by removing leukocytes to avoid the activation of the NF-κB pathway. Thus, PRP without concentrated leukocytes may be more suitable for the treatment of articular cartilage lesions.

Self-assembly of pifithrin-α-loaded layered double hydroxide/chitosan nanohybrid composites as a drug delivery system for bone repair materials.

Bone repair materials for the effective treatment of bone defects should simultaneously possess excellent biocompatibility and promote osteogenic differentiation. Herein, we prepared pifithrin-α-loaded layered double hydroxide/chitosan (PFTα-LDH-CS) nanohybrid composites for the first time according to the following steps: (i) the immersion of LDH nanoplates and PFTα in a CS solution; and (ii) the self-assembly synthesis of PFTα-LDH-CS nanohybrid composites after the pH value of the mixed solution was adjusted to 7.4. Interestingly, the LDH nanoplates with a thickness of ∼20 nm and width of ∼300 nm agglomerated together into flower-like shapes by self-assembly, and the CS was dispersed around the LDH nanoplates. The mesopores with the pore size of 3.95 nm among the LDH nanoplates served as channels for loading PFTα. Moreover, the CS around the LDH nanoplates increased the drug loading efficiency and drug sustained release property compared with the pure LDH nanoplates. The in vitro tests demonstrated that the human bone marrow-derived mesenchymal stem cells (hBMSCs) had good adhesion, spreading and proliferating on the LDH-CS and PFTα-LDH-CS, suggesting that both samples had the desired cytocompatibility. Note that the PFTα released from the PFTα-LDH-CS rapidly improved the cell proliferation, ALP activity, ECM mineralization and protein level of the Runt-related transcription factor 2 (RUNX2) and ß-catenin. The enhanced osteogenic differentiation of hBMSCs on the PFTα-LDH-CS may be attributed to the PFTα released from the abovementioned nanohybrid composites, which resulted in the accumulation of ß-catenin and activation of the ß-catenin-mediated transcription activity in the cell nucleus. Therefore, the PFTα-LDH-CS nanohybrid composites with excellent cytocompatibility and enhanced osteoinductivity have great applications for novel bone repair materials.

Strontium hydroxyapatite/chitosan nanohybrid scaffolds with enhanced osteoinductivity for bone tissue engineering.

For the clinical application of bone tissue engineering with the combination of biomaterials and mesenchymal stem cells (MSCs), bone scaffolds should possess excellent biocompatibility and osteoinductivity to accelerate the repair of bone defects. Herein, strontium hydroxyapatite [SrHAP, Ca10-xSrx(PO4)6(OH)2]/chitosan (CS) nanohybrid scaffolds were fabricated by a freeze-drying method. The SrHAP nanocrystals with the different x values of 0, 1, 5 and 10 are abbreviated to HAP, Sr1HAP, Sr5HAP and Sr10HAP, respectively. With increasing x values from 0 to 10, the crystal cell volumes and axial lengths of SrHAP become gradually large because of the greater ion radius of Sr2+ than Ca2+, while the crystal sizes of SrHAP decrease from 70.4nm to 46.7nm. The SrHAP/CS nanohybrid scaffolds exhibits three-dimensional (3D) interconnected macropores with pore sizes of 100-400µm, and the SrHAP nanocrystals are uniformly dispersed within the scaffolds. In vitro cell experiments reveal that all the HAP/CS, Sr1HAP/CS, Sr5HAP/CS and Sr10HAP/CS nanohybrid scaffolds possess excellent cytocompatibility with the favorable adhesion, spreading and proliferation of human bone marrow mesenchymal stem cells (hBMSCs). The Sr5HAP nanocrystals in the scaffolds do not affect the adhesion, spreading of hBMSCs, but they contribute remarkably to cell proliferation and osteogenic differentiation. As compared with the HAP/CS nanohybrid scaffold, the released Sr2+ ions from the SrHAP/CS nanohybrid scaffolds enhance alkaline phosphatase (ALP) activity, extracellular matrix (ECM) mineralization and osteogenic-related COL-1 and ALP expression levels. Especially, the Sr5HAP/CS nanohybrid scaffolds exhibit the best osteoinductivity among four groups because of the synergetic effect between Ca2+ and Sr2+ ions. Hence, the Sr5HAP/CS nanohybrid scaffolds with excellent cytocompatibility and osteogenic property have promising application for bone tissue engineering.

Novel Akt activator SC-79 is a potential treatment for alcohol-induced osteonecrosis of the femoral head.

Alcohol is a leading risk factor for osteonecrosis of the femoral head (ONFH). We explored the molecular mechanisms underlying alcohol-induced ONFH and investigated the protective effect of the novel Akt activator SC-79 against this disease. We found that ethanol inhibited expression of the osteogenic genes RUNX2 and OCN, downregulated osteogenic differentiation, impaired the recruitment of Akt to the plasma membrane, and suppressed Akt phosphorylation at Ser473, thereby inhibiting the Akt/GSK3ß/ß-catenin signaling pathway in bone mesenchymal stem cells. To assess SC-79's ability to counteract the inhibitory effect of ethanol on Akt-Ser73 phosphorylation, we performed micro-computerized tomography and immunofluorescent staining of osteopontin, osteocalcin and collagen type 1 in a rat model of alcohol-induced ONFH. We found that SC-79 injections inhibited alcohol-induced osteonecrosis. These results show that alcohol-induced ONFH is associated with suppression of p-Akt-Ser473 in the Akt/GSK3ß/ß-catenin signaling pathway in bone mesenchymal stem cells. We propose that SC-79 treatment to rescue Akt activation could be tested in the clinic as a potential therapeutic approach to preventing the development of alcohol-induced ONFH.

Logistic regression analysis of risk factors for femoral head osteonecrosis after healed intertrochanteric fractures.

PURPOSE: To evaluate the potential risk factors of the development of femoral head osteonecrosis after healed intertrochanteric fractures. METHODS: We retrospectively reviewed all patients who were operated upon with closed reduction and internal fixation for intertrochanteric fractures by our medical group from December 1993 to December 2012. Patients with healed fractures were identified. Age, gender, comorbidities favouring osteonecrosis, causes of injuries, fracture patterns, the location of the primary fracture line, time from injury to surgery, fixation methods, and the development of femur head osteonecrosis of these patients were summarised. Univariate and multivariate logistic regression analysis were performed to evaluate the correlation between potential risk factors and the development of femoral head osteonecrosis. RESULTS: A total of 916 patients with healed intertrochanteric fractures were identified. Femoral head osteonecrosis was found in 8 cases (0.87%). According to the results of univariate logistic regression, a more proximal fracture line, fixation with dynamic hip screws and age were found to be statistically significant factors. The results of multivariate logistic regression analysis indicated that the statistically significant predictors of femoral head osteonecrosis were younger age (odds ratio [OR] = 17.103; 95% confidence interval [CI], 1.988-147.111), a more proximal fracture line (OR = 31.439; 95% CI, 3.700-267.119) and applying dynamic hip screw as the internal fixation (OR = 11.114; 95% CI, 2.064-59.854). CONCLUSIONS: Regular follow-up is commended in young patients with a proximal fracture line who underwent closed reduction and internal fixation with dynamic hip screw, even though the bone had healed.

Exosomes Secreted by Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells Repair Critical-Sized Bone Defects through Enhanced Angiogenesis and Osteogenesis in Osteoporotic Rats.

Bone defects caused by trauma, severe infection, tumor resection and skeletal abnormalities are common osteoporotic conditions and major challenges in orthopedic surgery, and there is still no effective solution to this problem. Consequently, new treatments are needed to develop regeneration procedures without side effects. Exosomes secreted by mesenchymal stem cells (MSCs) derived from human induced pluripotent stem cells (hiPSCs, hiPSC-MSC-Exos) incorporate the advantages of both MSCs and iPSCs with no immunogenicity. However, there are no reports on the application of hiPSC-MSC-Exos to enhance angiogenesis and osteogenesis under osteoporotic conditions. HiPSC-MSC-Exos were isolated and identified before use. The effect of hiPSC-MSC-Exos on the proliferation and osteogenic differentiation of bone marrow MSCs derived from ovariectomized (OVX) rats (rBMSCs-OVX) in vitro were investigated. In vivo, hiPSC-MSC-Exos were implanted into critical size bone defects in ovariectomized rats, and bone regeneration and angiogenesis were examined by microcomputed tomography (micro-CT), sequential fluorescent labeling analysis, microfil perfusion and histological and immunohistochemical analysis. The results in vitro showed that hiPSC-MSC-Exos enhanced cell proliferation and alkaline phosphatase (ALP) activity, and up-regulated mRNA and protein expression of osteoblast-related genes in rBMSCs-OVX. In vivo experiments revealed that hiPSC-MSC-Exos dramatically stimulated bone regeneration and angiogenesis in critical-sized calvarial defects in ovariectomized rats. The effect of hiPSC-MSC-Exos increased with increasing concentration. In this study, we showed that hiPSC-MSC-Exos effectively stimulate the proliferation and osteogenic differentiation of rBMSCs-OVX, with the effect increasing with increasing exosome concentration. Further analysis demonstrated that the application of hiPSC-MSC-Exos+ß-TCP scaffolds promoted bone regeneration in critical-sized calvarial defects by enhancing angiogenesis and osteogenesis in an ovariectomized rat model.

New strategy to rescue the inhibition of osteogenesis of human bone marrow-derived mesenchymal stem cells under oxidative stress: combination of vitamin C and graphene foams.

To rescue the oxidative stress induced inhibition of osteogenesis, vitamin C (VC) was chemically modified onto three-dimensional graphene foams (3D GFs), then their regulation on osteogenesis of human bone marrow-derived mesenchymal stem cells (BM-MSCs) was studied. Combined action of VC + GF significantly decreased H2O2-induced oxidative stress, and rescued H2O2-inhibited cell viability, differentiation and osteogenesis of BM-MSCs in vitro. Further studies revealed that Wnt pathway may be involved in this protection of osteogenesis. Furthermore, an in vivo mouse model of BM-MSCs transplantation showed that VC + GF remarkably rescued oxidative stress inhibited calcium content and bone formation. The combination of VC and GF exhibited more pronounced protective effects against oxidative stress induced inhibition of osteogenesis, compared to monotherapy of VC or GF. Our study proposed a new strategy in stem cell-based therapies for treating bone diseases.

The Synthesis of Multisubstituted Pyrroles via a Copper-Catalyzed Tandem Three-Component Reaction.

An unprecedented nucleophilic addition/cyclization/aromatization cascade of basic chemicals, i.e., aromatic alkenes/alkynes, trimethylsilyl cyanide and N,N-disubstituted formamide, has been developed to give a series of multisubstituted pyrroles in moderate to good yields with high regioselectivities. This reaction not only reveals a new reaction mode for α-aminonitriles, but also provides a new and efficient cyclization pattern for the synthesis of multisubstituted pyrroles as well as their derivatives, which might facilitate related biological studies.

Enhanced antibacterial activity and osteoinductivity of Ag-loaded strontium hydroxyapatite/chitosan porous scaffolds for bone tissue engineering.

The properties of bone scaffolds, including biocompatibility, osteoinductivity and antibacterial activity, are of great importance for reconstruction of large bone defects and prevention of implant-associated infections. Herein, we develop an Ag-loaded strontium hydroxyapatite (SrHAP)/chitosan (CS) porous scaffold (Ag-SrHAP/CS) according to the following steps: (i) freeze-drying fabrication of a SrHAP/CS porous scaffold; and (ii) deposition of Ag nanoparticles on the above scaffold. In addition, HAP/CS and Ag-HAP/CS porous scaffolds are prepared under the same conditions without doping Sr element. All the HAP/CS, Ag-HAP/CS, SrHAP/CS and Ag-SrHAP/CS porous scaffolds provide a friendly environment for the adhesion, spreading and proliferation of human bone marrow mesenchymal stem cells (hBMSCs). The three-dimensional (3D) interconnected macropores with a pore size of 100-400 µm allow the spreading of hBMSCs throughout the whole scaffolds. Interestingly, the Sr ions and Ag ions released from the Ag-SrHAP/CS porous scaffolds significantly enhance their osteoinductivity and antibacterial activity, respectively. The Sr element in the SrHAP/CS and Ag-SrHAP/CS porous scaffolds increase the alkaline phosphatase (ALP) activity of hBMSCs, extracellular matrix (ECM) mineralization, and the expression levels of osteogenic-related genes BMP-2 and COL-I. Moreover, the Ag ions released from the Ag-HAP/CS and Ag-SrHAP/CS scaffolds can effectively inhibit the growth and attachment of Staphylococcus aureus (S. aureus, ATCC 25923). In conclusion, the Ag-SrHAP/CS porous scaffold possesses excellent biocompatibility, osteoinductivity and antibacterial activity, so it has great potential for application in bone tissue engineering to repair bone defects and avoid infections.

A Tin(IV) Chloride Promoted Tandem C-O Bond Cleavage/Nazarov Cyclization/Nucleophilic Addition Reaction of 1,1-Disubstituted Allylic Ethers toward the Synthesis of Multisubstituted Indenes.

A novel SnCl4-promoted tandem reaction toward multisubstituted indenes via a sequential C-O bond cleavage/Nazarov cyclization/nucleophilic addition reaction has been developed to afford a series of multisubstituted indenes with an all-carbon quaternary center in moderate to good yields.