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BACKGROUND: Osteoarthritis (OA) is an aging-related degenerative joint disorder marked by joint discomfort and rigidity. Senescent chondrocytes release pro-inflammatory cytokines and extracellular matrix-degrading proteins, creating an inflammatory microenvironment that hinders chondrogenesis and accelerates matrix degradation. Targeting of senescent chondrocytes may be a promising approach for the treatment of OA. Herein, we describe the engineering of an injectable peptide-hydrogel conjugating a stem cell-homing peptide PFSSTKT for carrying plasmid DNA-laden nanoparticles and Tanshinon IIA (pPNP + TIIA@PFS) that was designed to attenuate OA progression by improving the senescent microenvironment and fostering cartilage regeneration. RESULTS: Specifically, pPNP + TIIA@PFS elevates the concentration of the anti-aging protein Klotho and blocks the transmission of senescence signals to adjacent healthy chondrocytes, significantly mitigating chondrocyte senescence and enhancing cartilage integrity. Additionally, pPNP + TIIA@PFS recruit bone mesenchymal stem cells and directs their subsequent differentiation into chondrocytes, achieving satisfactory chondrogenesis. In surgically induced OA model rats, the application of pPNP + TIIA@PFS results in reduced osteophyte formation and attenuation of articular cartilage degeneration. CONCLUSIONS: Overall, this study introduces a novel approach for the alleviation of OA progression, offering a foundation for potential clinical translation in OA therapy.
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Condrócitos , Condrogênese , Glucuronidase , Hidrogéis , Proteínas Klotho , Células-Tronco Mesenquimais , Osteoartrite , Plasmídeos , Ratos Sprague-Dawley , Animais , Osteoartrite/terapia , Osteoartrite/tratamento farmacológico , Hidrogéis/química , Ratos , Condrócitos/metabolismo , Condrócitos/efeitos dos fármacos , Glucuronidase/metabolismo , Glucuronidase/farmacologia , Condrogênese/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Masculino , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Progressão da Doença , Nanopartículas/química , Humanos , DNA , Senescência Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacosRESUMO
Osteoarthritis (OA) is a common, degenerative joint disease characterized by articular cartilage degradation. Currently, clinical trials based on microRNA therapy have been performed to treat various diseases. However, no treatment has been found for arthritis. This study investigated the functions of miR-101 in cartilage degradation in vivo and evaluated the feasibility of using miR-101 as a therapeutic agent for OA. Mono-iodoacetate-induced arthritis (MIA) rats were used as an animal model of OA. miR-101 mimic or miR-101 inhibitor was injected into the rats' knees to evaluate its effects on cartilage degradation. Cartilage degradation aggravated at 14 days after the injection of miR-101 mimic. By contrast, miR-101 silencing reduced cartilage degradation. Moreover, the administration of miR-101 mimic is sufficient to cause cartilage degradation in the normal cartilage of rats. By contrast, miR-101 inhibitor could prevent this change. Microarray and qPCR were used to investigate the different expressed genes after injecting miR-101 mimic and miR-101 inhibitor in the rats' articular cartilage. Several cartilage degradation-related genes were selected and validated to function in cartilage degradation with miR-101. Our results demonstrated the therapeutic effect of miR-101 inhibition on cartilage degradation in MIA rats by regulating several cartilage degradation-related genes.
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Matriz Extracelular/metabolismo , Inativação Gênica , MicroRNAs/genética , Osteoartrite/terapia , Animais , Artrite Experimental/patologia , Artrite Experimental/terapia , Cartilagem Articular/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Vetores Genéticos , Iodoacetatos/química , Microscopia de Fluorescência , Análise de Sequência com Séries de Oligonucleotídeos , Osteoartrite/induzido quimicamente , Osteoartrite/patologia , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição SOX9/metabolismo , Líquido Sinovial/metabolismoRESUMO
Enhancing the regeneration of cartilage defects remains challenging owing to limited innate self-healing as well as acute inflammation arising from the overexpression of reactive oxygen species (ROS) in post-traumatic microenvironments. Recently, stem cell-derived exosomes (Exos) have been developed as potential cell-free therapy for cartilage regeneration. Although this approach promotes chondrogenesis, it neglects the emerging inflammatory microenvironment. In this study, a smart bilayer-hydrogel dual-loaded with sodium diclofenac (DC), an anti-inflammatory drug, and Exos from bone marrow-derived mesenchymal stem cells was developed to mitigate initial-stage inflammation and promote late-stage stem-cell recruitment and chondrogenic differentiation. First, the upper-hydrogel composed of phenylboronic-acid-crosslinked polyvinyl alcohol degrades in response to elevated levels of ROS to release DC, which mitigates oxidative stress, thus reprogramming macrophages to the pro-healing state. Subsequently, Exos are slowly released from the lower-hydrogel composed of hyaluronic acid into an optimal microenvironment for the stimulation of chondrogenesis. Both in vitro and in vivo assays confirmed that the dual-loaded bilayer-hydrogel reduced post-traumatic inflammation and enhanced cartilage regeneration by effectively scavenging ROS and reprogramming macrophages. The proposed platform provides multi-staged therapy, which allows for the optimal harnessing of Exos as a therapeutic for cartilage regeneration.
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Background: Developmental Dysplasia of the Hip (DDH) is a skeletal disorder where late-presenting forms often escape early diagnosis, leading to limb and pain in adults. The genetic basis of DDH is not fully understood despite known genetic predispositions. Methods: We employed Whole Genome Sequencing (WGS) to explore the genetic factors in late-presenting DDH in two unrelated families, supported by phenotypic analyses and in vitro validation. Results: In both cases, a novel de novo heterozygous missense mutation in RAF1 (c.193A>G [p.Lys65Glu]) was identified. This mutation impacted RAF1 protein structure and function, altering downstream signaling in the Ras/ERK pathway, as demonstrated by bioinformatics, molecular dynamics simulations, and in vitro validations. Conclusion: This study contributes to our understanding of the genetic factors involved in DDH by identifying a novel mutation in RAF1. The identification of the RAF1 mutation suggests a possible involvement of the Ras/ERK pathway in the pathogenesis of late-presenting DDH, indicating its potential role in skeletal development.
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Periprosthetic osteolysis induced by the ultrahigh-molecular-weight polyethylene (UHMWPE) wear particles is a major complication associated with the sustained service of artificial joint prostheses and often necessitates revision surgery. Therefore, a smart implant with direct prevention and repair abilities is urgently developed to avoid painful revision surgery. Herein, we fabricate a phosphatidylserine- and polyethylenimine-engineered niobium carbide (Nb2C) MXenzyme-coated micro/nanostructured titanium implant (PPN@MNTi) that inhibits UHMWPE particle-induced periprosthetic osteolysis. The specific mechanism by which PPN@MNTi operates involves the bioresponsive release of nanosheets from the MNTi substrate within an osteolysis microenvironment, initiated by the cleavage of a thioketal-dopamine molecule sensitive to reactive oxygen species (ROS). Subsequently, functionalized Nb2C MXenzyme could target macrophages and escape from lysosomes, effectively scavenging intracellular ROS through its antioxidant nanozyme-mimicking activities. This further achieves the suppression of osteoclastogenesis by inhibiting NF-κB/MAPK and autophagy signaling pathways. Simultaneously, based on the synergistic effect of MXenzyme-integrated coatings and micro/nanostructured topography, the designed implant promotes the osteogenic differentiation of bone mesenchymal stem cells to regulate bone homeostasis, further achieving advanced osseointegration and alleviable periprosthetic osteolysis in vivo. This study provides a precise prevention and repair strategy of periprosthetic osteolysis, offering a paradigm for the development of smart orthopedic implants.
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Nióbio , Osteogênese , Osteólise , Osteogênese/efeitos dos fármacos , Osteólise/patologia , Osteólise/prevenção & controle , Osteólise/metabolismo , Nióbio/química , Camundongos , Animais , Polietilenos/química , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Titânio/química , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismoRESUMO
Inflammatory responses play a central role in coordinating biomaterial-mediated tissue regeneration. However, precise modulation of dynamic variations in microenvironmental inflammation post-implantation remains challenging. In this study, the traditional ß-tricalcium phosphate-based scaffold is remodeled via ultrathin MXene-Ti3C2 decoration and Zn2+/Sr2+ ion-substitution, endowing the scaffold with excellent reactive oxygen species-scavenging ability, near-infrared responsivity, and enhanced mechanical properties. The induction of mild hyperthermia around the implant via periodic near-infrared irradiation facilitates spatiotemporal regulation of inflammatory cytokines secreted by a spectrum of macrophage phenotypes. The process initially amplifies the pro-inflammatory response, then accelerates M1-to-M2 macrophage polarization transition, yielding a satisfactory pattern of osteo-immunomodulation during the natural bone healing process. Later, sustained release of Zn2+/Sr2+ ions with gradual degradation of the 3D scaffold maintains the favorable reparative M2-dominated immunological microenvironment that supports new bone mineralization. Precise temporal immunoregulation of the bone healing process by the intelligent 3D scaffold enhances bone regeneration in a rat cranial defect model. This strategy paves the way for the application of ß-tricalcium phosphate-based materials to guide the dynamic inflammatory and bone tissue responses toward a favorable outcome, making clinical treatment more predictable and durable. The findings also demonstrate that near-infrared irradiation-derived mild hyperthermia is a promising method of immunomodulation.
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Inadequate initial osseointegration and consequent prosthesis loosening are the most severe complications after artificial arthroplasty. Proper immune responses are crucial for the successful implantation of artificial prostheses. Macrophages are central in osteoimmunomodulation because they exert distinct functions with highly plasticity. Herein, we developed an alkaline phosphatase (ALP) sensitive mussel-inspired coating on orthopedic implants for promoting osseointegration. First, the resveratrol-alendronate complexes were deposited on titanium implant surface through mussel-inspired interfacial interactions. Upon prosthesis implantation, macrophages first polarized towards M1 type to initiate inflammatory responses and bone regeneration. As osteogenesis progresses, increasing amounts of ALP secreted by osteoblasts was cleaved the resveratrol-alendronate complexes. Then, the released resveratrol further promoted osteogenic differentiation of BMSCs and induced locoregional macrophages M2 polarization. Our results demonstrated that the bioinspired osteoimmunomodulation coating remarkably facilitated the prosthesis-bone integration by spatiotemporally modulating macrophages switching from M1 to M2 polarization in response to a real-time healing signal during osteogenesis. In summary, the mussel-inspired osteoimmunomodulation coating technology may provide a new approach for promoting osseointegration after artificial arthroplasty.
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Fosfatase Alcalina , Osseointegração , Osteogênese , Resveratrol , Alendronato , Próteses e Implantes , Titânio/farmacologia , Propriedades de Superfície , Materiais Revestidos Biocompatíveis/farmacologiaRESUMO
Massive intra-articular infiltration of proinflammatory macrophages is a prominent feature of rheumatoid arthritis (RA) lesions, which are thought to underlie articular immune dysfunction, severe synovitis and ultimately joint erosion. Here we report an efferocytosis-informed nanoimitator (EINI) for in situ targeted reprogramming of synovial inflammatory macrophages (SIMs) that thwarts their autoimmune attack and reestablishes articular immune homeostasis, which mitigates RA. The EINI consists of a drug-based core with an oxidative stress-responsive phosphatidylserine (PtdSer) corona and a shell composed of a P-selectin-blocking motif, low molecular weight heparin (LMWH). When systemically administered, the LMWH on the EINI first binds to P-selectin overexpressed on the endothelium in subsynovial capillaries, which functions as an antagonist, disrupting neutrophil synovial trafficking. Due to the strong dysregulation of the synovial microvasculature, the EINI is subsequently enriched in the joint synovium where the shell is disassembled upon the reactive oxygen species stimulation, and PtdSer corona is then exposed. In an efferocytosis-like manner, the PtdSer-coroneted core is in turn phagocytosed by SIMs, which synergistically terminate SIM-initiated pathological cascades and serially reestablish intra-articular immune homeostasis, conferring a chondroprotective effect. These findings demonstrate that SIMs can be precisely remodeled via the efferocytosis-mimetic strategy, which holds potential for RA treatment.
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Artrite Reumatoide , Selectina-P , Camundongos , Animais , Selectina-P/metabolismo , Heparina de Baixo Peso Molecular , Articulações/metabolismo , Membrana Sinovial/metabolismoRESUMO
Tracking and eradicating Staphylococcus aureus in the periprosthetic microenvironment are critical for preventing periprosthetic joint infection (PJI), yet effective strategies remain elusive. Here, we report an implant nanoparticle coating that locoregionally yields bactericidal super chimeric antigen receptor macrophages (CAR-MΦs) to prevent PJI. We demonstrate that the plasmid-laden nanoparticle from the coating can introduce S. aureus-targeted CAR genes and caspase-11 short hairpin RNA (CASP11 shRNA) into macrophage nuclei to generate super CAR-MΦs in mouse models. CASP11 shRNA allowed mitochondria to be recruited around phagosomes containing phagocytosed bacteria to deliver mitochondria-generated bactericidal reactive oxygen species. These super CAR-MΦs targeted and eradicated S. aureus and conferred robust bactericidal immunologic activity at the bone-implant interface. Furthermore, the coating biodegradability precisely matched the bone regeneration process, achieving satisfactory osteogenesis. Overall, our work establishes a locoregional treatment strategy for priming macrophage-specific bactericidal immunity with broad application in patients suffering from multidrug-resistant bacterial infection.
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Receptores de Antígenos Quiméricos , Staphylococcus aureus , Animais , Camundongos , Osseointegração , Antibacterianos/farmacologia , Macrófagos/microbiologiaRESUMO
Osteoarthritis (OA) is a chronic condition caused by cartilage degradation, and there are currently no effective methods for preventing the progression of this disease; gene therapy is a relatively novel method for treating arthritis. Decreased collagen type II (Col2) expression within the cartilage matrix is an important factor for the development of OA, and Wnt3a serves a significant role in cartilage homeostasis. The present study assessed whether Wnt3a knockdown promoted Col2 expression in chondrocytes. Lentivirus-introduced small interfering RNA was used to knock down the expression of Wnt3a in primary rat chondrocytes, and then IL-1ß treatment was used to establish an OA chondrocyte model. The expression of target genes (Wnt3a, Col2, MMP-13 and ß-catenin) was analyzed using reverse transcription-quantitative PCR, western blotting and immunocytochemistry. There was significantly less MMP-13 and ß-catenin expression in the Wnt3a knockdown cells compared with the other controls. Col2 expression was significantly higher in the Wnt3a-knockdown cells compared with the control cells, indicating that knockdown of Wnt3a may promote Col2 expression. Consequently, Wnt3a was indicated to be an important factor in cartilage homeostasis, and Wnt3a knockdown may serve as a novel method for OA therapy.
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Periprosthetic bone defects are the most serious problem of revision total hip arthroplasty, which can easily lead to insufficient osteointegration between the prosthesis and host bone. Bone marrow mesenchymal stem cells (BMSCs) and a moderate inflammatory response at the prosthesis-bone interface play an important role in osteointegration. Here, we developed microarc oxide titanium implant loaded engineered exosomes (S-Exos) to promote osseointegration at the prosthesis-bone interface. First, Smurf1-shRNA was transferred into the BMSCs using a viral vector to prepare S-Exos, which were subsequently immobilized to the microarc oxide titanium implant surface with positively charged polyethyleneimine. The immobilized S-Exos could be slowly and uniformly released and subsequently phagocytosed by BMSCs and macrophages. Once the S-Exos were phagocytosed, they could simultaneously activate the BMP/Smad signaling pathway in the BMSCs and promote macrophage M2 polarization, both of which enhance osseointegration. Specifically, this S-Exos coating exhibits a dual effect of promoting osseointegration, including the osseointegration of BMSCs by activating the BMP/Smad signaling pathway and the macrophage M2 polarization promoting osseointegration. In summary, the construction of S-Exos modified microarc oxide titanium implants could provide a new method for promoting osteointegration between the prosthesis and host bone in revision total hip arthroplasty.
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Exossomos , Osseointegração , Osseointegração/fisiologia , Osteogênese , Titânio/farmacologia , Titânio/metabolismo , Exossomos/metabolismo , Polietilenoimina/metabolismo , RNA Interferente Pequeno/metabolismo , Óxidos/metabolismoRESUMO
Background: Surgeons continue to seek indicators for the diagnosis of peri-prosthetic joint infection (PJI), which is a serious complication after total joint arthroplasty (TJA). Many recent studies have assessed the value of d-dimer in diagnosing PJI because of the close relation between the d-dimer value and inflammation. However, the conclusions from different studies are still disputed. Methods: We searched for studies published from 2011 to March 2020 using online databases and screened studies based on the inclusion criteria. Diagnostic parameters of d-dimer, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were calculated, including the pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR) and the area under the curve (AUC). In addition, univariate meta-regression and subgroup analyses were performed to identify sources of heterogeneity. Results: A total of nine studies with 431 Patients with PJI were included. The pooled sensitivity, specificity, DOR, and AUC of d-dimer were 0.82 (95% confidence interval [CI], 0.73-0.89), 0.73 (95% CI, 0.58-0.83), 12 (95% CI, 5-30), and 0.85 (95% CI, 0.82-0.88), respectively. In addition, the sensitivity, specificity, and AUC of CRP were 0.78 (95% CI, 0.73-0.83), 0.80 (95% CI, 0.73-0.86) and 0.85 (95% CI, 0.81-0.87), respectively, whereas those of ESR were 0.68 (95% CI, 0.60-0.74), 0.83 (95% CI, 0.75-0.88), and 0.80 (95% CI, 0.76-0.83), respectively. Conclusions: d-dimer determination had similar performance to CRP and ESR in the diagnosis of PJI and may be a good addition to the current diagnostic criteria.
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Infecções Relacionadas à Prótese , Biomarcadores , Produtos de Degradação da Fibrina e do Fibrinogênio , Humanos , Infecções Relacionadas à Prótese/diagnóstico , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Electrospinning is a common technology to construct tissue engineering scaffolds for bone regeneration. However, pure electrospun scaffolds do not enrich seed cells or promote their osteogenic differentiation. Biological functionalization of tissue engineering scaffolds is currently a hot research topic. Therefore, in this study, the bone marrow-derived mesenchymal cells (BM-MSC)-specific affinity peptide E7 and a bone morphogenic protein 2 (BMP-2) mimetic peptide were concomitantly conjugated onto the surface of an electrospun scaffold to construct a functional PEB scaffold. Characterization of PEB scaffolds revealed that both E7 and BMP-2 mimetic peptides were successfully conjugated onto the surface of electrospun scaffolds. With regard to biological activity, the PEB scaffold could synchronously promote adhesion and osteogenic differentiation of BM-MSC as a result of the co-delivery of E7 and BMP-2 mimetic peptides, which proved superior compared with the other three scaffolds. Consequently, the PEB scaffold offers a new concept for the construction of bone tissue engineering scaffolds.
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Osteonecrosis of the femoral head (ONFH) is a common osteological disease. Treatment of ONFH prior to the collapse of the femoral head is critical for increasing therapeutic efficiency. Tissue engineering therapy using bone mesenchymal stem cells (BMSCs) combined with a scaffold is a promising strategy. However, it is currently unclear how to improve the efficiency of BMSC recruitment under such conditions. In the present study, a specific cyclic peptide for SpragueDawley rat BMSCs, CTTNPFSLC (known as C7), was used, which was identified via phage display technology. Its high affinity for BMSCs was demonstrated using flow cytometry and fluorescence staining. Subsequently, the cyclic peptide was placed on ßtricalcium phosphate (ßTCP) scaffolds using absorption and freezedrying processes. Adhesion, expansion and proliferation of BMSCs was investigated in vitro on the C7treated ßTCP scaffolds and compared with pure ßTCP scaffolds. The results revealed that C7 had a promoting effect on the adhesion, expansion and proliferation of BMSCs on ßTCP scaffolds. Therefore, C7 may be effective in future tissue engineering therapy for ONFH.
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Fosfatos de Cálcio , Adesão Celular , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese , Peptídeos Cíclicos/farmacologia , Alicerces Teciduais/química , Animais , Proliferação de Células , Necrose da Cabeça do Fêmur/terapia , Células-Tronco Mesenquimais/fisiologia , Peptídeos Cíclicos/uso terapêutico , Ratos , Ratos Sprague-DawleyRESUMO
Osteonecrosis of the femoral head (ONFH) is a refractory disease present worldwide. In the development of therapies for this disease, mesenchymal stem cells (MSC) are a promising candidate cell source in tissue engineering (TE) and regenerative medicine. MSCs harvested from bone marrow (BM) are the gold standard. A significant barrier for BMMSCbased therapies is the inability and decreased number of BMMSCs in the tissues of interest. The ability to recruit BMMSCs efficiently to defective or injured sites in tissues or organs, for example the necrotic area of the femoral head in vivo, has been a major concern. In the present study, a peptide sequence (CDNVAQSVC), termed D7, was identified through phage display technology using C57BL/6 mouse BMMSCs. Subsequent analysis suggested that the identified loopconstrained heptapeptide exhibited a high specific affinity for mouse BMMSCs. Due to this specific affinity for BMMSCs, the present study provides a selective method to improve MSCbased TE strategies for the treatment of ONFH.
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Medula Óssea/metabolismo , Medula Óssea/fisiologia , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/fisiologia , Peptídeos Cíclicos/metabolismo , Animais , Células da Medula Óssea/metabolismo , Células da Medula Óssea/fisiologia , Células Cultivadas , Necrose da Cabeça do Fêmur/metabolismo , Necrose da Cabeça do Fêmur/terapia , Camundongos , Camundongos Endogâmicos C57BL , Osteogênese/fisiologia , Osteonecrose/metabolismo , Osteonecrose/terapia , Engenharia Tecidual/métodosRESUMO
Mesenchymal stem cells (MSCs) are often used in orthopedic tissue engineering, and bone marrowderived mesenchymal stem cells (BMSCs) are currently considered the gold standard. One of the most important issues in MSCbased tissue engineering therapy is the low number of MSCs in pathological tissues. Achieving efficient recruitment of MSCs to defective or damaged tissues in vivo has been a difficult hurdle. The aim of the present study was to construct a biomaterial that can effectively recruit BMSCs to facilitate the repair of pathological tissues. So functional ßtricalcium phosphate (ßTCP) was synthesized using the BMSC affinity peptide DPIYALSWSGMA (DPI) adsorbed onto ßTCP through an adsorption/freezedrying strategy. C57BL/6 mousederived BMSCs were seeded onto the DPI peptidemodified ßTCP (ßTCPDPI); in vitro experiments demonstrated that ßTCPDPI enhanced BMSC adhesion and proliferation compared with unmodified ßTCP. Results from the present study indicated that functional ßTCP may be used as an ideal scaffold in tissue engineering and in regenerative medicine.
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Fosfatos de Cálcio/farmacologia , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Fragmentos de Peptídeos/química , Animais , Células Cultivadas , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Alzheimer's disease (AD) is a common neurodegenerative disorder in elderly people, and is associated with a heavy financial burden on our society. The use of serologic biomarkers is an attractive method to diagnose AD. Although the determination of blood-based biomarkers for AD has been explored in many studies, few practical diagnosis methods have been used in the clinic. In this work, we constructed a "chemical tongue" sensor array that is easy to use and based on four kinds of fluorescent gold nanoclusters (Au NCs) for discriminating between multiple proteins at nanomolar concentrations. The device utilizes a linear discrimination analysis based on fluorescence intensity response patterns. Using this chemical tongue sensor array, multiple proteins can be confidently identified even in complex biological systems, such as human urine. Most importantly, sera of AD patients could be effectively discriminated from those of osteoarthritis patients, or of healthy people. Also, the results obtained for the AD patients by the chemical tongue sensor array were validated by CSF determination. We conclude that the chemical tongue sensor array manufactured in this work paves the way for designing an auxiliary diagnosis method for AD that is less invasive and more convenient for the large-scale screening of patients.
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Doença de Alzheimer/diagnóstico , Técnicas Biossensoriais , Proteínas Sanguíneas/análise , Ouro/química , Nanopartículas Metálicas/química , Osteoartrite/diagnóstico , Análise Serial de Proteínas/métodos , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/sangue , Doença de Alzheimer/fisiopatologia , Biomarcadores/sangue , Estudos de Casos e Controles , Diagnóstico Diferencial , Ácidos Graxos/química , Feminino , Fluorescência , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite/sangue , Osteoartrite/fisiopatologia , Análise Serial de Proteínas/instrumentação , Espectrometria de Fluorescência , Compostos de Sulfidrila/químicaRESUMO
Ti6Al4V alloy is widely used for hip joint prostheses, however owing to its lack of biomimetic surface properties, it often suffers from poor osseointegration. It is well known that bone mesenchymal stem cells (BMSCs) play an important role in the osseointegration of the host bone and joint prostheses. One promising approach to improving the osseointegration of joint prostheses is to enrich the number of BMSCs at the periprosthetic site. Previous studies have reported that BMSC specific affinity peptide E7, can specifically enrich BMSCs. However, to date, few studies have reported the use of E7 in bone tissue engineering. In this study, we conjugated E7 peptide to Ti6Al4V alloy to fabricate a scaffold (BTS) to improve the biocompatibility of the alloy. E7 peptide efficiently improved the adhesion of BMSCs to Ti6Al4V alloy. In addition, the BTS scaffold was more conducive to osteogenesis than the RGD-functionalized and non-functionalized control scaffolds. The functional BTS scaffold could pave the way for designing functional joint prostheses, which promote osseointegration between the host bone and implant.
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Wear particle-induced periprosthetic osteolysis is the main cause of aseptic loosening of orthopaedic implants. The aim of this study is to determine the protective effect of quercetin (QUE) against titanium (Ti) particle-induced ERS-related apoptosis and osteolysis. In this study, RAW264.7 cells were pretreated with different concentrations (40, 80, and 160 µmol/l) of QUE for 30 min and then treated with Ti particle (5 mg/ml) for 24 h. Cell viability and apoptosis were determined using MTT assay and Annexin V-FITC apoptosis detection kit, respectively. Protein and mRNA expressions of ERS-related genes were examined by western blot and real-time PCR, respectively. The release of inflammatory cytokines was detected by ELISA. Then a mouse calvarial osteolysis model was established. Histological sections of calvaria were stained with H&E or TRAP. The results showed that Ti particle reduced cell viability and induced apoptosis in RAW264.7 macrophages. The cytotoxic effects of Ti particle were dramatically inhibited by QUE pretreatment. Interestingly, we found that QUE also significantly reduced Ti particle-induced up-regulation of the expression levels of PERK, IRE1, GRP78, CHOP, caspase-12 and caspase-3 and enhanced the down-regulation of Bcl-2. In addition, QUE decreased Ti particle-induced inflammatory cytokines release from RAW264.7 cells. Moreover, treatment with QUE markedly decreased osteoclast number. In a mouse calvarial osteolysis model, QUE inhibited Ti particle-induced osteolysis in vivo by inhibiting osteoclast formation and expressions of ERS-related genes. In conclusion, QUE can protect RAW264.7 cells from Ti particle-induced ERS-related apoptosis and suppress calvarial osteolysis in vivo.
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Calcitonin (CT) is an anti-absorbent, which has long been used for treatment of osteoporosis. However, little information is available about the effects of CT on osteoarthritis (OA). This study was mainly aimed to explore the effects of CT on the treatment of OA, as well as the underlying mechanisms. Chondrocytes were isolated from immature mice and then were incubated with lipopolysaccharide (LPS), CT, small interfering (si) RNA against bone morphogenetic protein (BMP)-2, and/or the inhibitors of MAPK/Wnt/NF-κB pathway. Thereafter, cell viability, apoptosis, nitric oxide (NO) and inflammatory factors productions, and expression levels of cartilage synthesis protein key factors, cartilage-derived morphogenetic protein (CDMP) 1, SRY (sex-determining region Y)-box 9 protein (SOX9), and MAPK/Wnt/NF-κB pathways key factors were determined. CT significantly reversed LPS-induced cell viability decrease, apoptosis increase, the inflammatory factors and NO secretion, the abnormally expression of cartilage synthesis proteins and the activation of MAPK/Wnt/NF-κB pathways (P<0.05). In addition, we observed that administration of the inhibitors of MAPK/Wnt/NF-κB pathways statistically further increased the levels of CDMP1 and SOX9 (P<0.05). Suppression of BMP-2 decreased the levels of CDMP1 and SOX9 and activated MAPK/Wnt/NF-κB pathways, and could partially abolish CT-modulated the expression changes in CDMP1 and SOX9, and MAPK/Wnt/NF-κB pathways key factors (P<0.05). The results showed that CT protects chondrocytes from LPS-induced apoptosis and inflammatory response by regulating BMP-2 and thus blocking MAPK/Wnt/NF-κB pathways.