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BACKGROUND: This study introduced an Augmented Reality (AR) navigation system to address limitations in conventional high tibial osteotomy (HTO). The objective was to enhance precision and efficiency in HTO procedures, overcoming challenges such as inconsistent postoperative alignment and potential neurovascular damage. METHODS: The AR-MR (Mixed Reality) navigation system, comprising HoloLens, Unity Engine, and Vuforia software, was employed for pre-clinical trials using tibial sawbone models. CT images generated 3D anatomical models, projected via HoloLens, allowing surgeons to interact through intuitive hand gestures. The critical procedure of target tracking, essential for aligning virtual and real objects, was facilitated by Vuforia's feature detection algorithm. RESULTS: In trials, the AR-MR system demonstrated significant reductions in both preoperative planning and intraoperative times compared to conventional navigation and metal 3D-printed surgical guides. The AR system, while exhibiting lower accuracy, exhibited efficiency, making it a promising option for HTO procedures. The preoperative planning time for the AR system was notably shorter (4 min) compared to conventional navigation (30.5 min) and metal guides (75.5 min). Intraoperative time for AR lasted 8.5 min, considerably faster than that of conventional navigation (31.5 min) and metal guides (10.5 min). CONCLUSIONS: The AR navigation system presents a transformative approach to HTO, offering a trade-off between accuracy and efficiency. Ongoing improvements, such as the incorporation of two-stage registration and pointing devices, could further enhance precision. While the system may be less accurate, its efficiency renders it a potential breakthrough in orthopedic surgery, particularly for reducing unnecessary harm and streamlining surgical procedures.
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Postmenopausal osteoporosis (PMOP) is a metabolic disorder characterized by the loss of bone density, which increases the risk of developing complications such as fractures. A pivotal factor contributing to the onset of PMOP is the diminished osteogenic differentiation capacity of bone marrow mesenchymal stem cells (BMSCs). MicroRNAs (miRNAs) play a substantial role in this process; however, their specific impact on regulating BMSCs osteogenesis remains unclear. Studies have evidenced a reduced expression of miR-18a-5p in PMOP, and concomitantly, our observations indicate an augmented expression of miR-18a-5p during the osteogenic differentiation of BMSCs. This investigation seeks to elucidate the regulatory influence of miR-18a-5p on BMSC osteogenic differentiation and the underlying mechanisms. In vitro experiments demonstrated that the overexpression of miR-18a-5p facilitated the osteogenic differentiation of BMSCs, while the downregulation of miR-18a-5p yielded converse outcomes. Mechanistically, We employed bioinformatics techniques to screen out the target gene Notch2 of miR-18a-5p. Subsequently, dual-luciferase reporter gene assays and rescue experiments substantiated that miR-18a-5p promotes BMSC osteogenic differentiation by suppressing Notch2. Finally, miR-18a-5p was overexpressed via adenovirus injection into the femoral bone marrow cavity, with results demonstrating its capability to enhance osteogenic differentiation and alleviate PMOP symptoms. Our findings disclose that miR-18a-5p fosters osteogenic differentiation of BMSC by inhibiting Notch2, thereby offering novel targets and strategies for PMOP treatment.
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Background: Diabetic bone defects present significant challenges for individuals with diabetes. While metformin has been explored for bone regeneration via local delivery, its application in treating diabetic bone defects remains under-explored. In this study, we aim to leverage 3D printing technology to fabricate a GelMA-Nanoclay hydrogel scaffold loaded with metformin specifically for this purpose. The objective is to assess whether the in situ release of metformin can effectively enhance osteogenesis, angiogenesis, and immunomodulation in the context of diabetic bone defects. Materials and methods: Utilizing 3D printing technology, we constructed a GelMA-Nanoclay-Metformin hydrogel scaffold with optimal physical properties and biocompatibility. The osteogenic, angiogenic, and immunomodulatory characteristics of the hydrogel scaffold were thoroughly investigated through both in vitro and in vivo experiments. Results: GelMA10%-Nanoclay8%-Metformin5mg/mL was selected as the bioink for 3D printing due to its favorable swelling rate, degradation rate, mechanical strength, and drug release rate. Through in vitro investigations, the hydrogel scaffold extract, enriched with metformin, demonstrated a substantial enhancement in the proliferation and migration of BMSCs within a high-glucose microenvironment. It effectively enhances osteogenesis, angiogenesis, and immunomodulation. In vivo experimental outcomes further underscored the efficacy of the metformin-loaded GelMA-Nanoclay hydrogel scaffold in promoting superior bone regeneration within diabetic bone defects. Conclusions: In conclusion, while previous studies have explored local delivery of metformin for bone regeneration, our research is pioneering in its application to diabetic bone defects using a 3D printed GelMA-Nanoclay hydrogel scaffold. This localized delivery approach demonstrates significant potential for enhancing bone regeneration in diabetic patients, offering a novel approach for treating diabetic bone defects. The translational potential of this article: Our study demonstrates, for the first time, the successful loading of the systemic antidiabetic drug metformin onto a hydrogel scaffold for localized delivery. This approach exhibits significant efficacy in mending diabetic bone defects, presenting a promising new avenue for the treatment of such conditions.
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Osteoporosis is a major public health problem with an urgent need for safe and effective therapeutic interventions. The process of shell formation in oysters is similar to that of bone formation in mammals, and oyster extracts have been proven to exert osteoprotective effects. Oyster mantle is the most crucial organ regulating shell formation, in which exosomes play an important role. However, the effects of oyster mantle-derived exosomes (OMEs) on mammalian osteoporosis and the underlying mechanisms remain unknown. The OMEs investigated herein was found to carry abundant osteogenic cargos. They could also survive hostile gastrointestinal conditions and accumulate in the bones following oral administration. Moreover, they promoted osteoblastic differentiation and inhibited osteoclastic differentiation simultaneously. Further mechanistic examination revealed that OMEs likely promoted osteogenic activity by activating PI3K/Akt/ß-catenin pathway in osteoblasts and blunted osteoclastic activity by inhibiting NF-κB pathway in osteoclasts. These favorable pro-osteogenic effects of OMEs were also corroborated in a rat femur defect model. Importantly, oral administration of OMEs effectively attenuated bone loss and improved the bone microstructure in ovariectomy-induced osteoporotic mice, and demonstrating excellent biosafety. The mechanistic insights from our data support that OMEs possess promising therapeutic potential against osteoporosis.
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Exosomas , Homeostasis , Osteoblastos , Osteogénesis , Osteoporosis , Ostreidae , Animales , Exosomas/metabolismo , Osteoporosis/metabolismo , Osteoporosis/tratamiento farmacológico , Osteoporosis/patología , Osteogénesis/efectos de los fármacos , Femenino , Ratones , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Homeostasis/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Diferenciación Celular/efectos de los fármacos , Ratas Sprague-Dawley , Huesos/efectos de los fármacos , Huesos/metabolismo , Huesos/patología , Exoesqueleto/química , Ratas , Ratones Endogámicos C57BL , Células RAW 264.7 , Ovariectomía , Fémur/efectos de los fármacos , Fémur/patología , Fémur/metabolismoRESUMEN
Bioprinting that can synchronously deposit cells and biomaterials has lent fresh impetus to the field of tissue regeneration. However, the unavoidable occurrence of cell damage during fabrication process and intrinsically poor mechanical stability of bioprinted cell-laden scaffolds severely restrict their utilization. As such, on basis of heart-inspired hollow hydrogel-based scaffolds (HHSs), a mechanical-assisted post-bioprinting strategy is proposed to load cells into HHSs in a rapid, uniform, precise and friendly manner. HHSs show mechanical responsiveness to load cells within 4 s, a 13-fold increase in cell number, and partitioned loading of two types of cells compared with those under static conditions. As a proof of concept, HHSs with the loading cells show an enhanced regenerative capability in repair of the critical-sized segmental and osteoporotic bone defects in vivo. We expect that this post-bioprinting strategy can provide a universal, efficient, and promising way to promote cell-based regenerative therapy.
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Bioimpresión , Regeneración Ósea , Hidrogeles , Ingeniería de Tejidos , Andamios del Tejido , Animales , Andamios del Tejido/química , Hidrogeles/química , Bioimpresión/métodos , Ingeniería de Tejidos/métodos , Humanos , Huesos , Ratones , Células Madre Mesenquimatosas/citología , Materiales Biocompatibles/química , Osteoporosis/terapiaRESUMEN
Due to persistent inflammation and limited osteogenesis, jawbone defects present a considerable challenge in regenerative medicine. Amelogenin, a major protein constituent of the developing enamel matrix, demonstrates promising capabilities in inducing regeneration of periodontal supporting tissues and exerting immunomodulatory effects. These properties render it a potential therapeutic agent for enhancing jawbone osteogenesis. Nevertheless, its clinical application is hindered by the limitations of monotherapy and its rapid release characteristics, which compromise its efficacy and delivery efficiency. In this context, calcium alginate hydrogel, recognized for its superior physicochemical properties and biocompatibility, emerges as a candidate for developing a synergistic bioengineered drug delivery system. This study describes the synthesis of an injectable calcium amelogenin/calcium alginate hydrogel using calcium alginate loaded with amelogenin. We comprehensively investigated its physical properties, its role in modulating the immunological environment conducive to bone healing, and its osteogenic efficacy in areas of jawbone defects. Our experimental findings indicate that this synthesized composite hydrogel possesses desirable mechanical properties such as injectability, biocompatibility, and biodegradability. Furthermore, it facilitates jawbone formation by regulating the bone-healing microenvironment and directly inducing osteogenesis. This research provides novel insights into the development of bone-tissue regeneration materials, potentially advancing their clinical application.
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Introduction: Bones are easily damaged. Biomimetic scaffolds are involved in tissue engineering. This study explored polydopamine (PDA)-coated poly lactic-co-glycolic acid (PLGA)-magnesium oxide (MgO) scaffold properties and its effects on bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation. Methods: PLGA/MgO scaffolds were prepared by low-temperature 3D printing technology and PDA coatings were prepared by immersion method. Scaffold structure was observed by scanning electron microscopy with an energy dispersive spectrometer (SEM-EDS), fourier transform infrared spectrometer (FTIR). Scaffold hydrophilicity, compressive/elastic modulus, and degradation rates were analyzed by water contact angle measurement, mechanical tests, and simulated-body fluid immersion. Rat BMSCs were cultured in scaffold extract. Cell activity on days 1, 3, and 7 was detected by MTT. Cells were induced by osteogenic differentiation, followed by evaluation of alkaline phosphatase (ALP) activity on days 3, 7, and 14 of induction and Osteocalcin, Osteocalcin, and Collagen I expressions. Results: The prepared PLGA/MgO scaffolds had dense microparticles. With the increase of MgO contents, the hydrophilicity was enhanced, scaffold degradation rate was accelerated, magnesium ion release rate and scaffold extract pH value were increased, and cytotoxicity was less when magnesium mass ratio was less than 10%. Compared with other scaffolds, compressive and elastic modulus of PLGA/MgO (10%) scaffolds were increased; BMSCs incubated with PLGA/MgO (10%) scaffold extract had higher ALP activity and Osteocalcin, Osteopontin, and Collagen I expressions. PDA coating was prepared in PLGA/MgO (10%) scaffolds and the mechanical properties were not affected. PLGA/MgO (10%)/PDA scaffolds had better hydrophilicity and biocompatibility and promoted BMSC osteogenic differentiation. Conclusion: Low-temperature 3D printing PLGA/MgO (10%)/PDA scaffolds had good hydrophilicity and biocompatibility, and were conducive to BMSC osteogenic differentiation.
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BACKGROUND: Bone mesenchymal stem cell (BMSC)-derived exosomes (B-exos) are attractive for applications in enabling alloantigen tolerance. An in-depth mechanistic understanding of the interaction between B-exos and dendritic cells (DCs) could lead to novel cell-based therapies for allogeneic transplantation. OBJECTIVES: To examine whether B-exos exert immunomodulatory effects on DC function and maturation. MATERIAL AND METHODS: After mixed culture of BMSCs and DCs for 48 h, DCs from the upper layer were collected to analyze the expression levels of surface markers and mRNAs of inflammation-related cytokines. Then, before being collected to detect the mRNA and protein expression levels of indoleamine 2,3-dioxygenase (IDO), the DCs were co-cultured with B-exos. Then, the treated DCs from different groups were co-cultured with naïve CD4+ T cells from the mouse spleen. The proliferation of CD4+ T cells and the proportion of CD4+CD25+Foxp3+ T cells were analyzed. Finally, the skins of BALB/c mice were transplanted to the back of C57 mice in order to establish a mouse allogeneic skin transplantation model. RESULTS: The co-culture of DCs with BMSCs downregulated the expression of the major histocompatibility complex class II (MHC-II) and CD80/86 costimulatory molecules on DCs. Moreover, B-exos increased the expression of IDO in DCs treated with lipopolysaccharide (LPS). The proliferation of CD4+CD25+Foxp3+ T cells increased when cultured with B-exos-exposed DCs. Finally, mice recipients injected with B-exos-treated DCs had significantly prolonged survival after receiving the skin allograft. CONCLUSIONS: Taken together, these data suggest that the B-exos suppress the maturation of DCs and increase the expression of IDO, which might shed light on the role of B-exos in inducing alloantigen tolerance.
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Exosomas , Células Madre Mesenquimatosas , Ratones , Animales , Exosomas/metabolismo , Trasplante Homólogo , Células Dendríticas , Factores de Transcripción Forkhead/metabolismo , Células de la Médula ÓseaRESUMEN
Background: In the bone marrow microenvironment of postmenopausal osteoporosis (PMOP), bone marrow mesenchymal stem cell (BMSC)-derived exosomal miRNAs play an important role in bone formation and bone resorption, although the pathogenesis has yet to be clarified. Methods: BMSC-derived exosomes from ovariectomized rats (OVX-Exo) and sham-operated rats (Sham-Exo) were co-cultured with bone marrow-derived macrophages to study their effects on osteoclast differentiation. Next-generation sequencing was utilized to identify the differentially expressed miRNAs (DE-miRNAs) between OVX-Exo and Sham-Exo, while target genes were analyzed using bioinformatics. The regulatory effects of miR-27a-3p and miR-196b-5p on osteogenic differentiation of BMSCs and osteoclast differentiation were verified by gain-of-function and loss-of-function analyses. Results: Osteoclast differentiation was significantly enhanced in the OVX-Exo treatment group compared to the Sham-Exo group. Twenty DE-miRNAs were identified between OVX-Exo and Sham-Exo, among which miR-27a-3p and miR-196b-5p promoted the expressions of osteogenic differentiation markers in BMSCs. In contrast, knockdown of miR-27a-3p and miR-196b-5p increased the expressions of osteoclastic markers in osteoclast. These 20 DE-miRNAs were found to target 11435 mRNAs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that these target genes were involved in several biological processes and osteoporosis-related signaling pathways. Conclusion: BMSC-derived exosomal miR-27a-3p and miR-196b-5p may play a positive regulatory role in bone remodeling.
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Remodelación Ósea , Células Madre Mesenquimatosas , MicroARNs , Animales , Ratas , Resorción Ósea/metabolismo , Diferenciación Celular/genética , Células Madre Mesenquimatosas/fisiología , MicroARNs/genética , Osteogénesis/genética , Femenino , Remodelación Ósea/genética , Exosomas/genéticaRESUMEN
Zinc is becoming one of the leading candidate materials for biodegradable orthopedic implants owing to its attractive properties in terms of degradation behavior and mechanical properties. However, the insufficient surface bio-activities postpone its clinical application. In this study, an organic-inorganic collagen entrapped calcium/zinc phosphates coating was constructed on Zn surface to lessen Zn2+ releasing rate and to leverage the surface osteogenic and angiogenic properties. Collagen molecules were immobilized onto Zn substrate and subsequently coordinated with calcium and zinc ions to promote the CaZnP inorganic phase growth, ensuing an intertwined collagen-CaZnP hybrid system. Consequently, the hybrid coating was highly coalesced and compact. Such high quality warranted the contained Zn2+ releasing in a tolerable rate favorable for cells viability. The collagen-CaZnP coated Zn showed remarkedly stronger osteogenicity as compared to the untreated Zn, ascertained by the MC3T3-E1 osteoblast cell proliferation and differentiation assays, such as alkaline phosphatase expression and calcium nodule formation results. In addition, this hybrid coating supported human umbilical vein endothelial cells (HUVECs) migration and tube formation. The enhanced osteogenic and angiogenic properties could be ascribed to the nature of collagen and calcium/zinc phosphate components, the hybrid micro/nano-structure as well as the ability of controlling the Zn2+ release of Zn substrate into a suitable concentration range. Our strategy provides a new avenue to surface modification of biodegradable metals for bone regenerative perspective.
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Osteogénesis , Zinc , Implantes Absorbibles , Calcio , Colágeno/farmacología , Células Endoteliales de la Vena Umbilical Humana , Humanos , Fosfatos/farmacología , Zinc/farmacologíaRESUMEN
Compared to traditional postoperative radiation and chemotherapy, immune checkpoint blockade (ICB) therapy demonstrates superiority by provoking own immune system to cure cancer completely even for some terminally ill patients. However, systemic administration of ICB is liable to cause severe immunity inflammation or immune storm. Here, an injectable, near infrared (NIR) responsive, multifunctional nanocomposite thermogel as a local ICB delivery system for cancer postsurgical therapy is proposed. By copolymerization of thermosensitive and zwitterionic monomer, the injectable thermogel with adjustable sol-gel transition temperature is obtained. Afterward, combined with functional mesoporous nanoparticles, the platform can absorb NIR light and transfer it into heat. The generated heat will promote retro Diels-Alder (D-A) reaction to degrade coating layer on nanoparticle, achieving NIR controlled ICB release. Furthermore, the local ICB delivery system is applied on an osteosarcoma postsurgical recurrence model and results indicate the platform with favorable biocompatibility can avoid early leakage of cargos and greatly increase drug content at tumor site. Besides, long-term controlled ICB release of the system effectively improves the amount of active T cells, resulting in excellent antitumor recurrence effect. Overall, this work suggests the local injectable nanocomposite thermogel is expected to be a promising tool for cancer postoperative therapy.
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Neoplasias Óseas , Nanocompuestos , Nanopartículas , Osteosarcoma , Humanos , Nanocompuestos/uso terapéutico , PolimerizacionRESUMEN
Integration of biological factors and hierarchical rigid scaffolds is of great interest in bone tissue engineering for fabrication of biomimetic constructs with high physical and biological performance for enhanced bone repair. Core/shell microspheres (CSMs) delivering bone morphogenetic protein-2 (BMP-2) and a strategy to integrate CSMs with 3D-printed scaffolds were developed herein to form a hybrid 3D system for bone repair. The scaffold was printed with polycaprolactone (PCL) and then coated with polydopamine. Shells of CSMs were electrosprayed with alginate. Cores were heparin-coated polylactic acid (PLA) microparticles fabricated via simple emulsion and heparin coating strategy. Assembly of microspheres and scaffolds was realized via a self-locking method with the assistance of controlled expansion of CSMs. The hybrid system was evaluated in the rat critical-sized bone defect model. CSMs released BMP-2 in a tunable manner and boosted osteogenic performance in vitro. CSMs were then successfully integrated inside the scaffolds. The assembled system effectively promoted osteogenesis in vitro and in vivo. These observations show the importance of how BMP-2 is delivered, and the core/shell microspheres represent effective BMP-2 carriers that could be integrated into scaffolds, together forming a hybrid system as a promising candidate for enhanced bone regeneration.
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Impresión Tridimensional , Andamios del Tejido , Animales , Heparina , Microesferas , Poliésteres , RatasRESUMEN
BACKGROUND: The most common complication of oblique lumbar interbody fusion (OLIF) is endplate fracture/subsidence. The aim of this study was to evaluate biomechanical stability in patients undergoing OLIF surgery with anterolateral screw fixation (ASF). METHODS: Based on a previously validated model technique, L4-L5 functional surgical models corresponding to the ASF and bilateral pedicle screw fixation (BPSF) methods were created. Finite element models were developed to compare the biomechanics of the ASF and BPSF groups. We retrospectively analyzed 18 patients with lumbar degenerative diseases who underwent OLIF with ASF in Shenzhen Hospital of Southern Medical University from April 2020 to April 2021. Intraoperative and postoperative complications were observed. RESULTS: Compared with the BPSF model, the maximum stresses of the L4 inferior endplate and L5 superior endplate were greatly increased in the ASF model. The contact surface between the vertebrae and screw (CSVS) in the ASF model produced nearly 100% more stress than the BPSF model at all moments. In clinical practice, after a 12-month follow-up, 7 adverse events were observed, including 3 cases of left thigh pain/numbness, 3 cases of cage subsidence, and 1 case of screw loosening. CONCLUSIONS: OLIF surgery with ASF could not reduce the maximum stresses on the endplate and CSVS, which may be a potential risk factor for cage subsidence and screw loosening.
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Tornillos Pediculares , Fusión Vertebral , Fenómenos Biomecánicos , Humanos , Vértebras Lumbares/cirugía , Región Lumbosacra/cirugía , Estudios Retrospectivos , Fusión Vertebral/métodosRESUMEN
Background: Osteosarcoma (OS) is the most common primary malignant bone tumour in children and adolescents, with rapid growth, frequent metastasis, and a poor prognosis, but its pathogenesis has not been fully elucidated. Exploring the pathogenesis of OS is of great significance for improving diagnoses and finding new therapeutic targets. Methods: Differentially expressed circRNAs (DECs), miRNAs (DEMs), methylated DNA sites (DMSs), and mRNAs (DEGs) were identified between OS and control cell lines. GSEA of DEGs and functional enrichment analysis of methylated DEGs were carried out to further identify potential biological processes. Online tools were used to predict the miRNA binding sites of DECs and the mRNA binding sites of DEMs, and then construct a circRNA-miRNA-mRNA network. Next, an analysis of the interaction between methylated DEGs was performed with a protein-protein interaction (PPI) network, and hub gene identification and survival analysis were carried out. The expression pattern of circRNA-miRNA-mRNA was validated by real-time PCR. Results: GSEA and functional enrichment analysis indicated that DEGs and methylated DEGs are involved in important biological processes in cancer. Hsa_circ_0001753/has_miR_760/CD74 network was constructed and validated in cell lines. Low expression levels of CD74 are associated with poor overall survival times and show good diagnostic ability. Conclusion: Methylated DEGs may be involved in the development of OS, and the hsa_circ_0001753/has_miR_760/CD74 network may serve as a target for the early diagnosis of and targeted therapy for OS.
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The ubiquitin-proteasome system (UPS) is an essential pathway that regulates the homeostasis and function of intracellular proteins and is a crucial protein-degradation system in osteoblast differentiation and bone formation. Abnormal regulation of ubiquitination leads to osteoblast differentiation disorders, interfering with bone formation and ultimately leading to osteoporosis. E3 ubiquitin ligases (E3) promote addition of a ubiquitin moiety to substrate proteins, specifically recognizing the substrate and modulating tyrosine kinase receptors, signaling proteins, and transcription factors involved in the regulation of osteoblast proliferation, differentiation, survival, and bone formation. In this review, we summarize current progress in the understanding of the function and regulatory effects of E3 ligases on the transcription factors and signaling pathways that regulate osteoblast differentiation and bone formation. A deep understanding of E3 ligase-mediated regulation of osteoblast differentiation provides a scientific rationale for the discovery and development of novel E3-targeting therapeutic strategies for osteoporosis.
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Osteosarcoma (OS) is the most common primary bone tumor worldwide. OS exhibits a range of aggressive behaviors, including early metastasis potential, rapid progression, poor clinical prognosis and insensitivity to chemoradiotherapy. Noncoding RNAs are transcripts that do not encode proteins. A significant number of studies published on OS have been focused on the aberrant expression of noncoding RNAs and their involvement in tumor initiation and progression. It has been confirmed that noncoding RNAs exert their regulatory functions at both the transcriptional and posttranscriptional level, which leads to tumor initiation or progression in OS. According to present knowledge, this review provides a stateoftheart overview of the functions and mechanisms of microRNAs, long noncoding RNAs and circular RNAs in terms of their involvement with OS. The review also covers their potential clinical application in the diagnosis, prognosis and treatment of OS. It is hoped that the information presented in this review on the involvement of noncoding RNAs in OS will lead to a more comprehensive understanding of OS and provide a useful perspective on the potential diagnostic and therapeutic applications of noncoding RNAs for patients with OS.
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Biomarcadores de Tumor/genética , Neoplasias Óseas/genética , Osteosarcoma/genética , ARN no Traducido/genética , Neoplasias Óseas/diagnóstico , Neoplasias Óseas/tratamiento farmacológico , Detección Precoz del Cáncer , Regulación Neoplásica de la Expresión Génica , Humanos , MicroARNs/genética , Osteosarcoma/diagnóstico , Osteosarcoma/tratamiento farmacológico , Pronóstico , ARN Circular/genética , ARN Largo no Codificante/genéticaRESUMEN
Dendritic cells release bioactive exosomes involved in immune regulation. Long noncoding RNAs (lncRNAs) are implicated in a number of immunoregulatory mechanisms. However, the roles of lncRNAs in dendritic cellderived exosomes remain to be elucidated. The present study aimed to investigate the roles of lncRNAs in exosomes derived from mature and immature dendritic cells and to find specific lncRNAs with immunoregulatory function. The expression profiles of lncRNAs in exosomes derived from bone marrow dendritic cells of C57 mice were illustrated. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses and Gene Set Enrichment Analysis were performed to identify potential targets correlated with immune regulation. In addition, lncRNAmiRNAmRNA networks were predicted using bioinformatics methods. Representative lncRNAs were further validated via reverse transcriptionquantitative PCR. A total of 437 lncRNAs were analyzed using RNAseq. Among these, the expression of ~87 lncRNAs was upregulated and 21 lncRNAs was downregulated in mature dendritic cellderived exosomes (Dex) compared with immature Dex. GO analyses indicated the involvement of upregulated lncRNAs in multiple biological functions, such as the immune system process, while downregulated lncRNAs were involved in poly(A) RNA binding. Analysis of the KEGG pathway identified the relationship of TNF signaling and ribosome pathway with upregulated lncRNAs and downregulated lncRNAs, respectively. The results of gene set enrichment analysis identified that three lncRNAassociated transcripts (Procr203, Clec4e202 and Traf1203) were highly associated with immunoregulatory functions including T helper cell differentiation and Janus kinaseSTAT signaling pathway. The results indicated the involvement of candidate lncRNAs in immunoregulation and suggested a new perspective on the modulation of lncRNAs in Dex.
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Células Dendríticas/metabolismo , Exosomas/genética , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Animales , Células de la Médula Ósea/metabolismo , Diferenciación Celular , Células Cultivadas , Biología Computacional , Células Dendríticas/citología , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Ontología de Genes , Redes Reguladoras de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Masculino , Ratones Endogámicos C57BL , MicroARNs/metabolismo , ARN Largo no Codificante/inmunología , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de SeñalRESUMEN
BACKGROUND: The most common complication of oblique lumbar interbody fusion (OLIF) is endplate fracture/subsidence. The mechanics of endplate fracture in OLIF surgery are still unclear. The aim of the present study was to evaluate the biomechanical stability in patients undergoing OLIF surgery with stand-alone (SA) and bilateral pedicle screw fixation (BPSF) methods. METHODS: A finite element model of the L1-L5 spinal unit was established and validated. Using the validated model technique, L4-L5 functional surgical models corresponding to the SA and BPSF methods were created. Simulations using the models were performed to investigate OLIF surgery. A 500-N compression force was applied to the superior surface of the model to represent the upper body weight, and a 7.5-Nm moment was applied to simulate the 6 movement directions of the lumbar spinal model: flexion and extension, right and left lateral bending, and right and left axial rotation. Finite element models were developed to compare the biomechanics of the SA and BPSF groups. RESULTS: Compared with the range of motion of the intact lumbar model, that of the SA model was decreased by 79.6% in flexion, 54.5% in extension, 57.2% in lateral bending, and 50.0% in axial rotation. The BPSF model was decreased by 86.7% in flexion, 77.3% in extension, 76.2% in lateral bending, and 75.0% in axial rotation. Compared with the BPSF model, the maximum stresses of the L4 inferior endplate and L5 superior endplate were greatly increased in the SA model. The L4 inferior endplate stress was increased to 49.7 MPa in extension, and the L5 superior endplate stress was increased to 47.7 MPa in flexion, close to the yield stress of the lamellar bone (60 MPa). CONCLUSIONS: OLIF surgery with BPSF could reduce the maximum stresses on the endplate, which might reduce the incidence of cage subsidence. OLIF surgery with the SA method produced more stress compared with BPSF, especially in extension and flexion, which might be a potential risk factor for cage subsidence.
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Análisis de Elementos Finitos , Vértebras Lumbares/cirugía , Región Lumbosacra/cirugía , Tornillos Pediculares , Fenómenos Biomecánicos/fisiología , Humanos , Vértebras Lumbares/patología , Región Lumbosacra/patología , Modelos Anatómicos , Rango del Movimiento Articular/fisiología , Fusión Vertebral/métodosRESUMEN
Three-dimensional (3D) bioprinting is an extremely convenient biofabrication technique for creating biomimetic tissue-engineered bone constructs and has promising applications in regenerative medicine. However, existing bioinks have shown low mechanical strength, poor osteoinductive ability, and lacking a suitable microenvironment for laden cells. Nanosilicate (nSi) has shown to be a promising biomaterial, due to its unique properties such as excellent biocompatibility, degrade into nontoxic products, and with osteoinductive properties, which has been used in bone bioprinting. However, the long term bone healing effects and associating risks, if any, of using nSi in tissue engineering bone scaffolds in vivo are unclear and require a more thorough assessment prior to practical use. Hence, a functional and biomimetic nanocomposite bioink composed of rat bone marrow mesenchymal stem cells (rBMSCs), nSi, gelatin and alginate for the 3D bioprinting of tissue-engineered bone constructs is firstly demonstrated, mimicking the structure of extracellular matrix, to create a conducive microenvironment for encapsulated cells. It is shown that the addition of nSi significantly increases the printability and mechanical strength of fabricated human-scale tissue or organ structures (up to 15 mm height) and induces osteogenic differentiation of the encapsulated rBMSCs in the absence of in vitro osteoinductive factors. A systematic in vivo research of the biomimetic nanocomposite bioink scaffolds is further demonstrated in a rat critical-size (8 mm) bone defect-repair model. The in vivo results demonstrate that the 3D bioprinted nanocomposite scaffolds can significantly promote the bone healing of the rat calvarial defects compared to other scaffolds without nSi or cells, and show rarely side effects on the recipients. Given the above advantageous properties, the 3D bioprinted nanocomposite scaffolds can greatly accelerate the bone healing in critical bone defects, thus providing a clinical potential candidate for orthopedic applications.