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INTRODUCTION: Fractures of the anterior process of the calcaneus (APC) are easily overlooked in clinical practice. Most patients have good to excellent clinical outcome after conservative treatment, while some patients may have persisting symptoms and unfavorable functional outcomes. The aim of this study was to identify the risk factors associated with unfavorable functional outcome after conservative treatment in APC fractures. METHODS: All patients presenting with APC fractures and receiving conservative treatment from April 2019 to April 2020 were retrospectively assessed. The primary outcome measurement was the ankle joint function assessed using Karlsson Scoring System at 2 years post-injury. The risk factors associated with unfavorable functional outcomes (Karlsson score ≤ 80) were evaluated by logistic regression analysis. RESULTS: In total, 84 patients were included with a mean age of 40 years. 26 (31%) patients presented with unfavorable functional outcome at 2 years post-injury. In multivariate logistic regression, concomitant fractures of talonavicular (TN) joints and older age were significantly associated with unfavorable functional outcome (p<0.05). Patients with concomitant fractures of TN joints had an odds ratio of 3.623 for unfavorable functional outcome. The optimal cutoff age for an unfavorable outcome was ≥ 47.5 years, with an odds ratio of 5.010. CONCLUSION: Most patients with APC fractures achieved good to excellent results when treated conservatively. Attention should be paid to those with concomitant fractures of TN joints and with age ≥ 47.5 years, which might lead to unfavorable functional recovery. LEVEL OF EVIDENCE: IV; case series.
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Traumatismos del Tobillo , Calcáneo , Traumatismos de los Pies , Fracturas Óseas , Traumatismos de la Rodilla , Humanos , Adulto , Persona de Mediana Edad , Calcáneo/diagnóstico por imagen , Calcáneo/lesiones , Estudios Retrospectivos , Fijación Interna de Fracturas/métodos , Extremidad Inferior , Resultado del TratamientoRESUMEN
Purpose: Delayed skin healing in diabetic wounds is a major clinical problem. The tRNA-derived small RNAs (tsRNAs) were reported to be associated with diabetes. However, the role of tsRNAs in diabetic wound healing is unclear. Our study was designed to explore the tsRNA expression profile and mine key potential tsRNAs and their mechanism in diabetic wounds. Methods: Skin tissues of patients with diabetic foot ulcers and healthy controls were subjected to small RNA sequencing. The role of candidate tsRNA was explored by loss- and gain-of-function experiments in HUVECs. Results: A total of 55 differentially expressed tsRNAs were identified, including 12 upregulated and 43 downregulated in the diabetes group compared with the control group. These tsRNAs were mainly concentrated in intercellular interactions and neural function regulation in GO terms and enriched in MAPK, insulin, FoxO, calcium, Ras, ErbB, Wnt, T cell receptor, and cGMP-PKG signaling pathways. tRF-Gly-CCC-039 expression was upregulated in vivo and in vitro in the diabetic model. High glucose disturbed endothelial function in HUVECs, and tRF-Gly-CCC-039 mimics further harmed HUVECs function, characterized by the suppression of proliferation, migration, tube formation, and the expression of Coll1a1, Coll4a2, and MMP9. Conversely, the tRF-Gly-CCC-039 inhibitor could attenuate high-glucose-induced endothelial injury to HUVECs. Conclusion: We investigated the tsRNAs expression profile in diabetic foot ulcers and defined the impairment role of tRF-Gly-CCC-039 in endothelial function in HUVECs. This study may provide novel insights into accelerating diabetic skin wound healing.
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Aims: The osteogenesis of human bone marrow mesenchymal stem cells (hBMSCs) plays a critical role in fracture healing. Osteogenic differentiation is regulated by a variety of post-translational modifications, but the function of protein palmitoylation in osteogenesis remains largely unknown. Methods: Osteogenic differentiation induction of hBMSCs was used in this study. RTâqPCR and immunoblotting assays (WB) were used to test marker genes of osteogenic induction. Alkaline phosphatase (ALP) activity, ALP staining and Alizarin red staining were performed to evaluate osteogenesis of hBMSCs. Signal finder pathway reporter array, co-immunoprecipitation and WB were applied to elucidate the molecular mechanism. A mouse fracture model was used to verify the in vivo function of the ZDHHC inhibitor. Key findings: We revealed that palmitic acid inhibited Runx2 mRNA expression in hBMSCs and identified ZDHHC16 as a potential target palmitoyl acyltransferase. In addition, ZDHHC16 decreased during osteogenic induction. Next, we confirmed the inhibitory function of ZDHHC16 by its knockdown or overexpression during osteogenesis of hBMSCs. Moreover, we illustrated that ZDHHC16 inhibited the phosphorylation of CREB, thus inhibiting osteogenesis of hBMSCs by enhancing the palmitoylation of CREB. With a mouse femur fracture model, we found that 2-BP, a general inhibitor of ZDHHCs, promoted fracture healing in vivo. Thus, we clarified the inhibitory function of ZDHHC16 during osteogenic differentiation. Significance: Collectively, these findings highlight the inhibitory function of ZDHHC16 in osteogenesis as a potential therapy method for fracture healing.
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Accelerating angiogenesis of diabetic wounds is crucial to promoting wound healing. Currently, vascular endothelial growth factor (VEGF), an angiogenesis-related bioactive molecule, is widely used in clinic to enhance wound angiogenesis, but it faces problems of inactivation and low utilization due to harsh microenvironment. Here, we developed a novel reactive oxygen species (ROS)-scavenging hydrogel aimed to polarize macrophages toward an anti-inflammatory phenotype, inducing efficient angiogenesis in diabetic wounds. This composite hydrogel with good biosafety and mechanical properties showed sustainable release of bioactive VEGF. Importantly, it could significantly reduce ROS level and rapidly improve wound microenvironment, which ensured the activity of VEGF in vitro and in vivo and successful healing eventually. At the same time, the composite hydrogel exhibited excellent antibacterial properties. In vivo results confirmed good anti-inflammatory, stimulated vascularization and accelerated wound healing attributed to the novel ROS-scavenging hydrogel, which might serve as a promising wound dressing in diabetic wound healing.
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Diabetes Mellitus , Hidrogeles , Humanos , Especies Reactivas de Oxígeno , Factor A de Crecimiento Endotelial Vascular/metabolismo , Cicatrización de Heridas , AntiinflamatoriosRESUMEN
BACKGROUND: Fractures of the lateral process of the talus (LTPF) are rare and only rarely are associated ligamentous injuries. The injury mechanism is commonly considered to be similar with ankle sprains, where excessive varus of the hindfoot leads to avulsion fractures of the lateral process of the talus. However, previous cadaveric studies have suggested that LTPF was more likely to be caused by eversion or external rotation force with dorsiflexion of the ankle. But no clinical evidence has been provided. CASE PRESENTATION: Two patients presented to the emergency department with ankle pain after ankle eversion or external rotation. Physical examination revealed tenderness and swelling on both medial and lateral sides of the ankles. Plain radiographs and computed tomography revealed LTPF and medial soft tissue swelling, and magnetic resonance imaging confirmed a discontinuity of the deltoid ligament in Case 1. Surgical exploration revealed rupture of the superficial layer of the deltoid ligaments with intact deep layer in both patients. Treatment included fixation of the lateral process of the talus with headless compression screws and repair of deltoid ligaments. Both patients achieved excellent clinical outcomes 1 year post injury. CONCLUSION: There are many possibilities of the injury mechanism of LTPF. These two cases provided clinical evidence that eversion or external rotation force, in addition to inversion, was also an important mechanism of LTPF.
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Traumatismos del Tobillo , Fracturas Óseas , Astrágalo , Adulto , Traumatismos del Tobillo/complicaciones , Traumatismos del Tobillo/diagnóstico por imagen , Traumatismos del Tobillo/cirugía , Articulación del Tobillo/cirugía , Fracturas Óseas/complicaciones , Fracturas Óseas/diagnóstico por imagen , Fracturas Óseas/cirugía , Humanos , Ligamentos Articulares/diagnóstico por imagen , Ligamentos Articulares/lesiones , Ligamentos Articulares/cirugía , Masculino , Astrágalo/diagnóstico por imagenRESUMEN
Wound repair remains a huge clinical challenge, which can cause bleeding, infection, and patient death. In our current research, a bioactive, injectable, multifunctional composite hydrogel doped with nanospheres was prepared with antibacterial and angiogenesis-promoting functions for the treatment of wounds. Amino groups in ε-polylysine (ε-EPL) undergo dynamic Schiff base reaction cross-linking with oxidized hyaluronic acid (OHA), and F127 exhibits unique temperature sensitivity to form an injectable thermosensitive hydrogel (FHE10), which can form a hydrogel to cover the wound at body temperature. Nanospheres (PNs) prepared using poly (glyceryl-sebacate-acrylate) (PGSA) were loaded into hydrogels (FHE10) for promoting wound repair. The prepared FHE10 exhibited rapid gelation, good injectable abilities, and showed resistance to the flourish of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). In vitro investigations showed that FHE10 had good hemocompatibility and cytocompatibility. FHE10@PNs exhibited good proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs) and human foreskin fibroblasts (HFF-1). Furthermore, FHE10@PNs significantly promoted reepithelialization and collagen deposition as well as micro-vascularization compared with the use of FHE10 or PNs alone, thereby accelerating the repair of wounds. In general, this study demonstrated that the multifunctional injectable composite hydrogel showed great potential in wound treatment.
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Diabetic ulcers, a difficult problem faced by clinicians, are strongly associated with an increase in cellular senescence. Few empirical studies have focused on exploring a targeted strategy to cure diabetic wounds by eliminating senescent fibroblasts (SFs) and reducing side effects. In this study, poly-l-lysine/sodium alginate (PLS) is modified with talabostat (PT100) and encapsulates a PARP1 plasmid (PARP1@PLS-PT100) for delivery to target the dipeptidyl peptidase 4 (DPP4) receptor and eliminate SFs. PARP1@PLS-PT100 releases encapsulated plasmids, displaying high selectivity for SFs over normal fibroblasts by targeting the DPP4 receptor, decreasing senescence-associated secretory phenotypes (SASPs), and stimulating the secretion of anti-inflammatory factors. Furthermore, the increased apoptosis of SFs and the disappearance of cellular senescence alleviates SASPs, accelerates re-epithelialization and collagen deposition, and significantly induces macrophage M2 polarization, which mediates tissue repair and the inflammatory response. This innovative strategy has revealed the previously undefined role of PARP1@PLS-PT100 in promoting diabetic wound healing, suggesting its therapeutic potential in refractory wound repair.
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Alginatos/metabolismo , Senescencia Celular/genética , Diabetes Mellitus Experimental/metabolismo , Dipeptidil Peptidasa 4/metabolismo , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Polilisina/análogos & derivados , Cicatrización de Heridas/genética , Animales , Células Cultivadas , Senescencia Celular/efectos de los fármacos , Diabetes Mellitus Experimental/genética , Dipeptidil Peptidasa 4/genética , Modelos Animales de Enfermedad , Nanosferas/metabolismo , Poli(ADP-Ribosa) Polimerasa-1/genética , Polilisina/metabolismo , Ratas , Ratas Sprague-Dawley , Cicatrización de Heridas/efectos de los fármacosRESUMEN
Bacterial infection and severe wound inflammation are two primary harmful problems that bring harm to the human body and may cause death when a large-scale skin defect occurs. Thus, developing an effective and quick wound healing strategy for curing skin damage and trauma is vital. This study has developed a multifunctional PLA/gelatin/ZnO nanofiber aerogel with a three-dimensional structure through electrospinning and freeze-drying technology for wound healing. It has validated that the nanofiber aerogel has an excellent antibacterial property and biocompatibility. Meanwhile, benefiting from its three-dimensional nanofiber structure, the PLA/gelatin/ZnO nanofiber aerogel possesses good water absorption and air permeability. In vivo experiments have determined that the PLA/gel/ZnO nanofiber aerogel scaffolds effectively promote skin infection's wound healing and enhance angiogenesis that is practical with increasing ZnO concentration.
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Nanofibras , Óxido de Zinc , Antibacterianos/uso terapéutico , Gelatina , Humanos , Cicatrización de HeridasRESUMEN
BACKGROUND: Diverse conclusions have been drawn regarding the association of homocysteine (HCY) deficiency and supplements of B vitamins with fracture incidence in older adults. The aim of this meta-analysis was to investigate the association of HCY and B vitamins (folate, vitamin B12, and B6) with fracture incidence in older adults and whether supplements of B vitamins reduce the risk of fracture. METHODS: The PubMed, Embase, and Cochrane library databases were systematically searched from their inception dates to 1 July 2019 to identify relevant published articles. Meta-analysis was performed to pool hazard ratios (HRs) or risk ratios (RRs) and 95% confidence intervals (CIs) using a random effects model. RESULTS: A total of 28 studies fulfilled the inclusion criteria. High serum HCY was an independent risk factor for fractures in older persons (HR =1.25, 95% CI: 1.12 to 1.40), but only at the highest quartile level (>15 µmol/L) (HR =1.71, 95% CI: 1.37 to 2.12), rather than the second and third quartile. Multiple sensitivity and subgroup analyses supported the consistency and stability of this result. A severe deficiency of folate, instead of vitamin B12 and B6, was found to increase the risk of fracture in older adults (HR =1.46, 95% CI: 1.06 to 2.02; 1.24, 95% CI: 0.79 to 1.95; 1.36, 95% CI: 0.90 to 2.06, respectively). For the interventional effect, there was no significant association of combined folate and vitamin B12, combined folate, vitamin B12 and B6, or single vitamin B6 supplementation with the decrease of fracture risk. DISCUSSION: This meta-analysis revealed that significantly elevated serum level of HCY is positively associated with fracture incidence in older adults, yet the necessity and threshold for intervention by B vitamins require further large-scale high-quality clinical trials to validate. PROSPERO IDENTIFIER: CRD42019122586.
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BACKGROUND: Cartilage injury and pathological degeneration are reported in millions of patients globally. Cartilages such as articular hyaline cartilage are characterized by poor self-regeneration ability due to lack of vascular tissue. Current treatment methods adopt foreign cartilage analogue implants or microfracture surgery to accelerate tissue repair and regeneration. These methods are invasive and are associated with the formation of fibrocartilage, which warrants further exploration of new cartilage repair materials. The present study aims to develop an injectable modified gelatin hydrogel. METHOD: The hydrogel effectively adsorbed proteoglycans secreted by chondrocytes adjacent to the cartilage tissue in situ, and rapidly formed suitable chondrocyte survival microenvironment modified by ε-poly-L-lysine (EPL). Besides, dynamic covalent bonds were introduced between glucose and phenylboronic acids (PBA). These bonds formed reversible covalent interactions between the cis-diol groups on polyols and the ionic boronate state of PBA. PBA-modified hydrogel induced significant stress relaxation, which improved chondrocyte viability and cartilage differentiation of stem cells. Further, we explored the ability of these hydrogels to promote chondrocyte viability and cartilage differentiation of stem cells through chemical and mechanical modifications. RESULTS: In vivo and in vitro results demonstrated that the hydrogels exhibited efficient biocompatibility. EPL and PBA modified GelMA hydrogel (Gel-EPL/B) showed stronger activity on chondrocytes compared to the GelMA control group. The Gel-EPL/B group induced the secretion of more extracellular matrix and improved the chondrogenic differentiation potential of stem cells. Finally, thus hydrogel promoted the tissue repair of cartilage defects. CONCLUSION: Modified hydrogel is effective in cartilage tissue repair.
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Agrecanos/química , Agrecanos/farmacología , Gelatina/química , Hidrogeles/química , Cicatrización de Heridas/efectos de los fármacos , Adsorción , Animales , Cartílago Articular/patología , Diferenciación Celular , Condrocitos/citología , Condrocitos/efectos de los fármacos , Matriz Extracelular , Humanos , Masculino , Ratones , Polilisina , Polímeros , Ratas , Ratas Sprague-Dawley , Ingeniería de Tejidos/métodosRESUMEN
BACKGROUND: Simultaneous dislocation of the radial head and distal radio-ulnar joint without fracture (Criss-Cross Injury) in an adult patient is rarely reported in previous studies. The pathological changes and injury patterns have not been clearly demonstrated. CASE PRESENTATION: A 26-year-old woman presented with acute pain of the right wrist and elbow after a fall from cycling. Physical examination revealed an unstable elbow and wrist joint. Plain radiographs showed volar dislocation of the radial head and dorsal dislocation of the distal radius without associated fracture, forming a criss-cross appearance of the ulna and radius on the lateral radiograph. MRI images confirmed partial rupture of the proximal interosseous membrane from its dorsal attachment on the radius, as well as partial rupture of the medial collateral ligament. Conservative treatment failed because the radiocapitellar joint and distal radio-ulnar joint could not be simultaneously reduced. Surgical exploration revealed a highly unstable radial head, but the annular ligament was found to be intact. Manual force was applied to reduce the radial head and a percutaneous K-wire was used to stabilize the proximal radioulnar joint with the forearm in full supination. After surgery, the elbow was immobilized in 90° flexion by a long arm cast for 4 weeks. The K-wire was removed at 6 weeks postoperatively. At 18 months postoperatively, the patient had regained a full range of flexion and extension, with normal supination and a slight limitation in pronation. CONCLUSIONS: The proximal IOM, especially the dorsal band, was injured in Criss-Cross injuries, while the central part of the IOM remained intact. This injury pattern distinguished itself from Essex-Lopresti injury, which mainly involves rupture of the central band of the IOM.
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Articulación del Codo/patología , Luxaciones Articulares/cirugía , Articulación de la Muñeca/patología , Adulto , Hilos Ortopédicos , Femenino , Humanos , Radiografía , Radio (Anatomía)/patología , Rango del Movimiento Articular , Rotura , Cúbito/patologíaRESUMEN
The control of massive bleeding and its related wound infection is the main challenge for both military and civilian trauma centres. In this study, a cationic superabsorbent hydrogel coordinated by mesoporous silica (CSH-MS) was synthesized by free-radical polymerization for both haemostasis and antibacterial use. The as-prepared CSH-MS has a rough surface, and its water absorption is approximately 5000%. The resultant CSH-MS1 could promote blood cell aggregation and facilitate plasma protein activation via haemadsorption, resulting in efficient blood clot formation. Furthermore, CSH-MS1 (with approximately 5.06% contents of MS) dramatically reduces bleeding time and reduces blood loss in a rat-tail amputation model. Moreover, the CSH-MSs exhibits good antibacterial activities, excellent cytocompatibility and negligible haemolysis. Therefore, CSH-MS can serve as a novel type of haemostatic material in clinical applications.
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Hemostáticos/farmacología , Hidrogeles/farmacología , Dióxido de Silicio/farmacología , Animales , Antibacterianos/farmacología , Coagulación Sanguínea/efectos de los fármacos , Cationes , Eritrocitos/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Hemostasis/efectos de los fármacos , Humanos , Hidrogeles/síntesis química , Hidrogeles/química , Masculino , Pruebas de Sensibilidad Microbiana , Porosidad , Conejos , Ratas Sprague-Dawley , Staphylococcus aureus/efectos de los fármacosRESUMEN
BACKGROUND: In the field of bone tissue engineering, there has been an increasing interest in biomedical materials with both high angiogenic ability and osteogenic ability. Among various osteogenesis materials, bioactive borosilicate and borate glass scaffolds possess suitable degradation rate and mechanical strength, thus drawing many scholars' interests and attention. OBJECTIVE: In this study, we fabricated bioactive glass scaffolds composed of borosilicate 2B6Sr using the Template-Method and incorporated Dimethyloxalylglycine (DMOG), a small-molecule angiogenic drug possessing good angiogenic ability, to improve bone regeneration. METHODS: The in-vitro studies showed that porous borosilicate bioactive glass scaffolds released slowly, a steady amount of DMOG and stimulated the proliferation and osteogenic differentiation of human bone marrow stromal cells hBMSCs. RESULTS: In-vivo studies showed that the borosilicate bioactive glass scaffolds could significantly promote new bone formation and neovascularization in rats' calvarial bone defects. CONCLUSION: These results indicated that DMOG-incorporated bioactive glass scaffold is a successful compound with excellent angiogenesis-osteogenesis ability, which has favorable clinical prospects.
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Compuestos de Boro/farmacología , Glicina/análogos & derivados , Neovascularización Patológica/tratamiento farmacológico , Osteogénesis/efectos de los fármacos , Silicatos/farmacología , Animales , Compuestos de Boro/química , Vidrio/química , Glicina/química , Glicina/farmacología , Humanos , Masculino , Neovascularización Patológica/patología , Imagen Óptica , Tamaño de la Partícula , Ratas , Ratas Sprague-Dawley , Silicatos/química , Propiedades de SuperficieRESUMEN
One of the hallmarks of osteoarthritis (OA) development is endochondral ossification, in which Runtrelated transcription factor2 (Runx2) is aberrantly expressed. Runx2 was previously identified to be regulated by microRNA2045p (miR2045p). The aim of the present study was to investigate the potential function of miR2045p regulating Runx2 during the development of OA and the underlying molecular mechanism. The expression levels of miR2045p and Runx2 were determined in tissue specimens. Rat OA models were established by transecting the anterior and posterior cruciate ligaments and removing the meniscus. Rats were treated with miR2045p agomir and miR2045p negative control (NC). All in vitro experiments were performed using rat primary chondrocytes and the SW1353 human bone chondrosarcoma cell line. It was identified that the expression of miR2045p was significantly decreased, whereas Runx2 was significantly increased, in human OA tissues compared with in nonOA tissues, and levels were inversely associated with each other in primary chondrocytes and chondrosarcoma cells. Overexpression of miR2045p decreased the proliferation of chondrocytes and SW1353 cells. Using a luciferase reporter assay, Runx2 was identified to be a direct target of miR2045p in chondrocytes and overexpressed miR2045p altered the expression of collagens II, X and matrix metalloproteinase (MMP)1 and MMP13 in primary chondrocytes and SW1353 cells. Histological analysis revealed that miR2045p treatment ameliorated the OAlike phenotype that is reflected by assessment of cartilage thickness and Mankin's score. Runx2 expression was gradually increased as the rats increased in age. At 10 weeks of miR2045p agomir treatment, altered expression levels of collagens II and X, cartilage oligomeric matrix protein fragment, aggrecan, MMP1 and MMP13 were observed in the treatment group compared with the NC group. In conclusion, the results of the present study indicated that miR2045p decreases chondrocyte proliferation and ameliorates the OAlike phenotype in rats with surgically induced OA by targeting Runx2.
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Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , MicroARNs/metabolismo , Osteoartritis/metabolismo , Anciano , Animales , Western Blotting , Proliferación Celular/genética , Proliferación Celular/fisiología , Supervivencia Celular/genética , Supervivencia Celular/fisiología , Condrocitos/metabolismo , Condrocitos/fisiología , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Femenino , Humanos , Inmunohistoquímica , Masculino , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 13 de la Matriz/genética , Metaloproteinasa 13 de la Matriz/metabolismo , MicroARNs/genética , Persona de Mediana Edad , Osteoartritis/genética , Ratas , Ratas Sprague-DawleyRESUMEN
Emerging evidence indicates that microRNAs (miRNAs, miRs) play diverse roles in the regulation of biological processes, including osteoblastic differentiation. In this study, we found that miR-383 is a critical regulator of osteoblastic differentiation. We showed that miR-383 was downregulated during osteoblastic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). Overexpression of miR-383 suppressed osteoblastic differentiation of BMSCs by downregulating alkaline phosphatase (ALP), matrix mineralization, and mRNA and protein levels of RUNX2 and OCN, whereas a knockdown of miR-383 promoted osteoblastic differentiation in vitro. The results of in vivo analysis indicated that inhibition of miR-383 expression enhanced the efficacy of new bone formation in a rat calvarial defect model. Mechanistic experiments revealed that special AT-rich-sequence-binding protein 2 (Satb2) was a direct and functional target of miR-383. Knockdown of Satb2 had inhibitory effects resembling those of miR-383 on the osteoblast differentiation of rat BMSCs. In addition, the positive effect of miR-383 suppression on osteoblastic differentiation was apparently abrogated by Satb2 silencing. Collectively, these results indicate that miR-383 plays an inhibitory role in osteogenic differentiation of rat BMSCs and may act by targeting Satb2.
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Diferenciación Celular/fisiología , Proteínas de Unión a la Región de Fijación a la Matriz/biosíntesis , Células Madre Mesenquimatosas/metabolismo , MicroARNs/biosíntesis , Osteoblastos/metabolismo , Osteogénesis/fisiología , Factores de Transcripción/biosíntesis , Animales , Masculino , Proteínas de Unión a la Región de Fijación a la Matriz/antagonistas & inhibidores , Ratas , Ratas Sprague-Dawley , Factores de Transcripción/antagonistas & inhibidoresRESUMEN
lncRNA-TUG1 (Taurine upregulated 1) is up-regulated and highly correlated with poor prognosis and disease status in osteosarcoma. TUG1 knockdown inhibits osteosarcoma cell proliferation, migration and invasion, and promotes apoptosis. However, its mechanism of action has not been well addressed. Growing evidence documented that lncRNA works as competing endogenous (ce)RNAs to modulate the expression and biological functions of miRNA. As a putative combining target of TUG1, miR-144-3p has been associated with the progress of osteosarcoma. To verify whether TUG1 functions through regulating miR-144-3p, the expression levels of TUG1 and miR-144-3p in osteosarcoma tissues and cell lines were determined. TUG1 was upregulated in osteosarcoma tissues and cell lines, and negatively correlated with miR-144-3p. TUG1 knockdown induced miR-144-3p expression in MG63 and U2OS cell lines. Results from dual luciferase reporter assay, RNA-binding protein immuno-precipitation (RIP) and applied biotin-avidin pull-down system confirmed TUG1 regulated miR-144-3p expression through direct binding. EZH2, a verified target of miR-144-3p was upregulated in osteosarcoma tissues and negatively correlated with miR-144-3p. EZH2 was negatively regulated by miR-144-3p and positively regulated by TUG1. Gain-and loss-of-function experiments were performed to analyze the role of TUG1, miR-144-3p and EZH2 in the migration and EMT of osteosarcoma cells. EZH2 over-expression partly abolished TUG1 knockdown or miR-144-3p overexpression induced inhibition of migration and EMT in osteosarcoma cells. In addition, TUG1 knockdown represses the activation of Wnt/ß-catenin pathway, which was reversed by EZH2 over-expression. The activator of Wnt/ß-catenin pathway LiCl could partially block the TUG1-knockdown induced osteosarcoma cell migration and EMT inhibition. In conclusion, our results showed that TUG1 plays an important role in osteosarcoma development through miRNA-144-3p/EZH2/Wnt/ß-catenin pathway.
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Neoplasias Óseas/genética , Proteína Potenciadora del Homólogo Zeste 2/genética , MicroARNs/genética , Osteosarcoma/genética , ARN Largo no Codificante/genética , Neoplasias Óseas/patología , Línea Celular Tumoral , Movimiento Celular , Proliferación Celular , Transición Epitelial-Mesenquimal , Regulación Neoplásica de la Expresión Génica , Humanos , Estadificación de Neoplasias , Osteosarcoma/patología , Regulación hacia Arriba , Vía de Señalización WntRESUMEN
We previously demonstrated that three-dimensional (3D) hydroxyapatite (HAP)-collagen (COL)-coated poly(ε-caprolactone) (PCL) scaffolds (HAP-COL-PCL) possess appropriate nano-structures, surface roughness, and nutrients, providing a favorable environment for osteogenesis. However, the effect of using 3D HAP-COL-PCL scaffolds incorporating BMSCs for the repair of bone defects in rats has been not evaluated. 3D PCL scaffolds coated with HAP, collagen or HAP/COL and incorporating BMSCs were implanted into calvarial defects. At 12 weeks after surgery, the rats were sacrificed and crania were harvested to assess the bone defect repair using microcomputed tomography (micro-CT), histology, immunohistochemistry and sequential fluorescent labeling analysis. 3D micro-CT reconstructed images and quantitative analysis showed that HAP-COL-PCL groups possessed better bone-forming capacity than HAP-PCL groups or COL-PCL groups. Fluorescent labeling analysis revealed the percentage of tetracycline labeling, alizarin red labeling, and calcein labeling in HAP-COL-PCL groups were all greater than in the other two groups (P < 0.05), and the result was confirmed by immunohistochemical staining and histological analysis of bone regeneration. This study demonstrates that 3D HAP-COL-PCL scaffolds incorporating BMSCs markedly enhance bone regeneration of bone defects in rats.