Exosomes derived from BMSCs in osteogenic differentiation promote type H blood vessel angiogenesis through miR-150-5p mediated metabolic reprogramming of endothelial cells.

Osteogenesis is tightly coupled with angiogenesis spatiotemporally. Previous studies have demonstrated that type H blood vessel formed by endothelial cells with high expression of CD31 and Emcn (CD31hi Emcnhi ECs) play a crucial role in bone regeneration. The mechanism of the molecular communication around CD31hi Emcnhi ECs and bone mesenchymal stem cells (BMSCs) in the osteogenic microenvironment is unclear. This study indicates that exosomes from bone mesenchymal stem cells with 7 days osteogenic differentiation (7D-BMSCs-exo) may promote CD31hi Emcnhi ECs angiogenesis, which was verified by tube formation assay, qRT-PCR, Western blot, immunofluorescence staining and µCT assays etc. in vitro and in vivo. Furthermore, by exosomal miRNA microarray and WGCNA assays, we identified downregulated miR-150-5p as the most relative hub gene coupling osteogenic differentiation and type H blood vessel angiogenesis. With bioinformatics assays, dual luciferase reporter experiments, qRT-PCR and Western blot assays, SOX2(SRY-Box Transcription Factor 2) was confirmed as a novel downstream target gene of miR-150-5p in exosomes, which might be a pivotal mechanism regulating CD31hi Emcnhi ECs formation. Additionally, JC-1 immunofluorescence staining, Western blot and seahorse assay results showed that the overexpression of SOX2 could shift metabolic reprogramming from oxidative phosphorylation (OXPHOS) to glycolysis to enhance the CD31hi Emcnhi ECs formation. The PI3k/Akt signaling pathway might play a key role in this process. In summary, BMSCs in osteogenic differentiation might secrete exosomes with low miR-150-5p expression to induce type H blood vessel formation by mediating SOX2 overexpression in ECs. These findings might reveal a molecular mechanism of osteogenesis coupled with type H blood vessel angiogenesis in the osteogenic microenvironment and provide a new therapeutic target or cell-free remedy for osteogenesis impaired diseases.

Nuciferine prevents bone loss by disrupting multinucleated osteoclast formation and promoting type H vessel formation.

Recently, type H vessels were reported to couple angiogenesis and osteogenesis during osteoclastogenesis, and tartrate-resistant acid phosphatase (Trap)+ preosteoclasts were found to secrete increased PDGF-BB to promote type H vessel formation. Therefore, utilization of type H vessels may be a strategy to treat diseases involving bone loss. In the present study, we found that nuciferine, a natural bioactive compound, has various effects, including inhibiting osteoclastogenesis and promoting type H vessel formation. Nuciferine inhibited osteoclastogenesis and bone resorption but increased the relative number of Trap+ preosteoclasts. Nuciferine restrained the expression of osteoclast-specific genes and proteins, promoted PDGF-BB production and potentiated related angiogenic activities by inhibiting the MAPK and NF-κB signaling pathways in vitro. We confirmed the bone-protective effects of nuciferine in ovariectomized mice and found that nuciferine treatment increased the PDGF-BB concentration and the number of type H vessels in the femur. In conclusion, our results demonstrated that nuciferine can decrease multinucleated osteoclast formation and promote type H vessel formation through preservation of Trap+ preosteoclasts via inhibition of the MAPK and NF-κB signaling pathways and may be an excellent agent for the treatment of diseases involving bone loss.

A Study to Compare the Efficacy of a Biodegradable Dynamic Fixation System With Titanium Devices in Posterior Spinal Fusion Between Articular Processes in a Canine Model.

The objective of this study was to apply a biodegradable dynamic fixation system (BDFS) for lumbar fusion between articular processes and compare the fusion results and biomechanical changes with those of conventional rigid fixation. Twenty-four mongrel dogs were randomly assigned to 2 groups and subjected to either posterior lumbar fusion surgery with a BDFS or titanium rods (TRs) at the L5-L6 segments. Six animals in each group were sacrificed at 8 or 16 weeks. Fusion conditions were evaluated by computed tomography (CT), manual palpation, biomechanical tests, and histological analysis. Biomechanical tests were performed at the L4-7 (for range of motion (ROM)) and L5-6 (for fusion stiffness) segments. Histological examination was performed on organs, surrounding tissues, and the fused area. The magnesium alloy components maintained their initial shape 8 weeks after the operation, but the meshing teeth were almost completely degraded at 16 weeks. The biomechanical analysis revealed an increased lateral bending ROM at 8 weeks and axial torsion ROM at 16 weeks. The L4-5 extension-flexion ROMs in the BDFS group were 2.29 ± 0.86 deg and 3.17 ± 1.08 deg at 16 weeks, respectively, compared with 3.22 ± 0.56 deg and 5.55 ± 1.84 deg in TR group. However, both groups showed similar fusion results. The BDFS design is suitable, and its degradation in vivo is safe. The BDFS can be applied for posterior lumbar fusion between articular processes to complete the fusion well. Additionally, the BDFS can reduce the decline in lateral motion and hypermotion of the cranial adjacent segment in flexion-extension motion.

Effectiveness of additional C2 decompression of the cervical spinal canal after cervical laminoplasty: a retrospective cohort study.

PURPOSE: This study aimed to assess the effects of additional C2 decompression of the cervical spinal canal on the postoperative outcomes after cervical laminoplasty in patients with cervical stenosis caused by ossification of the posterior longitudinal ligament (OPLL). MATERIALS AND METHODS: This retrospective cohort study included patients with cervical stenosis due to OPLL and treated between April 2014 and December 2015. The patients who underwent C2-7 (additional C2 decompression) and C3-7 posterior decompression were compared using the Japanese Orthopedic Association (JOA) scores, visual analog scale (VAS) scores, axial symptom scores, and intervals between the posterior margin of the vertebral body and the K-line. RESULTS: There were 36 and 24 patients in the additional C2 decompression and control groups, respectively. The JOA scores were higher in the additional C2 decompression group than the controls at 1 and 3 years (p < 0.05). Upper extremity motor function after the operation and at 1 and 3 years and lower extremity motor function after operation were improved in the additional C2 decompression group (all p < 0.05 vs. controls). VAS scores were lower in the additional C2 decompression group than controls at 1 year (p < 0.05). Axial symptom scores in the additional C2 decompression group were decreased postoperatively but increased at 1 and 3 years (p < 0.05 vs. controls). Finally, the posterior shift of the K-line in the additional C2 decompression group was significant (from 0.98 to 1.68 cm, p < 0.05). CONCLUSIONS: Additional C2 decompression might improve the effectiveness of cervical laminoplasty in patients with cervical stenosis caused by OPLL.

Cellular study of the LPS-induced osteoclastic multinucleated cell formation from RAW264.7 cells.

Despite the response to the receptor activator of nuclear factor κ-Β ligand (RANKL), a study has reported that lipopolysaccharide (LPS) could induce RAW264.7 linage osteoclastic differentiation. However, on the contrary, another study recently showed that the LPS-induced multinuclear cells from RAW264.7 did not express osteoclastic functions. Interestingly, in our previous study, we found that RAW264.7 cells pretreated with 10 ng LPS plus macrophage-colony stimulating factor did not show any effects for enhancing RANKL-induced osteoclastic cell differentiation. Therefore, in our current study, we aim to investigate the oteoclastogesis induction ability and efficacy of LPS in the RAW264.7 cell line and relevant molecular signaling. The osteoclastogenic activity of LPS-treated RAW264.7 linage was studied by bone resorption pits and fibrous actin study. Besides that, through polymerase chain reaction and western blot, we showed that the transcriptional factor c-Fos and Nfatc1 might be associated with LPS-induced osteoclastogenesis. Overall, the results of our current study showed positive proof for osteoclast generation from LPS-independent treatment, as well as established an optimal and efficient method for this process.

Evaluation of efficacy on RANKL induced osteoclast from RAW264.7 cells.

Established RAW264.7 cell lines for osteoclastic differentiation has been widely engaged in bone homeostasis research, however, the efficacy of RANKL independently stimulating has rarely been defined, because protocols were usually developed and modified by various laboratories. Otherwise, problematic issues are also lie in the cell's seeding density, RANKL stimulating time point, and distinguishing osteoclastogenesis ability of RANKL-treated RAW264.7 cells. Therefore, in the current study, we examined the efficacy of various concentrations of RANKL-treated RAW264.7 for its osteoclastic differentiation with or without pretreated other costimulators such as: LPS and/or M-CSF. The oteoclastogenesis ability of RANKL-treated RAW264.7 cells was demonstrated by bone resorption pit, F-actin, and osteoclastogenesis specific marker studies. Besides that, through tartrate-resistant acid phosphatase (TRAP) staining, we clarified to start the treatment with 30 ng/ml RANKL at 12 hr after seeded RAW264.7 with the density of 6.25 × 10 3 cells/cm 2 manifested an significantly increased number of multinucleated osteoclastic cells. Overall, our results establishing an optimal method for RANKL independently inducing RAW 264.7 cell osteoclastic differentiation, which could efficiently generate osteoclasts in vitro for significant advances in our understanding of bone biology.

Synovial fibroblast-targeting liposomes encapsulating an NF-κB-blocking peptide ameliorates zymosan-induced synovial inflammation.

Synovial fibroblasts (SFs) play a crucial role in the inflammatory process of rheumatoid arthritis (RA). The highly activated NF-κB signal in SFs is responsible for most of the synovial inflammation associated with this disease. In this study, we have developed an SF-targeting liposomal system that encapsulates the NF-κB-blocking peptide (NBD peptide) HAP-lipo/NBD. HAP-lipo/NBDs demonstrated efficient SF-specific targeting in vitro and in vivo. Our study also showed a significant inhibitory effect of HAP-lipo/NBD on NF-κB activation, inflammatory cytokine release and SF migration capability after zymosan stimulation. Furthermore, the systemic administration of HAP-lipo/NBDs significantly inhibited synovial inflammation and improved the pathological scores of arthritis induced by zymosan. Thus, these results suggest that an SF-targeting NF-κB-blocking strategy is a potential approach for the development of alternative, targeted anti-RA therapies.

Clinical and Radiographic Features of the Atlantoaxial Dislocation Associated With Kashin-Beck Disease.

OBJECTIVES: Keshin-Beck disease (KBD) is a particular type of osteoarthritis that affects many joints. However, the deformity of atlantoaxial joint has been rarely reported in KBD, and therefore its clinical and radiograph features have not been identified. METHODS: We reviewed data in 14 patients who were diagnosed with atlantoaxial dislocation (AAD) in KBD at our institution. The demographic data, clinical history, imaging data, operative data, and Japanese Orthopaedic Association score were collected for evaluation. RESULTS: The mean age at presentation was 50 ± 1.7 years old. The most common features of AAD in KBD were the osteoarthritis, characterized by hypertrophic dens and anterior arch of the atlas. The average inner anteroposterior diameter (IAPD) of C1 was 28 ± 3.5 mm and the average spinal canal diameter was 14 ± 3.3 mm, which were respectively lower than the control level. Five patients had severe C1 stenosis (IAPD < 26mm). Separated odontoid process, like os odontoideum, was seen 9 patients. The tip of dens fused to C1 was observed in 4 patients; 12 patients had high-riding vertebral artery; and 5 patients had severe C1 stenosis, and they underwent C1 laminectomy with C1-C2 interarticular fusion or occipital-cervical fusion. All the patients displayed neurologic improvement after surgery. CONCLUSIONS: The atlantoaxial level could be affected by KBD, which may lead to typical abnormalities and cause AAD. A C1 laminectomy with an C1-C2 interarticular fusion or occipital-cervical fusion is recommended for the patient with severe stenosis.

Fabrication of BMP-2-peptide-Deferoxamine- and QK-peptide-functionalized nanoscaffolds and their application for bone defect treatment.

The microenvironment in the healing process of large bone defects requires suitable conditions to promote osteogenesis and angiogenesis. Coaxial electrospinning is a mature method in bone tissue engineering (BTE) and allows functional modification. Appropriate modification methods can be used to improve the bioactivity of scaffolds for BTE. In this study, coaxial electrospinning with QK peptide (a Vascular endothelial growth factor mimetic peptide) and BMP-2 peptide-DFO (BD) was performed to produce double-modified PQBD scaffolds with vascularizing and osteogenic features. The morphology of coaxially electrospun scaffolds was verified by scanning electron microscopy (SEM) and transmission electron microscopy. Laser scanning confocal microscopy and Fourier transform infrared spectroscopy confirmed that BD covalently bound to the surface of the P and PQ scaffolds. In vitro, the PQBD scaffold promoted the adhesion and proliferation of bone marrow stromal cells (BMSCs). Both QK peptide and BD showed sustainable release and preservation of biological activity, enhancing the osteogenic differentiation of BMSCs and the migration of human umbilical vein endothelial cells and promoting angiogenesis. The combined ability of these factors to promote osteogenesis and angiogenesis is superior to that of each alone. In vivo, the PQBD scaffold was implanted into the bone defect, and after 8 weeks, the defect area was almost completely covered by new bone tissue. Histology showed more mature bone tissue and more blood vessels. PQBD scaffolds promote both angiogenesis and osteogenesis, offering a promising approach to enhance bone regeneration in the treatment of large bone defects.

Hsa-circ-0007292 promotes the osteogenic differentiation of posterior longitudinal ligament cells via regulating SATB2 by sponging miR-508-3p.

Ossification of the posterior longitudinal ligament (OPLL) is a disorder with multiple pathogenic mechanisms and leads to different degrees of neurological symptoms. Recent studies have revealed that non-coding RNA (ncRNA), including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), could influence the development of OPLL. Nevertheless, the molecular mechanisms linking circular RNAs (circRNAs) and the progression of OPLL is still unknown. The current research explored the expression profiles of OPLL-related circRNAs by microarray analysis, and applied qRT-PCR to validate the results. Subsequently, we confirmed the upregulation of hsa_circ_0007292 in OPLL cells by qRT-PCR and validated the circular characteristic of hsa_circ_0007292 by Sanger sequencing. Fluorescence in situ hybridization (FISH) unveiled that hsa_circ_0007292 was predominantly located in the cytoplasm. Functionally, gain-of-function and loss-of-function experiments showed that hsa_circ_0007292 promoted the osteogenic differentiation of OPLL cells. Mechanistically, the interaction of hsa_circ_0007292 and miR-508-3p was predicted and validated by bioinformatics analysis, dual-luciferase reporter assays, and Ago2 RNA immunoprecipitation (RIP). Similarly, we validated the correlation between miR-508-3p and SATB2. Furthermore, rescue experiments were performed to prove that hsa_circ_0007292 acted as a sponge for miR-508-3p, and SATB2 was revealed to be the target gene of miR-508-3p. In conclusion, our research shows that hsa_circ_0007292 regulates OPLL progression by the miR-508-3p/SATB2 pathway. Our results indicate that hsa_circ_0007292 can be used as a promising therapeutic target for patients with OPLL.

Nanohydroxyapatite/polyamide 66 crosslinked with QK and BMP-2-derived peptide prevented femur nonunion in rats.

Active interventions should be made to avoid delayed bone union and nonunion during fracture treatment. Nanohydroxyapatite/polyamide 66 (nHA/PA66), a simulated bioactive bone substitute with great biocompatibility and mechanical properties, has been widely used in bone regeneration. However, the limited bioactivity of nHA/PA66 has impeded its further application in tissue engineering. In this study, BMP-2-derived peptide and QK (a VEGF mimetic peptide) were dually grafted to PA66 polymer chains to prepare peptide-decorated HA/PA66-BMP-2-QK scaffolds to enhance bone formation after severe femoral fracture (periosteum scraped off) in SD rats. Fourier transform infrared spectroscopy (FTIR) confirmed that the BMP-2-derived peptide and QK were covalently bonded onto the surface of nHA/PA66. In vitro, BMP-2- and QK-modified scaffolds promoted the adhesion and proliferation ability of rBMSCs. After loading onto peptide-modified scaffolds, both BMP2-derived peptide and QK showed sustainable release and preserved bioactivity, improving the osteogenic differentiation ability of BMSCs. The combined ability of these factors to promote osteogenicity was better than that of a single peptide. Furthermore, the QK released from nHA/PA66-BMP-2-QK scaffolds improved the proliferation and tube formation ability of HUVECs. In vivo, femur nonunion in SD rats was successfully prevented by implanting HA/PA66-BMP-2-QK scaffolds into the fracture gap: the fracture line disappeared, the cortical bone showed continuity, the scaffolds were completely embedded and more vessels formed in the nonunion area than observed in other groups. Overall, the nHA/PA66-BMP-2-QK scaffolds simultaneously facilitated angiogenesis and osteogenesis, providing a promising method for reinforcing bone regeneration in nonunion treatment.

miRNA-187-5p Regulates Osteoblastic Differentiation of Bone Marrow Mesenchymal Stem Cells in Mice by Targeting ICAM1.

Osteoporosis (OP) is a common bone metabolic disease, the process of which is fundamentally irreversible. Therefore, the investigation into osteoblastic differentiation of bone marrow mesenchymal stem cells (BMSCs) will provide more clues for OP treatment. In the present study, we found that microRNA-187-5p (miR-187-5p) played a key role on osteoblastic differentiation, which was significantly upregulated during osteogenic differentiation of BMSCs in mice. Moreover, overexpression of miR-187-5p suppressed osteoblastic differentiation of BMSCs through increasing alkaline phosphatase (ALP), matrix mineralization, and levels of Osterix (OSX), and osteopontin (OPN) as well as runt-related transcription factor 2 (Runx2) in vitro. The results in vivo indicated that the upregulation of miR-187-5p enhanced the efficacy of new bone formation in the heterotopic bone formation assay. Luciferase reporter assay and western blot analysis revealed that miR-187-5p was involved in osteogenesis by targeting intracellular adhesion molecule 1 (ICAM-1). Furthermore, ICAM-1 silence inhibited osteoblastic differentiation of BMSCs. Taken together, our results suggested for the first time that miR-187-5p may promote osteogenesis by targeting ICAM-1, and provided a possible therapeutic target for bone metabolic diseases.

A novel dynamic fixation system with biodegradable components on lumbar fusion between articular processes in a canine model.

The objective of this study was to design a novel dynamic fixation system with biodegradable components, apply it for lumbar fusion between articular processes and compare the fusion results and biomechanical changes to those of conventional rigid fixation. The novel dynamic fixation system was designed using a finite element model, stress distributions were compared and 24 mongrel dogs were randomly assigned to two groups and subjected to either posterior lumbar fusion surgery with a novel dynamic fixation system or titanium rods at the L5-L6 segments. Lumbar spines were assessed in both groups to detect radiographic, manual palpation and biomechanical changes. Histological examination was performed on organs and surrounding tissues. In the novel dynamic fixation system, stress was mainly distributed on the meshing teeth of the magnesium alloy spacer. The magnesium alloy components maintained their initial shape 8 weeks after the operation, but the meshing teeth were almost completely degraded at 16 weeks. The novel dynamic fixation system revealed an increased lateral bending range of motion at 8 weeks; however, both groups showed similar radiographic grades, fusion stiffness, manual palpation and histological results. The novel dynamic fixation system design is suitable, and its degradation in vivo is safe. The novel dynamic fixation system can be applied for posterior lumbar fusion between articular processes and complete the fusion like titanium rods.

A modular programmed biphasic dual-delivery system on 3D ceramic scaffolds for osteogenesis in vitro and in vivo.

Single-factor delivery is the most common characteristic of bone tissue engineering techniques. However, bone regeneration is a complex process requiring multiple factors and specialized release mechanisms. Therefore, the development of a dual-delivery system allowing for programmed release kinetics would be highly desirable. Improvement of the molarity and versatility of the delivery system has rarely been studied. Herein, we report the development of a novel, modular programmed biphasic dual-release system (SCB), carrying a BMP2 and an engineered collagen I-derived recognition motif (Stath-DGEA), with a self-remodification feature on hydroxyapatite (HA)-based materials. The SCB system was loaded onto an additive manufactured (AM) scaffold in order to evaluate its bifactor osteogenic potential and its biphasic release behavior. Further, the biomechanical properties of the scaffold were studied by using the fluid-structure interaction (FSI) method. Section fluorescent labeling revealed that the HA scaffold has a relatively higher density and efficiency. Additionally, the results of the release and inhibition experiment suggested that the SCB system could facilitate the sustained release of therapeutic levels of two factors during the initial stage of implantation, thereby exhibiting a rapid high-dose release pattern at a specific time point during the second stage. The FSI prediction model indicated that the scaffold provides an excellent biomimetic mechanical and fluid dynamic microenvironment to promote osteogenesis. Our results indicated that incorporation of BMP2 with Stath-DGEA in the biphasic SCB system could have a synergetic effect in promoting the adhesion, proliferation, and differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro, under staged stimulations. Further, in vivo studies in both ectopic and orthotopic rat models showed that the SCB system loaded onto an AM scaffold could enhance osteointegration and osteoinduction throughout the osteogenic process. Thus, the novel synthetic SCB system described herein used on an AM scaffold provides a biomimetic extracellular environment that enhances bone regeneration and is a promising multifunctional, dual-release platform.

Chitosan-Gelatin Scaffolds Incorporating Decellularized Platelet-Rich Fibrin Promote Bone Regeneration.

Platelet-rich fibrin (PRF), which functions as a growth factor carrier, has been extensively used to promote soft and hard tissue repair. However, whether decellularized PRF (DPRF) maintains its bioactive effects is unknown. Chitosan/gelatin(C/G) base scaffolds display appropriate biocompatibility and mechanical properties, but they lack biological activity. Thus, the incorporation of DPRF into the C/G scaffold can theoretically improve both the bioactivity of the C/G scaffold and the strength of PRF. In this study, DPRF was prepared using a method combining repeated freeze-thawing and enzymatic digestion. Also, DPRF-loaded chitosan-gelatin scaffolds (C/G/DPRF) were fabricated, using C/G scaffolds as controls. The osteogenic potential of scaffolds was investigated in vitro and in vivo. Compared with the C/G scaffold, C/G/DPRF had a larger pore size (280.8 ± 11.7 µm vs 235.0 ± 11.6 µm; P < 0.05), increased water uptake ratio (13.90 ± 0.09 vs 11.05 ± 0.10; P < 0.05), and similar porosity (90.50 ± 0.87 vs 90.65 ± 0.67; P > 0.05) but reduced compressive modulus (0.81 ± 0.02 MPa vs 1.17 ± 0.05 MPa; P < 0.05). In vitro, C/G/DPRF scaffolds accelerated attachment, proliferation, and osteogenesis-related marker expression of bone marrow stem cells. In vivo, C/G/DPRF scaffolds led to enhanced bone healing and defect closure in a rat calvarial defect model. Thus, we concluded that DPRF remains bioactive and the prepared C/G/DPRF scaffold is a promising material for bone regeneration.

A three-dimensional (3D) printed biomimetic hierarchical scaffold with a covalent modular release system for osteogenesis.

Hydroxyapatite (HA) ceramics are well known for their biocompatibility, bioactivity, and osteoconductive nature. However, limited hierarchical structure and lack of ease in modularity hinder the widespread application of conventional HA ceramics. By using three-dimensional printing (3DP) techniques with multiple materials, including HA, complex biological and mechanical architecture of natural organisms can be achieved through biomimetics. In this study, we designed an osteoid, biomimetic, hierarchical, porous HA ceramic 3D printed scaffold (3DPs). Further incorporation of a covalent, modular, controlled release system (CMR), based on Watson-Crick's complementary oligonucleotides, and was added to carry a bone morphogenetic protein-2 (BMP2) peptide. The choice of a HA biomimetic scaffold housing BMP2 protein fragments was selected to successfully promote osteogenesis both in vitro and in vivo. Scanning electron microscopy, micro-computed tomography analysis and computer fluid dynamics simulations of the 3DPs showed a uniform biomimetic hierarchical structure and an effective interior permeability. Active molecules were found bound with high stability and modular to the scaffold surface via the CMR system. After 7 days of incubation under physiological conditions, approximately 90% of active factors remained bound. Compared to control groups, the 3DPs-CMR-BMP2 group significantly enhanced cell proliferation and adhesion. Moreover, the 3DPs-CMR-BMP2 group exhibited more extensive and sustained osteogenic effects through upregulated expression of osteogenic factors and enhanced calcium deposition, as compared to study and control groups. Furthermore, ectopic osteogenesis and a critical calvarial defect model confirmed that the 3DPs-CMR-BMP2 group significantly promoted in vivo bone healing versus control. Thus, our results showed that biomimetic hierarchical 3DPs with a CMR system successfully promote cell proliferation, adhesion, differentiation and osteogenesis, on a continuous cycle. The biomimetic hierarchical 3DPs with a CMR system offers a promising multi-functional, bone substitute material for treatment of patients with bone defects.

Dually optimized polycaprolactone/collagen I microfiber scaffolds with stem cell capture and differentiation-inducing abilities promote bone regeneration.

Micro-nano based fibrous scaffolds have been extensively studied in regenerative medicine. Bone marrow stem cells (BMSCs) and BMP2-derived peptides, two other important components for tissue engineering, have been successfully used for bone regeneration. However, a scaffold that specifically captures BMSCs and delivers BMP2-derived peptides to promote osteogenic differentiation of enriched BMSCs has not been reported. In this study, a microfiber scaffold was constructed by coaxial electrospinning technology using a polyvinylpyrrolidone/bovine serum albumin/BMP2-derived peptide compound as the core solution and a polycaprolactone/collagen I compound as the shell solution. The scaffolds were further functionalized by covalent grafting of a BMSC affinity peptide (E7) to develop a dual drug release system for the delivery of the BMP2-derived peptide and E7. Structural analysis indicated that the microfibers had a uniform diameter and homogeneous core-shell structure. Fourier transform infrared spectroscopy (FTIR) revealed that E7 was covalently bonded onto the surface of the fibers. In vitro, the E7-modified scaffolds promoted the initial adhesion of BMSCs and were more favorable for BMSC survival. Furthermore, the BMP2-derived peptide loaded in the E7-modified scaffolds was released in a sustained manner and retained bioactivity, significantly improving the osteogenic differentiation of BMSCs. In vivo, scaffolds loaded with the BMP2-derived peptide and E7 (PCME scaffolds) led to enhanced new bone formation and defect closure in a rat calvarial defect model. Overall, the PCME scaffold simultaneously facilitated all three of the essential elements needed for bone tissue engineering, providing a promising method for bone regeneration.

Modification of Human Umbilical Cord Blood Stem Cells Using Polyethylenimine Combined with Modified TAT Peptide to Enhance BMP-2 Production.

With the emerging role of umbilical cord blood-derived mesenchymal stem cells (hUCB-MSC) for bone regeneration and delivery of therapeutic proteins, there is an increasing need for effective gene delivery systems to modify such cells. mTAT, a TAT peptide sequence bearing histidine and cysteine residues, has been successfully used for intracellular gene delivery. Using a gWiz-GFP plasmid, we demonstrated that polyethylenimine combined with mTAT (mTAT/PEI) displayed good transfection efficacy in hUCB-MSC. hUCB-MSC transfected with mTAT/PEI were shown to express more BMP-2 protein and mRNA, indicating the feasibility of using the cells as a BMP-2 delivery system. Importantly, compared to PEI25, a "gold standard" nonviral transfection polymer, mTAT/PEI had limited toxicity to the cells. Furthermore, we demonstrated enhanced osteogenic activity in vitro for BMP-2 expressing hUCB-MSC. These results provide encouraging evidence for the potential use of mTAT/PEI to genetically modify hUCB-MSC as an approach to enhance tissue regeneration.

Sesamin inhibits IL-1ß-stimulated inflammatory response in human osteoarthritis chondrocytes by activating Nrf2 signaling pathway.

Sesamin, a bioactive component extracted from sesame, has been reported to exert anti-inflammatory and anti-oxidant effects. In this study, we evaluated the anti-inflammatory effects of sesamin on IL-1ß-stimulated human osteoarthritis chondrocytes and investigated the possible mechanism. Results demonstrated that sesamin treatment significantly inhibited PGE2 and NO production induced by IL-1ß. Sesamin inhibited MMP1, MMP3, and MMP13 production in IL-1ß-stimulated chondrocytes. Sesamin also inhibited IL-1ß-induced phosphorylation of NF-κB p65 and IκBα. Meanwhile, sesamin was found to up-regulate the expression of Nrf2 and HO-1. However, Nrf2 siRNA reversed the anti-inflammatory effects of sesamin. In conclusion, our results suggested that sesamin showed anti-inflammatory effects in IL-1ß-stimulated chondrocytes by activating Nrf2 signaling pathway.