Distinct mechanisms of iron and zinc metal ions on osteo-immunomodulation of silicocarnotite bioceramics.

The immunomodulatory of implants have drawn more and more attention these years. However, the immunomodulatory of different elements on the same biomaterials have been rarely investigated. In this work, two widely used biosafety elements, iron and zinc added silicocarnotite (Ca5(PO4)2SiO4, CPS) were applied to explore the routine of elements on immune response. The immune reactions over time of Fe-CPS and Zn-CPS were explored at genetic level and protein level, and the effects of their immune microenvironment with different time points on osteogenesis were also investigated in depth. The results confirmed that both Fe-CPS and Zn-CPS had favorable ability to secret anti-inflammatory cytokines. The immune microenvironment of Fe-CPS and Zn-CPS also could accelerate osteogenesis and osteogenic differentiation in vitro and in vivo. In terms of mechanism, RNA-seq analysis and Western-blot experiment revealed that PI3K-Akt signaling pathway and JAK-STAT signaling pathways were activated of Fe-CPS to promote macrophage polarization from M1 to M2, and its immune microenvironment induced osteogenic differentiation through the activation of Hippo signaling pathway. In comparison, Zn-CPS inhibited polarization of M1 macrophage via the up-regulation of Rap1 signaling pathway and complement and coagulation cascade pathway, while its osteogenic differentiation related pathway of immune environment was NF-κB signaling pathway.

Co-exchanged montmorillonite: a potential antibacterial agent with good antibacterial activity and cytocompatibility.

As a biocompatible material with rich resources and economic benefits, montmorillonite (MMT) has been widely used in the antibacterial field as a drug carrier and toxin adsorbent. In addition, the distinctive structure of MMT provides a possibility to tune its property in a wide range through ion-exchange. In this study, Co-montmorillonite (CoMMT) was prepared by the ion-exchanging method in a Co(NO3)2 solution and its antibacterial activity and cytocompatibility were investigated. The results showed that Co was introduced into MMT successfully and led to an increase in the interlayer spacing of MMT. Also, CoMMT showed a morphology of irregular aggregates consisting of stacked and intertwined lamellae with a uniform cobalt distribution. Besides, CoMMT had better dispersity and higher specific surface area than unmodified MMT. The antibacterial test results showed that CoMMT had good antibacterial activity against S. aureus and E. coli when the CoMMT concentration was higher than 0.2 mg mL-1 and 0.4 mg mL-1, respectively. The possible antibacterial mechanism of CoMMT was speculated and verified by a combination of SEM and EDS results. In addition, CoMMT showed no obvious cytotoxicity to MC3TC-E1 at the observed antibacterial concentration. These findings demonstrated that CoMMT with good biocompatibility and antibacterial activity could be used as a novel antibacterial agent for tissue engineering.

Osteoimmune reaction caused by a novel silicocarnotite bioceramic promoting osteogenesis through the MAPK pathway.

The host immune response to an implant is a key factor in determining the fate of bone grafts, which is thought to be a regulator of tissue regeneration. Figuring out the effects of the osteoimmune microenvironment on the osteogenesis of bone grafts can be a valuable strategy for their design and can further enhance the healing of bone defects. Our previous study demonstrated that the silicocarnotite (Ca5(PO4)2SiO4, CPS) bioceramic can significantly promote osteogenesis. The aim of this study is to investigate the immune reaction of CPS, the effects of the immune microenvironment on osteogenesis, and the related molecular mechanisms. Compared to hydroxyapatite (Ca10(PO4)6(OH)2, HA), the results showed that CPS could downregulate the pro-inflammatory phenotype and upregulate the anti-inflammatory phenotype, showing the lower levels of TNF-α and increased expression of IL-10. We further found that CPS could regulate the expression of NPPA, EDN1, and MMP9 in RAW 264.7 by RNA sequencing, which may be related to its superiority in osteogenesis. The osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs) was subsequently studied in a macrophage-conditioned medium pretreated with CPS, and the medium caused a significant promotion of the osteogenic differentiation of rBMSCs, demonstrating that CPS can generate a favorable immune microenvironment to promote rBMSCs differentiation. In terms of mechanism, CPS in the macrophage-conditioned medium promoted osteogenic differentiation through the MAPK pathway, including ERK1/2, JNK and P38. Our study demonstrated that osteogenic differentiation was influenced by the immune microenvironment generated via the implant, and also presented an effective tool for studying the mechanisms of macrophage polarization as well as functions.

Favorable osteogenic activity of iron doped in silicocarnotite bioceramic: In vitro and in vivo Studies.

BACKGROUND: Calcium phosphate silicate (Ca5(PO4)2SiO4 or CPS) is a promising bioceramic for bone grafting. Iron (Fe) is a trace element in the human body that has been reported to enhance the mechanical strength of CPS ceramics. However, the exact biofunctions of Fe, combined with another human trace element, viz. silicon (Si), in CPS and the optimal dose for Fe addition must be further investigated. METHODS: In vitro: the morphology, structure and cell adhesion were observed by SEM; the ability to promote osteogenic differentiation and mineralization was explored by ALP and alizarin red staining; the expression of osteogenic-specific genes and proteins was detected by PCR, WB and immunofluorescence. In vivo: Further exploration of bone regeneration capacity by establishing a skull defect model. RESULTS: In vitro, we observed increased content of adhesion-related proteins and osteogenic-related genes expression of Fe-CPS compared with CPS, as demonstrated by immunofluorescence and polymerase chain reaction experiments, respectively. In vivo micro-computed tomography images, histomorphology, and undecalcified bone slicing also showed improved osteogenic ability of Fe-CPS bioceramics. CONCLUSION: With the addition of Fe2O3, the new bone formation rate of the Fe-CPS scaffold after 12 weeks increased from 9.42% to 43.76%. Moreover, both in vitro and in vivo experimental outcomes indicated that Fe addition improved the CPS bioceramics in terms of their osteogenic ability by promoting the expression of osteogenic-related genes. Fe-CPS bioceramics can be employed as a novel material for bone tissue engineering on account of their outstanding new bone formation ability. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: This study suggests that Fe-CPS bioceramics can be employed as a novel material for bone tissue engineering on account of their outstanding new bone formation ability,which provides promising therapeutic implants and strategies for the treatment of large segmental bone defects.

Advanced protein adsorption properties of a novel silicate-based bioceramic: A proteomic analysis.

Silicate bioceramics have been shown to possess excellent cytocompatibility and osteogenic activity, but the exact mechanism is still unclear. Protein adsorption is the first event taking place at the biomaterial-tissue interface, which is vital to the subsequent cellular behavior and further influence the biomaterial-tissue interaction. In this work, the protein adsorption behavior of a novel CPS bioceramic was evaluated using the proteomics technology. The results showed that CPS adsorbed more amount and types of serum proteins than HA. FN1 and IGF1 proteins selected from proteomics results were validated by Western-blot experiment. Pathway analysis also revealed mechanistic insights how these absorbed proteins by CPS help mediate cell adhesion and promotes osteogenic activity. Firstly, the dramatically enhanced adsorption of FN1 could greatly promote cell adhesion and growth. Secondly, IGF1 was uniquely adsorbed on CPS bioceramic and IGF1 could activate Rap1 signaling pathway to promote cell adhesion. Thirdly, the increased adsorption of FN1, IGF1 and COL1A2 proteins on CPS explains its better ability on bone regeneration than HA. Fourthly, the increased adsorption of IGF1, CHAD, COL2A1 and THBS4 proteins on CPS explains its ability on cartilage formation. Lastly, the increased adsorption of immunological related proteins on CPS may also play a positive role in bone regeneration. In addition, CPS had a much better cell adhesion ability than HA, proving that more adsorbed proteins really had a positive effect on cell behavior. The more adsorbed proteins on CPS than HA might indicated a better bone regeneration rate at early stage of implantation.

Ferric oxide: A favorable additive to balance mechanical strength and biological activity of silicocarnotite bioceramic.

Ideal materials for bone regeneration should have not only a good bioactivity, but also a good mechanical strength to provide an initial support for new bone formation. How to get a balance between high mechanical property and good bioactivity is a challenging issue for bone regeneration materials. In the present work, a biocompatible additive Fe2O3 was selected to optimize the comprehensive properties of a novel calcium phosphate silicate (CPS) ceramic using a mechanical mixing method. The effects of Fe2O3 content on microstructure, bending strength, apatite formation ability and cytocompatibility of Fe-CPS bioceramics were investigated and the related mechanism was also discussed. The obtained Fe-CPS bioceramics showed enhanced mechanical and favorable bioactivity performances. Especially, the Fe-CPS bioceramic with 1.5 wt% Fe2O3 sintered at 1250 °C presented the highest bending strength of 91.9 MPa. While, Fe-CPS bioceramics still exhibited a good ability on apatite formation in simulated body fluid (SBF), and cytocompatibility test revealed that Fe-CPS bioceramics were favorable for cell adhesion and proliferation. All the results indicated that Fe-CPS bioceramics are promising candidate materials for bone regeneration at load bearing applications.

Osteoblastic and anti-osteoclastic activities of strontium-substituted silicocarnotite ceramics: In vitro and in vivo studies.

Osteoporosis bone defect is a refractory orthopaedic disease which characterized by impaired bone quality and bone regeneration capacity. Current therapies, including antiosteoporosis drugs and artificial bone grafts, are not always satisfactory. Herein, a strontium-substituted calcium phosphate silicate bioactive ceramic (Sr-CPS) was fabricated. In the present study, the extracts of Sr-CPS were prepared for in vitro study and Sr-CPS scaffolds were used for in vivo study. The cytocompatibility, osteogenic and osteoclastogenic properties of Sr-CPS extracts were characterized in comparison to CPS. Molecular mechanisms were also evaluated by Western blot. Sr-CPS extracts were found to promote osteogenesis by upregulating Wnt/ß-catenin signal pathways and inhibit osteoclastogenesis through downregulating NF-κB signal pathway. In vivo, micro-CT, histological and histomorphometric observation were conducted after 8 weeks of implantation to evaluate the bone formation using calvarial defects model in ovariectomized rats. Compared with CPS, Sr-CPS significantly promoted critical sized ovariectomy (OVX) calvarial defects healing. Among all the samples, Sr-10 showed the best performance due to a perfect match of bone formation and scaffold degradation rates. Overall, the present study demonstrated that Sr-CPS ceramic can dually modulate both bone formation and resorption, which might be a promising candidate for the reconstruction of osteoporotic bone defect.