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Li Q.,Peking University | Li Q.,National Engineering Laboratory for Digital and Material Technology of Stomatology | Zhou G.,Beihang University | Yu X.,Beihang University | And 3 more authors.
BioMedical Engineering Online | Year: 2015

Background: Bone substation grafts, such as hydroxyapatite (HA) and tricalciumphosphate (TCP), have been extensively used in clinical applications, but evidence suggests that they offer poor osteoinductive properties compared to allografts and autografts. In order to increase bone growth with such grafts, Bone Morphogenetic Protein 2 (BMP-2) was incorporated into a three dimensional reservoir. The purpose of the present study was to develop a novel drug delivery system which is capable of controlled release of BMP-2. Methods: DBB were prepared from bovine cancellous bone harvested from fetal bovine femur or tibia and then sinting at 1000°C. BMP-2-loaded chitosan (CS) microspheres were fabricated by cross-linking. Then the treated DBB powders were blended with chitosan microspheres solution. Finally, the composites were lyophilized with a freeze dryer to obtain the DBB/CMs scaffolds. X-ray diffractor (XRD), scanning electron microscopy (SEM) and Fourier transform infrared (FT-IR) were used to characterize the sample. The quantification of the delivery profile of BMP-2 was determined using an enzyme-linked immunosorbent assay (ELISA) kit. The in vitro assays were to characterize the biocompatibility of this composite. Results: In this study, BMP-2/Chitosan (CS) microspheres were successively loaded onto a deproteinized bovine bone (DBB) scaffold. The release profile of BMP-2 indicated an initial burst release followed by a more even sustained release. An in vitro bioactivity assay revealed that the encapsulated growth factor was biologically active. Conclusions: The cell culture assay suggest that the excellent biocompatibility of the DBB- BMP-2/CS. Therefore, this novel microsphere scaffold system can be effectively used in current tissue engineering applications. © Li et al.; licensee BioMed Central.

Liu Y.,Peking University | Chen T.,Peking University | Du F.,Peking University | Gu M.,Peking University | And 7 more authors.
Journal of Biomedical Nanotechnology | Year: 2016

In recent years, although several studies have demonstrated the potential of graphene-coated substrates in promoting attachment, proliferation and differentiation of osteoblasts and mesenchymal stem cells (MSCs), the effects of single-layer graphene on the osteogenic differentiation of human MSCs (hMSCs) remains unclear, especially in vivo. In this study, we transferred chemical vapor deposition (CVD) grown single-layer graphene to glass slides and observed its effects on adhesion, proliferation and osteogenic differentiation of human adipose-derived stem cells (hASCs) and human bone marrow mesenchymal stem cells (hBMMSCs) in vitro. Then, in vivo, we incubated hASCs and hBMMSCs on singlelayer graphene-coated smooth titanium (Ti) disks before implanting them into the back subcutaneous area of nude mice. We found that single-layer graphene accelerated cell adhesion to the substrate without influencing cell proliferation of hMSCs. Moreover, we present the first study that explores the epigenetic role of single-layer graphene in determining stem cell fate. By utilizing epigenetic approaches, we reveal that single-layer graphene promotes osteogenic differentiation of hMSCs both in vitro and in vivo, potentially by upregulating methylation of H3K4 at the promoter regions of osteogenesisassociated genes. Overall, our results highlight the potential of this material in implants and injured tissues in clinical applications. Copyright © 2016 American Scientific Publishers All rights reserved.

Zhang X.,Peking University | Zhang X.,Tsinghua University | Xu M.,Peking University | Song L.,Capital Medical University | And 8 more authors.
Biomaterials | Year: 2013

Combinations of calcium phosphate scaffolds and bioactive factors are promising niche-mimetic solutions for repairing large-sized bone defects. However, the importance of compatibility between scaffolds and bioactive factors on their osteogenic outcomes has been largely ignored. This study aimed to investigate the compatibility of calcinated antler cancellous bone (CACB) scaffolds with various bioactive factors including icariin (ICA), velvet antler polypeptides (VAP) or recombinant human bone morphogenetic protein-2 (rhBMP-2) as well as their combinational osteogenic potential invitro and invivo. Scanning electron microscopy and fourier transform infrared spectroscopy confirmed the uniform distribution and chemical stability of the reagents on CABC. Invitro release profiles showed relative steady release of ICA from ICA/CACB, burst VAP release from VAP/CACB, and minimal rhBMP-2 release from rhBMP-2/CACB composites. When compared with VAP and rhBMP-2, incorporation of ICA within CACB resulted in most increased cell attachment, proliferation, alkaline phosphatase activity, osteogenic gene expression, and mineralization of rat bone marrow mesenchymal stem cells. In rabbit mandible critical-sized defects, the most extensive osteogenesis and neovascularization were observed in the ICA/CACB group. Differences between the VAP/CACB and rhBMP-2/CACB groups were not apparent. Interestingly, low pro-inflammatory (TNF-α, IL-6) and high anti-inflammatory (IL-10) mRNA levels were observed at scaffold implantation sites which were in close association with amount of new bone formation. These findings highlight that the compatibility between scaffolds and bioactive factors should been taken into account when considering the formula of optimized bone defect repair. © 2013 Elsevier Ltd.

Wang T.-T.,Hebei United University | Zhang R.,Peking University | Wang L.,Hebei United University | Chen Y.,Peking University | And 3 more authors.
Diagnostic Pathology | Year: 2014

Background: The clinicopathologic characteristics of multiple ossifying fibroma (OF) are unclear due to the condition's rarity, making diagnosis challenging. Sporadic multiple OFs must be distinguished from hyperparathyroidism-jaw tumour syndrome (HPT-JT) related OF and other fibro-osseous lesions. Methods: Multiple OF cases were identified from ossifying fibroma cases. Clinical data including age, sex, anatomic site, radiographic features, clinical impression, treatment and available follow-up data as well as serum calcium, phosphorus, and parathyroid hormone (PTH) were recorded. GNAS and HRPT2 genetic mutations were examined in the two present cases. Case reports of sporadic multiple ossifying fibroma and HPT-JT-related OF were also reviewed. Results: The two present cases were confirmed as sporadic multiple OF, with no genetic GNAS and HRPT2 mutations found. The incidence of sporadic multiple ossifying fibroma was 2.0% (2/102). The total 18 sporadic multiform OF cases were characterized as followed: 13 (72.2%) female; 5 (27.8%) male; mean age 28.6 years; 2/16 (11.1%) cases only in the mandible; 4/18 (22.2%) cases only in the maxilla; and 12/18 (66.7%) cases in both the maxilla and mandible. Radiographically, the lesions were radiolucent in 5/18 (27.8%) cases and mixed density in 13/18 (72.2%) cases. Along with 24 cases of HPT-JT related OF were reviewed, sixteen (66.7%) patients were diagnosed with a single lesion, and 8 patients (33.3%) were diagnosed with multiple jaw lesions. Conclusions: Sporadic multiple OFs are very rare, but must be distinguished from HPT-JT related OF. We strongly recommend that patients diagnosed with multiple ossifying fibromas receive serum PTH testing and mutation screening of HRPT2. Virtual slides: http://www.diagnosticpathology.diagnomx.eu/vs/1194507146115753. © 2014 Wang et al.; licensee BioMed Central Ltd.

Liu Y.,Peking University | Liu Y.,National Engineering Laboratory for Digital and Material Technology of Stomatology | Zhang X.,Peking University | Jin X.,Peking University | And 7 more authors.
PLoS ONE | Year: 2014

Coating the surface of titanium implants or other bone graft substitute materials with calcium phosphate (Ca-P) crystals is an effective way to enhance the osteoconduction of the implants. Ca-P coating alone cannot confer pro-osteodifferentiation and antibacterial capabilities on implants; however, it can serve as a carrier for biological agents which could improve the performance of implants and bone substitutes. Here, we constructed a novel, bi-functional Ca-P coating with combined proosteodifferentiation and antibacterial capabilities. Different concentrations of metronidazole (MNZ) and simvastatin (SIM) were integrated into biomimetic Ca-P coatings on the surface of titanium disks. The biological effects of this bi-functional biomimetic coating on human bone marrow mesenchymal stem cells (hBMMSCs), human adipose derived stromal cells (hASCs), and Porphyromonas gingivalis were assessed in vitro. We observed that Ca-P coatings loaded with both SIM and MNZ display favorable release kinetics without affecting cell proliferation or attachment. In the inhibition zone test, we found that the bi-functional coating showed lasting antibacterial effects when incubated with Porphyromonas gingivalis for 2 and 4 days. Moreover, the osteodifferentiation of hBMMSCs and hASCs were increased when cultured on this bi-functional coating for 7 and 14 days. Both drugs were loaded onto the Ca-P coating at specific concentrations (10 -5 M SIM; 10-2 M MNZ) to achieve optimal release kinetics. Considering the safety, stability and low cost of SIM and MNZ, this novel bifunctional Ca-P coating technique represents a promising method to improve the performance of metal implants or other bone substitute materials, and can theoretically be easily translated to clinical applications. © 2014 Liu et al.

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