环磷酰胺诱导的小鼠骨髓细胞微核率的时间、剂量-效应观察

为研究环磷酰胺(CP)诱导的小鼠骨髓细胞微核率的时间-效应关系和剂量-反应关系,给小鼠一次腹腔注射不同剂量(按体质量计)CP(0、30、60、90 mg/kg)后,于不同时间(给药后12、24、36、486、0 h)、不同部位(胸骨与股骨)取材来观察小鼠骨髓细胞微核率的变化。结果表明,高、中、低三个剂量组的小鼠骨髓细胞微核率与0 mg/kg组相比较,其差异具有统计学意义(P<0.01),且4个剂量组之间两两比较也均有统计学意义(P<0.05),并表现出明显的剂量-反应关系;不同取样时间对小鼠骨髓细胞微核率的影响具有统计学意义(P<0.05),在36~48 h这个时间段能够观察到更高的微核率,通过线性回归的曲线拟合分析,以41 h取材最佳;胸骨取材与股骨取材之间的微核率差异无统计学意义(P>0.05)。环磷酰胺可作为诱导小鼠骨髓细胞微核的阳性对照,在一定范围内,具有剂量-反应关系和时间-效应关系。给小鼠一次腹腔注射量(按体质量计)60~90 mg/kg后36~48 h胸骨取材可取得较高微核率,结果满意。     

Osteogenesis of Osteoblast Seeded Polyurethane-Hydroxyapatite Scaffolds in Nude Mice

Summary: Biodegradable porous polyurethane (PU) scaffolds were used in a tissue engineering approach to create new bone. Two groups of elastomeric bioresorbable PU disks were seeded with osteoblasts and implanted into nude mice. One group had disks of pure PU while the other group had disks of PU- hydroxyapatite composite (PU-HA). After 5 weeks both groups showed radiographic and histologic evidence of significant bone formation. As the new bone formed it replaced the PU scaffolds. Although not statistically significant, there was a trend toward more bone formation in the PU-HA group. Bioresorbable PU shows promise for use in bone tissue engineering.     

Nanosilica-Anchored Polycaprolactone/Chitosan Nanofibrous Bioscaffold to Boost Osteogenesis for Bone Tissue Engineering

The strategy of incorporating bioactive inorganic nanomaterials without side effects as osteoinductive supplements is promising for bone regeneration. In this work, a novel biomass nanofibrous scaffold synthesized by electrospinning silica (SiO₂) nanoparticles into polycaprolactone/chitosan (PCL/CS) nanofibers was reported for bone tissue engineering. The nanosilica-anchored PCL/CS nanofibrous bioscaffold (PCL/CS/SiO₂) exhibited an interlinked continuous fibers framework with SiO₂ nanoparticles embedded in the fibers. Compact bone-derived cells (CBDCs), the stem cells derived from the bone cortex of the mouse, were seeded to the nanofibrous bioscaffolds. Scanning electron microscopy and cell counting were used to observe the cell adhesion. The Counting Kit-8 (CCK-8) assay was used. Alkaline phosphatase (ALP), Alizarin red staining, real-time Polymerase Chain Reaction and Western blot tests were performed to confirm the osteogenesis of the CBDCs on the bioscaffolds. The research results demonstrated that the mechanical property of the PCL together with the antibacterial and hydrophilic properties of the CS are conducive to promoting cell adhesion, growth, migration, proliferation and differentiation. SiO₂ nanoparticles, serving as bone induction factors, effectively promote the osteoblast differentiation and bone regeneration. This novel SiO₂-anchored nanofibrous bioscaffold with superior bone induction activity provides a better way for bone tissue regeneration.     

Multiple‐Level Porous Polymer Monoliths with Interconnected Cellular Topology Prepared by Combining Hard Sphere and Emulsion Templating for Use in Bone Tissue Engineering

A combination of hard sphere and high internal phase emulsion templating gives a platform for synthesizing hierarchically porous polymers with a unique topology exhibiting interconnected spherical features on multiple levels. Polymeric spheres are fused by thermal sintering to create a 3D monolithic structure while an emulsion with a high proportion of internal phase and monomers in the continuous phase is added to the voids of the previously constructed monolith. Following polymerization of the emulsion and dissolution of the templating structure, a down‐replicating topology is created with a primary level of pores as a result of fused spheres of the 3D monolithic structure, a secondary level of pores resulting from the emulsion's internal phase, and a tertiary level of interconnecting channels. Thiol‐ene chemistry with divinyladipate and pentaerythritol tetrakis(3‐mercaptopropionate) is used to demonstrate the preparation of a crosslinked polyester with overall porosity close to 90%. Due to multilevel porosity, such materials are interesting for applications in bone tissue engineering, possibly simulating the native sponge like bone structure. Their potential to promote ossteointegration is tested using human bone derived osteoblasts. Material–cell interactions are evaluated and they reveal growth and proliferation of osteoblasts both on surface and in the bulk of the scaffold.     

Biological Activity of an Injectable Biphasic Calcium Phosphate/PMMA Bone Cement for Induced Osteogensis in Rabbit Model

Polymethylmethacrylate (PMMA) bone cement is widely used in repair of vertebral fracture because of its good biomechanical properties and fast curing. However, the bioinertness of PMMA cement may cause interfacial loosening, fatigue, fracture, and ultimate failure. In this study, biphasic calcium phosphate (BCP) is introduced into PMMA cement to prepare an injectable composite bone cement (BCP_x/PMMA) and the content of BCP is optimized to achieve appropriate rate of absorption that matches the bone regeneration. The compressive strength of BCP_x/PMMA bone cement is found to comply with the International Standardization Organization standard 5833, and can promote biomineralization as well as adhesion, proliferation, and osteogenic differentiation of Sprague‐Dawley rat bone marrow mesenchymal stem cells in vitro. Furthermore, in vivo test performed on a rabbit radius defect model demonstrates that the presence of BCP can significantly improve the osteogenic efficacy of PMMA cement. Therefore, it is anticipated that BCP_x/PMMA bone cement, as a promising injectable biomaterial, is of great potential in bone tissue regeneration.     

Solid‐phase extraction of drugs with cuttlefish bone powder as a sorbent

We investigated cuttlefish bone powder for the solid‐phase extraction of naproxen, ibuprofen, and carbamazepine. The basic principles controlling the extraction are presented to aid in the choice of the nature and quantity of the extracting phase according to the sample matrix and the solute properties, based on the mechanisms of phase retention. Their retention mechanism is based on hydrogen bonding and electrostatic interactions. The results show a significant recovery rate for the three drugs, selectivity, and low cost. The method has successfully reduced the amount of tested pharmaceuticals with recoveries >87% at pH 4.     

Exploring the Proteomic Alterations from Untreated and Cryoablation and Irradiation Treated Giant Cell Tumors of Bone Using Liquid-Chromatography Tandem Mass Spectrometry

Giant cell tumors of bone (GCT) are benign tumors that show a locally aggressive nature and affect bones’ architecture. Recently, cryoablation and irradiation treatments have shown promising results in GCT patients with faster recovery and less recurrence and metastasis. Therefore, it became a gold standard surgical treatment for patients. Hence, we have compared GCT-untreated, cryoablation, and irradiation-treated samples to identify protein alterations using high-frequency liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Our label-free quantification analysis revealed a total of 107 proteins (p < 0.01) with 26 up-regulated (<2-folds to 5-fold), and 81 down-regulated (>0.1 to 0.5 folds) proteins were identified from GCT-untreated and treated groups. Based on pathway analysis, most of the identified up-regulated proteins involved in critical metabolic functions associated with tumor proliferation, angiogenesis, and metastasis. On the other hand, the down-regulated proteins involved in glycolysis, tumor microenvironment, and apoptosis. The observed higher expressions of matrix metalloproteinase 9 (MMP9) and TGF-beta in the GCT-untreated group associated with bones’ osteolytic process. Interestingly, both the proteins showed reduced expressions after cryoablation treatment, and contrast expressions identified in the irradiation treated group. Therefore, these expressions were confirmed by immunoblot analysis. In addition to these, several glycolytic enzymes, immune markers, extracellular matrix (ECM), and heat shock proteins showed adverse expressions in the GCT-untreated group were identified with favorable regulations after treatment. Therefore, the identified expression profiles will provide a better picture of treatment efficacy and effect on the molecular environment of GCT.     

Electrospun fibrous membranes of poly (lactic-co-glycolic acid) with β-tricalcium phosphate for guided bone regeneration application

Guided bone regeneration (GBR) have been proved great success for the treatment of bone defects in oral implantology. Bone defects are reconstructed by the use of a barrier membrane which prevents the invasion of soft tissue and create a space for guiding new bone growth into the bone defect. To develop GBR membranes with higher bioactivity, poly (lactic-co-glycolic acid) (PLGA) fibrous membranes containing β-tricalcium phosphate (β-TCP) were fabricated via electrospinning. The presence of 5 wt% and 10 wt% of β-TCP in the fibers improved mechanical properties. In vitro biocompatibility results have shown that all membranes are non-cytotoxic and the presence of β-TCP increased cell adhesion and proliferation. Furthermore, cell viability results demonstrated that the presence of 5 wt% β-TCP is advantageous for osteoblast proliferation. Therefore, the results suggest that PLGA with 5 wt% β-TCP fibrous membranes meet the requirement of morphological, physical, mechanical and bioactive properties for an effective GBR membrane.     

Intramarrow injection of β-catenin-activated,but not na?ve mesenchymal stromal cells stimulates self-renewal of hematopoietic stem cells in bone marrow

Bone marrow mesenchymal stromal cells (MSCs) have been implicated in the microenvironmental support of hematopoietic stem cells (HSCs) and often co-transplanted with HSCs to facilitate recovery of ablated bone marrows. However, the precise effect of transplanted MSCs on HSC regeneration remains unclear because the kinetics of HSC self-renewal in vivo after co-transplantation has not been monitored. In this study, we examined the effects of intrafemoral injection of MSCs on HSC self-renewal in rigorous competitive repopulating unit (CRU) assays using congenic transplantation models in which stromal progenitors (CFU-F) were ablated by irradiation. Interestingly, naïve MSCs injected into femur contributed to the reconstitution of a stromal niche in the ablated bone marrows, but did not exert a stimulatory effect on the in-vivo self-renewal of co-transplanted HSCs regardless of the transplantation methods. In contrast, HSC self-renewal was four-fold higher in bone marrows intrafemorally injected with β-catenin-activated MSCs. These results reveal that naïve MSCs lack a stimulatory effect on HSC self-renewal in-vivo and that stroma must be activated during recoveries of bone marrows. Stromal targeting of wnt/β-catenin signals may be a strategy to activate such a stem cell niche for efficient regeneration of bone marrow HSCs.     

Recent Advances in Nanomedicine for the Diagnosis and Treatment of Prostate Cancer Bone Metastasis

Patients with advanced prostate cancer can develop painful and debilitating bone metastases. Currently available interventions for prostate cancer bone metastases, including chemotherapy, bisphosphonates, and radiopharmaceuticals, are only palliative. They can relieve pain, reduce complications (e.g., bone fractures), and improve quality of life, but they do not significantly improve survival times. Therefore, additional strategies to enhance the diagnosis and treatment of prostate cancer bone metastases are needed. Nanotechnology is a versatile platform that has been used to increase the specificity and therapeutic efficacy of various treatments for prostate cancer bone metastases. In this review, we summarize preclinical research that utilizes nanotechnology to develop novel diagnostic imaging tools, translational models, and therapies to combat prostate cancer bone metastases.