拉曼光谱在骨组织研究中的应用

拉曼光谱具有快速、无损、能同时提供无机和有机成分信息,包括矿物晶格和胶原蛋白二级结构信息等特点,在骨组织研究中有着重要应用。本文结合本课题组工作,对骨组织的光谱背景处理、拉曼光谱在骨的力学性能、骨科疾病、成像研究以及活体检测等方面的应用进行了综述。     

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.     

Optimal internal architectures of femoral bone based on relaxation by homogenization and isotropic material design

The influence of the loading conditions on the trabecular architecture of a femur is investigated by using topology optimization methods. The response of the bone to physiological loads results in changes of the internal architecture of bone, reflected by a modification of internal effective density and mechanical properties. The homogenization based optimization model is developed for predicting optimal bone density distribution, wherein bone tissue is assumed to be a composite material consisting of a mixture of material and void. The homogenization scheme treats the geometric parameters of the microstructures and their orientation as design variables and homogenizes the properties in that microstructure, which is generally anisotropic. The penalization of the optimal material density then leads to a classical optimal structure which consists of regions with bone material and regions without bone material. The IMD (Isotropic Material Design) approach is next applied to determine the optimal elasticity tensor in terms of the bulk and shear moduli for the present loading applied to the femoral bone sample. IMD is able to provide both the external shape and topology together with the optimal layout of the isotropic moduli. Both topology optimization methods appear to be complementary. Simulations of the internal bone architecture of the human proximal femur results in a density distribution pattern with good consistency with that of the real bone.     

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.     

Fabrication of chitosan/collagen/hydroxyapatite scaffolds with encapsulated Cissus quadrangularis extract

Here, we demonstrated the fabrication of a composite scaffold (chitosan [CS], collagen [Col], and hydroxyapatite [HA]) with the incorporation of encapsulated Cissus quadrangularis (CQ) extract for tissue engineering applications. First, the crude extract of CQ loaded nanoparticles were synthesized via double emulsion technique using polycaprolactone (PCL) and polyvinyl alcohol (PVA) as oil and aqueous phases, respectively. Both PCL (20, 40, and 80 mg/mL) and PVA (0.5%, 1%, and 3% w/v) concentrations were varied to determine the optimum concentrations for CQ‐loaded nanoparticle preparation. The CQ‐loaded PCL nanoparticles (CQ‐PCL NPs), prepared with 20 mg/mL PCL and 0.5% (w/v) PVA, exhibited the smallest size of 334.22 ± 43.21 nm with 95.54 ± 1.49% encapsulation efficiency. Then, the CQ‐PCL NPs were incorporated into the CS/Col/HA scaffolds. These scaffolds were also studied for their ultrastructure, pore sizes, chemical composition, compressive modulus, water swelling, weight loss, and biocompatibility. The results showed that the addition of CQ‐PCL NPs into the scaffolds did not dramatically alter the ultrastructure and properties of the scaffolds, compared to CS/Col/HA scaffolds alone. However, incorporation of CQ‐PCL NPs in the scaffolds improved the release profile of CQ by preventing the initial burst release and prolonging the release rate of CQ. In addition, the CQ‐PCL NPs‐loaded CS/Col/HA scaffolds supported the attachment and proliferation of MC3T3‐E1 osteoblast cells.     

Micro‐Nanostructures of Cellulose‐Collagen for Critical Sized Bone Defect Healing

Bone tissue engineering strategies utilize biodegradable polymeric matrices alone or in combination with cells and factors to provide mechanical support to bone, while promoting cell proliferation, differentiation, and tissue ingrowth. The performance of mechanically competent, micro‐nanostructured polymeric matrices, in combination with bone marrow stromal cells (BMSCs), is evaluated in a critical sized bone defect. Cellulose acetate (CA) is used to fabricate a porous microstructured matrix. Type I collagen is then allowed to self‐assemble on these microstructures to create a natural polymer‐based, micro‐nanostructured matrix (CAc). Poly (lactic‐co‐glycolic acid) matrices with identical microstructures serve as controls. Significantly higher number of implanted host cells are distributed in the natural polymer based micro‐nanostructures with greater bone density and more uniform cell distribution. Additionally, a twofold increase in collagen content is observed with natural polymer based scaffolds. This study establishes the benefits of natural polymer derived micro‐nanostructures in combination with donor derived BMSCs to repair and regenerate critical sized bone defects. Natural polymer based materials with mechanically competent micro‐nanostructures may serve as an alternative material platform for bone regeneration.     

青鱼鳍骨、肋骨和鳃盖骨微观结构与力学性能的比较研究

本文从材料学角度定量分析了青鱼鳍骨、肋骨、鳃盖骨的微观结构、成分及力学特性。利用电子显微镜观察了鳍骨、肋骨、鳃盖骨的微观结构;通过X射线衍射、电镜能谱以及煅烧的骨灰综合分析,探讨了青鱼鳍骨、肋骨、鳃盖骨的有机、无机成分之间的密切联系;进行了青鱼鳍骨、肋骨、鳃盖骨的拉伸实验;利用显微硬度计对干、湿状态下的鳍骨、肋骨、鳃盖骨进行了硬度测试。结果显示,鳍骨无机物含量较多,鳃盖骨有机物较多,肋骨介于两者之间;矿化胶原蛋白束在鳍骨中的排列非常规则,在鳃盖骨中排列不是很规则,在肋骨中排列较规则;鳍骨中矿化胶原纤维束间孔洞非常少,骨质密实,而在腮盖骨中孔洞很多,骨质稀疏;干的鱼骨显微硬度均大于湿的鱼骨。结论是,鳍骨、肋骨、鳃盖骨的微观结构、成分及力学特性存在差异,其中鳍骨的力学性能明显强于鳃盖骨,肋骨介于二者之间,水份对青鱼鳍骨、肋骨、鳃盖骨的硬度有明显的影响。     

Surface Modification of Poly (ether ether ketone) with a Medlite C6 (ND-YAG Q-Switched) Skin Treatment Laser

Poly ether ether ketone (PEEK), a synthetic polymer, is expected to be useful as a biomaterial due to its appropriate mechanical, chemical, and biocompatibility properties. However, this polymer is biologically inert, requiring surface modification to improve its adhesion to bone cells for use as a bone substrate. Surface properties, such as roughness and hydrophilicity, are important factors in the adhesion of biomaterials to the surrounding tissue; therefore, in this study, laser treatment was performed for surface modification. The aim of the research described here was to investigate the effect of two laser parameters, fluency and wavelength, on the surface roughness and hydrophilicity to determine the optimum parameters for improving surface adhesion. The surface topography and average roughness (R_a) were investigated by atomic force microscopy (AFM). Surface morphology was also observed with an optical microscope, and the hydrophilicity of the surfaces was investigated with static contact angle tests. The results obtained showed that the samples treated at the wavelength of 532?nm with fluency of 8?J/cm², compared to fluencies of 4 and 12?J/cm², showed improved surface properties. However, in terms of radiation wavelength, the wavelength of 1064?nm at these three fluencies showed the most promising results for enhancing the surface properties of PEEK for bone implant applications.     

Digital volume correlation: Three-dimensional strain mapping using X-ray tomography

A three-dimensional extension of two-dimensional digital image correlation has been developed. The technique uses digital image volumes generated through high-resolution X-ray tomography of samples with microarchitectural detail, such as the trabecular bone tissue found within the skeleton. Image texture within the material is used for displacement field measurement by subvolume tracking. Strain fields are calculated from the displacement fields by gradient estimation techniques. Estimates of measurement precision were developed through correlation of repeat unloaded data sets for a simple sum-of-squares displacement-only correlation formulation. Displacement vector component errors were normally distributed, with a standard deviation of 0.035 voxels (1.22 m). Strain tensor component errors were also normally distributed, with a standard deviation of approximately 0.0003. The method was applied to two samples taken from the thigh bone near the knee. Strains were effectively measured in both the elastic and postyield regimes of material behavior, and the spatial patterns showed clear relationships to the sample microarchitectures.