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1.
In bone tissue engineering, a highly porous artificial extracellular matrix or scaffold is required to accommodate cells and guide the tissue regeneration in three-dimension. Calcium phosphate (CaP) ceramics are widely used for bone substitution and repair due to their biocompatibility, bioactivity, and osteoconduction. However, compared to alumina ceramics, either in the dense or porous form, the mechanical strength achieved for calcium phosphates is generally lower. In the present work, the major goal was to develop a tri-dimensional macroporous alumina scaffold with a biocompatible PVA/calcium phosphate coating to be potentially used as bone tissue substitute. This approach aims to combine the high mechanical strength of the alumina scaffold with the biocompatibility of calcium phosphate based materials. Hence, the porous alumina scaffolds were produced by the polymer foam replication procedure. Then, these scaffolds were submitted to two different coating methods: the biomimetic and the immersion in a calcium phosphate/polyvinyl alcohol (CaP/PVA) slurry. The microstructure, morphology and crystallinity of the macroporous alumina scaffolds samples and coated with CaP/PVA were characterized by X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM/EDX) analysis. Also, specific surface area was assessed by BET nitrogen adsorption method and mechanical behavior was evaluated by axial compression tests. Finally, biocompatibility and cytotoxicity were evaluated by VERO cell spreading and attachment assays under SEM. The morphological analysis obtained from SEM photomicrograph results has indicated that 3D macroporous alumina scaffolds were successfully produced, with estimated porosity of over 65% in a highly interconnected network. In addition, the mechanical test results have indicated that porous alumina scaffolds with ultimate compressive strength of over 3.0 MPa were produced. Concerning to the calcium phosphate coatings, the results have showed that the biomimetic method was not efficient on producing a detectable layer onto the alumina scaffolds. On the other hand, a uniform and adherent inorganic–organic coating was effectively formed onto alumina macroporous scaffold by the immersion of the porous structure into the CaP/PVA suspension. Viable VERO cells were verified onto the surface of alumina porous scaffold samples coated with PVA–calcium phosphate. In conclusion, a new method was developed to produce alumina with tri-dimensional porous structure and uniformly covered with a biocompatible coating of calcium phosphate/PVA. Such system has high potential to be used in bone tissue engineering.  相似文献   

2.
Bioceramic mixtures of tricalcium phosphate (TCP) and hydroxyapatite (HAp) are widely used for bone regeneration because of their excellent cytocompatibility, osteoconduction, and osteoinduction. Therefore, we hypothesized that incorporation of a mixture of TCP and HAp in microsphere-based scaffolds would enhance osteogenesis of rat bone marrow stromal cells (rBMSCs) compared to a positive control of scaffolds with encapsulated bone-morphogenic protein-2 (BMP-2). Poly(d,l-lactic-co-glycolic acid) (PLGA) microsphere-based scaffolds encapsulating TCP and HAp mixtures in two different ratios (7:3 and 1:1) were fabricated with the same net ceramic content (30 wt%) to evaluate how incorporation of these ceramic mixtures would affect the osteogenesis in rBMSCs. Encapsulation of TCP/HAp mixtures impacted microsphere morphologies and the compressive moduli of the scaffolds. Additionally, TCP/HAp mixtures enhanced the end-point secretion of extracellular matrix components relevant to bone tissue compared to the “blank” (PLGA-only) microsphere-based scaffolds as evidenced by the biochemical, gene expression, histology, and immunohistochemical characterization. Moreover, the TCP/HAp mixture groups even surpassed the BMP-2 positive control group in some instances in terms of matrix synthesis and gene expression. Lastly, gene expression data suggested that the rBMSCs responded differently to different TCP/HAp ratios presented to them. Altogether, it can be concluded that TCP/HAp mixtures stimulated the differentiation of rBMSCs toward an osteoblastic phenotype, and therefore may be beneficial in gradient microsphere-based scaffolds for osteochondral regeneration.  相似文献   

3.
A possible surgical technique in the replacement of a traumatized hip joint by a prosthesis system is to connect the acetabular component of the implant directly with pelvis bone tissue, without use of bone cement. It is possible to improve the osteointegration process and to ensure a better connection with bone tissue by coating the outside implant surface with a biocompatible ceramic. The best choice for a bioceramic coating is porous hydroxylapatite because its surface shows bonding-osteogenesis properties much higher than other materials. Here, a double HAp layer has been made by tape casting technology. The first layer was a high porous HAp ceramic with high osteophilic-osteoconductive characteristics. Because the scale of porosity was relatively insensitive to slurry composition and sintering temperature such a microstructure was produced using a particular technique described here. The second layer was dense HAp ceramic that resulted a substrate able to improve the mechanical properties of the brittle porous HAp layer. Several microstructure-designed ceramic coatings having the porous part with a controlled porosity can be obtained by tape casting using the same technique.  相似文献   

4.
采用有机泡沫浸渍工艺制备了高孔隙率的钙磷多孔陶瓷支架, 将多孔陶瓷样品浸于明胶溶液中渗涂得到陶瓷/明胶复合支架; 采用复合明胶涂层的方法对钙磷多孔陶瓷支架进行增强处理, 在不破坏多孔支架孔隙特征的情况下, 成功地在样品的孔壁上复合了明胶涂层。复合明胶涂层提高了样品的压缩强度和压缩模量, 与未涂覆样品相比, 涂敷样品受压时的应变特性发生了明显变化。尤其是渗涂5%明胶溶液的多孔样品, 在保持高孔隙率(82.8%)的条件下其压缩强度和压缩模量分别由原来的1.04MPa 和 0.105GPa增加到5.17MPa和0.325GPa。研究结果表明, 孔壁上复合明胶涂层可以有效地增强多孔陶瓷支架。   相似文献   

5.
In the present work, a novel route for the preparation of porous ceramic microspheres is described. Two ceramic powders, calcium-titanium-phosphate (CTP) and hydroxyapatite (HAp), were mixed with a sodium alginate solution that enabled the preparation of spherical particles, using the droplet extrusion method combined with ionotropic gelation in the presence of Ca2+. The spherical particles were subsequently sintered, to burn-off the polymer and obtain calcium-phosphate microspheres with a uniform size and an interconnected porous network. CTP microspheres with diameters ranging from 513 ± 24 μm to 792 ± 35 μm and with pores of approximately 40 μm were obtained. HAp microspheres presented diameters of 429 ± 46 μm and 632 ± 40 μm and pores of ca. 2 μm. Depending on the formulations tested, the structure of both calcium phosphates may become altered during the sintering process, suggesting that the ratio between the ceramic phase and the polymer solution is a critical parameter. Porous microspheres prepared using the described methodology are promising candidates as bone defect fillers and scaffolds for bone tissue regeneration.  相似文献   

6.
It is known that calcium-phosphate (Ca-P) coatings are able not only to improve the bone bonding behaviour of polymeric materials, but at the same time play a positive role on enhancing cell adhesion and inducing the differentiation of osteoprogenitor cells. Recently an innovative biomimetic methodology, in which a sodium silicate gel was used as a nucleative agent, was proposed as an alternative to the currently available biomimetic coating methodologies. This methodology is especially adequate for coating biodegradable porous scaffolds. In the present work we evaluated the influence of the referred to treatment on the mechanical properties of 50/50 (wt%) blend of corn starch/ethylene-vinyl alcohol (SEVA-C) based scaffolds. These Ca-P coated scaffolds presented a compressive modulus of 224.6 ± 20.6 and a compressive strength of 24.2 ± 2.20. Cytotoxicity evaluation was performed according ISO/EN 10993 part 5 guidelines and showed that the biomimetic treatment did not have any deleterious effect on L929 cells and did not inhibit cell growth. Direct contact assays were done by using a cell line of human osteoblast like cells (SaOS-2). 3 × 105 cells were seeded per scaffold and allowed to grow for two weeks at 37C in a humidified atmosphere containing 5% CO2. Total protein quantification and scanning electron microscopy (SEM) observation showed that cells were able to grow in the pre-mineralized scaffolds. Furthermore cell viability assays (MTS test) also show that cells remain viable after two weeks in culture. Finally, protein expression studies showed that after two weeks osteopontin and collagen type I were being expressed by SaOS-2 cells seeded on the pre-mineralized scaffolds. Moreover, alkaline phosphatase (ALP) activity was higher in the supernatants collected from the pre-mineralized samples, when compared to the control samples (non Ca-P coated). This may indicate that a faster mineralization of the ECM produced on the pre-mineralized samples was occurring. Consequently, biomimetic pre-mineralization of starch based scaffolds can be a useful route for applying these materials on bone tissue engineering.  相似文献   

7.
The development of cell therapy methods to confer osteogenic potential to synthetic bone replacement materials has become common during the last years. At present, in the bone tissue engineering field, two different approaches use patient own cultured osteogenic cells in combination with a scaffold material to engineer autologous osteogenic grafts. One of the approaches consists of seeding cells on a suitable biomaterial, after which the construct is ready for implantation. In the other approach, the seeded cells are further cultured on the scaffold to obtain in vitro formed bone (extracellular matrix and cells), prior to implantation. In the present study, we investigated the in vivo osteogenic potential of both methods through the implantation of porous hydroxyapatite (HA) scaffolds coated with a layer of in vitro formed bone and porous HA scaffolds seeded with osteogenic cells. Results showed that as early as 2 days after implantation, de novo bone tissue was formed on scaffolds in which an in vitro bone-like tissue was cultured, while it was only detected on the cell seeded implants from 4 days onwards. In addition, after 4 days of implantation statistical analysis revealed a significantly higher amount of bone in the bone-like tissue containing scaffolds as compared to cell seeded ones.  相似文献   

8.
The inability to maintain high concentrations of antibiotic at the site of infection for an extended period of time along with dead space management is still the driving challenge in treatment of osteomyelitis. Porous bioactive ceramics such as hydroxyapatite (HAp) and beta-tri calcium phosphate (β-TCP) were some of the alternatives to be used as local drug delivery system. However, high porosity and high interconnectivity of pores in the scaffolds play a pivotal role in the drug release and bone resorption. Ceftriaxone is a cephalosporin that has lost its clinical popularity. But has recently been reported to exhibit better bactericidal activity in vitro and reduced probability of resistance development, in combination with sulbactam, a β-lactamase inhibitor. In this article, a novel approach of forming HAp and pure β-TCP based porous scaffolds by applying together starch consolidation with foaming method was used. For the purpose, pure HAp and β-TCP were prepared in the laboratory and after thorough characterization (including XRD, FTIR, particle size distribution, etc.) the powders were used for scaffold fabrication. The ability of these scaffolds to release drugs suitably for osteomyelitis was studied in vitro. The results of the study indicated that HAp exhibited better drug release profile than β-TCP when drug was used alone indicating the high influence of the carrier material. However, this restriction got relaxed when a bilayered scaffold was formed using chitosan along with the drug. SEM studies along with EDAX on the drug-chitosan bilayered scaffold showed closest apposition of this combination to the calcium phosphate surface.  相似文献   

9.
Bone‐tissue engineering mandates the development of multi‐functional bioactive porous hydroxyapatite (HAp) scaffolds. Herein, microwave sintered HAp/ZnO and HAp/Ag composite scaffolds with ≈5–19% porosity are developed using 0–30 vol% graphite as a porogen. The mechanical properties of the porous scaffold are analyzed in detail, revealing that even being more porous, the reinforcement of ZnO (9% porosity, hardness of 2.8 GPa, and toughness of 3.5 MPa.m1/2) has shown to have better hardness and fracture toughness when compared to Ag (5% porosity, hardness of 1.6 GPa, and toughness of 2.6 MPa.m1/2). The flexural strength obtained experimentally are complemented with a finite‐element technique that adopts microstructural features in visualizing the effect of porosity on stress distribution. The antibacterial efficacy and cytocompatibility of these composites are validated by increased metabolic activity and conspicuous cell‐matrix interactions. The anticipation of the results reveal that HAp/ZnO (9% porosity) and HAp/Ag (5% porosity) composites can be used as a potential multi‐functional bone implant scaffolds.
  相似文献   

10.
Porous ceramics made of alumina and hydroxyapatite were created using a protein foaming method. Porosity and pore size distribution were successfully varied by means of chemical modification of the foaming protein Bovine serum albumin (BSA). The effectiveness of the BSA and of its chemical modifications as well as the influence of the dispersing agent were investigated using synchrotron tomography. Resulting porous ceramic materials were used as three-dimensional substrates for the cultivation of human peripheral stem cells. The cells proliferated and differentiated in culture. Five cell lines consistent with human blood cell lines were observed.  相似文献   

11.
Biomaterial surfaces and their nanostructures can significantly influence cell growth and viability. Thus, manipulating surface characteristics of scaffolds can be a potential strategy to control cell functions for stem cell tissue engineering. In this study, in order to construct a hydroxyapatite (HAp) coated genipin-chitosan conjugation scaffold (HGCCS) with a well-defined HAp nanostructured surface, we have developed a simple and controllable approach that allows construction of a two-level, three-dimensional (3D) networked structure to provide sufficient calcium source and achieve desired mechanical function and mass transport (permeability and diffusion) properties. Using a nontoxic cross-linker (genipin) and a nanocrystallon induced biomimetic mineralization method, we first assembled a layer of HAp network-like nanostructure on a 3D porous chitosan-based framework. X-ray diffraction (XRD) and high resolution transmission electron microscopy (HRTEM) analysis confirm that the continuous network-like nanostructure on the channel surface of the HGCCS is composed of crystalline HAp. Compressive testing demonstrated that the strength of the HGCCS is apparently enhanced because of the strong cross-linking of genipin and the resulting reinforcement of the HAp nanonetwork. The fluorescence properties of genipin-chitosan conjugation for convenient monitoring of the 3D porous scaffold biodegradability and cell localization in the scaffold was specifically explored using confocal laser scanning microscopy (CLSM). Furthermore, through scanning electron microscope (SEM) observation and immunofluorescence measurements of F-actin, we found that the HAp network-like nanostructure on the surface of the HGCCS can influence the morphology and integrin-mediated cytoskeleton organization of rat bone marrow-derived mesenchymal stem cells (BMSCs). Based on cell proliferation assays, rat BMSCs tend to have higher viability on HGCCS in vitro. The results of this study suggest that the fluorescent two-level 3D nanostructured chitosan-HAp scaffold will be a promising scaffold for bone tissue engineering application.  相似文献   

12.
The main principle of a bone tissue engineering (BTE) strategy is to cultivate osteogenic cells in an osteoconductive porous scaffold. Ceramic implants for osteogenesis are based mainly on hydroxyapatite (HA), since this is the inorganic component of bone. Rapid Prototyping (RP) is a new technology in research for producing ceramic scaffolds. This technology is particularly suitable for the fabrication of individually and specially tailored single implants. For tissue engineering these scaffolds are seeded with osteoblast or osteoblast precursor cells. To supply the cultured osteoblastic cells efficiently with nutrition in these 3D-geometries a bioreactor system can be used. The aim of this study was to analyse the influence of differently fabricated HA-scaffolds on bone marrow stromal cells. For this, two RP-techniques, dispense-plotting and a negative mould method, were used to produce porous ceramics. The manufactured HA-scaffolds were then cultivated in a dynamic system (bioreactor) with an osteoblastic precursor cell line. In our study, the applied RP-techniques give the opportunity to design and process HA-scaffolds with defined porosity, interconnectivity and 3D pore distribution. A higher differentiation of bone marrow stromal cells could be detected on the negative mould fabricated scaffolds, while cell proliferation was higher on the dispense-plotted scaffolds. Nevertheless, both scaffold types can be used in tissue engineering applications.  相似文献   

13.
This paper reviews advances in the fabrication of calcium phosphate materials for injured bone tissue regeneration. We examine the key features of rapid prototyping for the fabrication of porous ceramic scaffolds with tailored architectures, the technology of biopolymer-based composite materials reinforced with calcium phosphate particles, and the fabrication of porous scaffolds via cement route.  相似文献   

14.
Biomimetic porous scaffolds made of calcium phosphate mineral are promising structures to develop bone replacement materials. In order to fabricate scaffolds with a strut size of 450 μm, we used a stereolithographic technique which selectively polymerises photosensitive liquid resin by visible light to produce casting moulds for ceramic gelcasting. These moulds were filled with a water based thermosetting ceramic slurry which solidifies inside the mould. After burning the resin mould and sintering, hydroxylapatite structures with designed, fully interconnected macroporosity were obtained.The preosteoblastic cell line MC3T3-E1, derived from mouse calvariae, was used to test for biocompatibility in cell culture experiments. The cells were seeded on the scaffolds immersed in the culture medium and cultured for 2 weeks. Thereafter the cells on the scaffold were fixed and investigated by histological methods. The osteoblast-like cells were found to cover the whole external and internal surface of the scaffold, they were embedded in collagenous extracellular matrix. The cells had in particular the tendency to fill any crack or opening and to generally smooth the exposed surfaces.  相似文献   

15.
Tricalcium phosphate (TCP) ceramic is known in orthopedics to be a bioresorbable bone substitute. A porous TCP ceramic body also has high potential as a drug delivery system in bony defects. Porous alpha-TCP ceramic can be easily fabricated using conventional sintering of beta-TCP, since alpha-TCP is the thermodynamically stable phase at temperatures above 1 100 degrees C. However, the solubility of alpha-TCP is much higher than that of beta-TCP. Therefore, the dissolution of porous alpha-TCP progresses at a higher rate than bone repair. In the present study, we attempted to reduce the dissolution rate of porous alpha-TCP by employing an organic polymer coating. We fabricated porous alpha-TCP ceramic with a continuous 10-50 microm diameter pore structure by sintering a body made from a beta-TCP and potato starch slurry. The porous body obtained was coated with hydroxypropylcellulose (HPC), and then subjected to heat treatment. The chemical durability and mechanical properties of the body were examined before and after coating with the HPC. The dissolution of porous alpha-TCP in buffered solutions was reduced by coating with HPC and drying at 60 degrees C. The compressive strength of the porous alpha-TCP was also improved by coating with HPC. The results of in vivo experiments showed that some parts of the porous alpha-TCP ceramic coated with HPC remained in the canal of the tibia of a rabbit four weeks after implantation, whereas no residual was observed in a non-coated alpha-TCP ceramic. Coating with HPC was found to be effective for controlling bioresorption and improving the workability of porous alpha-TCP ceramic. The prepared porous alpha-TCP ceramic is expected to be useful as a novel material for bone fillers by incorporating it with drugs or osteoinductive factors.  相似文献   

16.
The adequate regeneration of large bone defects is still a major problem in orthopaedic surgery. Synthetic bone substitute materials have to be biocompatible, biodegradable, osteoconductive and processable into macroporous scaffolds tailored to the patient specific defect. Hydroxyapatite (HA) and tricalcium phosphate (TCP) as well as mixtures of both phases, biphasic calcium phosphate ceramics (BCP), meet all these requirements and are considered to be optimal synthetic bone substitute materials. Rapid prototyping (RP) can be applied to manufacture scaffolds, meeting the criteria required to ensure bone ingrowth such as high porosity and defined pore characteristics. Such scaffolds can be used for bone tissue engineering (BTE), a concept based on the cultivation of osteogenic cells on osteoconductive scaffolds. In this study, scaffolds with interconnecting macroporosity were manufactured from HA, TCP and BCP (60 wt% HA) using an indirect rapid prototyping technique involving wax ink-jet printing. ST-2 bone marrow stromal cells (BMSCs) were seeded onto the scaffolds and cultivated for 17 days under either static or dynamic culture conditions and osteogenic stimulation. While cell number within the scaffold pore system decreased in case of static conditions, dynamic cultivation allowed homogeneous cell growth even within deep pores of large (1,440 mm3) scaffolds. Osteogenic cell differentiation was most advanced on BCP scaffolds in both culture systems, while cells cultured under perfusion conditions were generally more differentiated after 17 days. Therefore, scaffolds manufactured from BCP ceramic and seeded with BMSCs using a dynamic culture system are the method of choice for bone tissue engineering.  相似文献   

17.
Collagen (Col) type I, as the major component of the bone extracellular matrix has been broadly studied for bone tissue engineering. However,inferior mechanical properties limit its usage for load bearing applications. In this research, freeze dried Col scaffolds are coated with graphene oxide (GO) through a covalent bond of the amine Col with the graphene carboxyl groups. The prepared scaffolds were then reduced using a chemical agent. Scanning electron microscopy exhibited a porous structure for the synthesized scaffolds with an approximate pore size of 100–220 ± 12 µm, which is in the suitable range for bone tissue engineering application. Reducing the GO coating improved the compressive modulus of the Col from 250 to 970 kPa. Apatite formation was also indicated by immersing the scaffolds in simulated body fluid after five days. The cytocompatibility of the scaffolds, using human bone marrow‐derived mesenchymal stem cells, was confirmed with MTT analysis. Alkaline phosphatase assay revealed that reducing the Col–GO scaffolds can effectively activate the differentiation of hBM‐MSCs into osteoblasts after 14 days, even without the addition of an osteogenic differentiation medium. The results of this study highlight that GO and its reduced form have considerable potential as bone substitutes for orthopaedic and dental applications.Inspec keywords: molecular biophysics, tissue engineering, biochemistry, cellular biophysics, graphene, biomedical materials, bone, proteins, scanning electron microscopy, porous materials, compressive strength, biomechanicsOther keywords: human bone marrow‐derived mesenchymal stem cells, reduced graphene oxide, bone extracellular matrix, inferior mechanical properties, load bearing applications, freeze‐dried Col scaffolds, amine Col groups, graphene carboxyl groups, bone tissue engineering, collagen type I, GO‐Col scaffolds, covalent bond, scanning electron microscopy, compressive modulus, apatite formation, cytocompatibility, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide analysis, alkaline phosphatase assay, osteogenic differentiation medium, dental applications, orthopaedic applications, porous structure, time 14.0 day, CO  相似文献   

18.
Novel hydroxyapatite-based hybrid materials with controlled porosity and good adhesion to silicon surfaces were designed as bio-package for MEMS with potential application for implants in the human body. These materials were prepared using synthetic Hydroxyapatite (HAp) powder with three different polymeric agglutinants. These porous materials have high ceramic content (up to 60 wt.% respect to resin) with a pore size between 100 and 350 microns and a pore volume fraction in the 25–60% range. These hybrid materials have high wearing resistance and hydrolytic stability. The samples were characterized mechanical and morphologically using XRD, SEM, densitometry, abrasion and mechanical tests.  相似文献   

19.
Bio-inspired materials with controlled topography have gained increasing interest in regenerative medicine, because of their ability to reproduce the physical features of natural extracellular matrix, thus amplifying certain biological responses both in vitro and in vivo, such as contact guidance and differentiation. However, information on the ability to adapt this high cell potential to 3D scaffolds, effective to be implanted in clinical bone defect, is still missing. Here, we examine the pattern of bone tissue generated within the implant in an ectopic model, seeding bone marrow progenitor cells onto PCL-MgCHA scaffolds. This composite material presented a porous structure with micro/nanostructured surfaces obtained by combining phase inversion/salt leaching and electrospinning techniques. Histological analysis of grafts harvested after 1-2-6 months from implantation highlights an extent of lamellar bone tissue within interconnected pores of fibre coated PCL-MgCHA composites, whereas uncoated scaffolds displayed sparse deposition of bone. Pure PCL scaffolds did not reveal any trace of bone for the overall 6 months of observation. In conclusion, we show that a structural modification in scaffold design is able to enhance bone regeneration possibly mimicking some physiological cues of the natural tissue.  相似文献   

20.
Synthesis and characterization of organic–inorganic macroporous hybrid scaffolds were investigated. The materials were prepared by combining 2-hydroxyethylmethacrylate (HEMA) and triethoxyvinylsilane (TEVS) chemically modified by Ca2 + and PO43 ? ions via sol–gel route. In this study we have constructed a sugar-based cracks-free three-dimensional (3D) network with interconnected porous architecture within the range of 150–300 μm and rough topography. The obtained results revealed that both topography and composition of prepared materials allow rapid growth of the bone-like apatite (HAp) layer on their surface after soaking in biological medium. Preliminary studies have shown that hybrids covered by HAp are non-cytotoxic and allow cell proliferation that make them a promising scaffolds in the field of bone regenerative medicine. The materials were mainly characterized by powder X-ray diffraction analysis (PXRD), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy–energy-dispersive spectroscopy (SEM–EDS) and transmission electron microscopy–energy-dispersive spectroscopy (TEM–EDS).  相似文献   

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