首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
The development of porous biodegradable scaffolds is of great interest in tissue engineering. In this regard, exploration of novel biocompatible materials is needed. Silk fibroin‐chondroitin sulfate‐sodium alginate (SF‐CHS‐SA) porous hybrid scaffolds were successfully prepared via lyophilization method and crosslinked by 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide‐ethanol treatment. According to the scanning electron microscopy studies, mean pore diameters of the scaffolds were in the range of 60–187 μm. The porosity percentage of the scaffold with SF‐CHS‐SA ratio of 70 : 15 : 15 (w/w/w %) was 92.4 ± 3%. Attenuated total reflectance Fourier transform infrared spectroscopy, X‐ray diffraction, and differential scanning calorimetry results confirmed the transition from amorphous random coil to crystalline β‐sheet in treated SF‐CHS‐SA scaffold. Compressive modulus was significantly improved in hybrid scaffold with SF‐CHS‐SA ratio of 70 : 15 : 15 (3.35 ± 0.15 MPa). Cytotoxicity assay showed that the scaffolds have no toxic effects on chondrocytes. Attachment of chondrocytes was much more improved within the SF‐CHS‐SA hybrid scaffold. Real‐time polymerase chain reaction analyses showed a significant increase in gene expression of collagen type II, aggrecan, and SOX9 and decrease in gene expression of collagen type I for SF‐CHS‐SA compared with SF scaffold. This novel hybrid scaffold can be a good candidate to be utilized as an efficient scaffold for cartilage tissue engineering. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41048.  相似文献   

2.
Poly(?‐caprolactone) (PCL)/gelatin (GE) nanofiber scaffolds with varying concentrations of lanthanum chloride (LaCl3, from 0 to 25 mM) were fabricated by electrospinning. The scaffolds were characterized by scanning electron microscopy, contact angle and porosity measurements, mechanical strength tests, and in vitro degradation studies. In vitro cytotoxicity and cell adhesion and proliferation studies were performed to assess the biocompatibility of the scaffolds, and in vivo wound healing studies were conducted to assess scaffold applications in the clinic. All prepared scaffolds were noncytotoxic, and the growth of adipose tissue–derived stem cells on LaCl3‐containing scaffolds was better than on the pure PCL/GE scaffold. Cell proliferation studies showed the greatest cell growth in the PCL/GE/LaCl3 scaffolds. Further, in vivo studies proved that the PCL/GE/LaCl3 scaffolds can promote wound healing. The results suggest that nanofiber scaffolds containing LaCl3 promote cell proliferation and have good biocompatibility, and thus potential for application in the treatment of skin wounds. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46672.  相似文献   

3.
Chitosan‐based scaffolds are widely studied in tissue regeneration because of their biocompatibility and biodegradability. Scaffolds are obtained by different techniques and can be modified with other polymers allowing controlling their properties. This article discusses the assembling of three‐dimensional chitosan porous scaffolds blended with gelatin. Gelatin was used to enhance cells attachment due to the presence of cell adhesion motifs, while improving mechanical strength. 2,5‐dimethoxy‐2,5‐dihydrofurane (DHF) was used as the crosslinking agent, because it allowed to control the reaction kinetics through temperature, time and DHF concentration. The results indicate that scaffolds morphology, pore sizes and distribution, compressive moduli and biodegradation in vitro with lysozyme, can be customized with variations of gelatin content and crosslinking degree. Scaffolds were neither cytotoxic nor genotoxic for human keratinocytes, exhibiting cell–substrate interactions. Our findings demonstrated that chitosan–gelatin scaffolds crosslinked with DHF, as a new crosslinking agent, are suitable in tissue engineering applications. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43814.  相似文献   

4.
As of 2010, 5.3 million orthopedic surgeries are performed each year, and this number is expected to increase to 6.2 million by 2020. On average, 27.7% of all orthopedic surgeries result in infection which often leads to osteomyelitis and the loss of supporting bone. In this study, we describe two synthetic bone grafts, or augmentation methods, for a biodegradable, silver nanoparticle (SNPs) containing antimicrobial scaffolds composed of pentaerythritol triacrylate‐co‐trimethylolpropane tris (3‐mercaptopropionate) (PETA) and hydroxyapatite (HA). This osteoinductive and degradable material is designed to stimulate proliferation of bone progenitor cells, and provide controlled release of antimicrobial components. The first method, denoted as the “incorporating method,” involves dissolving SNPs in ethanol, butanol, or isopropanol and directly incorporating the particles into the scaffold prior to polymerization. The second method, “coating method,” involves submerging fabricated scaffolds into their respective SNPs‐solution and mixing for 24 h. The coating method allowed better distribution and release of SNPs from the surface of the composites when exposed to extracellular media. The in vitro release of silver for both methods was quantified by inductively coupled plasma optical emission spectroscopy (ICP‐OES). The scaffolds made by means of the coating method showed increased release of silver with respect to time; no silver leached from the scaffolds formed by the incorporating method. Use of Alamar Blue assay demonstrated that the SNPs incorporation did not affect cell viability when tested with hASCs. The scaffolds formed by the coating method inhibited the proliferation of Staphylococcus aureus 99.5% and Escherichia coli by 99.9% within 24 h. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 41099.  相似文献   

5.
Considerable efforts have been devoted toward the development of electrospun scaffolds based on poly(ε‐caprolactone) (PCL) for bone tissue engineering. However, most of previous scaffolds have lacked the structural and mechanical strength to engineer bone tissue constructs with suitable biological functions. Here, we developed bioactive and relatively robust hybrid scaffolds composed of diopside nanopowder embedded PCL electrospun nanofibers. Incorporation of various concentrations of diopside nanopowder from 0 to 3 wt % within the PCL scaffolds notably improved tensile strength (eight‐fold) and elastic modulus (two‐fold). Moreover, the addition of diopside nanopowder significantly improved bioactivity and degradation rate compared to pure PCL scaffold which might be due to their superior hydrophilicity. We investigated the proliferation and spreading of SAOS‐II cells on electrospun scaffolds. Notably, electrospun PCL‐diopside scaffolds induced significantly enhanced cell proliferation and spreading. Overall, we concluded that PCL‐diopside scaffold could potentially be used to develop clinically relevant constructs for bone tissue engineering. However, the extended in vivo studies are essential to evaluate the role of PCL‐diopside fibrous scaffolds on the new bone growth and regeneration. Therefore, in vivo studies will be the subject of our future work. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134 , 44433.  相似文献   

6.
Specific pore structure is a vital essential for scaffolds applied in tissue engineering. In this article, poly(lactide‐co‐glycolide) (PLGA) scaffolds with a bimodal pore structure including macropores and micropores to facilitate nutrient transfer and cell adhesion were fabricated by combining supercritical CO2 (scCO2) foaming with particle leaching technique. Three kinds of NaCl particles with different scales (i.e., 100–250, <75, <10 μm) were used as porogens, respectively. In particular, heterogeneous nucleation occurred to modify scCO2 foaming/particle leaching process when NaCl submicron particles (<10 μm) were used as porogens. The observation of PLGA scaffolds gave a formation of micropores (pore size <10 μm) in the cellular walls of macropores (pore size around 100–300 μm) to present a bimodal pore structure. With different mass fractions of NaCl introduced, the porosity of PLGA scaffolds ranged from 68.4 ± 1.4 to 88.7 ± 0.4% for three NaCl porogens. The results of SEM, EDS, and in vitro cytotoxicity test of PLGA scaffolds showed that they had uniform structures and were compatible for cell proliferation with no toxicity. This novel scCO2 foaming/particle leaching method was promising in tissue engineering due to its ability to fabricate scaffolds with precise pore structure and high porosity. © 2016 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 43644.  相似文献   

7.
Hydroxyapatite (HA), the bone mineral and Cissus quadrangularis (CQ), a medicinal plant with osteogenic activity, are attaining increasing interest as a potential therapeutic agent for enhanced bone tissue regeneration. In the present study a synergistic effect of these two agents were analyzed by fabricating PCL‐CQ‐HA nanofibrous scaffolds by electrospinning and compared with PCL‐CQ and PCL (control) nanofibrous scaffolds. Morphology, composition, hydrophilicity, and mechanical properties of the electrospun PCL, PCL‐CQ, PCL‐CQ‐HA nanofibrous scaffolds were examined by Field emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FTIR), Contact angle and Tensile tests, respectively. The response of human foetal osteoblast cells on these scaffolds were evaluated using MTS assay, alkaline phosphatase activity, alizarin red staining, and osteocalcin expression for bone tissue regeneration. While the observed cellular response to both groups of scaffolds was better than for the control PCL scaffold, the PCL‐CQ‐HA nanofibrous scaffolds provided the most favorable substrate for cell proliferation and mineralization. The results showed that PCL‐CQ‐HA nanofibrous scaffolds had appropriate surface roughness for the osteoblast adhesion, proliferation, and mineralization comparing with other scaffolds. The observed investigation of physicochemical and biological properties suggests that the CQ‐HA loaded PCL nanofibrous scaffolds serve as a potential biocomposite material for bone tissue engineering. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 39835.  相似文献   

8.
Scaffold‐based interfacial tissue engineering aims to not only provide the structural and mechanical framework for cellular growth and tissue regeneration, but also direct cell behavior. Due to the disparity in composition of the osteochondral (cartilage and bone) interface, this work has developed a novel biomimetic biphasic nanocomposite scaffold integrating two biocompatible polymers containing tissue‐specific growth factor‐encapsulated core–shell nanospheres. Specifically, a poly(caprolactone) (PCL)‐based bone layer was successfully integrated with a poly(ethylene glycol) (PEG) hydrogel cartilage layer. In addition, a novel nanosphere fabrication technique for efficient growth factor encapsulation and sustained delivery via a wet coaxial electrospray technique was developed. Human bone marrow mesenchymal stem cell (hMSC) adhesion, osteogenic, and chondrogenic differentiation were evaluated. Our in vitro results showed significantly improved hMSC adhesion and differentiation in bone and cartilage layers, respectively. Studies have demonstrated promising results with novel biphasic nanocomposite scaffold for osteochondral tissue regeneration, thus, warranting further studies. © 2013 American Institute of Chemical Engineers AIChE J 60: 432–442, 2014  相似文献   

9.
Bioactive and biocompatible porous scaffold materials with adjustable pore structures and drug delivery capability are one of the key elements in bone tissue engineering. In this work, bioactive and biocompatible sodium alginate (SA)/hydroxyapatite (HAP) macroporous scaffolds are facilely and effectively fabricated based on 3D printing of the pre‐crosslinked SA/HAP hydrogels followed by further crosslinking to improve the mechanical properties of scaffolds. The pore structures and porosity (>80%) of the porous scaffolds can be readily tailored by varying the formation conditions. Furthermore, the in vitro biomineralization tests show that the bioactivity of the porous scaffolds is effectively enhanced by the addition of HAP nanoparticles into the scaffold matrix. Furthermore, the anti‐inflammatory drug curcumin is loaded into the porous scaffolds and the in vitro release study shows the sustainable drug release function of the porous scaffolds. Moreover, mouse bone mesenchymal stem cells (mBMSCs) are cultured on the porous scaffolds, and the results of the in vitro biocompatibility experiment show that the mBMSCs can be adhered well on the porous scaffolds. All of the results suggest that the bioactive and biocompatible SA/HAP porous scaffolds have great application potential in bone tissue engineering.  相似文献   

10.
In this study, novel poly(ε‐caprolactone) (PCL) composite scaffolds were prepared for bone tissue engineering applications, where gentamicin‐loaded β‐tricalcium phosphate (β‐TCP)/gelatin microspheres were added to PCL. The effects of the amount of β‐TCP/gelatin microspheres added to the PCL scaffold on various properties, such as the gentamicin release rate, biodegradability, morphology, mechanical strength, and pore size distribution, were investigated. A higher amount of filler caused a reduction in the mechanical properties and an increase in the pore size and led to a faster release of gentamicin. Human osteosarcoma cells (Saos‐2) were seeded on the prepared composite scaffolds, and the viability of cells having alkaline phosphatase (ALP) activity was observed for all of the scaffolds after 3 weeks of incubation. Cell proliferation and differentiation enhanced the mechanical strength of the scaffolds. Promising results were obtained for the development of bone cells on the prepared biocompatible, biodegradable, and antimicrobial composite scaffolds. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40110.  相似文献   

11.
Bioactive nanocomposite of nonstoichiometric apatite (ns‐AP) and poly(ε‐caprolactone) (PCL) was synthesized and its porous scaffold was fabricated. The results show that the hydrophilicity and cell attachment ratio on the composite surface improved with the increase of ns‐AP content in PCL. The composite scaffolds with 60 wt % ns‐AP content contained open and interconnected pores ranging in size from 200 to 500 μm and exhibit a porosity of around 80%. In addition, proliferation of MG63 cells on the composite scaffolds significantly increased with the increase of ns‐AP content, and the level of alkaline phosphatase (ALP) activity and nitric oxide (NO) production of the cells cultured on the composite scaffold were higher than that of PCL at 7 days, revealing that the composite scaffolds had excellent in vitro biocompatibility and bioactivity. The composite scaffolds were implanted into rabbit mandible defects, the results suggest that the introduction of ns‐AP into PCL enhanced the efficiency of new bone formation, and the ns‐AP/PCL composite exhibited in vivo good biocompatibility and osteogenesis. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2010  相似文献   

12.
《Ceramics International》2020,46(10):15725-15739
Tissue engineering has acquired remarkable attention as an alternative strategy to treat and restore bone defects during recent years. A scaffold is a fundamental component for tissue engineering, on which cells attach, proliferate and differentiate to form new desirable functional tissue. The composition, and structural features of scaffolds, including porosity and pore size, play a fundamental role in the success of tissue-engineered construct. This review summarizes the effect of porosity and pore size of bioceramic-based scaffolds on their mechanical properties and biological performances. The focus of this review is on scaffolds with porosities 40% and above. From the mechanical point of view, the degree of porosity is a more important factor than pore size and scaffolds with porosities greater than 40% were more likely to substitute trabecular bones. While for in vitro and in vivo performances, pore size appeared more influential feature and co-existence of macropores and micropores led to better bone formation.  相似文献   

13.
Bone tissue engineering using in situ forming 3D scaffolds can be an alternative to surgically treated scaffolds. This work aimed to develop in situ forming scaffolds using poly (lactic-co-glycolic acid) and a bone synthesizing drug (risedronate) with or without the porogenic agent (collagen). Hybrid scaffolds were formed through solvent-induced phase inversion technique and were morphologically evaluated using scanning electron microscopy (SEM). The effect of scaffolds on Saos-2 cell line viability using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide test besides their effect on cell growth using fluorescence microscope was assessed. Furthermore, alkaline phosphatase (ALP) activity as well as Ca2+ deposition on the scaffolds was evaluated. SEM images revealed the porous structure for collagen-based scaffolds. Saos-2 cell proliferation was significantly enhanced with risedronate-loaded scaffolds compared to those lacking the drug. Porous collagen-based scaffolds were more favorable for both the cell growth and the promotion of ALP activity. Furthermore, collagen-based scaffolds promoted the Ca2+ deposition compared to their counterparts without collagen. Such results suggest that collagen-based scaffolds offer excellent biocompatibility for bone regeneration, where this biocompatible nature of scaffold leads to the proliferation of cells that lead to the deposition of mineral on the scaffold. Such in situ forming 3D scaffolds provide a promising noninvasive approach for bone tissue engineering.  相似文献   

14.
A highly crosslinked composite dextran-based scaffold (named DexFoam) was tailored to overcome specific deficiencies of polymeric and ceramic bone scaffolds and to guarantee a bone-mimicking microenvironment for the proliferation of human mesenchymal stem cells in vitro. The creep resistance for up to 90% compressive stain, the capability to regain the original shape after deformation, and the good thermal stability in both physiological and “body limit” conditions make DexFoam a valid alternative to the currently available bone scaffolds. Histopathological evaluation for host reaction and tissue colonization of DexFoam scaffold, implanted subcutaneously in mice, demonstrated its in vivo biocompatibility and biodegradability.  相似文献   

15.
Naturally derived Hydroxyapatite (HAp) from fish scale is finding wide applications in the development of bone scaffold to promote bone regeneration. But porous HAp scaffold is fragile in nature making it unsuitable for bone repair or replacement applications. Thus, it is essential to improve the mechanical property of HAp scaffolds while retaining the interconnected porous structure for tissue ingrowth in vivo. In this study solvent casting particulate leaching technique is used to develop novel Puntius conchonius fish scale derived HAp bone scaffold by varying the wt.% of the HAp from 60 to 80% in PMMA matrix. Physico-chemical, mechanical, structural and bioactive properties of the developed scaffolds are investigated. The obtained results indicate that HAp-PMMA scaffold at 70?wt % HAp loading shows optimal properties with 7.26?±?0.45?MPa compressive strength, 75?±?0.8% porosity, 8.0?±?0.68% degradation and 190?±?11% water absorption. The obtained results of the scaffold can meet the physiological demands to guide bone regeneration. Moreover, in vitro bioactivity analysis also confirms the formation of bone like apatite in the scaffold surface after 28 days of SBF immersion. Thus, the developed scaffold has the potential to be effectively used in bone tissue engineering applications.  相似文献   

16.
There are numerous methods for producing scaffolds to be applied in bone tissue engineering. However, the best method of scaffold production is essential to consider, with respect to their chemical composition and mechanical and structural properties, so that debris is not produced when the scaffolds are evaluated in vitro or in vivo.The primary aim of the present investigation was to produce six novel β-TCP scaffold compositions, using sodium chloride as a porogen, with two different particle sizes, measuring 1–2 mm and 750 mm-1mm, and at varied concentrations (30, 50, and 70 wt %). Physical, chemical, mechanical, and in vitro characterizations were then performed on each scaffold composition, using artificial saliva, for 7 and 14 days, with promising results. The XRD diffractograms showed the formation of two new crystalline phases (NaCaPO4 and Ca5[PO4]3Cl) in the scaffolds, after their production. In addition, scaffold porosity, Young's modulus, and the maximum resistance of compression values were in the trabecular bone range and the in vitro test, using artificial saliva, was favorable in relation to scaffold bioactivity.  相似文献   

17.
The objective of this study was to develop novel porous composite scaffolds for bone tissue engineering through surface modification of polycaprolactone–biphasic calcium phosphate‐based composites (PCL–BCP). PCL–BCP composites were first fabricated with salt‐leaching method followed by aminolysis. Layer by layer (LBL) technique was then used to immobilize collagen (Col) and bone morphogenetic protein (BMP‐2) on PCL–BCP scaffolds to develop PCL–BCP–Col–BMP‐2 composite scaffold. The morphology of the composite was examined by scanning electron microscopy (SEM). The efficiency of grafting of Col and BMP‐2 on composite scaffold was measured by X‐ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). Both XPS and FTIR confirmed that Col and BMP‐2 were successfully immobilized into PCL–BCP composites. MC3TC3‐E1 preosteoblasts cells were cultivated on composites to determine the effect of Col and BMP‐2 immobilization on cell viability and proliferation. PCL–BCP–Col–BMP‐2 showed more cell attachment, cell viability, and proliferation bone factors compared to PCL–BCP‐Col composites. In addition, in vivo bone formation study using rat models showed that PCL–BCP–Col–BMP‐2 composites had better bone formation than PCL–BCP‐Col scaffold in critical size defect with 4 weeks of duration. These results suggest that PCL–BCP–Col–BMP‐2 composites can enhance bone regeneration in critical size defect in a rat model with 4 weeks of duration. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45186.  相似文献   

18.
《Ceramics International》2021,47(21):29535-29549
The employment of graphene and its derivatives, graphene oxide and reduced graphene oxide, is extending from bioimaging and fabrications of biosensors to drug delivery and tissue engineering in the biomedical area. Graphene family-incorporated scaffolds, used in bone tissue engineering and bone regenerative medicine, profit superior properties of these materials, such as enhanced mechanical properties, large surface area, and the existence of functional groups. At the same time, problems related to cytotoxicity and adverse immune response of graphene family are solved when they are applied to produce 3-dimensional scaffolds. The objective of this review is to focus on in vitro properties of scaffolds consisting of graphene or its derivatives, especially osteogenic and antibacterial properties, as well as the influence of graphene and its derivatives on in vivo performances of implanted bone scaffolds. The positive effect of graphene and its two derivatives on attachment, and cell proliferation, as well as in vitro osteogenic differentiation of different cells was undeniable. Besides, the synergetic outcome of using graphene family on the antibacterial feature of scaffolds, especially incorporation with the silver element, was effective. Moreover, successful treatment of critical-sized bone defects was reported during in vivo preclinical tests when graphene or its derivatives-incorporated scaffolds were used. However, the limited number of in vivo studies should be considered as one of the main shortcomings to use graphene as a promising candidate for treating bone defects. It is anticipated that the increased number of well-designed preclinical studies could improve the applications of graphene incorporated scaffolds in bone tissue engineering/regeneration, and find out explanations and appropriate solutions to possible long-term toxicity and nonbiodegradability of these materials.  相似文献   

19.
An excellent bioactive scaffold material which could induce and promote new bone formation is essential in the bone repair field. In this study, the bioactive material hydroxyapatite (HA) and the bone morphogenetic protein‐2 (BMP‐2) were added to poly‐l‐lactic acid (PLLA) using the electrospinning method. Scanning electron microscopy investigations performed on four different fiber scaffolds, PLLA, PLLA/HA, PLLA/BMP‐2 and PLLA/HA/BMP‐2, revealed that the fibers of all scaffolds are closely interwoven, and the presence of large interconnected voids between the fibers, resulting in a three‐dimensional porous network structure that was similar to the structure of the extracellular matrix of healthy bones. In the MG63 cell culture growth experiments, the composite scaffold material PLLA/HA/BMP‐2 showed a higher bioactivity than the other three scaffold materials. The four scaffold materials were implanted in rabbits’ tibia for 30 and 90 days. The results of the animal experiments indicate that the capability of the PLLA/HA/BMP‐2 composite to induce and promote bone tissue formation was better compared with PLLA/HA or PLLA/BMP‐2, suggesting that PLLA combined with HA/BMP‐2 is a promising material for bone tissue repair. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2015 , 132, 42249.  相似文献   

20.
In our previous study, a three‐dimensional gelatin/bioactive glass nanocomposite scaffold with a total porosity of about 85% and pore sizes ranging from 200 to 500 μm was prepared through layer solvent casting combined with lamination technique. The aim of this study was to evaluate in vitro biocompatibility and in vivo bone regeneration potential of these scaffolds with and without endothelial cells when implanted into a critical‐sized rat calvarial defect. MTT assay, SEM observation, and DAPI staining were used to evaluate cell viability and adhesion in macroporous scaffolds and results demonstrated that the scaffolds were biocompatible enough to support cell attachment and proliferation. To investigate the in vivo osteogenesis of the scaffold, blank scaffolds and endothelial/scaffold constructs were implanted in critical‐sized defects, whereas in control group defects were left untreated. Bone regeneration and vascularization were evaluated at 1, 4, and 12 weeks postsurgery by histological, immunohistochemical, and histomorphometric analysis. It was shown that both groups facilitated bone growth into the defect area but improved bone regeneration was seen with the incorporation of endothelial cells. The data showed that the porous Gel/BaG nanocomposite scaffolds could well support new bone formation, indicating that the proposed strategy is a promising alternative for tissue‐engineered bone defects.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号