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1.
This study developed a solvent merging/particulate leaching method for preparing three-dimensional porous scaffolds. Poly(L-lactic-co-glycolic acid) (PLGA) and sodium chloride particles were dry-mixed and cast into a special mold, through which a liquid could pass due to a pressure difference. An organic solvent was then poured into the mold to dissolve and merge the PLGA particles under negative pressure. A nonsolvent was conducted into the PLGA/salt composite to solidify and precipitate the merged PLGA matrix. Finally, a large amount of water was passed through the mold to leach out the salt particles so as to create a porous structure. The results revealed that a highly porous three-dimensional scaffold (>85 vol %) with a well interconnected porous structure could be achieved by this process. Porosity and the pore size of the scaffold were controlled using the ratio and the particle size of the added salt particles. A larger-volume scaffold was produced using a larger mold. This work provides a continuous and simple procedure for fabricating a bulk three-dimensional porous scaffold for tissue engineering.  相似文献   

2.
Biocompatible three-dimensional (3-D) porous scaffolds are of great interest for tissue engineering applications. We here present a novel combined freeze-drying/cross-linking process to prepare porous polysaccharide-based scaffolds. This process does not require an organic solvent or porogen agent. We unexpectedly found that cross-linking of biomacromolecules such as pullulan and dextran with sodium trimetaphosphate could be performed during freeze-drying. We have demonstrated that the freeze-drying pressure modulates the degree of porosity. High freeze-drying pressure scaffolds presented pores with a mean diameter of 55 ± 4 μm and a porosity of 33 ± 12%, whereas low freeze-drying pressure scaffolds contained larger pores with a mean diameter of 243 ± 14 μm and a porosity of 68 ± 3%. Porous scaffolds of the desired shape could be easily obtained and were stable in culture medium for weeks. In vitro viable mesenchymal stem cells were found associated with porous scaffolds in higher proportions than with non-porous scaffolds. Moreover, cells penetrated deeper into scaffolds with larger pores. This novel combined freeze-drying/cross-linking processing of polysaccharides enabled the fabrication of biocompatible scaffolds with controlled porosity and architectures suitable for 3-D in vitro culture and biomedical applications.  相似文献   

3.
In the present study, two biphasic calcium phosphate biomaterials (BCP) with HA/TCP ratios of 50/50 and 30/70 were obtained from a pure HA biomaterial. The biomaterials which showed the same three-dimensional geometry were implanted into corticocancellous costal defects of sheep. In the specimens of all three biomaterials, abundant bone formation, mineral dissolution from the biomaterial scaffolds, and active cellular resorption of the scaffolds was present after 6 and 12 months. Backscattered electron microscopy showed bone invasion into the pores of the scaffolds and micromechanical interlocking at the bone/biomaterial interface without intervening soft tissue. The pattern of bone formation and scaffold resorption was different for cortical and cancellous bone. No time-based effect, however, was observed. Overall, the BCP biomaterials had formed significantly more bone than the HA biomaterial. Also, scaffold resorption, which was followed by a replacement with newly formed bone, was significantly higher in the BCP biomaterials. Although no significant differences were observed between both BCP biomaterials, the present study had confirmed the assumption that HA/TCP compounding was suitable to improve bone formation and scaffold resorption in the investigated biomaterials and at the same time maintain the osteoconductive properties of the scaffolds.  相似文献   

4.
Hyaluronic acid (HA) plays a vital role in many tissues, influencing water content and mechanical function, and has been shown to have positive biological effects on cell behavior in vitro. To begin to determine whether these benefits can be accessed if HA is incorporated into collagen-based scaffolds for tissue engineering, HA-collagen composite matrices were prepared and selected properties evaluated. HA-collagen scaffolds were cross-linked with carbodiimide and loss rates of HA in culture medium assessed. Scaffold pore structures were evaluated by light and electron microscopy. Adult canine chondrocytes were grown in selected HA-collagen scaffolds to assess the effects of HA on cell behavior. Homogenous HA-collagen slurries were achieved when polyionic complexes were suppressed. HA was uniformly distributed through the scaffolds, which demonstrated honeycomb-like pores with interconnectivity among pores increasing as HA content increased. Virtually all of the HA added to the collagen slurry was incorporated into the composite scaffolds that underwent a 7-day cross-linking protocol. After 5 days in culture medium, the HA content in the scaffolds was 5-7% regardless of initial HA loading. After only 2 weeks in culture cartilaginous tissue was found in the chondrocyte-seeded HA-collagen scaffolds. This study contributes to the understanding of the effects of HA content, pH, and cross-link treatment on pore characteristics and degradation behavior essential for the design of HA-collagen scaffolds. The demonstration that these scaffolds can be populated by chondrocytes and support in vitro formation of cartilaginous tissue warrants further investigation of this material system for tissue engineering.  相似文献   

5.
Biomaterials made from synthetic polymers are becoming more pervasive in the medical field. Synthetic polymers are particularly advantageous as their chemical and mechanical properties can be easily tailored to a specific application. This work characterizes polymer scaffolds derived from the cyclic acetal monomer 5-ethyl-5-(hydroxymethyl)-β,β-dimethyl-1,3-dioxane-2-ethanol diacrylate (EHD). Both porous scaffolds and bilayer scaffolds based upon the EHD monomer were fabricated, and the resulting scaffolds' degradation and mechanical properties were studied. The results showed that by modifying the architecture of an EH scaffold, either by adding a porous network or a poly(ethylene glycol) (PEG) coating, the degradation and Young's modulus of the biomaterial can be significant altered. However, results also indicated that these architectural modifications can be accomplished without a significant loss in the flexural strength of the scaffold. Therefore, we suggest that porous EH scaffolds, and particularly porous EH-PEG bilayers, may be especially useful in dynamic tissue environments due to their advantageous architectural and mechanical properties.  相似文献   

6.
Fabrication of porous gelatin scaffolds for tissue engineering.   总被引:22,自引:0,他引:22  
H W Kang  Y Tabata  Y Ikada 《Biomaterials》1999,20(14):1339-1344
A novel method which employs water present in swollen hydrogels as a porogen for shape template was suggested for preparing porous materials. Biodegradable hydrogels were prepared through crosslinking of gelatin with glutaraldehyde in aqueous solution, followed by rinsing and washing. After freezing the swollen hydrogels, the ice formed within the hydrogel network was sublimated by freeze-drying. This simple method produced porous hydrogels. Irrespective of any rinsing and washing processes, water was homogeneously distributed into the hydrogel network, allowing the hydrogel network to uniformly enlarge and the ice to act as a porogen during the freezing process. Different porous structures were obtained by varying the freezing temperature. Hydrogels frozen in liquid nitrogen, had a two-dimensionally ordered structure, while the hydrogels prepared at freezing temperatures near -20 degrees C, showed a three-dimensional structure with interconnected pores. As the freezing temperature was lowered, the hydrogel structure gradually became more two-dimensionally ordered. These results suggest that the porosity of dried hydrogels can be controlled by the size of ice crystals formed during freezing. It was concluded that the present freeze-drying procedure is a bio-clean method for formulating biodegradable sponges of different pore structures without use of any additives and organic solvents.  相似文献   

7.
The aim of this study was to investigate the feasibility of fabricating porous crosslinked chitosan hydrogels in an aqueous phase using dense gas CO(2) as a foaming agent. Highly porous chitosan hydrogels were formed by using glutaraldehyde and genipin as crosslinkers. The method developed here eliminates the formation of a skin layer, and does not require the use of surfactants or other toxic reagents to generate porosity. The chitosan hydrogel scaffolds had an average pore diameter of 30-40 μm. The operating pressure had a negligible effect on the pore characteristics of chitosan hydrogels. Temperature, reaction period, type of biopolymer and crosslinker had a significant impact on the pore size and characteristics of the hydrogel produced by dense gas CO(2). Scanning electron microscopy and histological analysis confirmed that the resulting porous structures allowed fibroblasts seeded on these scaffolds to proliferate into the three-dimensional (3-D) structure of these chitosan hydrogels. Live/dead staining and MTS analysis demonstrated that fibroblast cells proliferated over 7 days. The fabricated hydrogels exhibited comparable mechanical strength and swelling ratio and are potentially useful for soft tissue engineering applications such as skin and cartilage regeneration.  相似文献   

8.
The aim of this study was to compare the ability of hard tissue regeneration of four types of stem cells or precursors under both in vitro and in vivo situations. Primary cultures of rat bone marrow, rat dental pulp, human bone marrow, and human dental pulp cells were seeded onto a porous ceramic scaffold material, and then either cultured in an osteogenic medium or subcutaneously implanted into nude mice. For cell culture, samples were collected at weeks 0, 1, 3, and 5. Results were analyzed by measuring cell proliferation rate and alkaline phosphatase activity, scanning electron microscopy, and real-time PCR. Samples from the implantation study were retrieved after 5 and 10 weeks and evaluated by histology and real-time PCR. The results indicated that in vitro abundant cell growth and mineralization of extracellular matrix was observed for all types of cells. However, in vivo matured bone formation was found only in the samples seeded with rat bone marrow stromal cells. Real-time PCR suggested that the expression of Runx2 and the expression osteocalcin were important for the differentiation of bone marrow stromal cells, while dentin sialophosphoprotein contributed to the odontogenic differentiation. In conclusion, the limited hard tissue regeneration ability of dental pulp stromal cells questions their practical application for complete tooth regeneration. Repeated cell passaging may explain the reduction of the osteogenic ability of both bone- and dentinal-derived stem cells. Therefore, it is essential to develop new cell culture methods to harvest the desired cell numbers while not obliterating the osteogenic potential.  相似文献   

9.
A method of compression-molding fine-powder blends of polycaprolactone (PCL) and poly(ethylene oxide) (PEO) and subsequently dissolving the PEO phase was investigated to prepare porous PCL scaffolds. Different mixing ratios of the two polymers from 20 to 70% PCL were used to study the effect of the mixing ratio on the morphology formation of the scaffold. The mixing ratio was found to play an important role in affecting the porosity of the scaffold and the size of pores. Murine embryonic stem cell derived osteogenic cells were utilized to test the suitability of these scaffolds in tissue engineering applications. The seeded cells were able to colonize and grow in these scaffolds. Based on the overall consideration of morphology, mechanical performance, and ability for cell attachment and proliferation, the scaffolds with approximately 30-40% PCL appear to be an appropriate choice for tissue engineering. These findings suggest that sacrificial compression-molding of PCL-PEO powder blends can be used in the generation of biocompatible scaffolds with controllable porosity and pore size and may be used for in vitro tissue engineering applications.  相似文献   

10.
Three-dimensional porous chitosan-polyvinyl pyrrolidone (PVP) scaffolds were fabricated for tissue engineering applications via liquid–liquid or liquid–solid phase separation. A mixture of an acidic aqueous solution with butanol as a non-solvent and a chitosan-PVP quaternary system were freeze-dried. We then studied the homogenous open pore structure and the minute pore distribution in order to improve the mass transfer and cell seeding efficiency while also obtaining the optimal ratio of PVP to provide high interconnectivity and to improve the open-pore structure. The properties of the porous chitosan-PVP scaffolds – including the microstructure, chemical release, water absorption properties, and cell proliferation tests were studied – and the results were compared against those obtained from conventional scaffolds. chitosan-PVP scaffolds with a porosity of over 70% were obtained, and the pore morphology on the surface and within the porous scaffolds showed the presence of homogenous open pores with excellent interconnectivity. As the PVP content increased, main pores (50–100 μm) and minute pores (4–10 μm) could be clearly observed. Also, the porous scaffold showed an improved efficiency for cell adhesion after the cells were cultured for 4 h. After 72 h, the cultured cells presented an increase in the cell proliferation and on the porous scaffolds. These results strongly suggest that the porous chitosan-PVP scaffolds can be widely used in tissue engineering, including for biopatches and artificial skin applications.  相似文献   

11.
Both surgical tympanoplasty and paper patch grafts are frequently procedured to heal tympanic membrane (TM) perforation or chronic otitis media, despite their many disadvantages. In this study, we report a new healing method of TM perforation by using three-dimensional (3D) porous chitosan scaffolds (3D chitosan scaffolds) as an alternative method to surgical treatment or paper patch graft. Various 3D chitosan scaffolds were prepared; and the structural characteristics, mechanical property, in vitro biocompatibility, and healing effects of the 3D chitosan scaffolds as an artificial TM in in vivo animal studies were investigated. A 3D chitosan scaffold of 5 wt.% chitosan concentration showed good proliferation of TM cells in an in vitro study, as well as suitable structural characteristics and mechanical property, as compared with either 1% or 3% chitosan. In in vivo animal studies, 3D chitosan scaffold were able to migrate through the pores and surfaces of TM cells, thus leading to more effective TM regeneration than paper patch technique. Histological observations demonstrated that the regenerated TM with the 3D chitosan scaffold consisted of three (epidermal, connective tissue, and mucosal) layers and were thicker than normal TMs. The 3D chitosan scaffold technique may be an optimal healing method used in lieu of surgical tympanoplasty in certain cases to heal perforated TMs.  相似文献   

12.
The present study investigated whether bone regeneration and biomaterial replacement would be improved by loading of biogenous biphasic biomaterial scaffolds (HA/TCP ratio 30/70) with rhBMP-2, and whether the placement of three barrier membranes differing in structure and porosity (prototyped SLA Ti specimens, GORE RESOLUT Adapt specimens, and titanized TiMESH light specimens) would have a synergistic effect. A rabbit calvarial model was used for the implantation studies. Histological specimens were obtained after 12 weeks and evaluated quantitatively for differences between the various material combinations. Loading of the biomaterials with rhBMP-2 significantly enhanced the amount of regenerated bone and caused a pronounced biomaterial replacement. While BMP-induced bone had formed uniformly over the surgical defects, bone regeneration in the absence of BMP depends on bone promotion from the margins of the defects toward the center. No positive effect on bone regeneration was seen for any of the placed barrier membranes. While the present study had shown that rhBMP-2 loading significantly increases bone regeneration using the investigated biomaterial, barrier-membrane placement may be useful in predetermining the final shape of the regenerative site but provides no additional beneficial impact on the amount and quality of the bone regeneration induced by rhBMP-2.  相似文献   

13.
Porous alginate/polyvinyl alcohol (PVA) hybrid scaffolds as bioartificial cell scaffolds were fabricated to improve cell compatibility as well as flexibility of the scaffolds. The alginate/PVA hybrid scaffolds with different PVA compositions up to 50 wt% were fabricated by a modified freeze-drying method including the physical cross-linking of PVA and the following chemical cross-linking of alginate. The prepared alginate/PVA hybrid scaffolds were characterized by morphology observations using scanning electron microscopy (SEM), the measurements of porosity and average pore sizes and the measurements of compressive strength and modulus. The scaffolds exhibited highly porous, open-cellular pore structures with almost the same surface and cross-sectional porosities (total porosities about 85%, regardless of PVA composition) and the pore sizes from about 290 microm to about 190 microm with increasing PVA composition. The alginate/PVA hybrid scaffolds were more soft and elastic than the control alginate scaffold without significant changes of mechanical strength. The scaffolds were examined for their in vitro cell compatibility by the culture of chondrocytes (human chondrocyte cell line) in the scaffolds and the following analyses by MTT assay and SEM observation. It was observed that the alginate/PVA scaffolds had better cell adhesion and faster growth than the control alginate scaffold. It seems that 30 wt% addition of PVA to alginate in the fabrication of the hybrid scaffolds is desirable for improving their flexibility and cell compatibility.  相似文献   

14.
The purpose of this study was to fabricate and characterize nanotubular structure on machined titanium (MA) and resorbable blast media (RBM) treated titanium by anodizing. The anodized MA and RBM were characterized with scanning electron microscopy, transmission electron microscopy, X-ray diffraction, energy disperse spectra, X-ray photoelectron spectra, and nano-indentation and scratch test. Highly ordered nanotubular layers of individually 100 nm in diameter and 500 nm in length approximately were formed regardless of the substrates. The nanotubular layers consisted mainly of amorphous TiO(2) with trace fluorine. The nanotubular surfaces on both the substrates significantly reduced water contact angles and elastic modulus compared with those prior to anodizing. The anodizing treatment significantly increased the surface roughness of the smooth MA, but significantly decreased the surface roughness of the roughened RBM. The critical delamination forces of the nanotubular layer were not obtained due to the limitation of surface roughness. The anodized RBM consisted of a nano-micro porous graded structure, or a nanotubular amorphous fluoride containing TiO(2) layer on top of micro-roughened titanium surface, which is expected to significantly improve the surface area that can be used to deliver drugs and growth factors and alleviate the interfacial elastic modulus mismatch as to enhance osseointegration when compared with conventional dental and orthopedic implant devices with smooth or acid etched surface.  相似文献   

15.
The aim of this study was to prepare poly-DL-lactide/polyethylene glycol (PDLLA/PEG) blends to improve medium absorption and cell proliferation in the three-dimensional (3-D) structure of their scaffolds. Carbon dioxide (CO2) was used as a foaming agent to create porosity in these blends. The results of Fourier transform infrared (FTIR) spectroscopy demonstrated that the blends were homogeneous mixtures of PDLLA and PEG. The peak shifts at 1092 and 1744 cm(-1) confirmed the presence of molecular interactions between these two compounds. Increasing the PEG weight ratio enhanced the relative crystallinity and hydrophilicity. The PDLLA/PEG blends (especially 80/20 and 70/30 weight ratios) exhibited linear degradation profiles over an incubation time of 8 weeks. The mechanical properties of PDLLA/PEG blends having less than 30 wt.% PEG were suitable for the fabrication of porous scaffolds. Increasing the concentration of PEG to above 50% resulted in blends that were brittle and had low mechanical integrity. Highly porous scaffolds with controllable pore size were produced for 30 wt.% PEG samples using the gas foaming technique at temperatures between 25 and 55 °C and pressures between 60 and 160 bar. The average pore diameters achieved by gas foaming process were between 15 and 150 μm, and had an average porosity of 84%. The medium uptake and degradation rate of fabricated PDLLA/PEG scaffolds were increased compared with neat PDLLA film due to the presence of PEG and porosity. The porous scaffolds also demonstrated a lower modulus of elasticity and a higher elongation at break compared to the non-porous film. The fabricated PDLLA/PEG scaffolds have high potential for various tissue-engineering applications.  相似文献   

16.
In this work, the fabrication and in vitro degradation of porous fumarate-based/alumoxane nanocomposites were evaluated for their potential as bone tissue engineering scaffolds. The biodegradable polymer poly (propylene fumarate)/propylene fumarate-diacrylate (PPF/PF-DA), a macrocomposite composed of PPF/PF-DA and boehmite microparticles, and a nanocomposite composed of PPF/PF-DA and surface-modified alumoxane nanoparticles were used to fabricate porous scaffolds by photo-crosslinking and salt-leaching. Scaffolds then underwent 12 weeks of in vitro degradation in phosphate buffered saline at 37 degrees C. The presence of boehmite microparticles and alumoxane nanoparticles in the polymer inhibited scaffold shrinkage during crosslinking. Furthermore, the incorporation of alumoxane nanoparticles into the polymer limited salt-leaching, perhaps due to tighter crosslinking within the nanocomposite. Analysis of crosslinking revealed that the acrylate and overall double bond conversions in the nanocomposite were higher than in the PPF/PF-DA polymer alone, though these differences were not significant. During 12 weeks of in vitro degradation, the nanocomposite lost 5.3% +/- 2.4% of its mass but maintained its compressive mechanical properties and porous architecture. The addition of alumoxane nanoparticles into the fumarate-based polymer did not significantly affect the degradation of the nanocomposite compared with the other materials in terms of mass loss, compressive properties, and porous structure. These results demonstrate the feasibility of fabricating degradable nanocomposite scaffolds for bone tissue engineering by photo-crosslinking and salt-leaching mixtures of fumarate-based polymers, alumoxane nanoparticles, and salt microparticles.  相似文献   

17.
利用NaCl作为致孔剂,采用干热交联壳聚糖与盐淅沥致孔法制备壳聚糖多孔支架,探索该支架制备方法的可行性和安全性.在研究中利用干热交联壳聚糖与盐淅沥致孔法制备了不同质量比(chitosan/NaCl)的壳聚糖多孔支架和冻干法制备的支架,并对所有支架的孔隙率、孔结构、力学性能等参数指标进行评价.结果表明,利用于热交联壳聚糖与盐淅沥致孔法制备过程中加入致孔剂(NaCl)越多,即NaCl/chitosan质量比越大,所构建多孔支架的平均孔径越大,孔隙率越高,抗拉性则下降.  相似文献   

18.
HA/TCP双相陶瓷与人成骨细胞生物相容性的体外实验研究   总被引:14,自引:2,他引:12  
体外复合培养条件下,通过观察人成骨细胞与HA/TCP的复合生长及监测材料对细胞生理功能的影响来评价材料的生物相容性。研究发现,复合培养过程中成骨细胞首先贴附于材料的表面,进而攀附于材料边缘切刻处及材料表层微孔的边缘,以后逐渐长入孔中。最后,材料几乎为细胞所覆盖。复合培养的成骨细胞与正常培养的成骨细胞一样,具有分泌大量胶原纤维、表现强烈碱性磷酸酶活性和形成矿化胞外基质等成骨表型。细胞能够与材料很好地复合生长、材料对细胞生理功能又无明显的影响表明HA/TCP与人成骨细胞具有良好的生物相容性。  相似文献   

19.
Biomaterial, an essential component of tissue engineering, serves as a scaffold for cell attachment, proliferation, and differentiation; provides the three dimensional (3D) structure and, in some applications, the mechanical strength required for the engineered tissue. Both synthetic and naturally occurring calcium phosphate based biomaterial have been used as bone fillers or bone extenders in orthopedic and reconstructive surgeries. This study aims to evaluate two popular calcium phosphate based biomaterial i.e., hydroxyapatite (HA) and tricalcium phosphate/hydroxyapatite (TCP/HA) granules as scaffold materials in bone tissue engineering. In our strategy for constructing tissue engineered bone, human osteoprogenitor cells derived from periosteum were incorporated with human plasma-derived fibrin and seeded onto HA or TCP/HA forming 3D tissue constructs and further maintained in osteogenic medium for 4 weeks to induce osteogenic differentiation. Constructs were subsequently implanted intramuscularly in nude mice for 8 weeks after which mice were euthanized and constructs harvested for evaluation. The differential cell response to the biomaterial (HA or TCP/HA) adopted as scaffold was illustrated by the histology of undecalcified constructs and evaluation using SEM and TEM. Both HA and TCP/HA constructs showed evidence of cell proliferation, calcium deposition, and collagen bundle formation albeit lesser in the former. Our findings demonstrated that TCP/HA is superior between the two in early bone formation and hence is the scaffold material of choice in bone tissue engineering.  相似文献   

20.
Novel three-dimensional scaffolds consisting of nano- and microsized hydroxyapatite (HA)/poly(epsilon-caprolactone) (PCL) composite were fabricated using a modified rapid-prototyping (RP) technique for bone tissue engineering applications. The size of the nano-HA ranged from 20 to 90 nm, whereas that of the micro-HA ranged from 20 to 80 microm. The scaffold macropores were well interconnected, with a porosity of 72-73% and a pore size of 500 microm. The compressive modulus of the nano-HA/PCL and micro-HA/PCL scaffolds was 3.187 +/- 0.06 and 1.345 +/- 0.05 MPa, respectively. The higher modulus of the nano-HA/PCL composite (n-HPC) was to be likely caused by a dispersion strengthening effect. The attachment and proliferation of MG-63 cells on n-HPC were better than that on the micro-HA/PCL composite (m-HPC) scaffold. The n-HPC was more hydrophilic than the m-HPC because of the greater surface area of HA exposed to the scaffold surface. This may give rise to better cell attachment and proliferation. Bioactive n-HA/PCL composite scaffold prepared using a modified RP technique has a potential application in bone tissue engineering.  相似文献   

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