聚DL-乳酸/磷酸盐复合多孔支架材料的制备及降解性能

采用溶液浇铸 -颗粒滤除法制备了聚 DL-乳酸 (PDL L A) ,聚 DL-乳酸 /羟基磷灰石 (PDL L A/ 2 0 wt%HA)、及聚 DL-乳酸 / β-磷酸三钙 (PDL L A/ 2 0 wt% β- TCP)复合多孔支架材料。研究了支架材料在体外降解中压缩强度、分子量、质量及水解液的 p H值变化规律。结果显示 :复合多孔支架中 HA和 β- TCP均匀分布在 PDL L A基质中 ,复合支架的孔隙率可达 84 % ,磷酸盐微粒的加入对多孔支架的孔隙率有一定的影响 ,但可提高多孔支架的压缩强度 ,并可中和 PDL L A降解所产生的酸性 ,延缓 PDL L A的降解速度。两者相比 ,HA的作用更为明显     

Biological performance of a new beta-TCP/PLLA composite material for applications in spine surgery: in vitro and in vivo studies

The objective of this research was to carry out an in vitro and in vivo study of the biological performance of PLLA/beta-TCP composite materials, to estimate the scope of their potential applications in bone surgery. Samples with increasing beta-TCP (0-60% w/w) contents were processed by injection molding. The in vitro study consisted of an evaluation of inflammatory potential by assaying the IL-1alpha secreted by monocytes, and then cell proliferation (counting) and phenotype expression (PAL and I collagen) in human osteogenous cells. The in vivo study was carried out using cylindrical implants of composite materials composed of composite materials containing 0 or 60% beta-TCP and pure beta-TCP, respectively. The implants were inserted in femoral sites in rabbits, using the Kathagen protocol. Each animal received a 60% implant, with either a 0 or a 100% implant in the contralateral femur, so that the materials could be compared with one another. Five animals were examined for each material and implantation period, giving a total of 30 animals. This study showed that adding increasing percentages of beta-TCP to a lactic acid polymer matrix stimulated the proliferation of human osteogenous cells and synthesis of the extracellular bone matrix in a dose-dependent manner. In vivo results indicate that, in comparison with pure PLA, tricalcium phosphate-containing composite materials had faster degradation kinetics, caused less inflammatory reaction, and promoted contact osteogenesis. The composite material containing 60% beta-TCP demonstrated a similar performance to pure tricalcium phosphate bone grafts in terms of osteogenesis, and is apparently compatible with the production of intra-osseous implants for situations representing high levels of mechanical strain.     

Architecture and properties of anisotropic polymer composite scaffolds for bone tissue engineering

Bone is a complex porous composite structure with specific characteristics such as viscoelasticity and anisotropy, both in morphology and mechanical properties. Bone defects are regularly filled with artificial tissue grafts, which should ideally have properties similar to those of natural bone. Open cell composite foams made of bioresorbable poly(L-lactic acid) (PLA) and ceramic fillers, hydroxyapatite (HA) or beta-tricalcium phosphate (beta-TCP), were processed by supercritical CO2 foaming. Their internal 3D-structure was then analysed by micro-computed tomography (microCT), which evidenced anisotropy in morphology with pores oriented in the foaming direction. Furthermore compressive tests demonstrated anisotropy in mechanical behaviour, with an axial modulus up to 1.5 times greater than the transverse modulus. Composite scaffolds also showed viscoelastic behaviour with increased modulus for higher strain rates. Such scaffolds prepared by gas foaming of polymer composite materials therefore possess suitable architecture and properties for bone tissue engineering applications.     

Biodegradable polymer/hydroxyapatite composites: surface analysis and initial attachment of human osteoblasts

Biodegradable polymer/hydroxyapatite (HA) composites have potential application as bone graft substitutes. Thin films of polymer/HA composites were produced, and the initial attachment of primary human osteoblasts (HOBs) was assessed to investigate the biocompatibility of the materials. Poly(epsilon-caprolactone) (PCL) and poly(L-lactic acid) (PLA) were used as matrix materials for two types of HA particles, 50-microm sintered and submicron nonsintered. Using ESEM, cell morphology on the surfaces of samples was investigated after 90 min, 4 h, and 24 h of cell culture. Cell activity and viability were assessed after 24 h of cell culture using Alamar blue and DNA assays. Surface morphology of the polymer/HA composites and HA exposure were investigated using ESEM and EDXA, respectively. ESEM enabled investigation of both cell and material surface morphology in the hydrated condition. Combined with EDXA it permitted chemical and visual examination of the composite. Differences in HA exposure were observed on the different composite surfaces that affected the morphology of attached cells. In the first 4 h of cell culture, the cells were spread to a higher degree on exposed HA regions of the composites and on PLA than they were on PCL. After 24 h the cells were spread equally on all the samples. The cell activity after 24 h was significantly higher on the polymer/HA composites than on the polymer films. There was no significant difference in the activity of the cells on the various composite materials. However, cells on PCL showed higher activity compared to those on PLA. A polymer surface exhibiting "point exposure" of HA appeared to provide a novel and favorable substrate for primary cell attachment. The cell morphology and activity results indicate a favorable cell/material interaction and suggest that PLA and PCL and their composites with HA may be candidate materials for the reconstruction of bony tissue. Further investigations regarding long-term biomaterial/cell interactions and the effects of acidic degradation products from the biodegradable polymers are required to confirm their utility.     

The primacy of octacalcium phosphate collagen composites in bone regeneration

We have engineered a scaffold constructed of synthetic octacalcium phosphate (OCP) and porcine collagen sponge (OCP/Col), and reported that OCP/Col drastically enhanced bone regeneration. In this study, we investigated whether OCP/Col would enhance bone regeneration more than beta-tricalcium phosphate (beta-TCP) collagen composite (beta-TCP/Col) or hydroxyapatite (HA) collagen composite (HA/Col). Discs of OCP/Col, beta-TCP/Col, or HA/Col were implanted into critical-sized defects in rat crania and fixed at 4 or 12 weeks after implantation. The newly formed bone and the remaining granules of implants in the defect were determined by histomorphometrical analysis, and radiographic and histological examinations were performed. Statistical analysis showed that the newly formed bone by the implantation of OCP/Col was significantly more than that of beta-TCP/Col or HA/Col. In contrast, the remaining granules in OCP/Col were significantly lower than those in beta-TCP/Col or HA/Col. Bone regeneration by OCP/Col was based on secured calcified collagen and bone nucleation by OCP, whereas bone regeneration by beta-TCP/Col or HA/Col was initiated by poorly calcified collagen and osteoconductivity by beta-TCP or HA. This study showed that the implantation of OCP/Col in a rat cranial defect enhanced more bone regeneration than beta-TCP/Col and HA/Col.     

Preparation,characterization, and in vitro degradation of bioresorbable and bioactive composites based on Bioglass-filled polylactide foams

Highly porous poly(D,L-lactide)/Bioglass composites scaffolds were prepared by thermally induced phase separation process of polymer solutions and subsequent solvent sublimation. A series of composite foams with different polymer/Bioglass weight ratios was prepared to study the influence of Bioglass content on the foam characteristics such as porous structure, density, and pore volume. The pore volume was decreased from 9.5 to 5.7 cm(3)/g when the Bioglass content was increased up to 40 wt %, but the overall pore morphology was not affected very much by changing the polymer/glass composition ratio. The composites foams were then incubated in phosphate-buffered saline at 37 degrees C to study the in vitro degradation of the polymer and to detect hydroxyapatite (HA) formation as an indication of their bioactivity. The addition of Bioglass to polymer foams increased the water absorption and weight loss as compared with pure polymer foams. However, the polymer molecular weight, determined by size exclusion chromatography, was found to decrease more rapidly and to a larger extent in absence of Bioglass. This delayed degradation rate in the composite foams was probably caused by the dissolution of alkaline ions from the Bioglass, resulting in a buffering effect of the incubation medium. After incubation for 7 days, HA was detected by X-ray diffractometry and Raman spectroscopy and confirmed by environmental scanning electron microscopy and energy-dispersive X-ray analysis. The porous composites developed here are promising materials for bone regeneration applications because the formation of HA on the surface of the pore walls should provide good environment for the adhesion and proliferation of osteoblasts and osteoprogenitor cells.     

Ectopic bone formation associated with mesenchymal stem cells in a resorbable calcium deficient hydroxyapatite carrier

Bone substitute materials can induce bone formation in combination with mesenchymal stem cells (MSC). The aim of the current study was to examine ectopic in vivo bone formation with and without MSC on a new resorbable ceramic, called calcium deficient hydroxyapatite (CDHA). Ceramic blocks characterized by a large surface (48 m2/g) were compared with beta-tricalcium phosphate (beta-TCP), hydroxyapatite (HA) ceramics (both ca. 0.5 m2/g surface) and demineralized bone matrix (DBM). Before implantation in the back of SCID mice carriers were freshly loaded with 2x10(5) expanded human MSC or loaded with cells and kept under osteogenic conditions for two weeks in vitro. Culture conditions were kept free of xenogenic supplements. Deposits of osteoid at the margins of ceramic pores occurred independent of osteogenic pre-induction, contained human cells, and appeared in 416 MSC/CDHA composites compared to 216 MSC/beta-TCP composites. ALP activity was significantly higher in samples with MSC versus empty controls (p<0.001). Furthermore, ALP was significantly (p<0.05) higher for all ceramics when compared to the DBM matrix. Compared to previous studies, overall bone formation appeared to be reduced possibly due to the strict human protocol. Ectopic bone formation in the novel biomaterial CDHA varied considerably with the cell pool and was at least equal to beta-TCP blocks.     

Effect of reinforcement particle size on in vitro behavior of beta-tricalcium phosphate-reinforced high-density polyethylene: a novel orthopedic composite

Beta-tricalcium phosphate-reinforced high-density polyethylene (beta-TCP/HDPE) is a new biomaterial, which was made to simulate bone composition and study its capacity to act like bony tissues. This material was produced by replacing mineral component and collagen soft tissue of bone with beta-TCP and HDPE, respectively. The biocompatibility of composite samples with different volume fractions of TCP (20, 30, and 40 vol %) and two different particle sizes (80-100 and 120-140 mesh size) was examined in vitro using the osteoblast cell line G-292 by proliferation, alkaline phosphatase (ALP) production, and cell adhesion assays. Cell-material interaction on the surface of the composites was observed by scanning electron microscopy (SEM). The effect of beta-TCP particle size on behavior of the osteoblast cell line was compared between two groups of the composite samples containing smaller and larger reinforcement particle sizes as well as with those of a negative control. In general, results showed that the composite samples containing larger particles supported a higher rate of proliferation and ALP production by osteoblast cells after 3, 7, and 14 days of incubation compared to the composite samples with smaller particle size and control. Furthermore, more cells were attached to the surface of composite samples containing larger particle size when compared to the smaller particle size composites (p<0.05). This number was nearly equal with numbers adhered on negative control [tissue culture polystyrene (TPS)] and significantly higher in comparison with composite control [polyethylene (PE)] (p<0.05). Adhered cells presented a normal morphology by SEM and many of the cells were seen to be undergoing cell division. These findings indicate that beta-TCP/HDPE composites are biocompatible, nontoxic, and in some cases, act to stimulate proliferation of the cells, ALP production, and cell adhesion when compared to the control counterparts. Furthermore, beta-TCP/HDPE samples with larger reinforcement particle size were shown to possess better biological properties.     

Magnesia-doped HA/beta-TCP ceramics and evaluation of their biocompatibility

The sintering behavior, mechanical properties and biocompatibility of magnesia (MgO)-doped HA/TCP biphasic ceramics were studied. Pure HA/TCP ceramics showed poor sinterability due to the phase transformation of beta- to alpha-TCP. MgO-doped HA/TCP ceramics showed high density without any phase transformation of beta-TCP up to 1300 degrees C, for MgO dopants incorporated into the beta-TCP preferably and increased thermal stability of beta-TCP. However, the addition of MgO higher than a critical content, suppressed grain growth of HA/TCP ceramics and lowered sinterability. The optimum amount of MgO doping was 1 wt%, which lead 99% relative density and higher mechanical properties than HA or beta-TCP ceramics. From in vitro test and in vivo test, 1 wt% MgO-doped HA/TCP ceramics showed a good biocompatibility without cytotoxicity. After implantation under the muscle of rabbits, beta-TCP phase was dissolved from the surface and a biological apatite covered the surface. These results proved that MgO addition increased drastically the sintering and mechanical properties of HA/beta-TCP ceramics without altering the biological safety and biocompatibility of the original composite.     

In vitro evaluation of biocompatibility of beta-tricalcium phosphate-reinforced high-density polyethylene; an orthopedic composite

Beta-tricalcium phosphate-reinforced high-density polyethylene (beta-TCP/HDPE) is a new biomaterial which was made as a copy of bone composition with the aim of replacement of bony tissues. The composite samples were prepared using medical grade TCP powder and granular polyethylene. The raw materials were first compounded and the resulting composite preforms were compression molded into desired shape. The biocompatibility of composite samples with different volume fractions of TCP (20, 30, and 40 vol %) was assessed by proliferation, alkaline phosphatase (ALP), and cell adhesion assays using G-292 osteoblast cells. Cell-material interaction on the surface of the composites was observed by scanning electron microscopy (SEM). The effect of beta-TCP/HDPE on the behavior of G-292 cells was compared with those of a composite and a negative control samples. Results showed the composite samples had a higher proliferation rate of G-292 cells in the presence of composite samples as compared to the composite control sample after 3, 7, and 14 days of incubation period. ALP production after incubation in the presence of composite samples was seen to peak on the day 7. The number of adhered cells on the composite samples was higher than the numbers adhered on composite and negative control samples after the above incubation periods. Morphology investigation of adhered cells by SEM indicated a normal morphology and also many of the cells were in the process of cell division. The above results indicate that beta-TCP/HDPE samples are biocompatible, nontoxic, and in some cases show an increase in the proliferation rate of the cells, ALP production, and cell adhesion as compared to the control counterparts.     

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) composite biomaterials for bone tissue regeneration: in vitro performance assessed by osteoblast proliferation, osteoclast adhesion and resorption, and macrophage proinflammatory response

The efficacy of composite materials for bone tissue engineering is dependent on the materials' ability to support bone regeneration whilst inducing a minimal inflammatory response. In this study we examined the in vitro osteogenic and inflammatory properties of poly(3-hydroxybutyrate-co-3-valerate) (PHBV) with various calcium phosphate-reinforcing phases: nano-sized hydroxyapatite (HA); submicron-sized calcined hydroxyapatite (cHA); and submicron-sized beta-tricalcium phosphate (beta-TCP), using bioassays of cultured osteoblasts, osteoclasts, and macrophages. Our study showed that the addition of a nano-sized reinforcing phase to PHBV, whilst improving osteogenic properties, also reduces the proinflammatory response. Proinflammatory responses of RAW264.7/ELAM-eGFP macrophages to PHBV were shown to be markedly reduced by the introduction of a reinforcing phase, with HA/PHBV composites having the lowest inflammatory response. Osteoclasts, whilst able to attach to all the materials, failed to form functional actin rings or resorption pits on any of the materials under investigation. Cultures of osteoblasts (MC3T3-E1) readily attached and mineralised on all the materials, with HA/PHBV inducing the highest levels of mineralization. The improved biological performance of HA/PHBV composites when compared with cHA/PHBV and beta-TCP/PHBV composites is most likely a result of the nano-sized reinforcing phase of HA/PHBV and the greater surface presentation of mineral in these composites. Our results provide a new strategy for improving the suitability of PHBV-based materials for bone tissue regeneration.     

In vivo evaluation of a novel porous hydroxyapatite to sustain osteogenesis of transplanted bone marrow-derived osteoblastic cells

Biosynthetic bone grafts are considered to contain one or more of three critical components: osteoprogenitor cells, an osteoconductive matrix, and osteoinductive growth factors. The basic requirements of the scaffold material are biocompatibility, mechanical integrity, and osteoconductivity. A major design problem is satisfying these requirements with a single composite. In this study, we hypothesize that one composite that combines bone marrow-derived osteoblasts and a novel mechanical reinforced porous hydroxyapatite with good biocompatibility and osteoconductivity (HA/BMO) can reach these requirements. A novel sintered porous hydroxyapatite (HA) was prepared by the following procedures. The HA slurry was foamed by adding polyoxyethylenelaurylether (PEI) and mixing. The pores were fixed by crosslinking PEI with diepoxy compounds and the HA porous body was sintered at 1200 degrees C for 3 h. The HA sintered porous body had a high porosity (77%), and was completely interconnected. Average pore diameter was 500 microm and the interconnecting path 200 microm in diameter. The compressive (17 MPa) and three-point bending (7 MPa) strengths were high. For in vivo testing, the 2-week subcultured HA/BMO (+) composites were implanted into subcutaneous sites of syngeneic rats until 8 weeks after implantation. These implants were harvested at different time points and prepared for the biochemical analysis of alkaline phosphatase activity (ALP) and bone osteocalcin content (OCN), and histological analysis. ALP and OCN in the HA/BMO group were much higher than those in the HA without BMOs control group 1 week after implantation (p < 0.001). Light microscopy revealed mature bone formation in the HA/BMO composite 4 weeks after implantation. In the SEM study, mineralized collagenous extracellular matrix was noted in HA/BMO composite 2 weeks after implantation with numbers of active osteoblasts. We conclude that the composite of the novel HA and cultured BMOs has osteogenic ability in vivo. These results provide a basis for further studies on the use of this composite as an implant in orthopaedic surgery.     

In vitro evaluation of novel bioactive composites based on Bioglass-filled polylactide foams for bone tissue engineering scaffolds

Highly porous poly(DL-lactic acid) (PDLLA) foams and Bioglass-filled PDLLA composite foams were characterized and evaluated in vitro as bone tissue engineering scaffolds. The hypothesis was that the combination of PDLLA with Bioglass in a porous structure would result in a bioresorbable and bioactive composite, capable of supporting osteoblast adhesion, spreading and viability. Composite and unfilled foams were incubated in simulated body fluid (SBF) at 37 degrees C to study the in vitro degradation of the polymer and to detect hydroxyapatite (HA) formation, which is a measure of the materials' in vitro bioactivity. HA was detected on all the composite samples after incubation in SBF for just 3 days. After 28 days immersion the foams filled with 40 wt % Bioglass developed a continuous layer of HA. The formation of HA for the 5 wt % Bioglass-filled foams was localized to the Bioglass particles. Cell culture studies using a commercially available (ECACC) human osteosarcoma cell line (MG-63) were conducted to assess the biocompatibility of the foams and cell attachment to the porous substrates. The osteoblast cell infiltration study showed that the cells were able to migrate through the porous network and colonize the deeper regions within the foam, indicating that the composition of the foams and the pore structures are able to support osteoblast attachment, spreading, and viability. Rapid formation of HA on the composites and the attachment of MG-63 cells within the porous network of the composite foams confirms the high in vitro bioactivity and biocompatibility of these materials and their potential to be used as scaffolds in bone tissue engineering and repair.     

An amphiphilic degradable polymer/hydroxyapatite composite with enhanced handling characteristics promotes osteogenic gene expression in bone marrow stromal cells

Electrospun polymer/hydroxyapatite (HA) composites combining biodegradability with osteoconductivity are attractive for skeletal tissue engineering applications. However, most biodegradable polymers such as poly(lactic acid) (PLA) are hydrophobic and do not blend with adequate interfacial adhesion with HA, compromising the structural homogeneity, mechanical integrity and biological performance of the composite. To overcome this challenge, we combined a hydrophilic polyethylene glycol (PEG) block with poly(d,l-lactic acid) to improve the adhesion of the degradable polymer with HA. The amphiphilic triblock copolymer PLA–PEG–PLA (PELA) improved the stability of HA–PELA suspension at 25 wt.% HA content, which was readily electrospun into HA–PELA composite scaffolds with uniform fiber dimensions. HA–PELA was highly extensible (failure strain >200% vs. <40% for HA–PLA), superhydrophilic (～0° water contact angle vs. >100° for HA–PLA), and exhibited an 8-fold storage modulus increase (unlike deterioration for HA–PLA) upon hydration, owing to the favorable interaction between HA and PEG. HA–PELA also better promoted osteochondral lineage commitment of bone marrow stromal cells in unstimulated culture and supported far more potent osteogenic gene expression upon induction than HA–PLA. We demonstrate that the chemical incorporation of PEG is an effective strategy to improve the performance of degradable polymer/HA composites for bone tissue engineering applications.     

Mixing conditions for cell scaffolds affect the bone formation induced by bone engineering with human bone marrow stromal cells, beta-tricalcium phosphate granules, and rhBMP-2

Bone regenerative medicine via tissue engineering is expected to be an alternative treatment for conventional autogenous bone graft, as it is less invasive. One of the best triads for bone engineering is bone marrow stromal cells, calcium phosphate ceramics, and bone morphogenetic protein (BMP). However, the optimal mixing conditions for BMP-induced osteoblasts and ceramic granules remain unclear. Therefore, we investigated the effect of the mixing conditions for cell scaffolds on the bone-forming potential. The cells were mixed with beta-tricalcium phosphate (beta-TCP) granules followed by osteoblast induction with recombinant human BMP-2 (rhBMP-2) (first mixture), or were first induced with rhBMP-2 on plastic dishes and then mixed with the beta-TCP granules (last mixture) just prior to the operation. Both the first and last mixtures were transplanted into nude mice subcutaneously, with the amount of bone formation analyzed histomorphometrically. In addition, cell numbers and alkaline phosphatase (ALP) activity before transplantation was determined in both the mixtures. In vitro analyses revealed that cell numbers were greater in the last mixture, whereas ALP activity was greater in the first mixture. In vivo analyses revealed that the first mixture was much more osteogenic than the last mixture with respect to new bone formation and osteocalcin synthesis. These data suggest that cell-scaffold mixing conditions have a significant influence on the bone-forming capacity via bone engineering and that first mixture might be the optimal condition for rhBMP-2-induction of human osteoblasts.     

聚DL-乳酸/羟基磷灰石复合材料修复长骨缺损的实验研究

目的 :评价羟基磷灰石 (HA)复合聚DL 乳酸 (PDLLA )制备的材料体内成骨能力。方法 :将PDLLA和PDLLA/HA( 2 0wt % )材料采用盐结晶颗粒沥滤法制成三维多孔材料 ,45例 1cm兔桡骨去骨膜缺损分为三组 ,分别植入 2种材料和作空白对照 ,术后 2 ,4,8,12周行X线、组织学及扫描电镜观察骨生成状况 ,8、12周行生物力学测试 (三点折弯强度 )。结果 :泡沫状PDLLA/HA ( 2 0wt % )材料比纯PDLLA成骨更好 (P <0 .0 5 ) ,实验组与对照组相比差异有显著性 (P <0 .0 5 )。结论 :PDLLA具有良好的生物相容性 ,制成多孔状具有较好的骨传导性能 ,HA( 2 0wt % )的加入促进了多孔PDLLA的骨传导能力 ,提高了骨生成的质量。PDLLA/HA( 2 0wt % )复合材料是一种有临床应用前景的骨移植材料。     

Polylactic acid-phosphate glass composite foams as scaffolds for bone tissue engineering

Phosphate glass (PG) of the composition 0.46(CaO)-0.04(Na(2)O)-0.5(P(2)O(5)) was used as filler in poly-L-lactic acid (PLA) foams developed as degradable scaffolds for bone tissue engineering. The effect of PG on PLA was assessed both in bulk and porous composite foams. Composites with various PG content (0, 5, 10, and 20 wt %) were melt-extruded, and either compression-molded or foamed through supercritical CO(2). Dynamic mechanical analysis on the bulk composites showed that incorporating 20 wt % PG resulted in a significant increase in storage modulus. Aging studies in deionized water in terms of weight loss, pH change, and ion release inferred that the degradation was due to PG dissolution, and dependent on the amount of glass in the composites. Foaming was only possible for composites containing 5 and 10 wt % PG, as an increase in PG increased the foam densities; however, the level of porosity was maintained above 75%. PLA-T(g) in the foams was higher than those obtained for the bulk. Compressive moduli showed no significant reinforcement with glass incorporation in either expansion direction, indicating no anisotropy. Biocompatibility showed that proliferation of human fetal bone cells was more rapid for PLA compared to PLA-PG foams. However, the proliferation rate of PLA-PG foams were similar to those obtained for foams of PLA with either hydroxyapatite or beta-tricalcium phosphate.     

碳化硅对成骨细胞增殖和分化的影响

目的研究碳化硅(SiC)的体外生物相容性,为探索SiC作为骨缺损修复材料的可行性奠定基础。方法实验组为实质SiC,对照组为致密羟基磷灰石(HA)。将取自大鼠胎鼠颅骨的成骨细胞分别接种于2组材料表面。分别采用扫描电子显微镜、四甲基偶氮唑盐(MTT)法、碱性磷酸酶(ALP)含量测定技术、流式细胞仪检测细胞周期,观察分析两种材料表面上的细胞的增殖和分化能力。结果扫描电子显微镜显示:原代成骨细胞在两种材料表面伸展良好,细胞在材料表面伸出很多伪足,并且在材料表面可见明显的细胞外基质沉积。两组材料上细胞的MTT值从1、3、5、7 d随观察时间逐渐升高,并且各个时间点实质SiC均高于致密HA,两者之间的差异有统计学意义(P<0.05)。两组材料上细胞的ALP含量从1、3、5、7 d随观察时间逐渐升高;在1、3 d时,致密HA高于实质SiC,在5、7 d时,致密HA低于实质SiC,但是两组之间的差异均无统计学意义(P>0.05)。1、3、5 d两组材料的细胞增值指数(PI)值逐渐升高,高峰出现在第5天,第7天时下降,两组材料在各个时间点的PI均接近,差异无统计学意义(P>0.05)。结论实质SiC具有与致密HA相似的良好的细胞相容性。     

Migration of Co-cultured Endothelial Cells and Osteoblasts in Composite Hydroxyapatite/Polylactic Acid Scaffolds

Regeneration of bone in large segmental bone defects requires regeneration of both cortical bone and trabecular bone. A scaffold design consisting of a hydroxyapatite (HA) ring surrounding a polylactic acid (PLA) core simulates the structure of bone and provides an environment for indirect and direct co-culture conditions. In this experiment, human umbilical vein endothelial cells (EC) and normal human primary osteoblasts (OB) were co-cultured to evaluate cell migration and interactions within this biphasic composite scaffold. Both cell types were able to migrate between the different material phases of the scaffold. It was also observed that OB migration increased when they were co-cultured with ECs, whereas EC migration decreased in co-culture. The results show that co-culture of ECs and OBs in this composite biphasic scaffold allows for migration of cells throughout the scaffold and that pre-seeding a scaffold with ECs can increase OB infiltration into desired areas of the scaffold.