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1.
目的:通过建立兔股骨缺损的动物实验模型,对采用等温化学气相沉积法和等离子喷涂技术所制备的石墨化炭/炭复合材料+羟基磷灰石涂层(C/C+HA)复合骨植入材料进行骨植入实验的的生物相容性进行评价,探索该复合材料作为植入机体骨组织的可行性依据.方法:采用骨科钻在实验动物股骨髁上钻孔的方法建立骨缺损的动物实验模型,将待研究比较的实验材料分别植入实验动物的股骨髁内,持续观察8周,在术后第2、4、8周时应用X线照片、组织学染色和扫描电镜技术,分别观察所研究材料在机体内对骨缺损愈合及其对机体的影响,进行组间比较和相关性分析.结果:石墨化炭/炭复合材料+羟基磷灰石涂层(C/C+HA)复合骨植入材料的骨植入实验生物相容性良好,材料与骨组织结合牢固,界面中成骨细胞生长明显,且炭颗粒脱落现象少,未见炎症细胞浸润.植入动物体内的材料在植入期未引起机体局部的炎症浸润反应且表面脱落的碳颗粒在机体组织中也未引起局部严重的炎症反应.在实验动物植入材料后的连续8周观察期中,组织学观察显示:表面涂有HA的炭/炭复合材料对骨组织形态改建上表现良好,其与骨组织接界处所形成的纤维结缔组织膜层厚度明显比未涂HA的材料要小,与骨组织结合更为紧密和牢固;碳颗粒出现脱落游离的现象明显减少.结论:在炭/炭复合材料表面涂以HA生物涂层对骨的形态改建和促进骨小梁生长等方面具有良好的作用,是一种具有发展潜力的骨修复材料.  相似文献   

2.
磷酸三钙涂层镁合金材料的细胞相容性研究   总被引:1,自引:0,他引:1  
目的:制备磷酸三钙(β-TCP)涂层镁合金材料,评价材料表面的特性及体外的细胞生物适应性。方法 电化学法制备β-TCP涂层镁合金材料(β-TCP-Mg-AI-Zn),观测金属材料表面微观结构特性和能谱分析,小鼠颅骨源成骨细胞与材料直接接触培养,荧光染色观察材料表面细胞生长状况,检测成骨细胞增殖和碱性磷酸酶(ALP)活性。结果 β-TCP涂层Mg-AI-Zn材料表面呈多孔状,材料表面含有镁、钙和磷等元素;成骨细胞与材料直接接触培养24 h及48 h后,材料表面有大量的成骨细胞粘附、伸展、汇合;与Mg-AI-Zn材料比较,β-TCP-Mg-AI-Zn材料明显地促进细胞增殖、显著地增加成骨细胞中ALP活性 (P<0.05)。结论 β-TCP涂层改善了Mg-AI-Zn镁合金材料表面特性及体外的细胞相容性,有望成为新一代可降解医用金属材料。  相似文献   

3.
王健  吴松  陈腾飞  朱东波  王靖  周天宝 《生物磁学》2011,(14):2615-2619
目的:通过建立兔股骨缺损的动物实验模型,对采用等温化学气相沉积法和等离子喷涂技术所制备的石墨化炭/炭复合材料+羟基磷灰石涂层(C/C+HA)复合骨植入材料进行骨植入实验的的生物相容性进行评价,探索该复合材料作为植入机体骨组织的可行性依据。方法:采用骨科钻在实验动物股骨髁上钻孔的方法建立骨缺损的动物实验模型,将待研究比较的实验材料分别植入实验动物的股骨髁内,持续观察8周,在术后第2、4、8周时应用X线照片、组织学染色和扫描电镜技术,分别观察所研究材料在机体内对骨缺损愈合及其对机体的影响,进行组间比较和相关性分析。结果:石墨化炭/炭复合材料+羟基磷灰石涂层(C/C+HA)复合骨植入材料的骨植入实验生物相容性良好,材料与骨组织结合牢固,界面中成骨细胞生长明显,且炭颗粒脱落现象少,未见炎症细胞浸润。植入动物体内的材料在植入期未引起机体局部的炎症浸润反应且表面脱落的碳颗粒在机体组织中也未引起局部严重的炎症反应。在实验动物植入材料后的连续8周观察期中,组织学观察显示:表面涂有HA的炭/炭复合材料对骨组织形态改建上表现良好,其与骨组织接界处所形成的纤维结缔组织膜层厚度明显比未涂HA的材料要小,与骨组织结合更为紧密和牢固;碳颗粒出现脱落游离的现象明显减少。结论:在炭/炭复合材料表面涂以HA生物涂层对骨的形态改建和促进骨小梁生长等方面具有良好的作用,是一种具有发展潜力的骨修复材料。  相似文献   

4.
目的:柞蚕丝素(tussah silk fibroin,TSF)和羟基磷灰石(hydroxyapatite,HA)均具有良好的生物活性和生物相容性,是组织工程研究的热点i,但结构单一及微米级的材料所表现出的性能简单,不能满足人们对生物材料支架性能的要求,本课题将两者按不同比例进行复合,探讨不同皮芯比例羟基磷灰石/柞蚕丝素(HA/TSF)的骨仿生纳米纤维的生物学性能。方法:首先利用同轴静电纺丝技术,以TSF水溶液为皮,HA水溶液为芯,制备不同皮芯比例的HA/TSF骨仿生纳米纤维,然后将人成骨肉瘤细胞(MG-63)种植在不同皮芯比例的HA/TSF纳米纤维上。在不同的时间点分别通过倒置显微镜、扫描电镜观察细胞形态学变化;通过四甲基偶氮噻唑蓝比色(Four methyl azo thiazole blue colorimetric, MTT)法、碱性磷酸酶(alkaline phosphatase,ALP)活性检测法观察细胞在材料表面的增殖和分化,从多角度来评价材料的生物学性能。结果:通过形态学观察,SEM观察以及MTT检测,发现除空白对照组外,各组样品均显示良好的生物相容性,均能促进细胞的黏附、增殖,尤以HA/TSF为2:1时最明显;通过MG-63细胞的ALP活性检测,发现当HA/TSF比例为2:1时,最能促进细胞ALP活性的表达,有利于诱导成骨细胞的分化。结论:皮芯结构的HA/TSF骨仿生纳米纤维具有良好的生物学性能,且二者在自然界来源丰富,价格便宜,为临床骨组织缺损修复的应用奠定了一定的实验基础  相似文献   

5.
目的比较纳米细菌(nanobacteria,NB)与羟基磷灰石(hydroxyapatite,HA)对成骨细胞的细胞毒作用,进一步探讨引起细胞毒作用的机制。方法实验分为正常对照组、HA组、NB组,其中HA组和NB组悬液浓度均为2麦氏浊度(M),对照组只加培养基,分别作用成骨细胞,通过CCK-8试剂盒检测其对细胞的增殖抑制作用;采用Hoechest33258荧光染色和Annexin V-FITC/PC双标法流式细胞仪(flow cotymetry)检测细胞凋亡率;倒置显微镜和透射电镜观察细胞形态的变化;Western blot法检测钙化相关蛋白BMP-2、凋亡相关蛋白Bax的表达。结果 CCK-8结果显示:NB对人成骨细胞C3H10有细胞增殖抑制作用,而HA则没有。Hoechst33258荧光染色和双标法流式细胞仪结果显示:NB对人成骨细胞C3H10主要通过促进细胞凋亡发挥细胞毒作用,而HA对人成骨细胞C3H10无细胞毒性作用。在倒置显微镜下观察,作用48 h后的人成骨细胞C3H10形态无明显变化。透射电镜结果显示:NB可引起人成骨细胞C3H10发生凋亡,出现凋亡小体;而HA组和对照组无明显变化。Western blot结果显示:NB组的钙化相关蛋白BMP-2和凋亡相关蛋白Bax均高于对照组;而HA组和对照组比较,HA组钙化相关蛋白BMP-2高于对照组,而凋亡相关蛋白Bax无明显变化。结论与对照组比较,纳米细菌能引起人成骨细胞C3H10细胞毒性作用,主要是通过促进细胞凋亡发挥细胞毒作用,能促进钙化相关蛋白BMP-2的表达,凋亡相关蛋白Bax的表达升高也预示着纳米细菌可促进细胞发生凋亡;而羟基磷灰石对人成骨细胞C3H10无细胞毒性作用,可促进钙化相关蛋白BMP-2的表达,但不能促进人成骨细胞C3H10发生凋亡。  相似文献   

6.
FGF-2对人骨髓间充质干细胞增殖和向成骨细胞分化的影响   总被引:4,自引:0,他引:4  
探讨体外培养条件下,成纤维细胞生长因子-2(FGF-2)和地塞米松(Dex)对第7代人骨髓间充质干细胞(MSCs)增殖和向成骨细胞分化的作用以及两者联合使用的效应。MSCs经含FGF-2或/和Dex的培养液作用后,于不同时间采用MTT法测定细胞增殖情况;对硝基苯磷酸(pNPP)法测定碱性磷酸酶(ALP)活性;ELISA法测定骨钙蛋白(OC)含量;茜素红S染色法对沉积的钙盐进行染色。发现:(1)FGF-2组细胞的生长速度为对照组的1.31倍,Dex/FGF-2组细胞的生长速度为FGF-2组的1.12倍。(2)Dex组的ALP活性、OC含量和细胞外基质钙盐沉积分别为对照组的17.0倍、2.12倍和10.56倍,并能形成成熟的羟基磷灰石(HA)结晶和骨结节;FGF-2组的ALP活性比对照组降低了76.7%,虽然OC含量、钙盐沉积增加,但不能形成成熟的HA结晶和骨结节;FGF-2对Dex诱导的ALP活性增加和HA结晶形成有拮抗作用。由此证明:(1)FGF-2可促进MSCs的增殖,Dex对MSCs的增殖无明显作用;Dex能增强FGF-2对MSCs的促增殖效应。(2)Dex可使MSCs分化为成熟的成骨细胞,是一个有效的成骨细胞分化诱导剂;FGF-2可使MSCs分化为未成熟的成骨细胞;FGF-2拮抗Dex诱导MSCs分化为成熟的成骨细胞。  相似文献   

7.
为了研究珍珠质自然涂层钛种植体表面的体外生物相容性,将珍珠质自然涂层的钛片与MC3T3E1成骨样细胞复合培养以观察细胞的生长、增殖和分化.分别以羟基磷灰石涂层钛片和没有涂层的纯钛片作为对照组,以MC3T3E1细胞单纯培养作为空白组,分别培养3天,5天和7天,通过倒置相差显微镜和扫描电镜观察细胞生长情况,流式细胞技术检测细胞增殖活性,金氏比色法检测碱性磷酸酶(ALP)活性以及蛋白质印迹(Western blotting)法测定转化生长因子-β1(TGF-β1)表达水平.结果发现,细胞在珍珠质周围能形成良好附着,在其表面生长丰满.细胞培养第3天,第5天和第7天时,珍珠质表面的细胞增殖指数分别为(35.9±2.5)%、(69.7±3.3)%和(58.2±2.6)%,ALP活性分别为(6.123±2.917)U/g、(17.486±1.986)U/g和(23.987±1.372)U/g.第5天和7天时,实验组的细胞增殖指数、ALP活性和TGF-β1表达水平显著高于对照组和空白组(P<0.05).珍珠质自然涂层钛表面有利于MC3T3E1细胞的生长、增殖和分化,表明了珍珠质涂层能提高种植体表面的生物相容性,有可能会促进种植后的骨整合.  相似文献   

8.
目的:探讨三碘甲腺原氨酸(T3)对大鼠成骨细胞增殖的影响。方法:采用骨组织块法原代分离培养新生SD大鼠颅骨成骨细胞,然后配置含不同浓度(10-7mol/L、10-8mol/L、10-9mol/L)T3的培养基,与大鼠成骨细胞共培养4d,采用噻唑蓝(MTT)法检测细胞增殖能力,采用细胞化学染色法测定成骨细胞ALP活性。结果:T3可促进大鼠成骨细胞增殖,并呈剂量依赖性;随T3浓度的增高,成骨细胞表达ALP活性增强(P<0.05)。结论:10-7mol/L~10-9mol/L浓度的T3可通过刺激成骨细胞的增殖,对预防和治疗骨质疏松发挥一定作用。  相似文献   

9.
[目的]引入界面增强剂磺化聚醚醚酮,合成新型聚醚醚酮/磺化聚醚醚酮/羟基磷灰石三元复合材料,研究该材料对成骨细胞MG-63氧化作用影响。[方法]将成骨细胞MG-63和各材料共同培养,测量材料对细胞产生丙二醛和活性氧化自由基水平,通过扫描电镜观察三元复合材料微观结构。[结果]三元复合材料上成骨细胞MG-63产生的丙二醛和活性氧化自由基浓度低于其他材料,HA质量百分比达到30%时,成骨细胞MG-63产生的丙二醛和活性氧化自由基浓度与在模拟人体正常生长条件下的数据接近,统计量P 0. 05,氧化应激作用差异相对不明显,微观下该材料具备类骨的微孔结构和孔隙。[结论]该材料对成骨细胞MG-63氧化作用低于聚醚醚酮/羟基磷灰石材料,类骨的微孔结构能够增加细胞接触面积和有利于营养运输与骨诱导作用。  相似文献   

10.
目的:探讨三碘甲腺原氨酸(T3)对大鼠成骨细胞增殖的影响.方法:采用骨组织块法原代分离培养新生SD大鼠颅骨成骨细胞,然后配置含不同浓度(10-7mol/L、10-8mol/L、10-9mol/L )T3的培养基,与大鼠成骨细胞共培养4d,采用噻唑蓝(MTT)法检测细胞增殖能力,采用细胞化学染色法测定成骨细胞ALP活性.结果:T3可促进大鼠成骨细胞增殖,并呈剂量依赖性;随T3浓度的增高,成骨细胞表达ALP活性增强(P<0.05).结论:10-7mol/L~10-9mol/L浓度的T3可通过刺激成骨细胞的增殖,对预防和治疗骨质疏松发挥一定作用.  相似文献   

11.
High pure and crystalline Hydroxyapatite (HA) coatings on titanium alloy were prepared by hydrothermal synthesis (HS) of plasma-sprayed (PS) precursors from brushite powders (HS-HA). In vitro and in vivo tests were done to evaluate its biological property. The HS-HA coating was compared with the current PS-HA coating. Cultures of the primary osteoblasts on these two HA coatings showed similar cell attachment, proliferation and alkaline phosphatase (ALP) expression. The cell morphology on the coatings was demonstrated by scanning electron microscopy (SEM). The cell spread well at 1 day after seeding culture and the extracellular matrix was secreted after 14 days culture. Histomorphometric analysis was conducted on samples implanted in femoral bone of four dogs for 1 and 3 months, and bone-implant contact percentage was evaluated by light microscopy. The calcium and phosphate distribution on the interface of bone-implant was analysed by SEM and electron dispersive X-ray (EDX) analysis. The results show the osteoconduction of HS-HA coated implants.  相似文献   

12.
Natural bone consists of hard nanostructured hydroxyapatite (HA) in a nanostructured protein-based soft hydrogel template (ie, mostly collagen). For this reason, nanostructured HA has been an intriguing coating material on traditionally used titanium for improving orthopedic applications. In addition, helical rosette nanotubes (HRNs), newly developed materials which form through the self-assembly process of DNA base pair building blocks in body solutions, are soft nanotubes with a helical architecture that mimics natural collagen. Thus, the objective of this in vitro study was for the first time to combine the promising attributes of HRNs and nanocrystalline HA on titanium and assess osteoblast (bone-forming cell) functions. Different sizes of nanocrystalline HA were synthesized in this study through a wet chemical precipitation process following either hydrothermal treatment or sintering. Transmission electron microscopy images showed that HRNs aligned with nanocrystalline HA, which indicates a high affinity between both components. Some of the nanocrystalline HA formed dense coatings with HRNs on titanium. More importantly, results demonstrated enhanced osteoblast adhesion on the HRN/nanocrystalline HA-coated titanium compared with conventional uncoated titanium. Among all the HRN/nanocrystalline HA coatings tested, osteoblast adhesion was the greatest when HA nanometer particle size was the smallest. In this manner, this study demonstrated for the first time that biomimetic HRN/nanocrystalline HA coatings on titanium were cytocompatible for osteoblasts and, thus, should be further studied for improving orthopedic implants.  相似文献   

13.
We investigated the effects of different implant biomaterials on cultured canine bone marrow stromal cells (BMSC) undergoing differentiation into osteoblasts (dBMSC). BMSC were isolated from canine humerus by marrow aspiration, cultured and differentiated on calcium phosphate scaffold (CPS), hydroxyapatite, hydroxyapatite in gel form and titanium mesh. We used the MTT method to determine the effects of osteogenic media on proliferation. The characteristics of dBMSC were assessed using alizarin red (AR), immunocytochemistry and osteoblastic markers including alkaline phosphatase/von Kossa (ALP/VK), osteocalcin (OC) and osteonectin (ON), and ELISA. The morphology of dBMSC on the biomaterials was investigated using inverted phase contrast microscopy and scanning electron microscopy. We detected expression of ALP/VK, AR, OC and ON by day 7 of culture; expression increased from day 14 until day 21. CPS supported the best adhesion, cell spreading, proliferation and differentiation of BMSCs. The effects of the biomaterials depended on their surface properties. Expression of osteoblastic markers showed that canine dBMSCs became functional osteoblasts. Tissue engineered stem cells can be useful clinically for autologous implants for treating bone wounds.  相似文献   

14.
Therapeutic strategies for bone regeneration involve the selection of suitable biomaterials, growth factors, and cell types to mimic the cellular microenvironment where molecular and mechanical signals control the reconstruction of bone tissue. The immobilization of basic fibroblast growth factor (FGF-2) on powdered silicon-substituted hydroxyapatite (Si-HA) allows to prepare a biofunctional biomaterial able to interact with bone cells in a very specific way. The biological activity of FGF-2/Si-HA, evaluated in Saos-2 osteoblasts and MC3T3-E1 preosteoblasts through the PLCγ and MAPK/ERK signal transduction pathways, shows that FGF-2 immobilized on Si-HA provides the right signals to cells stimulating crucial intracellular mechanisms of osteoblast proliferation and differentiation.  相似文献   

15.
Inorganic polyphosphate has been expected to accelerate bone regeneration. However, there are limited evidences to prove that polyphosphate adsorbed on the surface of a hydroxyapatite plate enhances calcification of cultured osteoblasts. In this study, we examined the effect of polyphosphate adsorbed onto the surface of a hydroxyapatite plate on the attachment, proliferation, differentiation, and calcification of osteoblasts. After hydroxyapatite plates were soaked in solutions of polyphosphate, the plate surfaces were analyzed by scanning electron microscopy and toluidine blue staining to confirm adsorption of polyphosphate. The hydroxyapatite plates were further subjected to the measurements of surface roughness, water contact angle, and the binding capacity of calcium ions. Cell culture experiments were carried out using MC3T3-E1 pre-osteoblastic cells. It was found that soaking a hydroxyapatite plate in a polyphosphate solution gave rise to an increase in surface roughness and reduction in water contact angle in a concentration-dependent manner, suggesting the adsorption of polyphosphate onto the surface of a hydroxyapatite plate. It was further observed that surface-adsorbed polyphosphate exhibited an inhibitory effect on cell adhesion and proliferation. In contrast, cell differentiation was promoted on hydroxyapatite plates with adsorbed polyphosphate, when assessed from expression of differentiation marker genes including alkaline phosphatase, osteopontin, and osteocalcin. In addition, calcification of the culture was enhanced on hydroxyapatite plates with relatively low density of adsorbed polyphosphate. Our results as a whole provided an evidence to show that there is a narrow window with regard to the surface density of adsorbed polyphosphate for the enhancement of osteoblast calcification.  相似文献   

16.
Several studies are currently ongoing to construct synthetic bone-like materials with composites of natural and polymeric materials with HA (hydroxyapatite). The present study aims to fabricate composite nanofibrous substrate of Chit/HA (chitosan/HA - 80:25) prepared by dissolving in TFA/DCM (trifluoroacetic acid/dichloromethane) (70:30, w/w) for 5 days and electrospun to fabricate a scaffold for bone tissue engineering. HA (25 wt %) was sonicated for 30 min to obtain a homogenous dispersion of nanoparticles within the Chit (80 wt %) matrix for fabricating composite nanofibrous scaffold (Chit/HA). The nanofibres of Chit and Chit/HA were obtained with fibre diameters of 274 ± 75 and 510 ± 198 nm, respectively, and characterized by FESEM (field emission scanning electron microscopy) and FTIR (Fourier transform infrared). The interaction of hFOBs (human fetal osteoblasts) and nanofibrous substrates were analysed for cell morphology (FESEM), mineralization [ARS (Alizarin Red-S) staining], quantification of minerals and finally identified the elements present in Chit/HA/osteoblasts by EDX (energy-dispersive X-ray) analysis. EDX analysis confirmed that the spherulites contain calcium and phosphorus, the major constituents in calcium phosphate apatite, the mineral phase of the bone. Mineralization was increased significantly (P<0.001) up to 108% in Chit/HA compared with Chit nanofibres. These results confirmed that the electrospun composite Chit/HA nanofibrous substrate is a potential biocomposite material for the proliferation and mineralization of hFOBs required for enhanced bone tissue regeneration.  相似文献   

17.
We have produced Bioglass coatings for Orthopedic implants by using a novel coating technique, CoBlast. The two resultant surfaces, designated BG and hydroxyapatite (HA)/BG, were compared with their HA counterpart, OsteoZip in terms of osteoblastic cell attachment, adhesion, proliferation, differentiation, and growth factor production. BG and HA/BG were demonstrated by goniometry to be more hydrophilic than OsteoZip. This corresponded to enhanced protein adsorption, cell attachment, and cell adhesion documented by both quantitative and qualitative assessments. BG and HA/BG surfaces had a significant initial release of Si and Ca ions, and this was consistent with elevated cell proliferation and basic fibroblast growth factor levels. However, OsteoZip, being similar to HA/BG, exhibited better osteogenic differentiation than BG did, shown by augmented differentiation marker activity at both protein and mRNA levels. Sandwich ELISA was used to quantify angiopoietin and inducible nitric oxide synthase which are involved in peri‐prosthetic angiogenesis and aseptic loosening of total hip replacement, respectively. Both Bioglass‐derived coatings provide superior initial osteoconductivity to OsteoZip, and HA/Bioglass composite coating outruns in long‐term osteogenic differentiation and prognostic bioprocesses. The novel coatings discovered in this study have significant potential in providing both orthopedic and therapeutic functions. Biotechnol. Bioeng. 2011;108: 454–464. © 2010 Wiley Periodicals, Inc.  相似文献   

18.
The novel non-thermal CoBlast process has been used recently to create a hydroxyapatite coating on metallic substrates with improved biological response compared to an uncoated implant. In this study, we compared the biological effect of coatings deposited by this process and the industrial standard technique - plasma-spray. Physicochemical properties of these two coatings have been found to be significantly different in that CoBlast HA is less rough but more hydrophilic than the plasma-spray HA as evidenced by data obtained from profilometry and goniometry. Mesenchymal stem cell attachment and adhesion are enhanced on CoBlast HA. Analysis by a combination of EDX and ICP suggests that the higher crystallinity retained by the CoBlast HA result in slower coating dissolution. Detailed in vitro evaluation reveals that plasma-spray HA might induce slightly faster cell proliferation and earlier osteogenic differentiation, but CoBlast HA becomes equivalent to it by the late osteogenic stage. PCR array facilitated the identification of differentially regulated genes involved in various functional aspects of in vitro osteogenesis by the CoBlast HA coating. The expression level of the functional protein products of these genes are in agreement with the PCR data. Coating metallic screws with HA significantly improves the in vivo osseointegration. By measuring of removal force using torque measurement instrument and analyzing the patterns found in X-ray images it is demonstrated that the two HA coatings elicit comparable osseointegration. Using simulated impaction model, CoBlast HA is shown to maintain better performance in cell attachment and mineralization than plasma-spray HA, especially following significant impactions. This might indicate a potentially greater osteoconductivity of CoBlast HA coating in shear-stress associated surgical applications. Collectively, it was demonstrated that CoBlast HA is an effective alternative to plasma-spray HA coating and a promising replacement for specialized surgical applications.  相似文献   

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