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1.
目的探讨骨形态发生蛋白(Bone morphogenetic protein,BMP)9-2双表达定向诱导多潜能干细胞C3H10向成骨细胞分化的情况。方法将C3H10细胞分为4组:BMP9-2、BMP2、BMP9和GFP组,将4种重组腺病毒分别感染C3H10细胞,通过碱性磷酸酶(Alkaline phosphatase,ALP)染色、定量测定及钙茜素红染色,观察BMP9-2双表达对C3H10细胞成骨分化的定向诱导作用。结果 BMP9-2双表达能诱导C3H10细胞ALP的表达,其染色活性高于BMP2组1.6倍,BMP9组2.5倍,GFP组4倍;其BMP9-2双表达定量表达A520值在病毒感染后5、7、9 d均高于BMP2、BMP9和GFP组;BMP9-2双表达能诱导C3H10细胞钙盐的沉积,感染14 d后,钙茜素红染色可见钙化结节,其染色活性高于BMP2组1.4倍,BMP9组1.3倍,GFP组5.2倍。结论 BMP9-2双表达能定向诱导C3H10细胞成骨分化,其成骨活性强于单一成骨诱导因子BMP2和BMP9。 相似文献
2.
《中国生物制品学杂志》2016,(3)
目的探讨重组腺病毒介导的FOXO/MDA-7对乳腺癌MDA-MB-231细胞凋亡的影响及其机制。方法将MDA-MB-231细胞分为Ad-FOXO/MDA-7组、Ad-FOXO组、Ad-MDA-7组和空白对照组,采用Real-time PCR、q RTPCR和Western blot法检测各组细胞中FOXO和MDA-7基因的转录及蛋白表达水平;MTT法检测细胞的增殖活力;流式细胞术检测细胞的凋亡情况;Transwell法检测细胞的侵袭能力;Western blot法检测细胞中Ras、Raf、MEK和ERK蛋白的表达情况。结果与其他各组相比,转染Ad-FOXO/MDA-7后,MDA-MB-231细胞中FOXO和MDA-7的表达明显增强,细胞的增殖活力和侵袭能力明显被抑制(P0.05),凋亡率明显升高,细胞中Ras、Raf、MEK和ERK蛋白的表达也明显被抑制。结论重组腺病毒Ad-FOXO/MDA-7能明显促进乳腺癌细胞的凋亡,为乳腺癌的基因治疗提供了新的思路。 相似文献
3.
目的探讨DN-ALK2对人乳腺癌细胞MDA-MB-231增殖、迁移和侵袭能力的影响。方法 RT-PCR法检测MDA-MB-231细胞中ALK2基因mRNA的转录;分别以腺病毒DN-ALK2和RFP感染MDA-MB-231细胞,并设空白对照组,通过MTT法、平板集落形成试验、细胞划痕试验及Transwell侵袭试验检测细胞的增殖、集落形成、迁移及侵袭能力。结果 MDA-MB-231细胞中内源性表达ALK2。腺病毒DN-ALK2和RFP感染MDA-MB-231细胞36 h后,荧光表达量一致,约为70%。MDA-MB-231/DN-ALK2细胞中高表达DN-ALK2。与MDA-MB-231/RFP组相比,MDA-MB-231/DN-ALK2组细胞的增殖活力、集落形成率及划痕愈合率均显著降低(P<0.05),穿膜细胞数也明显减少(P<0.05);而MDA-MB-231/RFP组与空白对照组比较,差异无统计学意义(P>0.05)。结论DN-ALK2可以在体外抑制乳腺癌MDA-MB-231细胞的增殖、迁移与侵袭。 相似文献
4.
目的探讨经典Wnt信号通路关键节点β-catenin对骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)诱导间充质干细胞(mesenchymal stem cells,MSCs)成骨分化的影响。方法用重组腺病毒介导BMP9在C3H10T1/2细胞中过表达,联用β-catenin重组腺病毒上调β-catenin的表达,并通过RNA干扰抑制β-catenin的表达。分析C3H10T1/2细胞碱性磷酸酶(alkaline phosphatase,ALP)活性的变化;RT-PCR检测细胞成骨分化相关基因骨桥蛋白(osteopontin,OPN)和骨钙蛋白(osteocalcin,OC)基因mRNA的转录水平;茜素红S染色检测细胞的钙盐沉积。结果 BMP9单独作用能诱导C3H10T1/2细胞向成骨方向分化,并增强细胞ALP活性;单独的β-catenin无成骨诱导作用,但可剂量依赖性地增强BMP9诱导的C3H10T1/2细胞的ALP活性,并促进BMP9诱导的细胞OPN和OC基因mRNA的转录水平及钙盐沉积;抑制β-catenin表达可显著降低BMP9诱导的C3H1OT1/2细胞的ALP活性(P0.05),下调OPN和OC基因mRNA的转录水平,并抑制钙盐沉积。结论经典Wnt信号通路可能通过β-catenin协同BMP9诱导C3H10T1/2细胞成骨分化,且BMP9诱导的成骨分化可能需要通过Wnt/β-catenin途径来实现。 相似文献
5.
目的探讨乌司他丁(Ulinastatin,UTI)和泰索帝(Taxotere,TXT)对体外培养的人乳腺癌细胞MDA-MB-231中u-PA、uPAR、ERK表达的影响。方法将MDA-MB-231(ER-)细胞分为4组:UTI组(UTI 800 U/ml)、TXT组(TXT 3.7μg/ml)、UTI+TXT组(UTI 800 U/ml+TXT 3.7μg/ml)、对照组(等量生理盐水)。给药后24 h,分别采用荧光定量RT-PCR检测各组细胞中uPA、uPAR、ERK基因mRNA的水平,Western blot法检测各组细胞中uPA、uPAR、p-ERK1/2蛋白的表达水平。结果 UTI组和UTI+TXT组MDA-MB-231(ER-)细胞中uPA和uPAR基因mRNA的水平均明显低于对照组(P<0.05),而TXT组中二者的表达水平均明显高于对照组(P<0.01),各组间ERK基因mRNA的水平差异无统计学意义(P>0.05);UTI组和UTI+TXT组中uPA、uPAR和p-ERK1/2蛋白的表达水平均明显低于对照组(P<0.01),而TXT组中3种蛋白的表达水平均明显高于对照组(P<0.05)。结论UTI可抑制MDA-MB-231细胞中uPA、uPAR、p-ERK的表达,而TXT可上调三者的表达。 相似文献
6.
目的研究姜黄素对转化生长因子-β1(Transforming growth factor-beta 1,TGF-β1)诱导的人乳腺癌MDA-MB-231细胞基质金属蛋白酶-9(Matrix metallopeptidase-9,MMP-9)表达及其侵袭能力的影响,探讨姜黄素在防治TGF-β1诱导的乳腺癌侵袭转移中可能的作用机制。方法采用CCK-8法检测姜黄素对MDA-MB-231细胞的细胞毒性作用。将MDA-MB-231细胞分为阴性对照组(无血清RPMI1640培养基/DMSO)、TGF-β1处理组(无血清RPMI1640培养基/DMSO+10 ng/ml TGF-β1),姜黄素+TGF-β1处理组(无血清RPMI1640培养基+5、7.5、10μmol/L姜黄素+10 ng/ml TGF-β1)。处理24 h后,采用侵袭小室试验观察细胞的侵袭能力;处理不同时间后,采用Western blot法检测细胞MMP-9、p-Smad2、p-ERK1/2以及p-p38的表达,明胶酶谱法检测各组细胞上清液中MMP-9的活性变化;以ERK特异性抑制剂PD98059、p38特异性抑制剂SB202580、姜黄素和/或TGF-β1处理细胞48 h,采用Western blot和明胶酶谱法检测MMP-9的表达。结果低浓度姜黄素(≤10μmol/L)对MDA-MB-231细胞无明显的细胞毒性作用;姜黄素呈剂量依赖性明显减少了TGF-β1诱导的细胞穿膜数量(P<0.001);姜黄素可显著抑制TGF-β1诱导的MDA-MB-231细胞MMP-9、p-Smad2、p-ERK1/2及p-p38蛋白的表达,且呈浓度、时间依赖性(P<0.05);姜黄素随浓度的增加,对TGF-β1诱导的MDA-MB-231细胞MMP-9活性的抑制作用明显增强(P<0.05);PD98059抑制MMP-9蛋白表达及活性的作用与姜黄素相似,而SB202580对MMP-9无明显影响。结论姜黄素可能通过TGF-β/Smad、TGF-β/ERK信号通路下调TGF-β1诱导的MMP-9的表达及其活性,从而抑制肿瘤的侵袭能力。 相似文献
7.
目的探讨人骨形态发生蛋白9(Human bone morphogenetic protein 9,hBMP9)对人骨肉瘤细胞MG63和U2OS的抑制作用及其机制。方法用重组腺病毒AdBMP9分别感染MG63和U2OS细胞,并设空白对照组(不加任何处理因素)和AdGFP感染对照组,免疫细胞化学法(Immunocytochemistry,ICC)和Western blot法检测感染后两种细胞中hBMP9的表达水平;MTT和台盼蓝拒染活细胞计数法检测细胞的增殖活力;Hoechst/PI荧光双染法检测细胞的凋亡情况;划痕愈合试验检测细胞的迁移能力;ICC法检测Wnt/β-catenin信号途径中β-catenin的表达。结果AdBMP9感染的两种细胞中hBMP9的表达水平均明显高于空白对照组和AdGFP感染组(P<0.05);hBMP9表达的上调可抑制MG63和U2OS细胞的增殖,且呈时间依赖性(P<0.01),并使两种细胞的凋亡率明显增加(P<0.01),迁移能力明显下降(P<0.01),β-catenin的表达量明显减少(P<0.01)。结论 hBMP9可能通过下调Wnt/β-catenin信号途径活性,抑制骨肉瘤细胞的增殖、迁移,并促进其凋亡。 相似文献
8.
《中国生物制品学杂志》2013,(9)
目的探讨骨形态发生蛋白9(Bone morphogenetic proteins 9,BMP9)对食管鳞状细胞癌细胞增殖、克隆、迁移、侵袭及细胞周期的影响。方法用AdGFP和AdBMP9分别感染3株食管鳞状细胞癌细胞株ECA109、KYSE150和KYSE180,并设不感染病毒的3种细胞作为对照,采用RT-PCR法检测AdBMP9感染的3株细胞中BMP9基因mRNA的转录水平,MTT法检测病毒感染细胞096 h各组细胞的增殖活力,平板克隆法检测各组细胞的克隆形成能力,划痕试验检测各组细胞的迁移能力,Transwell小室试验检测各组细胞的侵袭能力,并对3组ECA109细胞进行细胞周期的检测。结果 AdBMP9感染的3株细胞中BMP9基因mRNA的转录水平均明显提高。与AdGFP感染和未感染病毒的细胞相比,3种细胞在过表达BMP9后,增殖活力明显降低(P<0.05);细胞克隆形成能力均下降(下降率达70%以上),且克隆细胞团明显变小;细胞迁移和侵袭能力均下降;AdBMP9感染的ECA109细胞处于G1期的细胞比例明显上升,S期比例明显下降(P<0.05)。结论 BMP9可明显抑制食管鳞癌细胞的增殖、克隆、迁移和侵袭能力,并将细胞周期阻滞于G1期。 相似文献
9.
目的探讨cAMP-PKA-CREB信号通路在骨形态发生蛋白9(bone morphogenetic protein 9,BMP9)诱导小鼠间充质干细胞(mesenchymal stem cells,MSCs)C3H10T1/2成骨分化过程中的作用及其机制。方法将C3H10T1/2细胞分别加入不同浓度的cAMP-PKA-CREB信号通路抑制剂H89(1、2.5、5和10μmol/L),检测其对碱性磷酸酶(alkaline phosphatase,ALP)活性的影响;通过ALP定量和钙盐沉积试验分别检测H89对BMP9诱导C3H10T1/2细胞早期和晚期成骨分化的影响;经Western blot法检测H89对C3H10T1/2细胞中磷酸化CREB、骨钙素(Osteocalcin,OCN)和成骨关键转录因子Runx2表达水平的影响;通过Wentern blot及荧光素酶活性的检测,观察H89对经典信号通路BMPs-smad1/5/8的影响。结果随着H89浓度的增加,对BMP9诱导的C3H10T1/2细胞ALP的抑制作用明显增强(P0.05),且呈剂量依赖性;ALP定量和钙盐沉积试验结果表明,H89可明显抑制BMP9诱导的C3H10T1/2细胞早期及晚期成骨分化;H89可显著抑制BMP9诱导的C3H10T1/2细胞中磷酸化CREB、OCN及Runx2蛋白的表达(P0.05),与AdBMP9组比较,H89对经典BMPs-smad1/5/8信号通路无明显影响(P0.05)。结论阻断cAMP-PKA-CREB信号通路可抑制BMP9诱导的MSCs C3H10T1/2的成骨分化,为BMP9的临床应用奠定了理论基础。 相似文献
10.
目的探讨慢病毒介导的人乳头瘤病毒(Human papilloma virus,HPV)16型E6基因shRNA在体内对荷瘤裸鼠宫颈癌生长的抑制作用。方法将BALB/c-nu裸鼠随机分为空白对照组、干扰组和无义干扰组,经皮下接种宫颈癌Caski细胞(2×106个),移植瘤直径达0.5 cm时,空白对照组于瘤体局部注射PBS,干扰组和无义干扰组分别于瘤体局部注射靶向HPV16型E6基因的shRNA-PLL3.7干扰慢病毒和无义干扰慢病毒(3×108TU/ml),2周后检测瘤体大小和重量的变化,采用免疫组化法检测肿瘤组织中HPV16型E6、P53和P21蛋白的表达。结果与无义干扰组和空白对照组比较,干扰组裸鼠体内肿瘤明显缩小,瘤体重量明显降低(P<0.000 1),无义干扰组与空白对照组比较,差异无统计学意义(P>0.05);干扰组肿瘤组织中HPV16型E6蛋白的表达被抑制,P53和P21蛋白的表达水平明显升高。结论慢病毒介导的HPV16型E6基因shRNA能有效抑制动物体内宫颈癌的生长,具有潜在的开发应用前景。 相似文献
11.
The knowledge of human bone viscoelasticity is an important issue for defining semirigid calcified tissues implants. A very sensitive technique was used to investigate bone viscoelasticity: the thermally stimulated creep method. A study of demineralized human bone was performed to determine the molecular origin of bone viscoelasticity. The thermally stimulated creep spectra of bone and demineralized bone, at the hydrated state, present a similar shape with one main retardation mode located at −133 and −120°C, respectively. This mode is shifted toward higher temperatures after dehydration, revealing the existence of another mode at around −155°C. The analysis of elementary spectra of bone and demineralized bone has shown that retardation times follow an Arrhenius equation, and that two compensation phenomena are observed with comparable compensation parameters. The first compensation phenomenon, which corresponds to the main retardation mode, was attributed to motions of water molecules located inside the collagen triple helix. The second compensation phenomenon, which reveals the existence of another relaxation mode at higher temperatures, was assigned to movements of hydrophilic side chains bound to water molecules. As for the mode observed at around −155°C, it was associated with motions of aliphatic side chains. Overall, bone viscoelasticity originates from the organic matrix. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 79: 2527–2533, 2001 相似文献
12.
Porous conical implants made by sintering micro-spheres made from bioactive and bio-compatible glasses have been tested mechanically as well as in vivo by inserting the implant through the cortical bone into the bone marrow. The behaviour is compared to a reference implant made by sintering micro-spheres of metallic titanium. Due to capillary forces the implant pores were filled with bone marrow fluid when inserted. An extended bone ingrowth occurred in the cones of bioactive glass, in the titanium ones only in the cortical area. Further, the factors influencing the formation of a chemical bond between glass and living tissue are discussed. 相似文献
13.
《Ceramics International》2016,42(13):14910-14917
The synthetic bone powder was studied as a raw materials for bone china, completely replacing natural bone ash raw materials. The physical and thermal properties of samples obtained by the two bone powders were tested and comparatively studied. Performance tests included pyroplastic deformation, flexural strength, bulk density, sintering shrinkage, water absorption, transmittance, thermal expansion coefficient and the thermal shock resistance. The phase composition and morphology were investigated by X-ray diffraction (XRD) and scanning electron microscopy (SEM), respectively. The results indicated that using synthetic bone powder could shorten the preparation time, reduce the sintering temperature and result in high-quality bone china. The pyroplastic deformation decreased from 9.14% to 7.92%, the three-point flexural strength increased from 123 MPa to 191 MPa, the light transmittance (at a 2-mm thickness) increased from 6.7% to 11.2%, the thermal expansion coefficient decreased from 8.24×10−6 °C−1 to 7.69×10−6 °C−1, and the thermal shock resistance increased from 140 °C to 180 °C. A continuous interface layer without cracks was produced by using the synthetic bone powder. 相似文献
14.
《应用陶瓷进展》2013,112(3):110-116
AbstractCommercial bone china microstructures are far from equilibrium consisting of distinct regions of lath-like (0·4–10 μm) anorthite and spheroidal (1–3 μm) β-tricalcium phosphate (β-TCP) nodules embedded in a heterogeneous composition glass along with isolated irregular (≤30 μm) α-quartz crystals. The composition and morphology of the phases formed on firing suggest that anorthite crystallised in clay relicts by the reaction of metakaolin with CaO, β-TCP crystallised from decomposition of bone ash, and that the liquid formed on vitrification has variable composition depending on the composition of adjacent phases. P2 O5was never detected in the glass suggesting that any P2 O5 that dissolves in the liquid on firing is transient. The cracks sometimes observed in and around β-TCP clusters arise from thermal expansion mismatch between β-TCP and the surrounding glass and anorthite. 相似文献
15.
《Ceramics International》2023,49(5):7888-7904
Monetite (MC) is a type of calcium phosphate cement (CPC). It has various morphologies, continuous degradation and absorption properties; however, its high porosity will affect its mechanical properties. There is minimal research on MC is immature. Amorphous calcium phosphate (ACP) is widely used and exhibits good biocompatibility; however, it is unstable. In this study, MC was combined with ACP at different weight ratios to form new types of bone cements. The mechanical properties, biocompatibility, and inductive ability for osteogenesis and osteoclasts of different MC/ACP composite bone cements were evaluated. The biocompatibility and effects on osteogenic and osteoclast differentiation of MC/ACP composite bone cements were investigated in vitro using mouse bone marrow mesenchymal stem cells (mBMMSCs) and mouse monocyte/macrophage cell line RAW 264.7. RAW 264.7 cells can differentiate into osteoclasts by osteoclast differentiation. The results indicated that the overall performance of the MC/ACP composite bone cement was better than that of the MC or ACP alone. Compared to the other groups, the biocompatibility of MC75 (75 wt% MC and 25 wt% ACP) was optimal; it was able to induce mBMMSCs osteogenesis to a greater extent. MC75 was the least favorable for the proliferation, migration, and differentiation of osteoclasts. The mechanical properties, setting time and injectability of MC75 meet clinical application requirements. This study demonstrated that MC75 is a promising bone cement for repairing bone defects. 相似文献
16.
《Ceramics International》2020,46(10):16465-16471
Pore size and connectivity of artificial bone scaffolds play key role in regulating cell ingrowth and vascularization during healing. The objective of this study was to develop a novel process for preparing 3D pore-interconnected open-cell bone substitutes with varying pore sizes. This was achieved by thermal-induced expansion, drying, then sintering the mixture of biphasic calcium phosphate (BCP) and a thermal responsive porogen comprising chitosan (CS) and hydroxypropyl methyl cellulose (HPMC). The interpolymer complexes (IPCs) of CS/HPMC were prepared and investigated by FT-IR. The mixtures of IPCs/BCP were heated up to 100 °C for analyzing their thermal expansion properties. This resulted in ~13% and ~42% volume increment for IPC-1/BCP and IPC-2/BCP, respectively, while ~230% volume increased in the case of IPC-3/BCP (therefore chosen for sintering bone blocks). Heating rate-dependent (0.20–0.25 °C/min range) sintering profiles for IPC-3/BCP were utilized to produce BCP bone blocks. Gasification of IPC during sintering resulted in the formation of interconnected porous structures, and the morphology was investigated by SEM, revealing varying sizes ranging from 106 ± 13 μm to 1123 ± 75 μm. The pore size range of bone blocks from 235 ± 46 μm to 459 ± 76 μm portrayed significantly high MC3T3-E1 cell viability with prominent filopodial extensions, and elongated cells, depicting efficient biocompatibility. Therefore, the process for preparing porous interconnected 3D bone blocks were feasible, thereby serving as an alternative for potential bone tissue engineering applications. 相似文献
17.
《Ceramics International》2023,49(12):19578-19594
Bone defects have attracted much attention for a long time and seriously affect the function of the motor system. At present, the application of biological materials and biological scaffolds implanted in the defect site to promote the healing of bone defects is the main treatment method for bone defect repair. In recent years, the emergence of two-dimensional materials has brought new opportunities for biological materials. As a two-dimensional nanomaterial based on ceramics, MXene has unique physical and chemical properties, such as electrical conductivity, hydrophilicity, and antibacterial and photothermal effects, which make it a very broad application prospect in bone defect biomaterials. This review will start from the pathophysiological changes of bone defects and intervention factors of bone defect repair, introduce in detail the preparation and modification methods, physical and chemical properties and biological characteristics of the two-dimensional material MXene, and review the application status and research progress of MXene in bone defect repair and bone tissue regeneration. This provides a reference for the further application of MXene in bone defect repair. 相似文献
18.
Wu CC Li XB Han TS Li P Liu GW Wang WZ Wang Z 《International journal of molecular sciences》2012,13(3):3431-3443
The objective of our study was to evaluate whether feeding pseudopurpurin affects bone mineral density and bone geometry architecture in rats. Pseudopurpurin was extracted, analyzed and purified using an HLPC-ESI-MS. Rats were given 0% and 0.5% pseudopurpurin powder in their diet. Femurs of rats were examined at 0.5, 1 and 2 months after pseudopurpurin feeding. Compared with rats in the group 0%, the bone mineral density, and the calcium, magnesium, zinc and manganese concentrations in the rats femur in the group 0.5% increased significantly at 1 month and 2 months after pseudopurpurin feeding. Analytical results of micro-computed tomography showed that the group 0.5% displayed an increase in the trabecular volume fraction, trabecular thickness and trabecular number of the distal femur at 1 and 2 months after pseudopurpurin feeding, and the mean thickness, inner perimeter, outer perimeter, and area of the femur diaphysis were significantly increased at 2 months after pseudopurpurin feeding compared with the group 0%. In parallel, the trabecular separation and structure model index of the distal femur were decreased, compared with the group 0% at 1 and 2 months after pseudopurpurin feeding. In the 0.5% and 0% groups, there was no damage to kidney and liver by histopathology analysis. The long-term feeding of pseudopurpurin is safe for rats. The feeding of 0.5% pseudopurpurin which has specific chemical affinities for calcium for bone improvement and level of bone mineral density, enhances the geometry architecture compared with the 0% group. 相似文献
19.
《Ceramics International》2020,46(1):353-364
Bone grafting materials for repair of alveolar bone deficits have improved markedly in recent years, increasing the applicability and success of oral implantology. The long-term success rate of dental implant surgery is strongly dependent on the quality and stability of residual bone tissue. Therefore, reconstruction of resorbed alveolar bone is a challenge for clinicians. In the present study, we have developed bioactive bone regeneration particles (BRPs) using amorphous calcium phosphate and 58S bioglass as raw materials. The structural characteristics, biocompatibility, and osteoinductivity of these BRPs were compared to commercially available bovine spongy bone (BSB) without organic components. X-ray diffractometry (XRD) and scanning electron microscopy (SEM) showed that BRPs were composed of β-tricalcium phosphate (β-TCP) and calcium silicate in the form of hexagonal crystals, while BSB was mainly hydroxyapatite (HA) arranged in orderly nano-sized crystals. The viability of human bone marrow mesenchymal stem cells (hBMSCs) cultured in BRP-containing medium was roughly equal to that of hBMSCs in control medium. Moreover, hBMSCs in BRP medium exhibited greater proliferation rates, substrate attachment, alkaline phosphatase (ALP) activity, alizarin red staining intensity, and expression levels of osteogenic-related genes (COL-I, OCN, Runx-2, ALP, BSP) than hBMSCs in BSB medium, indicating the superior osteoinductivity of BRPs. Silicon ions released from BRPs during cell culture were crucial for these enhanced biological properties. BRPs also demonstrated superior osteoconduction and osteoinduction properties for bone defect repair, suggesting promise for alveolar bone repair surgery. 相似文献