Assessment of tobacco heating system 2.4 on osteogenic differentiation of mesenchymal stem cells and primary human osteoblasts compared to conventional cigarettes

BACKGROUND Cigarette smoking(CS)is the most common method of consuming tobacco.Deleterious effects on bone integrity,increased incidence of fractures,and delayed fracture healing are all associated with CS.Over 150 of the 6500 molecular species contained in cigarette smoke and identified as toxic compounds are inhaled by CS and,via the bloodstream,reach the skeletal system.New technologies designed to develop a reduced-risk alternative for smokers are based on electronic nicotine delivery systems,such as e-cigarettes and tobacco heating systems(THS).THS are designed to heat tobacco instead of burning it,thereby reducing the levels of harmful toxic compounds released.AIM To examine the effects of THS on osteoprogenitor cell viability and function compared to conventional CS.METHODS Human immortalized mesenchymal stem cells(n=3)and primary human preosteoblasts isolated from cancellous bone samples from BG Unfall Klinik Tübingen(n=5)were osteogenically differentiated in vitro with aqueous extracts generated from either the THS 2.4“IQOS”or conventional“Marlboro”cigarettes for up to 21 d.Cell viability was analyzed using resazurin conversion assay(mitochondrial activity)and calcein-AM staining(esterase activity).Osteogenic differentiation and bone cell function were evaluated using alkaline phosphatase(AP)activity,while matrix formation was analyzed through alizarin red staining.Primary cilia structure was examined by acetylatedα-tubulin immunofluorescent staining.Free radical production was evaluated with 2’,7’-dichlorofluoresceindiacetate assay.RESULTS Our data clearly show that THS is significantly less toxic to bone cells than CS when analyzed by mitochondrial and esterase activity(P<0.001).No significant differences in cytotoxicity between the diverse flavors of THS were observed.Harmful effects from THS on bone cell function were observed only at very high,non-physiological concentrations.In contrast,extracts from conventional cigarettes significantly reduced the AP activity(by two-fold)and matrix mineralization(four-fold)at low concentrations.Additionally,morphologic analysis of primary cilia revealed no significant changes in the length of the organelle involved in osteogenesis of osteoprogenitor cells,nor in the number of ciliated cells following THS treatment.Assessment of free radical production demonstrated that THS induced significantly less oxidative stress than conventional CS in osteoprogenitor cells.CONCLUSIONTHS was significantly less harmful to osteoprogenitor cells during osteogenesisthan conventional CS. Additional studies are required to confirm whether THS isa better alternative for smokers to improve delays in bone healing followingfracture.

EFFECTS OF APATITE CERAMICS ON CELL GROWTHS AND DNA SYNTHESES IN HUMAN OSTEOBLAST LIKE CELL

Sintered strontium hydroxyapatite (Sr-HAP) and hydroxapatite (HAP) werekinds or apatite ceramics as substitutes of bone tissues.They should be safety to human body.Sr-HAP and HAP were co-cultured with human osteoblast like cells for 2, 4,and 6 days invitro. Effect of Sr-HAP and HAP on cell growth numbers with thymidine incorporationtest. In order to estimated cytotoxicities of materials,relative cell growth rate (RGR) werecalculated from data and score of cytotoxicties were assaied. Results showed cell growth numbers and rate of 3H-thymidine incorporation in Sr-HAP and HAP groups were the same asthat of blank control group. Values were increased with the increase of co-cultured time.There were no inhibition of cell growth and DNA syntheses in human osteoblast like cells. Accoding to RGR and score of cytotoxicity degree,all values were qurlified with sdandard ofbiomaterials. Results suggested these qurlified with sdandard or biomaterials. Results suggested these apatite ceramics had no obvious cytotoxicities.

Human adipose derived stem cells are superior to human osteoblasts (HOB) in bone tissue engineering on a collagen-fibroin-ELR blend

The ultrastructure of the bone provides a unique mechanical strength against compressive, torsional andtensional stresses. An elastin-like recombinamer (ELR) with a nucleation sequence for hydroxyapatitewas incorporated into films prepared from a collagen-silk fibroin blend carrying microchannel patternsto stimulate anisotropic osteogenesis. SEM and fluorescence microscopy showed the alignment ofadipose-derived stem cells (ADSCs) and the human osteoblasts (HOBs) on the ridges and in the groovesof microchannel patterned collagen-fibroin-ELR blend films. The Young's modulus and the ultimatetensile strength (UTS) of untreated films were 0.58 ± 0.13 MPa and 0.18 ± 0.05 MPa, respectively. After 28days of cell culture, ADSC seeded film had a Young's modulus of 1.21 ± 0.42 MPa and UTS of0.32 ± 0.15 MPa which were about 3 fold higher than HOB seeded films. The difference in Young'smodulus was statistically significant (p: 0.02). ADSCs attached, proliferated and mineralized better thanthe HOBs. In the light of these results, ADSCs served as a better cell source than HOBs for bone tissueengineering of collagen-fibroin-ELR based constructs used in this study. We have thus shown theenhancement in the tensile mechanical properties of the bone tissue engineered scaffolds by usingADSCs.

EFFECTS OF PARATHYROID HORMONE AND ESTRADIOL ON PROLIFERATION AND FUNCTION OF HUMAN OSTEOBLASTS FROM FETAL LONG BONE AN IN VITRO STUDY

The effect of parathyroid hormone (PTH) (0.01 nM-10 nM) and 17 -estradiol (E2, 1 nmol-10 nM) alone or in combination on 3H thymidine incorporation, alkaline phosphatase and adenylate cyclase activities were investigated in human fetal osteoblasts using serum-free monolayer primary cultures. The results showed that PTH inhibited cell proliferation while E, promoted it. On alkaline phosphatase activity, PTH showed a complex results while E, were slightly inhibitory. PHT-E2 combination suggested that E2 could alter the effect of PTH alone, also potentiated the anabolic and antagonize the catabolic effects of PTH on bone formation.

重组人乳铁蛋白通过促进成骨细胞增殖加速大鼠骨折愈合

目的研究重组人乳铁蛋白(rhLF)对模型大鼠股骨骨折愈合及大鼠原代成骨细胞增殖的影响。方法将雄性大鼠随机分为3组:单纯骨折组(SF组)、胶原膜处理组(CF组)和rhLF复合胶原膜处理组(CLF组)。通过X线胶片及HE和Masson染色组织学观察比较骨折愈合情况。培养的24 h新生SD大鼠原代成骨细胞以不同浓度rhLF进行处理,MTT法检测成骨细胞增殖。结果CLF组在术后1、2、3周时骨痂较SF组及CF组体积增加更明显(P<0.05),4周时组间对比差异无统计学意义。CLF组软骨痂和硬骨痂增殖更明显,并率先出现成熟骨小梁和板层骨,骨痂成熟度高。与对照组比较,第1天100 mg/L和500 mg/L rhLF组对体外培养的大鼠原代成骨细胞,吸光度(A)值有明显降低(P<0.05)。第5天及第10天时随rhLF浓度增加,A值明显升高(P<0.05)。结论在体内rhLF具有加速骨折愈合作用,表现为骨痂形成快、体积大、成熟度高。它也可以促进体外培养的成骨细胞增殖,主要表现在成骨细胞快速增殖阶段。

人诱导多能干细胞成骨分化的研究进展

骨骼疾病如骨质疏松、骨关节炎等已成为亟待解决的人类健康问题,细胞治疗及组织工程技术被认为是理想的治疗方法之一。人诱导多能干细胞(human induced pluripotent stem cells,hiPSCs)具备体外长期自我更新和分化所有三胚层来源体细胞的独特功能,已成为目前最有前景的成骨细胞来源。因此,需要构建成分明确的hiPSCs体外成骨向诱导分化体系,获得符合临床应用要求的成骨样细胞。许多团队在促进hiPSCs向成骨分化成熟的直接路径和经间充质干细胞的间接路径方面取得了实质性的进展,本文针对这两类成骨分化路径及其应用现状进行综述,以期为骨再生技术提供参考。现有研究借助拟胚体法和单层诱导法,基于生物材料,构建可支持hiPSCs体外培养和成骨向诱导分化体系。然而,目前的研究主要存在成分不明确,分化效率低等局限,基于特定化合物严格调控的分阶段式和三维定向诱导体系是未来的主要研究方向。

成骨诱导人牙周膜干细胞来源外泌体促进炎症微环境下人牙周膜干细胞成骨分化

背景:成骨诱导间充质干细胞来源外泌体具有较强的成骨分化能力,但是在炎症微环境下对人牙周膜干细胞成骨分化的影响尚不明确。目的:探究成骨诱导人牙周膜干细胞来源外泌体在炎症微环境下对人牙周膜干细胞成骨分化的影响。方法:收集离体牙并分离培养人牙周膜干细胞,成骨诱导3 d后提取外泌体。将人牙周膜干细胞分为4组:对照组加入成骨诱导培养基,外泌体组加入含5μg/mL外泌体的成骨诱导培养基,炎症模型和炎症模型+外泌体组以1μg/mL脂多糖处理24 h构建细胞炎症微环境,炎症模型组在脂多糖处理后加入成骨诱导培养基,炎症模型+外泌体组在脂多糖处理后加入含5μg/mL外泌体的成骨诱导培养基。通过茜素红以及碱性磷酸酶染色法检测各组人牙周膜干细胞的成骨分化能力;实时荧光定量PCR与免疫印迹法检测各组人牙周膜干细胞中Runt相关转录因子2、骨桥蛋白、成骨细胞特异性转录因子Osterix(OSX)和wnt通路相关蛋白β-catenin的表达。结果与结论:(1)与对照组相比,炎症模型组碱性磷酸酶染色相对面积、矿化结节染色相对面积以及Runt相关转录因子2、骨桥蛋白、OSX的表达量显著降低(P <0.05);(2)与炎症模型组相比,炎症模型+外泌体组碱性磷酸酶染色相对面积、矿化结节染色相对面积以及Runt相关转录因子2、骨桥蛋白、OSX的表达显著升高(P <0.05);(3)与对照组相比,炎症模型组wnt通路相关蛋白β-catenin表达量显著增加(P <0.05);与炎症模型组相比,炎症模型+外泌体组β-catenin表达量显著降低(P <0.05)。结果表明,成骨诱导人牙周膜干细胞来源外泌体可促进炎症微环境下人牙周膜干细胞的成骨分化,其作用机制可能与wnt/β-catenin信号通路有关。

Study of Tissue Engineered Dental Implants of human alveolar Bone-Evaluations on their osteogenic activities invitro

The development of the activated cellular bony implant, in light of the principle on tissue engineering, has brought about a new era to the fields of dental maxillofacial implantation. The present study separated the osteoblast like cells from human alveolar bone and seeded them into 3 types of biodegradable scaffold to form the complexes and then evaluated their osteogenic activities in vitro, in order to acquire experimental data that are essential to future clinical practice of this new type of therapeutical procedure in oral and maxillofacial surgery. Material and methods: Human alveolar bone origin cells were separated from alveolar bone around the third impacted teeth of 3 patients by enzyme digestion and went on cultures with α MEM containing β glycerophosphate and Dexamethasone at 5% CO2 ,37℃ for 21 28 days. Confirmed osteoblasts like cells were then seeded onto 3 types of degradable biomaterials of polyglycolic acid scaffold, collagen sponge, and L lactic acid/ε caprolactone to form cell matrices complexes. The 3 types of complex were continued to culture for 21 28 days in vitro at the same conditions with the single layer cultured cells. The cell proliferation, morphological changes, ALPase activity and mineral nodules formation on scaffolds were measured and observed at 3 days intervals to evaluate the affinities & the osteogenic activities of the human alveolar osteoblast like cells in the 3 different complexes. Result and discussion: The results indicated that the cultured human alveolar bone origin cells from 3 patients could successfully express the osteoblasts phenotype in single layered culturing in vitro after stimulated by β glycerophosphate and Dexamethasone. It has been shown that the cultured osteoblast like cells seeded on PGAS matrix had the highest attachmental, proliferative and osteogenic activities, suggesting a good bio affinity between the human alveolar osteoblast like cells and the PGAS matrix. The statistical analysis (ANOVA) showed that there were significant differences between PGAS osteoblasts complex and CLGS or LACT complexes on osteogenic activities. (P<0.05). It was also noticed that cultured human alveolar osteoblasts seeded in biodegradable materials had a delayed peak period on cell proliferation and PLAase production ,suggesting the osteoblasts seeded on scaffolds need a period of time to adjust themselves before they can normally proliferate and expres their phenotypes. Conclusion: PGAS osteoblasts complex is worth to be further developed into a tissue engineered cellular artificial bony implant for reconstructing the oral maxillofacial bony defects in a more effective way in the future.

人胎儿成骨细胞系hFOB1.19在骨组织工程中的应用

背景:现阶段广泛使用的动物成骨细胞模型的生物学特性与人细胞差距较大;原代人成骨细胞分离培养难提纯、培养代次有限;骨肉瘤细胞有异常生长的风险,都不能完全满足研究需求。1995年,有研究利用猴空泡病毒40转染了人胎儿四肢细胞,筛选得到一组具有分化为较早期成骨祖细胞——永生化人成骨细胞系hFOB1.19。目的:综述hFOB1.19的基本特性、培养方法及作为骨组织工程材料细胞模型的应用。方法:在中国知网、万方数据、SinoMed、PubMed、Web of Science、Medicine及Cochrane Library数据库中查找从1995年至2022年5月的相关文献,中文检索词为“人胎儿成骨细胞系1.19、人成骨细胞系、永生化细胞、猴空泡病毒40、骨组织工程、骨替代材料、生物材料研究”,英文检索词为“hFOB1.19,Human osteoblast cell line,Immortalized cells,Monkey vacuolating virus 40,Bone tissue engineering,Bone substitute materials,Biomaterials research”,筛选后得到73篇文献进行综述。结果与结论:(1)hFOB1.19在形态、表型、核型、生物学特性及分化潜能方面与体内成骨细胞高度相似,拥有快速且稳定的生长能力。(2)hFOB1.19已广泛应用于细胞黏附、增殖、分化、矿化、基因表达及蛋白合成方面,以检验骨组织工程材料的生物相容性及成骨性能。(3)hFOB1.19能够弥补动物细胞来源不同、骨肉瘤细胞异常增殖可能、原代成骨细胞分离难及传代次数少的缺点,在骨组织工程实验应用中性能稳定,增殖良好,但因培养条件特殊,细胞传代次数有限及生物安全性等方面的问题,目前在骨组织工程材料领域应用尚且不够广泛。(4)当前,骨组织工程材料发现速度快,但开发过程中缺乏系统全面的研究体系及统一的研究标准,文章总结hFOB1.19在不同研究中的应用,旨在为研究者们提供方法选择引导并推动相关标准完善。

纳秒脉冲电场刺激促进人真皮成纤维细胞成骨的实验研究

目的 探究超短纳秒脉冲电场(Nanosecond Pulsed Electric Field,nsPEF)是否能够促进成纤维细胞向成骨细胞的表型转化。方法 用0、5 kV/cm的场强刺激成纤维细胞,观察其在普通培养基和三向分化诱导培养基中生长情况,蛋白印迹测定细胞内调控生长及分化Hippo通路下游信号分子水平,并通过体内注射细胞-凝胶复合体进行裸鼠体内成骨实验。结果 5kV/cm nsPEF促进了成纤维细胞的增殖,并促进了成骨、成软骨分化,但对成脂分化影响不显著。低能量脉冲电场刺激抑制了Hippo通路下游各信号分子磷酸化水平,促进了裸鼠体内成骨,灰度分析显示,YAP1及LATS1的磷酸化蛋白与非磷酸化蛋白比值在5 kV/cm的脉冲电场刺激后明显降低,提示HIPPO信号通路受到抑制(P<0.05)。结论 低强度nsPEF刺激有效促进了成纤维细胞的增殖和表型转化,并在裸鼠模型上证实了可行性和有效性。

阿伦磷酸钠对人成骨细胞增殖的影响

目的 :改良成人成骨细胞消化培养方法 ,研究阿伦磷酸钠对成人成骨细胞增殖的影响。方法 :手术中获取病人的髂骨块 ,酶消化法得到成骨细胞 ,传 4代后以 1× 10 5/ml的浓度种于 2块 96孔板中。MTT方法检测阿伦酸钠对成骨细胞增殖的影响。结果 :高于 10 4M浓度的阿伦磷酸钠明显抑制成骨细胞的增殖 ,浓度低于 10 5M时 ,成骨细胞的增殖不受影响。结论

人成骨细胞体外培养、鉴定及护骨素表达的研究

目的 用酶消化法从正常人松质骨中分离成骨细胞 ,进行体外培养和功能鉴定 ,观察用 1 7β 雌二醇 (1 7β E2 )和雌激素受体拮抗剂ICI1 82 780处理后 ,成骨细胞中护骨素 (OPG)的表达变化。方法 用胰酶 -胶原酶消化法从正常人松质骨中分离培养成骨细胞 ,I型胶原染色用VanGie son法 ,钙化结节用茜素红染色 ,用钙钴法显示成骨细胞中碱性磷酸酶的表达 ,骨钙素和OPG基因的表达用RT -PCR检测 ,OPG蛋白用WesternBlot检测。结果 用酶消化法分离的人成骨细胞 ,在体外培养时可维持合成I型胶原、碱性磷酸酶、骨钙素 ,并形成钙化结节等功能 ;体外培养的人成骨细胞表达OPG ,1 7β E2 上调其表达 ,并能被雌激素受体拮抗剂ICI1 82 780阻断。结论 用酶消化法进行人成骨细胞培养 ,方便易行 ,可培养大量高纯度人成骨细胞供研究使用 ;1 7β E2 上调OPG表达 ,而绝经后雌激素缺乏时 ,OPG的表达相应减少 ,可能是骨质疏松的发病因素之一。

体外培养人成骨细胞骨相关基因表达

目的 探讨体外培养人成骨细胞骨相关基因表达情况。方法 应用组织块移行生长法分离培养人成骨细胞 ,活体观察与碱性磷酸酶染色观察其生物学特性 ,采用RT PCR方法检测培养 14d的二代人成骨细胞基因COL1,BGP ,OPG ,ON ,IGF 1,TGF β的表达情况 ,并用GAPDH予以矫正。结果 相差倒置显微镜观察可见细胞呈三角形、四角形或不规则形 ,5 0 %～ 90 %的细胞碱性磷酸酶呈阳性 ,培养 14d的二代人成骨细胞检测到基因COL1,BGP ,OPG ,ON ,IGF 1,TGF β的mRNA表达。结论 体外培养人成骨细胞可表达COL1,BGP ,OPG ,ON ,IGF 1,TGF

铁调素对成骨细胞骨相关基因表达的影响

目的研究铁调素对人成骨细胞(hFOB1.19)Ⅰ型胶原(COL1)、骨保护素(OPG)、骨钙素(BGP)基因表达的影响。方法成骨细胞在34℃下进行体外培养,以不同浓度铁调素(50 nmol/L、100 nmol/L、200 nmol/L、400 nmol/L)干预72 h后,收集细胞,分别抽取总RNA,采用半定量RT-PCR法检测成骨细胞中COL1、OPG、BGP mRNA表达的变化。结果 RT-PCR检测显示不同浓度铁调素干预后,各组COL1、OPG、BGP mRNA均有表达;不同浓度组的COL1、OPG、BGP mRNA表达光密度比值不同,组间密度比值比较存在统计学意义(P<0.05),但铁调素在400 nmol/L时抑制OPG的表达(P<0.05)。结论 铁调素可上调成骨细胞(hFOB1.19)COL1、OPG、BGP mRNA表达,铁调素浓度增加转录水平逐渐增加,显示有浓度依赖性。

补肾中药骨灵片对人成骨细胞功能和p38活化原蛋白激酶通路的影响

目的探讨含补肾中药骨灵片大鼠血清对人成骨细胞功能和对p38活化原蛋白激酶通路的影响。方法取人髂骨松质骨,采用改良胰酶-胶原酶消化法分离培养人成骨细胞;根据血清药理学的方法制备不同浓度(低、中、高剂量)的骨灵片血清,并以生理盐水处理的大鼠血清为阴性对照,将制备好的血清加入第二代的人成骨细胞培养液中培养。采用Western-blot及Real time RT-PCR的方法观察各组成骨细胞护骨素和护骨素配体基因表达和磷酸化p38含量,用MTT法测定各组成骨细胞增殖率,利用茜素红染色法测定矿化结节形成的数量。结果含骨灵片(低、中、高剂量)药液血清处理的大鼠血清均能上调成骨细胞护骨素基因表达,下调护骨素配体基因表达,刺激成骨细胞的增殖,增加矿化结节形成的数量,刺激p38磷酸化。结论补肾中药骨灵片可能通过p38活化原蛋白激酶通路调控护骨素以及护骨素配体的表达,促进成骨细胞增殖以及矿化结节形成,抑制骨破坏,维持骨平衡。

生长因子网络调节对骨形成作用的研究Ⅴ.不同生长因子交互应用对人胚成骨样细胞增殖及分化的影响

目的：考察将转化生长因子β（ＴＧＦ－β）、胰岛素样生长因子Ⅱ（ＩＧＦ－Ⅱ）、碱性成纤维细胞生长因子（ｂＦＧＦ）及血小板衍生性生长因子（ＰＤＧＦ）４种生长因子两两交互应用对人胚成骨样细胞增殖与分化的影响。方法：在不同的实验间期检测成骨样细胞３Ｈ－胸腺嘧啶脱氧核苷、３Ｈ－脯氨酸的掺入量和碱性磷酸酶的含量。结果：ＰＤＧＦ、ｂＦＧＦ、ＩＧＦ－Ⅱ和ＴＧＦ－β４种生长因子交互联合应用，均对促进体外培养的人胚成骨样细胞的增殖和分化功能具有协同作用。结论：具有协同作用的生长因子联合应用，可提高其促进骨形成的生物效应。

钙离子对人成骨细胞迁移与成骨分化的影响

目的研究不同浓度Ca^(2+)对人成骨细胞迁移与成骨分化的影响,探讨促进迁移与成骨合适的Ca^(2+)浓度及相关机制。方法设置Ca^(2+)浓度,Transwell检测成骨细胞迁移;CCK-8法评估成骨细胞增殖;逆转录聚合酶链反应(RTPCR)检测细胞成骨分化相关基因的表达;茜素红染色检测成骨分化生成的矿化结节。钙敏感受体(CaSR)拮抗剂拮抗后,观察Ca^(2+)对人成骨细胞迁移与成骨分化的影响。结果在迁移实验中,2、4、6 mmol·L^(-1)的Ca^(2+)在3个时间点(8、16、24 h)都能明显地促进人成骨细胞的迁移,10 mmol·L^(-1)的Ca^(2+)在8 h时明显抑制迁移。2～10 mmol·L^(-1)Ca^(2+)能促进人成骨细胞的增殖、成骨分化与矿化,8、10 mmol·L^(-1)的Ca^(2+)诱导的矿化作用更明显。Ca SR拮抗降低Ca^(2+)诱导的人成骨细胞迁移与成骨分化作用。结论低浓度Ca^(2+)有利于人成骨细胞迁移,高浓度Ca^(2+)有利于人成骨细胞分化,4、6 mmol·L^(-1)的Ca^(2+)能较明显地同时诱导人成骨细胞迁移与成骨分化,Ca^(2+)-Ca SR通路参与相应的信号传导。

成骨细胞特异性识别多肽对人成骨细胞增殖和矿化影响的实验研究

目的观察成骨细胞特异性识别多肽对人颅骨成骨细胞(HCO)增殖和矿化作用的影响。方法体外常规培养人颅骨成骨细胞,分为5个实验组(10^(-4)、10^(-5)、10^(-6)、10^(-7)、10^(-8)mol/L)以及空白对照组,MTT法检测细胞的增殖,紫外分光光度法检测细胞碱性磷酸酶(ALP)的表达,茜素红染色及半定量分析检测细胞矿化程度,RT-QPCR法检测细胞骨钙素(OCN)mRNA的表达。结果此浓度范围内的特异性多肽对成骨细胞增殖有促进作用(P<0.05),最大效应浓度是10^(-5)mol/L;能增加ALP活性(P<0.05),最大效应浓度是10^(-4)mol/L。通过茜素红染色观察和半定量分析,14、21 d实验组钙盐沉积量高于对照组,半定量分析结果显示浓度10^(-5)mol/L时,吸光度(OD)值最高,但差异无统计学意义;RT-QPCR法显示14、21 d实验组OCN mRNA表达量均高于对照组(P<0.05)。结论 10^(-8)~10^(-4)mol/L浓度范围的成骨细胞特异性识别多肽能够促进人颅骨成骨细胞的增殖和矿化,且在10^(-5)mol/L浓度促进成骨细胞增殖及矿化相关蛋白表达最为显著。

阿仑膦酸钠对人成骨样细胞增殖和分化的影响

目的 :研究阿仑膦酸钠对人成骨样细胞增殖和分化的影响。方法 :进行OS 732人成骨样细胞生长曲线的绘制、MTT测定、形态学观察、碱性磷酸酶 (ALP)测定、ALP染色等。结果 :①d3时 ,与对照组相比 ,10 - 6 mol·L- 1 浓度组的细胞数目增加 2 2 .6 % ;而 10 - 5mol·L- 1 浓度组的细胞数目明显低于对照组。d 6时 10 - 6 ,10 - 7,10 - 8mol·L- 1 三种浓度组细胞数目比对照组分别增加 2 4 .7% ,2 4 .7%和 31.0 %。 10 - 5mol·L- 1 浓度组的细胞数目仍明显低于对照组。MTT测定得到了类似的结果。②加药后d4 ,10 - 6 ,10 - 7,10 - 8mol·L- 1 三种浓度组的细胞与空白对照组相比细胞数目多、形态大而规则 ,染色质深而致密 ,核质比小。③ALP染色结果显示 ,10 - 6 ,10 - 7mol·L- 1 浓度组细胞ALP颗粒染色明显深于对照组 ,并且ALP颗粒在胞质中分布均匀。ALP的定量测定结果表明 ,10 - 6 ,10 - 8mol·L- 1 浓度组细胞ALP水平明显高于对照组。结论 :阿仑膦酸钠对体外培养的OS 732人成骨样细胞具有促增殖和分化作用 ,并且这种作用具有最适合的浓度。

纳米陶瓷支架的研制及其生物相容性评价

目的 制备纳米多孔羟基磷灰石 (HA)和磷酸三钙 (TCP)支架 ;研究新型陶瓷支架的生物相容性。方法 用Sel- gel法和直接沉淀法分别制备纳米级的HA及TCP粉体 ,采用有机泡沫浸渍法制备多孔陶瓷支架 ;采用人骨膜成骨细胞 (periosteal derivedosteoblastcells ,POB)与两种陶瓷支架的体外复合培养 ,观察细胞在材料上的生长及功能表达。结果 制备出具有良好孔隙结构和一定强度的纳米多孔陶瓷支架材料 ;人POB在HA和TCP表面良好的生长 ,并发挥成骨功能。结论 以HA、TCP的纳米级粉体为原料 ,采用有机泡沫浸渍法可制备出具有良好孔隙结构和较好力学强度的多孔陶瓷支架 ;新型多孔陶瓷支架有良好的生物相容性。