Immunomodulatory activity of polycaprolactone nanoparticles with calcium phosphate salts against Leishmania infantum infection

Objective:To prepare and characterize polycaprolactone(PCL)nanoparticles loaded with sonicator fragmented(SLA)and freeze-thaw Leishmania antigens(FTLA)and to investigate the in vitro immunogenicity of antigen-encapsulated nanoparticles with calcium phosphate adjuvant.Methods:The water/oil/water binary emulsion solvent evaporation method was used to synthesize antigen-loaded PCL nanoparticles.Particles were characterized by scanning electron microscopy and zeta potential measurements.Their cytotoxicity in J774 macrophages in vitro was determined by MTT analysis.In addition,the amount of nitric oxide and the level of cytokines produced by macrophages were determined by Griess reaction and ELISA method,respectively.The protective effect of the developed formulations was evaluated by determining the infection index percentage in macrophages infected with Leishmania infantum.Results:Compared to the control group,SLA PCL and FTLA PCL nanoparticles with calcium phosphate adjuvant induced a 6-and 7-fold increase in nitric oxide,respectively.Additionally,the vaccine formulations promoted the production of IFN-γand IL-12.SLA PCL and FTLA PCL nanoparticles combined with calcium phosphate adjuvant caused an approximately 13-and 11-fold reduction in infection index,respectively,compared to the control group.Conclusions:The encapsulation of antigens obtained by both sonication and freeze-thawing into PCL nanoparticles and the formulations with calcium phosphate adjuvant show strong in vitro immune stimulating properties.Therefore,PCL-based antigen delivery systems and calcium phosphate adjuvant are recommended as a potential vaccine candidate against leishmaniasis.

Calcium phosphate nanoparticles show an effective activation of the innate immune response in vitro and in vivo after functionalization with flagellin

For subunit vaccines,adjuvants play a key role in shaping the magnitude,persistence and form of targeted antigen-specific immune response.Flagellin is a potent immune activator by bridging innate inflammatory responses and adaptive immunity and an adjuvant candidate for clinical application.Calcium phosphate nanoparticles are efficient carriers for different biomolecules like DNA,RNA,peptides and proteins.Flagellin-functionalized calcium phosphate nanoparticles were prepared and their immunostimulatory effect on the innate immune system,i.e.the cytokine production,was studied.They induced the production of the proinflammatory cytokines IL-8 （Caco-2 cells） and IL-1β（bone marrow-derived macrophages; BMDM） in vitro and IL-6 in vivo after intraperitoneal injection in mice.The immunostimulation was more pronounced than with free flagellin.

Novel rechargeable calcium phosphate nanoparticle-containing orthodontic cement

White spot lesions （WSLs）, due to enamel demineralization, occur frequently in orthodontic treatment. We recently developed a novel rechargeable dental composite containing nanoparticles of amorphous calcium phosphate （NACP） with long-term calcium （Ca） and phosphate （P） ion release and caries-inhibiting capability. The objectives of this study were to develop the first NACP- rechargeable orthodontic cement and investigate the effects of recharge duration and frequency on the efficacy of ion re-release. The rechargeable cement consisted of pyromellitic glycerol dimethacrylate （PMGDM） and ethoxylated bisphenol A dimethacrylate （EBPADMA）. NACP was mixed into the resin at 40% by mass. Specimens were tested for orthodontic bracket shear bond strength （SBS） to enamel, Ca and P ion initial release, recharge and re-release. The new orthodontic cement exhibited an SBS similar to commercial orthodontic cement without CaP release （P〉 0.1）. Specimens after one recharge treatment （e.g., 1 min immersion in recharge solution repeating three times in one day, referred to as ＂1 min 3 times＂） exhibited a substantial and continuous re-release of Ca and P ions for 14 days without further recharge. The ion re-release did not decrease with increasing the number of recharge/re-release cycles （P〉 0.1）. The ion re-release concentrations at 14 days versus various recharge treatments were as follows： 1 min 3 times〉3 min 2 times〉 1 min 2 times〉6 min 1 time〉3 min 1 time〉 1 min 1 time. In conclusion, although previous studies have shown that NACP nanocomposite remineralized tooth lesions and inhibited caries, the present study developed the first orthodontic cement with Ca and P ion recharge and long-term release capability. This NACP-rechargeable orthodontic cement is a promising therapy to inhibit enamel demineralization and WSLs around orthodontic brackets.

Effects of quaternary ammonium chain length on the antibacterial and remineralizing effects of a calcium phosphate nanocomposite

Composites containing nanoparticles of amorphous calcium phosphate （NACP） remineralize tooth lesions and inhibit caries. A recent study synthesized quaternary ammonium methacrylates （QAMs） with chain lengths （CLs） of 3-18 and determined their effects on a bonding agent. This study aimed to incorporate these QAMs into NACP nanocomposites for the first time to simultaneously endow the material with antibacterial and remineralizing capabilities and to investigate the effects of the CL on the mechanical and biofilm properties. Five QAMs were synthesized： DMAPM （CL3）, DMAHM （CL6）, DMADDM （CL12）, DMAHDM （CL16）, and DMAODM （CL18）. Each QAM was incorporated into a composite containing 20% NACP and 50% glass fillers. A dental plaque microcosm biofilm model was used to evaluate the antibacterial activity. The flexural strength and elastic modulus of nanocomposites with QAMs matched those of a commercial control composite （n = 6; P 〉 0.1）. Increasing the CL from 3 to 16 greatly enhanced the antibacterial activity of the NACP nanocomposite （P 〈 0.05）; further increasing the CL to 18 decreased the antibacterial potency. The NACP nanocomposite with a CL of 16 exhibited biofilm metabolic activity and acid production that were 10-fold lesser than those of the control composite. The NACP nanocomposite with a CL of 16 produced 2-log decreases in the colony-forming units （CFU） of total microorganisms, total streptococci, and mutans streptococci. In conclusion, QAMs with CLs of 3-18 were synthesized and incorporated into an NACP nanocomposite for the first time to simultaneously endow the material with antibacterial and remineralization capabilities. Increasing the C/reduced the metabolic activity and acid production of biofilms and caused a 2-log decrease in CFU without compromising the mechanical properties. Nanocomposites exhibiting strong anti-biofilm activity, remineralization effects, and mechanical properties are promising materials for tooth restorations that inhibit caries.

One-year water-ageing of calcium phosphate composite containing nano-silver and quaternary ammonium to inhibit biofilms

Dental composites are commonly used restorative materials; however, secondary caries due to biofilm acids remains a major problem. The objectives of this study were （1） to develop a composite containing quaternary ammonium dimethacrylate （QADM）, nanoparticles of silver （NAg）, and nanoparticles of amorphous calcium phosphate （NACP）, and （2） to conduct the first investigation of the mechanical properties, biofilm response and acid production vs water-ageing time from 1 day to 12 months. A 4 x 5 design was utilized, with four composites （NACP-QADM composite, NACP-NAg composite, NACP-QADM-NAg composite, and a commercial control composite）, and five water-ageing time periods （1 day, and 3, 6, 9, and 12 months）. After each water- ageing period, the mechanical properties of the resins were measured in a three-point flexure, and antibacterial properties were tested via a dental plaque biofilm model using human saliva as an inoculum. After 12 months of water-ageing, NACP-QADM- NAg had a flexural strength and elastic modulus matching those of the commercial control （P〉 0.1）. Incorporation of QADM or NAg into the NACP composite greatly reduced biofilm viability, metabolic activity and acid production. A composite containing both QADM and NAg possessed a stronger antibacterial capability than one with QADM or NAg alone （P〈0.05）. The anti-biofilm activity was maintained after 12 months of water-ageing and showed no significant decrease with increasing time （P〉0.1）. In conclusion, the NACP-QADM-NAg composite decreased biofilm viability and lactic acid production, while matching the load- bearing capability of a commercial composite. There was no decrease in its antibacterial properties after 1 year of water-ageing. The durable antibacterial and mechanical properties indicate that NACP-QADM-NAg composites may be useful in dental restorations to combat caries.

Microwave assisted synthesis & properties of nano HA-TCP biphasic calcium phosphate

Biphasic calcium phosphate （BCP） nanopowders were synthesized by using microwave and non-microwave irradiation assisted processes. The synthesized powders were pressed under a pressure of 90 MPa, and then were sintered at 1000-1200℃ for 1 h. The mechanical properties of the samples were investigated. The formed phases and microstructures were characterized by X-ray diffraction （XRD） and scanning electron microscopy （SEM）. The results showed that the synthesis time was shorter, along with a more homogeneous microstructure, when the microwave irradiation assisted method was applied. The compression strength and the Young＇s modulus of the samples synthesized with microwave irradiation were about 60 MPa and 3 GPa, but those of the samples synthesized without microwave irradiation were about 30 MPa and 2 GPa, respectively. XRD patterns of the microwave irradiation assisted and non-microwave irradiation assisted nanopowders showed the coexistence of hydroxyapatite （HA） and lricalcium phosphate （TCP） phases in the system.

Preparation and functional study of pH-sensitive amorphous calcium phosphate nanocarriers

Recently,multifunctional nanoparticles have shown great prospects in cancer treatment,which have the ability to simultaneously deliver the drug,image and target tumor cells.In this paper,we designed a luminescent nanoparticles platform based on hydrothermal hyaluronic acid/amorphous calcium phosphate(HA-FCNs/ACP)with multifunctional properties for drug delivery,bio-imaging,and targeting treatment.HA-FCNs/ACP shows an ability to load curcumin(Cur)with pH-sensitive responsive drug release behavior and excellent biocompatibility.HA-FCNs/ACP dispersed in the cytoplasm through the overexpressed CD44 receptor that is actively targeted into human lung cancer cells(A549 cells).Meanwhile,the viability of A549 cells was significantly inhibited in vitro.The prepared HA-FCNs and HA-FCNs/ACP both exhibit excellent targeted bioimaging performance on cancer cells.Hence,the as-prepared nanoparticles have promising applications in treating tumor disease.

Inorganic phosphate in the development and treatment of cancer:A Janus Bifrons?

Inorganic phosphate(Pi) is an essential nutrient to living organisms. It is required as a component of the energy metabolism,kinase/phosphatase signaling and in the formation and function of lipids,carbohydrates and nucleic acids and,at systemic level,it plays a key role for normal skeletal and dentin mineralization. Pi represents an abundant dietary element and its intestinal absorption is efficient,minimally regulated and typically extends to approximately 70%. Maintenance of proper Pi homeostasis is a critical event and serum Pi level is maintained within a narrow range through an elaborate network of humoral interactions and feedback loops involving intestine,kidney,parathyroid gland and bone,and depends on the activity of a number of hormones,including parathyroid hormone,1,25-dihydroxy vitamin D,and fibroblast growth factor 23 as major regulators of Pi homeostasis. Notably,Pi intake seemingly continues to increase as a consequence of chronic high-phosphorus(P) diets deriving from the growing consumption of highly processed foods,especially restaurant meals,fast foods,and convenience foods. Several recent reports have generated significant associations between high-P intake or high-serum Pi concentration and morbidity and mortality. Many chronic diseases,including cardiovascular diseases,obesity and even cancer have been proposed to be associated with high-P intakes and high-serum Pi concentrations. On the other hand,there is also evidence that Pi can have antiproliferative effects on some cancer cell types,depending on cell status and genetic background and achieve additive cytotoxic effects when combined with doxorubicin,illustrating its potential for clinical applications and suggesting that up-regulating Pi levels at local sites for brief times,might contribute to the development of novel and cheap modalities for therapeutic intervention in some tumours. Overall,the influence of Pi on cell function and the possible relationship to cancer have to be fully understood and investigated further.

Redispersible and stable amorphous calcium phosphate nanoparticles functionalized by an organic bisphosphate

Although much effort has been focused on the preparation of stable amorphous calcium phosphate （ACP） nanoparticles in aqueous solution, the redispersibility and long-term stability of ACP nanoparticles in aqueous solution remains an unresolved problem. In this work, stable colloidal ACPs were prepared by using an organic bisphosphonate （BP） as a sterically hindered agent in aqueous solution. The harvested calcium phosphate nanoparticles were characterized by inductively coupled plasma atomic emission spectrometry （ICP-AES）, Fourier transform infrared （FTIR）, X-ray diffraction （XRD）, dynamic light scattering （DLS） and transmission electron microscopy （TEM）. ICP-AES, FTIR and XRD results suggested the particles were ACP. DLS and TEM results indicated that the size of the ACP nanoparticles were in the range of 60 nm with a spherical morphology. The resulting calcium phosphate nanoparticles retained its amorphous nature in aqueous solution for at least 6 months at room temperature due to the stabilizing effect of the organic bisphosphonate. Moreover, the surface of the ACP nanoparticles adsorbed with the organic bisphosphate used showed good redispersibility and high colloid stability both in organic and aqueous solutions.

Preparation of porous chitosan-poly(acrylic acid)-calcium phosphate hybrid nanoparticles via mineralization

In this work, the preparation of chitosan-poly(acrylic acid)-calcium phosphate hybrid nanoparticles (CS-PAA-CaP NP) based on the mineralization of calcium phosphate (CaP) on the surface of chito-san-poly (acrylic acid) nanoparticles (CS-PAA NPs) was reported. CS-PAA-CaP NPs were achieved by directly adding ammonia to the aqueous solution of CS-PAA nanoparticles or by thermal decomposi-tion of urea in the aqueous solution of CS-PAA nanoparticles, resulting in the mineralization of CaP on the surface of CS-PAA NPs. Through these two routes, especially using urea as a pH-regulator, the precipitation of CS-PAA NPs, a common occurrence in basic environment, was avoided. The size, morphology and ingredient of CS-PAA-CaP hybrid nanoparticles were characterized by dynamic light scattering (DLS), transmission electron microscope (TEM), scanning electron microscope (SEM), thermogravimetry analysis (TGA) and X-ray diffractometer (XRD). When urea was used as the pH regulator to facilitate the mineralization during the thermal urea decomposition procedure, regular CS-PAA-CaP hybrid nanoparticles with a porosity-structural CaP shells and 400―600 nm size were obtained. TGA result revealed that the hybrid NPs contained approximately 23% inorganic component, which was consistent with the ratio of starting materials. The XRD spectra of hybrid nanoparticles in-dicated that dicalcium phosphate (DCP: CaHPO4) crystal was a dominant component of mineralization. The porous structure of the CS-PAA-CaP hybrid NPs might be greatly useful in pharmaceutical and other medical applications.

一种新型基因传输载体-磷酸钙纳米颗粒用于肿瘤基因治疗的研究

目的研究新型非病毒载体磷酸钙纳米颗粒的生物学特性,评估其在肿瘤基因治疗中应用的前景。方法化学方法制备,氯化钙修饰纳米颗粒,用琼脂糖凝胶电泳分析磷酸钙纳米颗粒与DNA的结合效率及对DNA的保护作用,用绿色荧光蛋白基因作报告基因,将磷酸钙纳米颗粒为基因载体转染鼻咽癌(CNE-2)细胞和在动物体内转染肿瘤细胞;以及将磷酸钙纳米与自杀基因yCDglyTK结合,并在体外对鼻咽癌进行基因治疗。结果电镜观察证实磷酸钙纳米颗粒进入细胞内,磷酸钙纳米颗粒与DNA结合后,能对DNA起保护作用;磷酸钙纳米颗粒作为基因转染的载体,将绿色荧光蛋白基因导入CNE-2细胞,用荧光显微镜观察到高水平的绿色荧光蛋白表达;体内转染结果显示,纳米介导的基因在肿瘤组织中有很好的表达;以磷酸钙纳米为载体介导自杀基因yCDglyTK在体外治疗鼻咽癌的实验中显示,在加入5-FC后大量的CNE-2细胞出现死亡。结论磷酸钙纳米颗粒具有优良的生物学特性,是有应用前景的基因转染和基因治疗载体之一。

纳米磷酸钙作为猪瘟多肽疫苗佐剂的研究

为了探索纳米磷酸钙(NCaP)作为猪瘟多肽疫苗佐剂的可行性,本研究利用猪瘟多肽疫苗作为抗原,制备纳米磷酸钙吸附猪瘟多肽疫苗。将制备好的疫苗免疫兔后,分别检测兔抗体水平和攻毒后体温变化情况。检测结果显示,纳米磷酸钙对猪瘟多肽吸附率达70%;免疫后纳米磷酸钙佐剂组抗体水平低于弗氏佐剂,但与游离猪瘟多肽疫苗组相比,有较大提高;除弗氏佐剂组外,所有实验兔均有体温升高现象,纳米磷酸钙佐剂组体温升高的时间与空白组相比推迟16 h。结果表明,NCaP作为佐剂相对游离猪瘟多肽可提高其免疫效果,但其诱导的抗体水平低于弗氏佐剂组。

联合应用纳米金和磷酸钙骨水泥支架促进牙髓干细胞的增殖和成骨分化

目的:磷酸钙骨水泥(calcium phosphate cement,CPC)支架以其良好的生物相容性和骨引导活性而广泛应用于组织工程骨缺损的修复中,但是其骨引导活性有限。因此考虑联合应用纳米金(gold nanoparticles,Au NPs)以促进牙髓干细胞的增殖和成骨分化。方法:以CPC支架作为基底接种牙髓干细胞,在培养基中添加5μg/m L Au NPs进行干细胞培养,以常规培养基组为对照。用透射电镜检测Au NPs的形貌。以荧光显微镜检测细胞接种4 h后的黏附状态,以CCK-8检测细胞增殖情况,检测细胞碱性磷酸酶的活性,并进行茜素红染色以说明矿化物形成情况。结果:Au NPs为均匀的球形,直径20 nm左右。细胞接种后4 h,实验组和对照组的细胞黏附状态没有显著差异。CPC上Au NPs培养基中的细胞14 d时数量多于对照组(P<0.05);且Au NPs培养基中细胞的碱性磷酸酶活性显著增高(P<0.05),矿化物的形成也显著增多(P<0.05)。结论:CPC支架联合Au NPs可显著促进牙髓干细胞的增殖和成骨分化。

用于幽门螺杆菌疫苗佐剂的磷酸钙纳米颗粒的制备

目的制备可作为疫苗佐剂的磷酸概纳米颗粒。方法以Ca(OH)2和H3PO4为原料用共沉淀法制备磷酸钙纳米颗粒,所制得的产物经XRD表征,用透射电子显微镜观察产物的形貌。将该磷酸钙纳米颗粒与幽门螺杆菌重组尿素酶B(rUreB)和重组黏附素A(rHpaA)混合后经灌胃免疫小鼠。结果所制得的磷酸钙纳米颗粒平均粒径在20 nm左右,该磷酸钙纳米颗粒显著提高了rUreB和rHpaA的免疫保护作用。结论该磷酸钙纳米颗粒可作为幽门螺杆菌疫苗的佐剂。

癌胚抗原启动子调控融合自杀基因yCDglyTK载体的构建及其应用研究

目的构建靶向性基因治疗载体pcDNA3.1(-)CVyCDglyTK,研究癌胚抗原(CEA)启动子是否能控制新型融合自杀基因yCDglyTK在CEA阳性结肠癌细胞中专一性表达和杀伤作用。方法采用PCR、RT-PCR、融合PCR、酶切、连接等技术构建由CEA启动子、CMV增强子驱动的融合自杀基因pcDNA3.1(-)CVyCDglyTK表达载体和分别由CEA启动子和巨细胞病毒(CMV)增强子驱动的融合自杀基因pcDNA3.1(-)CEAyCDglyTK、pcDNA3.1(-)CMVyCDglyTK表达载体,以磷酸钙纳米为载体分别转染CEA阳性的人结肠癌细胞株LOVO细胞和CEA阴性的Hela细胞,采用RT-PCR、免疫荧光法检测感染细胞中yCDglyTK基因的表达,并用MIT法检测感染后细胞对5-氟胞嘧啶(5-FC)的敏感性。结果LOVO细胞在感染以上三种质粒表达载体后均有yCDglyTKmRNA表达,且对5-FC的敏感性明显增强,Hela细胞在纳米-pcDNA3.1(-)CMVyCDglyTK复合物感染后有yCDglyTKmRNA表达,对5-FC的敏感性增强,而在纳米-pcDNA3.1(-)CEAyCDglyTK及纳米-pcDNA3.1(-)CVyCDglyTK复合物感染后则没有yCDglyTKmRNA表达,5-FC对其亦无杀伤作用。结论该实验构建的由CEA启动子、CMV增强子驱动的融合自杀基因yCDglyTK靶向性基因治疗载体,能使融合自杀基因在CEA阳性细胞中专一性表达,从而达到靶向治疗肿瘤的目的。

核-壳型脂质体基因载体的研究进展

脂质体作为一种重要的非病毒基因载体,在核酸和药物转运方面有着巨大的优势和潜力。近年来脂质体基因载体的研究基础上,主要介绍了脂质体-聚阳离子-DNA复合载体(LPD)和脂质体-磷酸钙纳米粒子复合载体(LCP)两种核-壳型脂质体基因载体,展望了核-壳型脂质体基因载体的发展方向及其在基因治疗领域的应用前景。

以多层磷酸钙纳米颗粒为载体的O型口蹄疫病毒VP1和VP2基因重组疫苗的构建

构建以多层纳米颗粒为载体的O型口蹄疫病毒基因重组疫苗。在GenBank中查询编码O型口蹄疫病毒(FMDV)VP1 141-160氨基酸和VP2 1-33氨基酸的核苷酸序列,合成该两段基因的重组基因并命名为VP1-VP2基因。再通过PCR扩增VP1和VP2基因,并将VP1、VP2及VP1-VP2基因分别插入到真核表达载体pcDNA3.1中,构建3种重组真核表达质粒(pcDNA3.1-VP1、pcDNA3.1-VP2和pcDNA3.1-VP1-VP2),并分别对重组质粒进行酶切、PCR鉴定及测序分析。将构建的重组质粒制备成多层磷酸钙纳米颗粒,检测其粒径、稳定性及体外转染效率。重组质粒酶切结果显示,基因片段大小与预期相符;测序显示与GenBank中相应基因序列同源性为100%。制备的多层纳米颗粒粒径约为530nm;在4℃过夜保存后,溶液均匀透明;体外转染HEK293细胞的转染效率在9.6%左右。成功构建了以多层磷酸钙纳米颗粒为载体的O型口蹄疫病毒VP1和VP2基因重组质粒,为进一步研究该基因疫苗奠定了基础。

偏磷酸钙纳米粒子的细胞相容性

背景:偏磷酸钙具有优异的细胞相容性能和降解性能及细胞亲和性,人骨髓间充质干细胞可以在多孔偏磷酸钙孔洞内生长和增殖,但有关偏磷酸钙纳米粒子的研究较少。目的:制备纳米级偏磷酸钙微粒,通过流式细胞术快速检测不同浓度纳米级偏磷酸钙微粒对人骨髓间充质干细胞凋亡的影响。方法:采用湿法球磨法制备偏磷酸钙纳米粒子,通过扫描电镜和透射电镜观察纳米粒子的形貌,通过X射线衍射分析确定纳米粒子的晶体结构。将偏磷酸钙纳米粒子混入CYAGON OricellTM基础培养基,使得偏磷酸钙纳米粒子的质量浓度分别为10,1,0.1 mg/L,将其与人骨髓间充质干细胞共培养7 d,通过流式细胞术分析偏磷酸钙纳米粒子质量浓度与细胞凋亡的关系。结果与结论:采用湿法球磨法成功制备了偏磷酸钙纳米粒子,直径为10-30 nm,粒径分布较均匀,分散性较好,但晶体形状不规则;X射线衍射分析晶相检测其主晶相为β-Ca(PO3)2晶体。10 mg/L质量浓度组细胞G0/G1和G2/M比例高于1,0.1 mg/L质量浓度组(P<0.01);10 mg/L质量浓度组细胞早期、中晚期、总细胞凋亡率高于1,0.1 mg/L质量浓度组(P<0.01);说明偏磷酸钙纳米粒子对人骨髓间充质干细胞的增殖有影响,当其质量浓度从1 mg/L增加至10 mg/L后,细胞凋亡率显著增加。

磷酸钙盐药物载体的初步研究

采用化学沉淀法制备了磷酸钙盐的纳米离子，用透射电镜观察其形态和粒径大小；利用吸附载上甲硝唑原料药，紫外分光光度计测定载药量，进行体外释药试验。结果表明，磷酸钙盐粒子平均粒径30-60nm，载药量8％，载药后具有一定的缓释作用。

人鼻咽癌CNE-2细胞转染方法的比较研究

目的比较三种非病毒载体转染方法转染人鼻咽CNE-2细胞。方法分别采用脂质体转染法、电穿孔转染法、磷酸钙纳米颗粒转染法将pEGFP-N1质粒DNA转染人鼻咽癌CNE-2细胞,转染24 h后荧光显微镜观察EGFP的表达,计算转染率;MTT实验比较各转染方法对CNE-2细胞的细胞毒性。结果脂质体转染组的转染率为(51.9±2.8)%;电穿孔转染组的转染率为(42.8±2.6)%;磷酸钙纳米颗粒转染组的转染率为(48.1±2.3)%。经统计学分析,三种方法转染CNE-2细胞的转染率相互之间有显著的差异(P<0.001);MTT结果显示,磷酸钙纳米颗粒转染法的细胞毒性最小。结论磷酸钙纳米颗粒转染法转染效率相对较高,而细胞毒性小,可作为转染CNE-2细胞的首选转染方法。