利用重组人骨形态发生蛋白-2诱导椎间盘成骨的实验研究

目的确定在兔子的椎间盘内注射重组人骨形态发生蛋白-2（recombinant human bone morphogenetic protein-2，rhBMP-2）诱导椎体间融合的可行性。方法将24只成年新西兰大白兔，随机分为2组，每组12只。用微量注样器将含有rhBMP-2200μg的生理盐水溶液20μl和等量的生理盐水分别注射到成年新西兰大白兔的L4～5椎间盘的髓核内。术后10、30、60及90天进行X线照相和组织学检查。结果注射椎间盘未见免疫排斥反应。实验组可见纤维环和软骨终板成骨并在相邻椎体间形成骨桥。对照组的椎间盘内未见骨形成。结论利用注射的方法，rhBMP-2可诱导椎间盘成骨，达到椎体间融合的目的。     

注射型rhBMP-2骨修复材料对兔桡骨骨缺损修复的实验研究

探讨以纤维蛋白胶(fibrin sealant,FS)为载体,复合重组BMP-2(rhBMP-2)的注射型骨修复材料,修复兔桡骨节段性骨缺损的能力,为其临床应用提供实验依据。将实验动物分为:实验组(FS rhBMP-2)、对照组(FS)。于28只新西兰白兔右侧桡骨干处造成1.5cm缺损,然后严密缝合皮下组织及皮肤,将各组材料经正常皮肤注射骨缺损处,术后4、8、16、24周进行放射学、组织学和骨密度等方法检查,对其成骨效应进行研究。结果表明:实验组(FS rhBMP-2)骨缺损区在成骨活跃程度、骨密度和再生髓腔结构等方面均显著优于对照组(FS),使骨缺损得到了彻底修复(P<0.01);对照组不能产生骨性愈合。由此说明以纤维蛋白胶为载体复合rhBMP-2的注射型骨修复材料具有高效的骨修复能力。     

rhBMP-2／无定形磷酸钙纳米缓释微粒成骨活性的实验研究

目的探讨无定形磷酸钙（ACP）对rhBMP-2的缓释作用，检测rhBMP-2／ACP纳米缓释微粒的成骨活性。方法用扫描电镜观察已制备rhBMP-2／ACP缓释微粒的大小、形态：测定rhBMP-2的体外释放情况并描绘曲线；用MTT法检测缓释微粒对兔骨髓间充质干细胞（MSC）增殖情况的影响：用碱性磷酸酶（ALP）试剂盒检测细胞ALP活性以反映缓释微粒对其分化的影响．并与ACP的作用进行比较；将缓释微粒植入大鼠股部肌袋．通过X线、组织形态学观察评价缓释微粒的异位成骨能力。结果缓释微粒大小为100nm左右．具有典型的无定形球形面貌。开始时为快速释放期．随后呈缓慢持续释放。缓释微粒能显著促进MSC的增殖和分化，植入大鼠股部肌袋12周．材料大部分降解．有明显的骨形成。结论ACP可作为rhBMP-2合适的缓释载体材料．生成的纳米缓释微粒具有良好的降解性能和成骨活性。     

复合rhBMP-2的注射型成骨材料修复犬桡骨骨缺损的实验研究

探讨以纤维蛋白胶(fibrin sealant,FS)为载体,复合rhBMP-2\bFGF的注射型骨修复材料,修复犬桡骨节段性缺损的作用,为其临床应用提供实验依据.实验分为:实验组(FS bFGF rhBMP-2)、对照组(FS).于12只成年健康家犬右侧桡骨中上段造成2cm缺损,制成犬桡骨2cm骨缺损实验模型,然后严密缝合皮下组织及皮肤,将实验材料经伤口周围正常皮肤注射到骨缺损处,术后4、8、16、24w进行放射学检查,术后24w,标本进行组织学和骨密度检查,研究其成骨效应.结果表明:实验组骨缺损区在成骨活跃程度、骨密度和再生髓腔结构等方面均显著优于对照组,使骨缺损得到了成功的修复(P＜0.01).以纤维蛋白胶为载体复合rhBMP-2和bFGF的注射型骨修复材料,具有高效的骨修复能力,对犬的骨缺损有很好的修复效果.     

rhBMP-2/聚乳酸与聚乙醇酸共聚物载药微球的制备及成骨活性研究

目的制备一种具有良好降解性和成骨活性可注射的rhBMP-2载体材料。方法采用复乳-溶剂蒸发技术制备携载rhBMP-2的聚乳酸与聚乙醇酸共聚物P(LGA)微球。测定材料的制备参数及其特性,包括材料的形貌、载药率、释药速度,并将载药微球植入鼠股部肌袋,通过X线、组织学评价载体材料的异位成骨能力。结果载药微球粒径为(253±64)μm,载药率0.52%±0.14%,载药微球rhBMP-2体外释放24h时为15.2%±0.8%,随后呈持续缓慢释放,28d时总计达48.6%±5.3%。载药微球植入鼠股部肌袋4周,材料周围有明显的骨形成。结论载有rhBMP-2的PLGA微球具有良好的缓释效果和生物活性,是一种较为理想的生长因子载体材料和释放系统。     

β-TCP降解和rhBMP-2诱导成骨的关系

探讨β-TCP的降解率是否可以影响rhBMP-2的诱导成骨量.将不同降解率的β-TCP1和β-TCP2分别与rhBMP-2复合制备成不同降解率的两种复合物,β-TCP1/rhBMP-2和β-TCP2/rhBMP-2.然后分别种植于小鼠股部肌袋中.种植后分别进行组织学检查、组织形态计量、碱性磷酸酶和降解量测定.两种复合物种植后均有软骨和骨形成,成骨量随时间推移而增加.但是,种植后8周时β-TCP1/rhBMP-2组成骨量显著高于β-TCP2/rhBMP-2组(P＜0.05).种植后4周以前两组的ALP水平均随时间推移而升高(P＜0.05),但是,在4～8周间则无明显差异.此外,种植后8周时β-TCP1/rhBMP-2的降解量显著高于β-TCP2/rhBMP-2(P＜0.05).β-TCP的降解率可以影响BMP的诱导成骨量.     

rhBMP-2增强腱骨界面愈合的生物力学研究

目的:探讨rhBMP-2增强移植腱骨隧道界面愈合能力。方法:取成年新西兰兔同侧半腱肌肌腱作为自体移植材料,建立70只新西兰兔双侧前交叉韧带重建动物模型。实验组在移植腱-骨隧道界面注射填充以纤维蛋白胶作为载体的rhBMP-2,对照组仅填充纤维蛋白胶,完全空白对照组则不作任何填充。分别于术后第2、4和8周取材,进行生物力学检测。结果:实验组最大载荷在2、4、8周较对照组分别增强13.53%、82.13%、93.23%。实验组刚度在4、8周较对照组分别增强71.91%、121.99%,差异在统计学上显著(P<0.05)。结论:rhBMP-2可以在术后早期提高腱骨界面的最大载荷和刚度,增强了腱骨界面的结合力,促进腱骨界面愈合。     

重组人骨形态发生蛋白2修饰的组织工程化骨修复眼眶骨折的实验研究

目的 探讨组织工程化骨修复眼眶骨折缺损的治疗效果.方法 体外构建以自体骨髓基质干细胞(BMSC)为种子细胞、可降解吸收的生物材料聚乳酸羟基乙酸聚合物(PLGA)为载体、重组人骨形态发生蛋白2(rhBMP-2)为生长因子的组织工程化骨,将实验动物分为对照组(植入PLGA/rhBMP-2复合物)和实验组(植入组织工程化骨),观察术后1个月、3个月和6个月伤口愈合情况、并发症及眼眶外观、CT影像学和组织学变化.结果 术后所有动物伤口愈合良好,无并发症和眼球凹陷.CT三维成像显示术后3个月实验组的缺损范围[(25.1±6.8) mm2]小于对照组[(55.3±7.7)mm2];术后6个月,实验组的眼眶骨折缺损消失,而对照组仍存在.组织学结果显示,术后1个月即可观察到实验组植入区边缘植入物开始缓慢吸收,少量成骨细胞沿支架长人材料内,而对照组未观察到;术后3个月可见实验组形成条带状新生骨长入将其分割包绕呈交叉排列,材料降解吸收明显高于对照组;术后6个月实验组植人材料完全被降解吸收,同时被新生骨组织取代,植入物与自身骨组织紧密结合,融为一体.而对照组仅部分降解吸收.结论 重组人骨形态发生蛋白2修饰的组织工程化骨具有较强的传导成骨和诱导成骨活性,生物相容性好,材料可完全降解,为骨组织取代,对眼眶骨折缺损具有较好的修复效果.     

BMP_2活性多肽/鼠尾Ⅰ型胶原复合物植入异位成骨的实验研究

用动物实验的方法验证鼠尾Ⅰ型胶原支架与自行研制合成的BMP2活性多肽复合后诱导异位成骨的能力与作用。自行提取鼠尾Ⅰ型胶原以及与BMP2活性多肽复合,冻干后低真空模式下电镜观察胶原结构。实验分2组。对照组:单纯鼠尾Ⅰ型胶原组;实验组:BMP2活性多肽/鼠尾Ⅰ型胶原复合物组。将12只大白鼠随机分成2组,每只大白鼠右侧大腿作2 cm的切口,制备股四头肌肌袋模型,将上述材料分别植入肌袋。术后第3周和第6周分别作放射学检查(X-ray,CT),第6周将所有大白鼠处死作组织学(HE染色)检查。鼠尾Ⅰ型胶原冻干后孔径大小合适与BMP2活性多肽复合后无明显改变。术后第3周和第6周放射学检查可见实验组有明显的钙化影形成,且第6周成骨范围要明显大于第3周时所见,而对照组无成骨现象。术后第6周组织学观察可见实验组植入区有成骨细胞和大量新骨形成,而对照组仅见炎性细胞改变。鼠尾Ⅰ型胶原是一种较好的载体支架材料,自行研制合成的BMP2活性多肽与其复合后,具有较强的异位诱导成骨能力。     

两种壳聚糖载rhBMP-2纳米微球对SD大鼠体内异位成骨的影响

目的　通过SD大鼠异位成骨实验来探究重组人骨形态发生蛋白-2/壳聚糖/硫酸葡聚糖(rhBMP-2/CS/DS) 复合微球和重组人骨形态发生蛋白-2/壳聚糖 (rhBMP-2/CS) 微球对SD大鼠体内异位成骨的影响。 方法　随机将36只SD大鼠平均分为三组（n=12）,分别为A组 (rhBMP-2), B组(rhBMP-2/CS), C组(rhBMP-2/CS/DS)。制备股四头肌肌袋模型后,分别将三种材料植入股四头肌肌袋肌间隙中。分别在4,8和12周时大体观察植入区组织硬度,每组处死4只大鼠后取出异位骨块,并切取异位骨化的组织行micro-CT扫描及 Mimics软件三维重建;检测各组织块骨体积分数（bone volume fraction,BVF）、骨小梁厚度（trabecular thichness,Tb.Th）、骨密度（bone mineral density,BMD）;并行组织学观察和ALP活性、钙含量检测。 结果 4周时,A、B、C三组植入区周围组织质地均稍硬,三者并无明显区别;8周和12周时,三组植入区硬度明显增加,且C组比A、B组质地更硬。4周时,HE染色可见三组有少量骨组织形成,但不明显;B、C两组BVF、Tb.Th、BMD,碱性磷酸酶（ALP）活性、钙含量均高于A组;B、C两组以上指标差异无统计学意义。8、12周时,HE染色可见到三组骨组织逐渐增多,并逐渐成熟,且B、C两组可见到比A组更成熟的骨组织,C组骨组织比B组更成熟;B、C两组BVF、Tb.Th、BMD,ALP活性、钙含量均高于A组,C组以上指标均高于B组。 结论　rhBMP-2/CS/DS纳米缓释微球的成骨效果明显强于rhBMP-2/CS纳米微球和单独rhBMP-2,其可能在骨组织工程领域有较好的运用前景。     

Implantation of green tea catechin α-tricalcium phosphate combination enhances bone repair in rat skull defects

The purpose of the present study is to investigate effects of the combination of epigallocatechin-3-gallate (EGCG) and α-tricalcium phosphate (α-TCP) on bone regenerative capacity in a bilateral rat calvarial bone defect model. Materials and methods: Bilateral 5-mm-diameter calvarial defects were created in adult male Wistar rats and filled with preparations of EGCG (0, 0.1, 0.2, 0.4 mg) combined with α-TCP particles. This was done by dissolving EGCG in 100% ethanol (50 μL/14 mg) and dropping under sterile condition. The control group was left unfilled (n = 8). The animals were sacrificed at 2 and 4 weeks. Radiological images were taken, and histological analysis was done. Six animals from control (0 mg EGCG + α-TCP) group and (0.2 mg EGCG+ α-TCP) group were labeled with fluorescent dyes and histomorphometrically analyzed (n = 6) at 2 and 4 weeks. Results: Histomorphometric analysis revealed that the combination of EGCG and α-TCP at doses of 0.1 and 0.2 mg yielded significantly more new bone formation than untreated control group at 2 and 4 weeks (p > 0.05). Mineral apposition rate at 0.2-TCP group was enhanced compared with the one of the positive control α-TCP group at 4 weeks (p > 0.05). Conclusion: The combination of α-TCP particles and 0.2 mg EGCG stimulates maximum bone regeneration in rat calvarial defects, and this combination would be potentially effective as bone graft material.     

rhBMP-2m对化疗损伤小鼠的修复治疗作用

目的 :探讨重组人骨形成蛋白 2 (rhBMP 2m)对环磷酰胺 (CTX)致骨髓损伤小鼠的治疗作用。方法 :18只小鼠随机分为 3组 :即CTX注射组、BMP治疗组和PBS对照组 ,每组 6只小鼠。CTX组和BMP治疗组小鼠 ,一次性腹腔注射 2 0 0mg/kgCTX建立骨髓损伤的模型。注射CTX第 2天 ,BMP治疗组每只小鼠腹腔注射 0 .5mg的rhBMP 2m开始治疗。观察 3组小鼠外周血白细胞数的变化。分别在第 5天和第 8天 ,用流式细胞仪 (FCM )检测骨髓有核细胞中DNA的含量及细胞周期的变化 ,同时进行粒单细胞集落形成 (CFU GM)细胞培养。结果 :注射CTX后第 4天 ,BMP治疗组和CTX注射组的外周血白细胞数均降至最低点 ,然后逐渐回升 ,两组相比较无显著差异 (P >0 .0 5 )。注射CTX后第 5天 ,BMP治疗组骨髓有核细胞中 ,G0 /G1期细胞的比例较CTX组明显提高 ,细胞的坏死及凋亡率明显下降 (P <0 .0 1)。第 8天 ,BMP治疗组的骨髓有核细胞数较CTX组增加明显 (P <0 .0 1)。结论 :BMP对CTX所致小鼠骨髓的损伤具有修复治疗作用     

骨髓间充质干细胞藻酸钙移植物与复合降解膜修复皮肤缺损

背景：一直以来学者们认为皮肤移植是修复大面积组织缺损最有效的方法,但都存在供体来源限制和免疫排斥反应等问题,因此,加速真皮的构建在皮肤组织工程中极为重要。 目的：观察骨髓间充质干细胞藻酸钙凝胶、碱性成纤维细胞生长因子复合缓释降解膜修复皮肤缺损的效果。 方法：新西兰大耳白兔15只,取其自体髂骨骨髓,体外分离出骨髓间充质干细胞进行培养、扩增、传代并纯化备用;制作皮肤全层缺损模型3处,随机植入自体骨髓间充质干细胞藻酸钙凝胶、碱性成纤维细胞生长因子复合降解膜(实验组),自体骨髓间充质干细胞藻酸钙凝胶(对照1组),不含骨髓间充质干细胞的藻酸钙凝胶(对照2组),术后均用羊膜覆盖创面。术后7,14,21 d取新生创面组织,进行苏木精-伊红染色、免疫组织化学染色及图像分析仪测定。 结果与结论：实验组皮下真皮组织增生明显,其成纤维细胞、血管及胶原纤维较多,特别是术后14,21 d表皮增生较快,其覆盖范围较大,术后21 d,新生表皮大多重建呈多层结构,明显优于对照1组和对照2组。由此可见,骨髓间充质干细胞藻酸钙凝胶和碱性成纤维细胞生长因子复合降解膜植入皮肤缺损局部,加速了真皮组织的再生修复,从而促进表皮的再生和重建。   中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

The role of rhFGF-2 soaked polymer membrane for enhancement of guided bone regeneration

Abstract

The purposes of this study are to confirm the role of Fibroblast Growth Factor-2 (FGF-2) in bone regeneration by adding various concentrations of FGF-2 to the collagen membrane and applying it to the Biphasic Calcium Phosphate (BCP) bone graft site for guided bone regeneration, to explore the potential of collagen membrane as FGF-2 carrier, and to determine the optimum FGF concentration for enhancement of bone regeneration. Four bone defects of 8 mm in diameter were created in 18 New Zealand rabbit calvaria. After BCP bone graft, graft material was covered with collagen membranes adding various concentration of FGF-2. The concentration of FGF-2 was set at 1.0, 0.5, 0.1 mg/ml, and same amount of saline was used in the control group. To confirm the bone regeneration over time, six New Zealand rabbits were sacrificed each at 2, 4, and 12 weeks, and the amounts of new bone and residual bone graft material were analyzed by histologic and histomorphometric analysis. Qualitative analyses are also conducted through immunohistochemistry, Tetrate-resistant acid phosphatase (TRAP) stain and Russell-Movat pentachrome stain. As the healing period increased, the formation of new bone increased and the amount of residual graft material decreased in all experimental groups. Immunohistochemistry, TRAP staining and pentachrome staining further showed that the addition of FGF-2 promoted bone regeneration in all experimental groups. It was also confirmed that polymer collagen membrane can be used as a useful carrier of FGF-2 when enhanced early stage of new bone formation is required.     

New bone formation in a bone defect associated to dental implant using absorbable or non-absorbable membrane in a dog model

The aim of this research was to determine the bone formation capacity in fenestration defects associated with dental implants using absorbable and non-absorbable membranes. Six dogs were used in the study. In both tibias of each animal 3 implants were installed, and around these 5 mm circular defects were created. The defects were covered with absorbable membranes (experimental group 1), non-absorbable membranes (experimental group 2), and the third defect was not covered (control group). At 3 and 8 weeks post-surgery, the animals were euthanized and the membranes with the bone tissue around the implants were processed for histological analysis. The statistical analysis was conducted with Tukey’s test, considering statistical significance when p<0.1. Adequate bone repair was observed in the membrane-covered defects. At 3 weeks, organization of the tissue, bone formation from the periphery of the defect and the absence of inflammatory infiltrate were observed in both experimental groups, but the defect covered with absorbable membrane presented statistically greater bone formation. At 8 weeks, both membrane-covered defects showed adequate bone formation without significant differences, although they did in fact present differences with the control defect in both periods (p>0.1). In the defects without membrane, continuous connective tissue invasions and bone repair deficiency were observed. There were no significant differences in the characteristics and volume of the neoformed bone in the defects around the implants covered by the different membranes, whereas the control defects produced significantly less bone. The use of biological membranes contributes to bone formation in three-wall defects.     

Osseous tissue reaction around hydroxyapatite block implanted into proximal metaphysis of tibia of rat with collagen-induced arthritis

This study was conducted to investigate the influence of osteoporosis on new bone formation around a hydroxyapatite (HA) block implanted into the proximal metaphysis of the tibia of rats with collagen-induced arthritis (CIA). Ten rats were immunized with an emulsion of bovine type II collagen and Freund's complete adjuvant (arthritis group). Another 10 rats, which were not immunized were used as the control group. Seventeen days after immunization, HA block was implanted into the proximal metaphysis of the tibia. Four weeks after implantation, all rats were killed. The serum level of tetrate-resistant acid phosphatase (TRAP), bone mineral density (BMD) in the proximal metaphysis of the tibia and the affinity index in the arthritis group were 28.0+/-3.5 IU/ml, 130.3+/-28.7 mg/cm2 and 77.6+/-10.8%, respectively, and those in the control group were 24.6+/-5.5 IU/ml, 175.9+/-30.5 mg/cm2 and 56.3+/-14.8%. The serum level of TRAP was higher (P < 0.05) and BMD was lower (P < 0.005) in the arthritis group. The amount of new bone formation around the HA block was larger (affinity index, P < 0.05) in the arthritis group than in the control group. These findings suggest that bone formation around HA block might be enhanced even in conditions associated with highly activated bone resorption and bone formation, such as arthritis.     

Preparation of beta-tricalcium phosphate microsphere-hyaluronic acid-based powder gel composite as a carrier for rhBMP-2 injection and evaluation using long bone segmental defect model

The specific objective of this study was to evaluate whether rhBMP-2-loaded bio-scaffolds can be used as effective rhBMP-2 carriers in the implantation of bone defect sites or poor bone quality in host bone. The rhBMP-2 release pattern test showed slow results in both groups, and a 1:9 ratio composition with a high water-absorption rate was selected for in vivo study. All animals euthanized after 9 weeks. The new bone formation and bone quantity and quality of fibular samples were examined. The results showed that the rhBMP-2 powder gel composite improved the new bone formation in the cortical bone and the marrow space. The length of new bone formation ratio of the rhBMP-2 loaded composite group was significantly higher than the powder gel group. The composite of powder gel seems to be a nice carrier, and slow release of rhBMP-2 can promote new bone formation in a segmental cortical bone defect after implantation.     

The use of injectable sonication-induced silk hydrogel for VEGF(165) and BMP-2 delivery for elevation of the maxillary sinus floor

Sonication-induced silk hydrogels were previously prepared as an injectable bone replacement biomaterial, with a need to improve osteogenic features. Vascular endothelial growth factor (VEGF(165)) and bone morphogenic protein-2 (BMP-2) are key regulators of angiogenesis and osteogenesis, respectively, during bone regeneration. Therefore, the present study aimed at evaluating in situ forming silk hydrogels as a vehicle to encapsulate dual factors for rabbit maxillary sinus floor augmentation. Sonication-induced silk hydrogels were prepared in?vitro and the slow release of VEGF(165) and BMP-2 from these silk gels was evaluated by ELISA. For in?vivo studies for each time point (4 and 12 weeks), 24 sinus floors elevation surgeries were made bilaterally in 12 rabbits for the following four treatment groups: silk gel (group Silk gel), silk gel/VEGF(165) (group VEGF), silk gel/BMP-2 (group BMP-2), silk gel/VEGF(165)/BMP-2 (group V?+?B) (n?=?6 per group). Sequential florescent labeling and radiographic observations were used to record new bone formation and mineralization, along with histological and histomorphometric analysis. At week 4, VEGF(165) promoted more tissue infiltration into the gel and accelerated the degradation of the gel material. At this time point, the bone area in group V?+?B was significantly larger than those in the other three groups. At week 12, elevated sinus floor heights of groups BMP-2 and V?+?B were larger than those of the Silk gel and VEGF groups, and the V?+?B group had the largest new bone area among all groups. In addition, a larger blood vessel area formed in the remaining gel areas in groups VEGF and V?+?B. In conclusion, VEGF(165) and BMP-2 released from injectable and biodegradable silk gels promoted angiogenesis and new bone formation, with the two factors demonstrating an additive effect on bone regeneration. These results indicate that silk hydrogels can be used as an injectable vehicle to deliver multiple growth factors in a minimally invasive approach to regenerate irregular bony cavities.     

Biodentine enhances the proliferation and differentiation of osteoblasts through upregulating bone morphogenetic protein-2北大核心

BACKGROUND: Biodentine is a new kind of biocompatible calcium silicate dental restoration material. It can not only increase the secretion of growth factors in dental pulp cells, but also induce the mineralization of dentin. However, the research of Biodentine in bone repair is few. OBJECTIVE: To investigate the effects of Biodentine on the proliferation and differentiation of bone morphogenetic protein-2 interfered mouse osteoblast precursor cell lines by constructing mouse osteoblast precursor cell lines in which bone morphogenetic protein-2 was interfered. METHODS: The 0.1, 1, 10, and 100 g/L Biodentine extract was applied to mouse osteoblast precursor cells for 24 hours. The optimal concentration of 10 g/L was selected by CCK-8 and alkaline phosphatase for the following experiments. Biodentine extract was used to treat mouse osteoblast precursor cells for 1, 3, 5 and 7 days, and the non-intervention group was used as the control group. The cell proliferation and osteogenic differentiation were observed by CCK-8 assay and alkaline phosphatase activity test. Mouse osteoblast precursor cell lines were constructed by interfering with bone morphogenetic protein-2. The proliferation and osteogenic differentiation of cells were observed by CCK-8 assay and alkaline phosphatase activity test. The mouse osteoblast precursor cell lines interfering with bone morphogenetic protein-2 were constructed. The Biodentine group was added with 10 g/L Biodentine extract, and the non-intervention group was set as the control. After 24 and 48 hours of treatment, the proliferation and osteogenic differentiation of the cells were observed by CCK-8 assay and alkaline phosphatase activity test. After 24 hours of treatment, the expression levels of alkaline phosphatase mRNA and protein were detected by real-time PCR and western blot assay. RESULTS AND CONCLUSION: (1) 10 g/L Biodentine extract could promote the proliferation and osteogenic differentiation of mouse osteoblast progenitor cells at 1, 3, 5 and 7 days. (2) Interfering with bone morphogenetic protein-2 could inhibit the proliferation and osteogenic differentiation of mouse osteoblast progenitor cells. (3) The cell proliferation and osteogenic differentiation abilities of Biodentine group were higher than those of control group at 24 and 48 hours (P < 0.05), and the expression levels of alkaline phosphatase mRNA and protein of Biodentine group were higher than those of the control group (P<0.05). (4) The results showed that 10 g/L Biodentine could enhance the proliferation and osteogenic differentiation of osteoblasts through bone morphogenetic protein-2. © 2022, Publishing House of Chinese Journal of Tissue Engineering Research. All rights reserved.     

H2-Bl基因转染对小鼠血管平滑肌细胞生物学行为的影响

目的:H2-Bl基因转染小鼠血管平滑肌细胞(VSMC),观察其凋亡、增殖和抗外周血单个核细胞(PBMC)杀伤性等生物学行为的变化,探讨借鉴母胎免疫耐受模型防治心脏移植术后免疫排斥反应的机制。方法:0.5mg/L空质粒、0.5mg/LH2-Bl质粒、1.0mg/LH2-Bl质粒转染小鼠血管平滑肌细胞后,采用实时荧光定量PCR、流式细胞术、酶标仪检测不同时间质粒的转染效率、H2-Bl基因mRNA的表达量、转染VSMC的凋亡与增殖情况以及PBMC对靶细胞的毒性作用。结果:荧光定量PCR结果显示,基因组的H2-Bl基因mRNA表达量明显高于对照组(P<0.001)。H2-Bl基因促进VSMC凋亡,转染24h后0.5mg/LH2-Bl质粒组、1.0mg/LH2-Bl质粒组与空白对照比较,差异均有统计学意义(P<0.05,P<0.01)。H2-Bl基因抑制VSMC增殖,转染24h和48h后0.5mg/LH2-Bl质粒组、1.0mg/LH2-Bl质粒组与对照组比较差异均有统计学意义(P<0.05,P<0.01)。细胞毒性试验在转染24h后,0.5mg/LH2-Bl质粒组、1.0mg/LH2-Bl质粒组细胞毒性均较空白对照明显降低(P<0.05,P<0.01)。结论:pEGFP-N1-H2B1质粒载体转染小鼠VSMC后,能够有效促进其凋亡,抑制其增殖,降低PBMC对靶细胞的毒性作用,诱导免疫耐受。