应用3D适形打印制备网状复合体对兔颅骨缺损的修复作用及安全性研究

目的:探讨采用3D适形打印技术制备的羟基磷灰石/聚乳酸网状复合体在兔颅骨缺损中的修复作用及安全性。方法:以24只新西兰兔为研究对象,以羟基磷灰石/聚乳酸为材料,采用3D适形打印技术制备网状复合体,于兔颅骨顶部制成两个颅骨全层缺损,分别为孔A(左)和孔B(右),孔A(阳性对照组)以自体颅骨为修复材料,孔B(实验组)以复合体为修复材料,观察缺损修复区域的形态学、影像学(X线及CT扫描)及组织学检查结果。结果:植入后24周时,形态学显示:阳性对照组可见致密的骨组织修复,与缺损边缘界限不清,实验组中支架孔隙内纤维组织由新生骨质取代,且新生骨成熟度较提高,材料表面有部分吸收。CT扫描观察显示:冠状面上,阳性对照组缺损修复区域与周围正常骨组织融合为一体,实验组修复材料与缺损边缘融合紧密,与周围正常骨组织结合良好,部分边缘结合不连贯。组织学观察显示:实验组材料部分降解,材料间隔可见新生骨小梁。研究中无实验动物死亡,皮肤切口处缝合良好,无皮下积液,无移植物脱出、红肿感染等情况出现。结论:以3D适形打印技术制备的羟基磷灰石/聚乳酸复合体对兔颅骨缺损有较好的修复作用,能促进缺损区域新骨的形成和生长,且安全性较高。     

自体ADSCs复合PRF修复家兔耳软骨缺损的实验研究

目的:将未诱导的自体脂肪干细胞(ADSCs)与富血小板纤维蛋白(PRF)复合,作为一种全新的软骨修复材料,探讨其对家兔耳软骨全层缺损修复的可行性.方法:取家兔10只,于每只家兔耳部做4处软骨全层缺损,随机分为A、B、C、D组,A组,作为空白对照;B组植入自体ADSCs;C组植入自体PRF;D组植入自体ADSCs与PRF的复合物.分别于术后1月、2月、3月取材,进行大体及HE染色观察,并使用IPP6.0软件对软骨生成量进行半定量分析.结果:HE染色显示,3月后,A组几乎无新生软骨生成,B、C、D三组软骨生成量依次增多,D组尤为明显.IPP6.0统计结果显示,移植物植入3月后,A组软骨缺损修复率为(1.68±0.17)％,B组为(15.4±0.91.)％,C组为(32.0±2.76)％,D组为(85.77±4.88)％.各组间有显著统计学差异,与HE染色结果相符.结论:未诱导的自体ADSC s复合自体PRF作为一种全新的软骨修复材料,可以有效的修复家兔耳软骨全层缺损,具有潜在的临床应用价值.     

牙周基础治疗和牙周再生术联合正畸治疗对广泛型侵袭性牙周炎合并牙槽骨缺损患者的临床研究

摘要 目的：观察牙周基础治疗和牙周再生术联合正畸治疗对广泛型侵袭性牙周炎合并牙槽骨缺损患者的临床疗效及对血清炎症因子和牙龈沟细菌微生态的影响。方法：分析我院2017年8月~2018年8月期间接收的83例广泛型侵袭性牙周炎合并牙槽骨缺损患者的临床资料。根据治疗方式的不同将患者分为A组（40例,牙周基础治疗和牙周再生术治疗）和B组（43例,牙周基础治疗和牙周再生术联合正畸治疗）。观察两组疗效、血清炎症因子、牙龈沟细菌微生态、牙周指标及牙槽骨密度、牙槽骨缺损高度。结果：B组的临床总有效率高于A组（P<0.05）。B组治疗结束后龈沟出血指数（SBI）、牙周探诊深度（PD）、临床附着丧失（AL）、牙菌斑指数（PLI）、牙龈指数（GI）低于A组（P<0.05）。B组治疗结束后牙槽骨缺损高度低于A组,牙槽骨密度高于A组（P<0.05）。B组治疗结束后血清白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、白介素-8（IL-8）水平低于A组（P<0.05）。B组治疗结束后杆菌、球菌、丝状菌、弯曲菌检出率低于A组,梭状菌、螺旋体检出率高于A组（P<0.05）。结论：广泛型侵袭性牙周炎合并牙槽骨缺损患者采用牙周再生术、牙周基础治疗联合正畸治疗,疗效显著,可有效改善牙周状况及牙槽骨缺损情况,降低血清炎症因子水平,但该联合治疗方案会对口腔的细菌微生态产生破坏作用。因此,在接受该方案治疗时,应注意口腔清洁,尽量减轻对牙龈沟细菌微生态的影响。     

骨碎补结合组织工程软骨促进软骨再生的实验研究

目的:评估骨碎补结合组织工程软骨治疗对实验兔软骨缺陷模型软骨再生的疗效。方法:将h IGF-1基因转染MSCs,并与脱细胞真皮基质(ADM)构建组织工程软骨。24只新西兰白兔随机分为A、B、C、D四组,A、C组进行自体软骨移植,B、D组进行改建的细胞-ADM移植。C、D组用40%骨碎补汤喂养4周,150 m L/d。第12周处死实验动物,分离缺损关节软骨部位,蜡块包埋染色,通过总体形态评价软骨再生组织。采用组织学评分评估再生软骨质量。采用甲苯胺蓝染色评价缺损部位产生软骨糖胺聚糖的情况。结果:与B组比较,C组和D组的新生软骨覆盖度、新骨髓的颜色、缺损边缘和表面粗糙度均显著提高(P0.05);再生软骨的组织学评分软骨表面评分显著改善(P0.05)。C组与D组具有比其他组更好的基质、细胞分布和表面指数。并且有较厚的透明样软骨组织,具有正常的糖胺聚糖产生。表明该治疗方法可以通过再生透明样软骨且没有不良事件来减少软骨缺陷。结论:工程软骨结合骨碎补治疗可显著改善兔膝关节软骨缺损修复的质量,为临床治疗软骨病变提供重要理论依据。     

负压封闭引流对兔颅骨外露创面愈合效果的实验研究

目的:探讨负压封闭引流技术(VSD)对兔颅骨外露缺损创面愈合的治疗效果。方法:选取成年新西兰大白兔76只,平均分为四组并建立兔颅骨外露实验模型。其中,A组(19只):于兔颅骨上方制作直径为2.0cm的圆形创面,保留骨膜,采用-120mmHg负压引流和常规换药治疗;B组(19只):实验动物处理同A组,仅采用常规换药治疗;C组(19只):在兔颅骨上制作直径2.0cm的圆形创面,剔除骨膜,治疗方法同A组;D组(19只):实验动物处理同C组,治疗方法同B组。每组各抽取10只,观察创面愈合率和创面愈合时间;其余9只分别在第7天、10天、20天、30天进行取材检测,分析疗效机制。结果:A组创面愈合时间为19.40±1.65天,B组为24.00±2.31天;C组为25.40±4.43天,D组为30.00±5.50天。运用VSD治疗和常规治疗创面愈合时间比较有统计学意义(P0.05)。结论:VSD治疗兔骨外露缺损创面能有效缩短创面愈合时间,促进血管再生,胶原蛋白合成。     

软骨细胞联合BMP/bFGF移植对关节软骨损伤的修复作用

目的：探讨同种异体软骨细胞移植联合骨形态发生蛋白（BMP）/碱性成纤维细胞生长因子（bFGF）对关节软骨损伤的修复作用。方法：取24只14周龄成年大白兔,随机分为A、B、C、D组,每组6只,于双侧膝关节软骨处制作软骨缺损模型,A组采用软骨细胞移植联合应用BMP/bFGF处理,B组采用单纯软骨细胞移植,C组采用单纯BMP/bFGF修复,D组采用磷酸盐缓冲液（PBS）作为阴性对照,于处理后8、12、24周行形态学、电镜观察及组织学评分。结果：8周时,A组关节修复面与周围结合紧密,可见大量软骨细胞出现,电镜下有软骨基质形成;B、C组仅有少量软骨细胞;D组未见修复。12周时,A组关节修复面与周围组织界限模糊,软骨细胞增殖活跃,电镜下可见成熟软骨基质;B、C组修复块周围有肉芽组织生成,电镜下可见未成熟的软骨基质出现;D组可见肉芽组织形成。24周时,A组修复面周围组织融合,电镜下软骨细胞纵行排列;B、C组关节面修复不完全,电镜下软骨细胞分布不均;D组见大量肉芽组织形成。24周时,A组组织学评分（1.87±0.65）,明显低于B组（3.49±0.71）、C组（3.43±0.83）组和D组（13.45±0.97）,差异均有统计学意义（P〈0.05）,B、C组均明显低于D组,差异有统计学意义（P〈0.05）,B、C组之间比较无明显差异。结论：软骨细胞联合BMP/bFGF移植能够促进软骨生长,提高软骨损伤的修复质量。     

rhBMP-2诱导异位软骨修复重建兔气管缺损的实验研究

目的：探讨重组人骨形成蛋白-2（rhBMP-2）作为激活物诱导异位软骨修复并重建免气管缺损的可行性。方法：取24只新西兰大白兔,制备气管前壁软骨1／3缺损模型。随机分为A、B组,每组12只,A组为实验组,在气管缺损处前壁颈前肌肉修补,多点注射rhBMP-2;B组于气管软骨缺损部位直接颈前肌群修补。术后观察动物一般情况,于4、8、12周取材进行大体观察、HE染色观察重建区域情况。结果：术后A组动物均存活至实验完成,B组因气道感染及气道分泌物堵管潴留致使实验兔死亡,其余动物出现皮下气肿,呼吸不畅等情况。组织学观察A组有明显的新生软骨细胞及少量软骨样组织,可见结缔组织包绕,周围肌肉组织完整,排列整齐,未见明显坏死组织,有少量淋巴细胞浸润。B组未见软骨组织生成,可见大量肉芽组织增生,结缔组织排列紊乱,伴少量坏死组织,大量淋巴浸润。结论：rhBMP-2可通过注射到颈前肌肉修补肌群中诱导软骨细胞和软骨样组织生成,减轻炎症反应,联合颈前肌瓣修复重建气管缺损能充分维持修复重建后的气道形态,具有减少术后皮下气肿、气管狭窄的作用,有望用于临床修复重建气管纽织缺损。     

缺氧条件下建立SD大鼠牙周炎动物模型的研究

目的:在模拟高原缺氧条件下建立稳定的缺氧SD大鼠牙周炎动物模型。方法:40只SD大鼠,雌雄各半,采用随机数字表法分为4组:常氧对照组(A组),常氧牙周炎组(B组),缺氧对照组(C组),缺氧牙周炎组(D组),每组10只。B、D组大鼠结扎牙颈部、高糖饮食、口腔涂菌及注射激素,A、C组不作任何处理,正常饮食。C、D组喂养的同时进入模拟海拔5000 m的低压氧舱。8 w后检测牙龈指数GI、菌斑指数PLI、牙周袋深度PD、牙槽骨吸收度ABL及组织病理学观察,对模型进行评估。结果:GI、PD及ABL指数在B组与A组,D组与C组,C组与A组,D组与B组比较均有统计学差异(P0.05)。PLI指数在B组与A组,D组与C组比较有统计学差异(P0.05),在C组与A组,D组与B组比较无统计学差异(P0.05)。HE染色见A、C组无明显改变,B、D组牙龈糜烂,结合上皮向根方增殖迁移,牙周膜间隙增宽。结论:将采用结扎牙颈部、高糖饮食、口腔涂菌及注射激素综合方法作用的SD大鼠放入模拟海拔5000 m的低压氧舱,建立模拟高原缺氧大鼠牙周炎模型具有可行性,为高原牙周病的防治奠基了实验基础。     

骨质中硫酸软骨素类蛋白多糖的类型和特征

采用骨质蛋白质的三步 (盐酸胍 - EDTA-盐酸胍 )提取法 ,较完全地提取兔长骨和人牙槽骨骨质中各类蛋白多糖 ( PGs) ,并采用凝胶过滤和离子交换柱层析等方法进行纯化 ,再用单克隆抗体 ( MAb2 B6、MAb3B3和 MAb1 B5)检测、分析其中 PGs的类型和性质 .结果表明 ,兔长骨中 PGs的主要类型为 DS类 ( 4 5k D)、C6S类 ( 2 0 0 k D)、C4S类 ( 4 5k D)和 COS类 ( 2 0 0 k D) PG;人牙槽骨中则主要含 DS类 PG( 4 5k D) ,和少量 COS类 PG( 4 5k D和 1 1 0 k D) ,未发现 C4S类 PG.根据此结果可以推测 ,兔长骨以混合方式 (软骨成骨和类骨质成骨 )骨化 ,而人牙槽骨则以类骨质成骨为主 .两者骨质结构和损伤后修复方式可能也有一定的差异 .     

聚己内酯/壳聚糖神经导管复合骨髓间充质干细胞促进大鼠坐骨神经损伤修复的研究

目的:观察聚己内酯/壳聚糖神经导管复合骨髓间充质干细胞修复大鼠坐骨神经缺损的效果。方法:将24只SD大鼠随机分为4组,制备右侧坐骨神经5mm缺损模型,A组聚己内酯/壳聚糖神经导管复合骨髓间充质干细胞移植组;B组聚己内酯神经导管复合骨髓间充质干细胞移植组;C组壳聚糖神经导管复合骨髓间充质干细胞移植组;D组自体神经移植组。术后每2周进行坐骨神经功能指数检测,12周时行电生理、腓肠肌湿重恢复率、组织学观察和免疫组织化学检测。结果:坐骨神经功能指数显示,A组运动功能恢复速度较B、C组快,但比D组慢。A组电生理和腓肠肌湿重恢复率的检测结果与C、D组相比无统计学意义(P0.05),但优于B组(P0.05)。组织学观察,A组再生神经纤维排列密集。S-100免疫组织化学结果表明A组有大量雪旺细胞增生。结论:聚己内酯/壳聚糖神经导管复合骨髓间充质干细胞能够促进周围神经损伤修复,效果与壳聚糖神经导管、自体神经相同,优于聚己内酯神经导管。     

The Effect of Brain-Derived Neurotrophic Factor on Periodontal Furcation Defects

This study aimed to observe the regenerative effect of brain-derived neurotrophic factor (BDNF) in a non-human primate furcation defect model. Class II furcation defects were created in the first and second molars of 8 non-human primates to simulate a clinical situation. The defect was filled with either, Group A: BDNF (500 µg/ml) in high-molecular weight-hyaluronic acid (HMW-HA), Group B: BDNF (50 µg/ml) in HMW-HA, Group C: HMW-HA acid only, Group D: empty defect, or Group E: BDNF (500 µg/ml) in saline. The healing status for all groups was observed at different time-points with micro computed tomography. The animals were euthanized after 11 weeks, and the tooth-bone specimens were subjected to histologic processing. The results showed that all groups seemed to successfully regenerate the alveolar buccal bone, however, only Group A regenerated the entire periodontal tissue, i.e., alveolar bone, cementum and periodontal ligament. It is suggested that the use of BDNF in combination with a scaffold such as the hyaluronic acid in periodontal furcation defects may be an effective treatment option.     

深低温冻存组织工程化软骨在关节软骨缺损修复的应用

目的：探讨经深低温冻存组织工程化软骨修复关节软骨缺损的可行性。方法：分离收集3周龄新西兰大白兔关节软骨细胞进行体外培养,接种于PGA三维支架材料上,复合物体外培养1周后冻存,冻存1个月后解冻、复苏及体外培养,1周后接种于已建立的双侧兔膝关节软骨缺损模型的膝关节软骨缺损处,并设对照组。分别于手术后4周、8周、12周行大体标本及组织观察。结果：大体观察结果表明,实验组与对照组缺损处均由软骨组织修复;组织学观察可以见到实验组和对照组关节软骨缺损处有密集的软骨细胞,均有软骨生成及基质分泌,两组差异无统计学意义。结论：应用深低温冻存组织工程化软骨修复关节软骨缺损的方法是有效可行的,为其进一步临床应用提供了实验依据。     

Application of autologous periosteal cells for the regeneration of class III furcation defects in Beagle dogs

The aim of this study was to evaluate the healing of class III furcation defects following transplantation of autogenous periosteal cells combined with β-tricalcium phosphate (β-TCP). Periosteal cells obtained from Beagle dogs’ periosteum explant cultures, were inoculated onto the surface of β-TCP. Class III furcation defects were created in the mandibular premolars. Three experimental groups were used to test the defects’ healing: group A, β-TCP seeded with periosteal cells were transplanted into the defects; group B, β-TCP alone was used for defect filling; and group C, the defect was without filling materials. Twelve weeks post surgery, the tissue samples were collected for histology, immunohistology and X-ray examination. It was found that both the length of newly formed periodontal ligament and the area of newly formed alveolar bone in group A, were significantly increased compared with both group B and C. Furthermore, both the proportion of newly formed periodontal ligament and newly formed alveolar bone in group A were much higher than those of group B and C. The quantity of cementum and its percentage in the defects (group A) were also significantly higher than those of group C. These results indicate that autogenous periosteal cells combined with β-TCP application can improve periodontal tissue regeneration in class III furcation defects.     

大鼠心肌缺血再灌注模型构建的改良

目的改良大鼠心肌缺血再灌注模型的构建方法,增强实用性。方法32只SD雄性大鼠随机分为四组（A组：伪手术组;B组：缺血组;C组：缺血30 min再灌注组;D组：缺血60 min再灌注组）。缺血过程在人工机械通气条件下完成,术前术中监测心电图变化,模型成功72 h后抽血进行心肌酶谱分析,并观察左室压变化。结果造模成功率为91%。B、C、D组各心肌酶含量明显高于A组（P〈0.05）,C、D组低于B组（P〈0.05）。B、C、D组左室压值低于A组（P〈0.05）,C、D组高于B组（P〈0.05）。结论改进后的模型制备方法简便易行,成功率高,评价指标符合临床应用。     

Bone regeneration by BMP-2 enhanced adipose stem cells loading on alginate gel

Adipose stem cells (ASCs) have the potential to differentiate into a variety of cell lineages both in vitro and in vivo. In this study, ASCs were harvested from normal Sprague–Dawley (SD) rats and transfected by BMP-2 gene before they were loaded on alginate. The ability of bone regeneration was determined in rat critical-size cranial defects. An 8-mm diameter defect was created in the calvarias of 36 rats; these rats were divided into three groups. In experimental group, the defects were filled with alginate gel combined with BMP-2 transfected ASCs; in negative control group, the defects were filled with alginate gel mixed with normal ASCs; in blank controls, the defects were filled with cell-free alginate gel. Four rats of each group were killed and the cranial defect sites were observed at 4, 8 and 16 weeks after surgery. There was complete repair of cranial defects in experimental group using the alginate gel loading BMP-2 transfected ASC, but only partial repair in negative controls and in the blank control. The engineering approach combining BMP-2 enhanced ASCs with alginate gel can therefore stimulate bone regeneration and repair for the large size bone defects.     

姜黄素衍生物B06对2型糖尿病大鼠心肌的保护作用及其机制探讨

目的：研究姜黄素衍生物B06对2型糖尿病大鼠心肌的保护作用及机制。方法：雄性sD大鼠35只,随机均分为正常对照组（NC组）、高脂组（肿组）、高脂治疗组（rT组）、糖尿病组（DM组）和糖尿病治疗组（DT组）（n=7）,后四组高脂喂养4周后,DM组及七rr组用低剂量链脲佐菌素（srz）诱导糖尿病,FTr组和町组用0．2mg／kg·d的姜黄素衍生物B06灌胃。用生化方法测大鼠血糖和血脂浓度,用ELISA法测血胰岛素水平,并计算胰岛素抵抗指数,用光镜和电镜观察心肌形态,用Westernblot法测心肌腺苷酸活化蛋白激酶a（AMPKa）和磷酸化腺苷酸活化蛋白激酶a（p-AMPKa）的蛋白表达。结果：HF组及DM组血糖、血脂、血胰岛素、胰岛素抵抗指数升高,经B06治疗后均下降;HF组及DM组AMPKa和p-AMPKa表达下降,经／306治疗后升高;DM纽心肌间质胶原纤维增多,心肌细胞内线粒体扩张,经B06治疗后病变减轻。结论：B06可缓解2型糖尿病大鼠心肌病变,AMPKa和p-AMPKa表达升高可能参与其中。     

米格列奈与瑞格列奈分别联合二甲双胍在新诊断2型糖尿病的临床价值比较

目的：探讨米格列奈和瑞格列奈在治疗新诊断2型糖尿病的临床价值比较。方法：将2010年1月至2012年12月间我院新诊断的120例2型糖尿病患者随机分为A、B两组,其中A组患者60例,采用瑞格列奈联合二甲双胍进行治疗,B组患者60例,采用米格列奈联合二甲双胍进行治疗。对两组患者血糖水平及12周后相应指标进行统计分析。结果：治疗3天后两组患者的空腹血糖及餐后血糖水平明显低于治疗前,B组患者血糖降低幅度大于A组（P〈0．05）;治疗12周后,两组空腹血糖及餐后血糖水平显著减小,B组减小程度高于A组;患者空腹及餐后胰岛素水平有一定的增加,B组增加程度高于A组;B组患者HbAlc的降低程度高于A组;两组同时还可以降低HOMA胰岛素的抵抗指数以及增大胰岛素的敏感指数,其中B组效果好于A组;B组患者出现低血糖的人次少于A组。结论：米格列奈与瑞格列奈分别联合二甲双胍可以短期降低新诊断2型糖尿病患者的血糖水平,在中期治疗后米格列奈联合二甲双胍可以更有效的改善患者糖尿病的各项指标。     

Evaluating the bone regeneration in calvarial defect using osteoblasts differentiated from adipose-derived mesenchymal stem cells on three different scaffolds: an animal study

The aim of this study was to investigate the effect of three different scaffolds on the viability and differentiation of adipose-derived mesenchymal stem cells (ADMSCs) to osteoblast for bone regeneration of calvarial defect in rabbit model. Adipose was harvested from the nape of 12 rabbits by direct surgery or hollow-tip cannula. Two standardized circular calvarial defects (case and control), 8 mm in diameter each, were created in all the animals. The animals were divided into 3 different groups. In group 1 (G1), the defect was filled with polyamide + ADMSC. In group 2, poly lactic-co-glycolic acid + ADMSC was used. In group 3, decellularized amniotic membrane + ADMSC was applied. In the control defect, the non-seeded scaffolds were applied for filling the defect. Decellularized pericardial scaffolds were used as a membrane on the scaffolds. The animals were euthanized 2, 4, and 8 weeks of operation and new bone formation was assessed by different analyses. Immunohistochemical (IHC) staining with osteopontin and osteocalcin antibodies was also performed. After 2 weeks of wound healing, minimal bone regeneration was detected in all groups. Almost complete defect closure was observed in all experimental groups after 8 weeks of operation, with the greatest defect closure in the animals treated with polyamide scaffolds as compared to biopsies obtained from control defects and other experimental groups. The maximal tensile load was higher in G1, 4 and 8 weeks postoperatively, suggesting the usefulness of polyamide + ADMSC for bone regeneration in calvarial defects. Results of the IHC staining demonstrated a significant difference between seeded and non-seeded scaffold in both short- and long-term follow-ups (P < 0.05). In addition, a significant difference was observed in enhancement of IHC staining of both markers in polyamide group (seeded or non-seeded) 4 and 8 weeks postoperatively in comparison with other scaffolds. It was concluded that bone regeneration in critical calvarial defect was more successful in seeded polyamide.