携载rhBMP-2微球的新型可注射自凝固复合人工骨的制备及特性研究

[目的]制备一种具有良好降解性和成骨活性、可注射的自凝固新型骨修复材料。[方法]制备携载rhBMP-2的聚乳酸与聚乙醇酸共聚物（PLGA）微球，并将其与rhBMP-2／磷酸钙骨水泥（CPC）复合，制备出rhBMP-2／PLGA微球／CPC复合人工骨。探讨了材料的特性，包括形貌、固化时间、抗压强度及反映材料体外降解速度的指标一体外降解液Ca、P浓度变化，测定复合材料rhBMP乏的释药速度及体外诱导MSCs细胞成骨分化的能力。[结果]与单纯CPC-rhBMP-2相比，复合材料的固化时间少量增加，抗压强度下降明显。体外降解速度及体外释药明显提高，释放的rhBMP-2具有骨诱导活性。[结论]rhBMP-2／PLGA微球／磷酸钙骨水泥新型复合人工骨是具有良好应用前景的骨修复材料。     

曲安奈德PLGA微球对兔上腭部创面瘢痕形成影响的实验研究

目的:探讨曲安奈德PLGA微球对创面瘢痕愈合中瘢痕生成的抑制作用,研究局部应用曲安奈德的最佳给药途径。方法:3组共24只兔的上腭部创面模型,分别应用曲安奈德PLGA微球以及曲安奈德悬浊液,在瘢痕形成不同时间后切取瘢痕标本进行HE、VG染色及酶组织化学染色并电镜观察其组织形态学改变来分析曲安奈德不同给药途径的差异。结果:三组兔上腭创面愈合时间不同,表面颜色、质地不一样。电镜、常规染色以及免疫组织化学染色皆说明曲安奈德PLGA微球有明显的瘢痕抑制作用,且效果高于曲安奈德悬浊液。结论:曲安奈德PLGA微球能抑制瘢痕的增殖过程,使瘢痕组织纤维化过程明显减轻。合并手术一次性给药治疗瘢痕有很好的临床价值。     

以PLGA为支架的组织工程脂肪的实验研究

目的：研究以PLGA为支架材料形成的组织工程脂肪作为软组织填充材料的可行性。方法：实验组：取兔的脂肪间充质干细胞，行体外扩增培养至第3代，达到一定数量后成脂诱导1周，接种于PLGA支架上，继续诱导1周，置于兔背部肩胛骨两侧皮下，分别于4、8周从大体和组织学方面观察脂肪形成情况；对照组：单纯以未接种细胞的支架移植于兔背部肩胛骨两侧皮下，于相同时间点进行检测。结果：实验组植入4周后，该复合物在支架中有脂肪形成，并可见细胞突入支架材料中；8周后支架材料继续降解，脂肪组织形成明显，伴有少量血管长入。对照组支架材料逐渐崩解，未见脂肪组织形成。结论：以PLGA为支架材料形成的组织工程脂肪作为软组织填充材料提供了一个好的思路。     

Adsorption of Salmon Calcitonin to PLGA Microspheres

Purpose. The interaction of salmon calcitonin (sCT) and poly (d,l-lactide-co-glycolide) was detected during preparation and evaluation of microspheres. The purpose of this study was to quantitate the extent and nature of the interaction. Methods. Blank microspheres were prepared by an aqueous emulsification solvent extraction technique. Adsorption studies were carried out at six concentrations of sCT and three concentrations of microspheres. Adsorption isotherms were constructed using the Langmuir and Freundlich treatments. Results. Adsorption at 1 mg/ml sCT concentration resulted in almost complete depletion of the peptide from the adsorption medium with the time to reach maximum adsorption decreasing with increasing microsphere concentration. At sCT concentrations below 100 µg/ml, a true equilibrium occurred in 1 hour or less while at higher concentrations (up to 350 µg/ml), a transient equilibrium was reached in 1 to 2 hours, followed by further adsorption of the peptide. The adsorption followed the Langmuir isotherm at concentrations below 200 µg/ml, indicating formation of a monolayer. Multilayer interaction, described by the Freundlich isotherm, occurred at higher concentrations and resulted in complete depletion of sCT from the adsorption medium. The affinity constant during monolayer formation was 0.09 and the plateau surface concentration was 5.1 µg/mg. The multilayer peptide-peptide adsorption showed a lower affinity (0.025) but higher capacity (24 µg/mg) than the monolayer peptide-polymer adsorption. Conclusions. The results show that poly (d,l-lactide-co-glycolide) microspheres have a high adsorption capacity for sCT which must be considered in formulating a controlled delivery product of this peptide.     

胎儿骨髓间充质干细胞复合PLGA支架的形态学观察

目的 探讨胎儿骨髓间充质干细胞(fetal bone marrow mesenehymal stem cells，fBMSCs)与低热高压法制作的聚丙交酯-乙交酯(PLGA)支架材料复合培养。构建组织工程化骨的可行性。方法 预制PLGA支架材料，取5月胎儿肱骨和股骨骨髓，用单核细胞分离液分离fBMSCs，含双抗的低糖DMEM原代和传代培养，倒置显微镜下观察细胞生物学特性，用F1TC标记的抗CDl05、CD44和用PE标记的抗CD34、CD29对第4代细胞进行流式细胞鉴定，并与PLGA支架材料复合培养l周后行扫描电镜观察。结果原代及传代培养的fBMSCs增殖较迅速，第4代CD105阳性细胞占84．08％，CD29阳性细胞占88．77％，CD44阳性细胞占89．53％。复合PLGA支架材料细胞生长状态良好。结论 骨髓间充质干细胞(bone mesenchymal stem cells，BMSCs)是骨组织工程适宜的种子细胞，与低热高压制作的PLGA支架材料复合可体外构建组织工程骨。     

以冻干骨为平台重建组织工程化关节软骨的研究

目的:探索在兔冻干异体骨关节平台上利用组织工程的方法再造人工关节软骨的可行性。方法:实验分成四组,运用组织工程方法将不同构成的同种异体兔软骨细胞-支架材料复合物种植于兔冻干异体骨关节平台上,植入实验兔体内,3月后取材观察体内成软骨情况。结果:仅种植兔软骨细胞的冻干骨关节支架上未见新生软骨形成。粘合有种植兔软骨细胞的PLGA膜片的冻干骨支架关节腔内见新生类软骨样物出现。自体软骨层孔洞缺损模型使用软骨细胞-PLGA膜片复合物可修复软骨缺损。结论:软骨细胞-PLGA膜片复合物实验动物关节内可形成软骨样物;但在冻干骨关节平台上尚不能形成有功能意义的关节软骨层。     

PLGA为佐剂的OVA纳米疫苗皮下注射可预防小鼠过敏反应性气道炎症和调节Th1/Th2反应

目的 探讨聚乳酸.羟基乙酸共聚物[poly（D,L-lactic-co-glycolic）acid,PLGA]包裹的卵清蛋白（OVA）纳米癌苗（POM）对哮喘小鼠的免疫治疗效果.方法 包裹不同剂量（低、中、高）的OVA纳米粒子和对照（OVA、空白纳米粒子、PBS）通过皮下注射给予小鼠,再用OVA进行致敏和激发,通过肺组织学、支气管肺泡灌洗液（BALF）细胞计数、测定BALF和脾细胞培养上清液中细胞因子的含量,观察小鼠呼吸道炎症和免疫学改变.结果 肺部组织学和BALF中细胞计数结果显示,与PBS对照组相比,OVA治疗组、中剂量和高剂量OVA纳米组的肺部嗜酸性浸润显著减轻,BALF中总细胞和嗜酸性细胞显著减少.卸胞因子测定结果显示,与PBS对照组相比,中、高剂量OVA纳米组的BALF和脾细胞培养上清液中IFN-γ显著升高,Ⅱ,4水平显著降低.OVA治疗组中IL-4水平显著下降,而IFN-γ水平无显著差异.结论 OVA纳米疫苗可预防哮喘嗜酸性气道炎症,其可能的机制之一是调节了过敏性哮喘的Th1/Th2失平衡反应.     

心肌细胞吞噬聚乳酸-乙醇酸纳米粒子的形态学实验

目的　探讨心肌细胞吞噬聚乳酸—乙醇酸 ( polylactic polyglycolicacid ,PLGA)纳米粒子的形态学机理 ,制备组织细胞吞噬纳米粒子的相关模型 ,为纳米粒子作为包载各类药物或基因载体提供实验形态学依据。方法 :将制备的PLGA纳米粒子加入生理盐水中 ,超声振荡成注射用纳米粒子悬浮液后 ,注射至活体家兔心肌组织内 ,4 8小时后取材切片、电镜下观察。结果 :心肌细胞胞质内及胞核内可见大量被吞噬的纳米粒子 ,并且出现了PLGA纳米粒子被吸收、降解的迹象。结论 :心肌细胞完全可以通过吞噬方式大量摄取 ;PLGA纳米粒子 ,将PLGA纳米粒子作为向心肌细胞内转运某种药物或基因的载体是完全可行的。     

Effects of EGF and bFGF on Irradiated Parotid Glands

Radiotherapy is common treatment for head-and-neck cancer, during which the salivary glands are often included within the radiation field. The most common side effect of this treatment is the development of oral dryness (xerostomia). This study considers the administration of epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF or FGF2) at physiological concentrations before and after irradiation in order to repair radiation-induced damage in salivary gland cells. As a preliminary examination of the efficacy of this approach we have characterized the effects of EGF and bFGF on the apoptotic response of 15-Gy irradiated rat salivary glands in vitro. Also, we have developed a controlled-release delivery system to effectively administer the growth factor to the gland since local delivery is essential to avoid unwanted protection of cancer cells. In vitro administration of bFGF prior to and immediately after irradiation partially protected (44%) the rat parotid gland. EGF did not show any significant radioprotective effect on parotid glands after a single 15-Gy irradiation dose. Encapsulation, storage and release of bFGF from biodegradable 50/50 PLGA microspheres did not affect the functionality of the growth factor in vitro.