无细胞骨胶原基质的理化性能和组织相容性研究

制备了一种无细胞骨胶原基质(Acellular bone collagen matrix,ABCM)和ABCM-PDLLA复合材料。对材料的各种理化性能进行测试,并通过酶联免疫吸附(Enzyme linked immunosorbent assay,ELISA)和兔桡骨骨干缺损修复实验评价材料的组织相容性。结果表明材料中主要的骨矿成分和有机成分分别是碳酸盐羟基磷灰石和胶原,脂肪和细胞成分被完全脱除;ABCM材料具有较好的力学强度和微孔结构,加入PDLLA有助于提高材料的力学性能;ELISA检测显示材料引起的免疫反应持续时间较短,加入PDLLA可降低免疫反应。体内移植实验表明材料主要以传导成骨方式实现骨的修复和替代。总之,两种材料均具有较好的组织相容性,ABCM-PDLLA具有更好的力学性能和较低的免疫原性,更适合用作骨移植材料或骨组织工程支架材料。     

巴布剂基质制备研究

目的研究出一种比较理想的巴布剂基质。方法选用多种高分子材料和保湿剂组合、试验，以粘着力、赋形性和重复粘贴性为指标，考察其质量。结果制备出了一种以丙烯酸树脂Ⅱ、Ⅲ号为粘性剂，聚乙二醇、甘油为保湿剂，硫酸镁作交联荆较为理想的巴布剂基质。结论巴布剂是以水溶性高分子材料为基质的一种新型外用贴剂。具有粘性适中、不过敏、可反复贴用、作用时间长，药物穿透性好等优点。     

可溶性MA104细胞牛轮状病毒受体的制备及鉴定

可溶性ＭＡ１０４细胞牛轮状病毒受体的制备及鉴定訾自强，孙文敏，徐燕，任中原在病毒致病过程中，病毒和细胞表面的病毒受体的结合是感染的第一步。因此，病毒受体的作用是很重要的。然而，研究病毒受体，如受体的理化性状、结构、功能的精细分析，首先要将受体分子从细...     

脱细胞神经基质的制备方法的研究进展

面神经损伤的修复是口腔颌面外科难题之一,筛选促进神经再生理想的移植物,是解决这一难题的关键。自体神经移植被认为是修复面神经损伤的“金标准”,但可引起多种并发症,且自体神经材料来源有限,常无法满足临床需要。近年来,     

医用壳聚糖膜的制备与性能研究

作者以天然高分子壳聚成膜材料，制备了医用壳聚糖膜。经性能测试和体外降解实验表明：该膜具有较好的力学性能和可降解性，可用于外科手术以防止术后粘连。     

不同基质刚度三维丝素支架制备及其性能研究

目的 使不同浓度丝素蛋白(silk fibroin, SF)与胶原蛋白(collagen, COL)发生氢键结合,通过联合凝胶体系,制备出一种刚度可调的复合支架,并对其理化性质进行表征。方法 分别将不同质量SF加入海藻酸钠溶液中溶解,然后分别加入不同浓度COL溶液和一定质量碳酸钙(CaCO₃)粉末。取出混合溶液,加入一定质量葡萄糖酸内酯(gluconic acid lactone, GDL)粉末,成形后得到SC1、SC2、SC3组水凝胶,冷冻干燥后得到不同SF支架。结果 成功制备刚度可调的SF支架,SC1、SC2、SC3组压缩模量分别为(17.31±2.73)、(24.12±1.81)、(32.54±1.81) kPa。观察其内部结构,发现SC1～SC3组材料孔隙越来越小,数量越来越少,材料亲水性能越来越好。结论 通过改变SF与COL溶液浓度可以制备具有不同基质刚度三维多孔支架,且SF和COL的浓度与SF支架材料的压缩模量、吸水率、保水率、溶胀率成正比,与孔隙率成反比。研究结果有望为构建可用于诱导间充质干细胞成骨分化的适宜基质刚度骨支架提供理论指导。     

新型壳聚糖纳米氧化铁磁珠的制备和理化性能测定

目的 研制一种新型壳聚糖纳米氧化铁磁珠,并检测其理化性质,为更加优化的免疫磁珠技术奠定基础.方法 采用凝聚沉淀的方法,通过在纳米氧化铁磁珠表面包被一层壳聚糖分子,制备出壳聚糖纳米氧化铁磁珠.并对磁珠的粒径大小、显微特征、壳聚糖外壳和磁珠内核的关系以及壳聚糖磁珠的理化性能进行检测、分析.结果 制作过程简便、易行,制作出的壳聚糖纳米氧化铁磁珠粒径均匀,成球性与稳定性好.结论 成功制备了新型纳米壳聚糖氧化铁磁珠,为靶向药物载体、细胞分离、核酸提取等应用提供了实验材料.     

煅烧牛松质骨的制备、理化性能及生物相容性研究

探索一种新的植骨材料— β-磷酸三钙 (β- TCP)型煅烧牛松质骨的制备方法 ,研究其孔径、孔隙率、材料的成分、抗压和抗折强度等理化性能 ,并通过溶血试验、微核试验、全身急性毒性试验、局部刺激试验、热源检测试验、体内植入试验等研究表明 ,β- TCP型煅烧牛松质骨具有一定的强度 ,合适的孔径 ,微孔互相连通 ,有可靠的生物安全性 ,可作为植骨材料 ,用于临床。     

人造血液基质聚合牛血红蛋白的抗原性研究

目的 通过动物实验,推导人造血液基质聚合牛血红蛋白（poly-BHb)输入人体后可能引起的免疫反应。方法 制备兔抗人Hb、兔Hb和poly-BHb的抗体,研究对兔而言这3种Hb的抗原性的强弱和它们之间抗原性的交叉程度。将大量poly-BHb溶液输入家兔体内,观察家兔的免疫反应。通过计算机模拟,比较天然人、兔、牛Hb之间氨基酸序列、空间结构和抗原位点的差异。结果 天然牛Hb与人Hb分子的构象几乎一致,poly-BHb与人Hb有高度相似的抗原性。poly-HBb反复大量输入家兔休丙未能测出抗体。结论 从结果初步推论,poly-BHb溶液有限次数输入人体,引起免疫反应的可能性较小,但尚需要更多的试验及人体安全性试验加以证实。     

NaOH消蚀法制备胎儿脱细胞真皮基质的研究

目的：探索NaOH消蚀法对胎儿真皮基质的成分及其形态结构的影响。方法：使用NaOH消蚀法制备胎儿皮肤的脱细胞真皮基质(ADM)，应用组织化学和免疫组织化学染色方法检测细胞的余留及细胞外基质成分的破坏情况，应用扫描电镜观察ADM的构筑。结果：NaOH消蚀处理16h即可去除真皮细胞成分。制备的ADM韧性好，勿需戊二醛交联。弹性纤维的含量有所减少而胶原纤维形态完整，排列正常，保留了基本的真皮构筑。HLA—DR呈阴性，Fibronectin(FN)呈弱阳性，Laminin(LN)和Ⅳ型胶原均呈阴性。结论：NaOH消蚀法简易经济，去除细胞成分较彻底，所制备的ADM韧性好，胶原纤维形态完整，排列正常，但对基膜的成分及结构破坏较重。     

The Effects of Collagen Fragments on the Extracellular Matrix Metabolism of Bovine and Human Chondrocytes

Cartilage matrix degradation generates collagen type II fragments. The objective of this study is to explore the possibility that these collagen fragments may be part of an endogenous metabolic feedback. Initially, collagen fragments were extracted from normal or osteoarthritic cartilage, as part of a matrix fragment preparation. Later, collagen fragments were generated by digestion of bovine collagen type II with bacterial collagenase (col2f). These fragments were added to cultures of isolated chondrocytes (bovine and human) and cartilage explants (human). In a dose-dependent manner, col2f caused inhibition of cell attachment to collagen, inhibition of collagen synthesis, and induction of matrix degradation. In addition, when col2f were added to human cartilage explants, an induction of gelatinase activity was detected in the media. These data sets present first evidence that degradation products of collagen may be directly involved in the regulation of cartilage homeostasis.     

高纯度猪软骨II型胶原的制备与检测

探讨以猪透明软骨为原材料制备高纯度II型胶原的方法,为批量化生产提供依据.以猪透明软骨为原料,采用盐酸胍抽提蛋白多糖,酸性条件下酶解,中间过程经多步纯化去除杂胶原、降解及变性产物,最后经Sepharose H.P.阴离子柱层析纯化制备出产品并以Sigma公司产品作对照.SDS-PAGE电泳、氨基酸成份分析和紫外最大吸收光谱的结果表明,所提取的II型胶原性质符合文献报导,纯度比Sigma公司的产品高.所提取的II型胶原为高纯度的II型胶原,由于采用肉用猪作原料,来源丰富,成本低,适合批量化的产品开发.     

生物性骨载体的制备

对生物性骨载体（Ｂｉｏｌｏｇｉｃａｌ Ｂｏｎｅ Ｃａｒｒｉｅｒ，ＢＢＣ）的制备方法，成分分析、动物体内植入试验、生物力学强度，超微结构，交叉抗原性，临床应用等一系列研究做了详尽的报道，阐述了为尽量降低ＢＢＣ抗原性而采取综合处理方法的合理性，并分析归纳出ＢＢＣ作为骨替代材料的优点；（１）良好的生物相容性；（２）合适的力学强度；（３）天然的多孔结构和合适的孔隙率；（４）能为宿主吸收替代；（５）来源广泛     

细胞外基质材料在骨组织工程中的研究进展

由疾病、外部创伤等原因引起的大骨骼缺损的治疗需要通过骨移植手术,寻找安全易得的替代骨已经成为临床上的重要课题,近年来快速发展的组织工程骨为解决这一难题提供了一种新的途径。支架材料作为组织工程的核心要素,其表面性状、结构,机械性能和生物学性能均能调控细胞的各种生命活动和体内组织的修复再生。细胞外基质由于其天然性、低免疫排斥性和优异的生物相容性等特点,已被广泛用作再生医学的支架材料。通过回顾近些年来细胞外基质材料在骨组织工程中的应用,阐述多种细胞外基质材料的构建修饰方法及其体外、体内的生物学效应,并对其在骨再生领域的应用前景进行展望。     

Age-Related Changes in the Synthesis of Matrix Macromolecules by Bovine Articular Cartilage

Calf and mature cow articular cartilage was labeled in vitro with [³⁵S]SO₄ and [³H]glycine and kinetics of incorporation of both isotopes by cartilage fragments was determined by scintillation spectroscopy. The cartilage fragments were then extracted in sequence with 4M GuHCl (Guanidium chloride) and pepsin. The pepsin digest was adjusted to 1.3 M NaCl and pepsin-solubilized collagen salted out. The 4M GuHCl extract, collagen and pepsin-resistent residue were then freeze-dried. The 4M GuHCl extract was further fractionated by DEAE (Diethylaminoethyl) 52 ion exchange chromatography to obtain protein and PG (Proteoglycan) fractions. The protein fraction was also characterised by SDS-PAGE and PG fraction by Sepharose C1–2B chromatography under associative conditions in the presence and absence of an exogenous HA (Hyaluronic acid). The GAG (Glycosaminoglycan) side chains of the PG samples were analysed by Sephadex G-200 column chromatography and their composition determined by paper chromatography after chondroitinase ABC digestion.

Linear incorporation of both isotopes was observed from 1 to 18 hours of incubation and roughly equal amounts of f³⁵S]SO₄ counts were found on per cell bases in both cartilages although less [³H]glycine was incorporated by cow chondrocytes. It was also found that calf chondrocytes synthesize much greater proportion of the collagen whereas the cow cells synthesize PGs of smaller hydrodynamic sizes, bearing shorter GAG side chains that are enriched in KS (Keratan sulfate) and Ch-6S (Chondroitin-6 sulfate isomer). A failure of cow ³⁵S-PGs monomers to interact with an exogenous HA in the presence of other extracted components was also demonstrated.

The relevance of these findings for the mechanism of cartilage damage in aging and osteoarthritis is discussed.     

The Osteoinductive Activity of Bone Morphogenetic Protein (BMP) Purified by Repeated Extracts of Bovine Bone

Native bone morphogenetic proteins (BMPs) extracted from bone have been used clinically to stimulate bone regeneration and repair. However, preparation of purified BMP is a laborious process. This study investigated the yield, activity and cost effectiveness of repeatedly extracting the same bone matrix to produce purified BMP.

While repeated extraction was able to increase the yield 62% the activity of the partially purified BMP in later extracts decreased both in vitro and in vivo. This decline in activity appears to be due to an increase in non-BMP contaminants, such as collagen, in the extracts. When the first three extracts were combined and processed together activity was equivalent to that of the first extract. A simple analysis based on the cost of reagents used and the time required for purification indicates that separate processing of the extracts is inefficient while combining the first and second extracts and processing them together would result in a small cost saving.

Based on this study we would recommend that the demineralized bone matrix be extracted no more than twice and that the extracts be combined for further processing.     

微波辅助脱钙技术在猪脱钙骨基质制备中的应用研究

以猪后腿骨干骺端为对象,考察微波辅助脱钙技术在脱钙骨基质制备过程中的作用.以脱钙骨的脱钙率考察微波对脱钙效果的影响;利用Micro-CT和显微镜观察脱钙骨的大体三维结构;利用Micro-CT测量其孔隙率,利用显微镜及Digimizer3.1软件测量其孔径大小;利用物性测试仪测量脱钙骨的压缩模量以表征其力学性能.结果表明,脱钙骨脱钙率的测量结果显示,与传统脱钙法相比,微波制备法仅需2h就可使脱钙率达到100％,脱钙时间可缩短82％;且制备的脱钙骨具有良好的孔隙率和互相连通的孔径结构,孔隙率达到85％以上,孔径大小为( 345±76) μm,与传统制备方法无显著差异(P＞0.05);并具有良好的力学性能,在脱除90％以上的钙后脱钙骨的压缩模量仍可达到7.11 N/mm2.微波技术可以有效提高脱钙骨基质的制备效率,且制备效果良好,为微波技术在脱钙骨制备领域的应用提供了试验依据.