用流式细胞术评价一种新型人工肝材料的生物相容性

目的通过用流式细胞术(FCM)观察丙稀酰胺接枝改性聚丙烯膜 (PP-g-AAm)的生物相容性,来评价FCM在检测医用生物材料的生物相容性中的作用.方法:用材料浸提液培养L929细胞24 h后用FCM检测细胞增殖周期; 用改性前、后的膜材料分别与新提取的PRP(富含血小板血浆)和PBMC(末梢血单个核细胞)孵育培养后,用FCM分别检测血小板和PBMC的激活标志物CD62P、CD63和CD69.结果:PP-g-AAm 组的PI与阴性组及空白对照组比较差异无统计学意义(P=0.063,P=0.053),而与阳性对照组比较差异有统计学意义(P=0.002).PP-g-AAm膜组CD62P、CD63及CD69的表达率明显少于对照组(P=0.042, P=0.004,P=0.013).结论:PP-g-AAm无细胞毒性并具有良好的生物相容性,FCM在生物材料的生物相容性评价中有着广泛的应用价值.     

壳聚糖的生物相容性*

背景：壳聚糖是惟一一种被广泛应用于生物医学工程领域的碱性、带有正电荷的天然多糖,其生物相容性是决定这些应用价值的关键。 目的：综述了壳聚糖的生物相容性,包括组织相容性、血液相容性和力学相容性。 方法：由第一作者检索1990/2011 PubMed数据库、中国知网数据库及万方数据库有关壳聚糖及其衍生物在生物医学上的应用和生物相容性等方面的文献。 结果与结论：壳聚糖作为可生物降解高分子材料具有良好的组织相容性及与人体组织相匹配所需要的力学相容性,被逐渐应用于人工皮肤、手术缝合线、眼科修复、人工骨骼、牙齿修复、肿瘤治疗等方面。但壳聚糖的促凝血作用使其血液相容性很差,目前很多研究关注于寻找解决这一问题的方法,改善其血液相容性,扩展其在生物医学工程上的应用领域,使其更加安全有效地与人体心血管系统直接接触。 关键词：壳聚糖;组织相容性;血液相容性;力学相容性;生物材料 doi:10.3969/j.issn.1673-8225.2012.12.034     

血管组织工程生物支架材料细胞生物相容性实验研究

目的采用新方法研究天然生物支架材料胶原/透明质酸膜,明胶海绵的细胞相容性。方法应用WST-1法测定平滑肌细胞与材料的粘附率,增殖力,^3H-TDR掺入法测定DNA合成率,BrdU细胞标记免疫组化鉴定,分析组织工程生物支架材料的细胞相容性。结果WST-1法测定,^3H-TDR掺入法测定DNA合成率,BrdU细胞标记结果表明：胶原/透明质酸膜与平滑肌细胞的粘附率最高,细胞的增殖和代谢状况较好,明胶海绵较低。结论应用WST-1法测定,^3H-TDR掺入法测定DNA合成率,BrdU细胞标记方法研究细胞相容性方法简便可行;天然复合生物材料胶原/透明质酸膜具有较理想的细胞相容性。     

新型生物医学材料——类金刚石膜的研究进展

类金刚石因具有良好的细胞相容性，血液相容性及高耐磨性高硬度等特点，而成为一种很有发展前景的生物膜材料。本综述了20世纪90年代以来类金刚石的理化性质，生物学方面的研究进展，并对未来的发展进行了展望。     

新型生物医学材料——类金刚石膜的研究进展

类金刚石因具有良好的细胞相容性、血液相容性及高耐磨性高硬度等特点 ,而成为一种很有发展前景的生物膜材料。本文综述了 2 0世纪 90年代以来类金刚石的理化性质、生物学方面的研究进展 ,并对未来的发展进行了展望。     

基于连笔直写的微结构血管支架体外测评

目的 评价基于连笔直写的生命体微结构成形技术制备的管状支架的力学性能及其生物相容性.方法 对该血管支架采用径向顺应性、缝合强度、爆破压力等力学性能检测;通过溶血率、体外动态凝血实验、血小板黏附实验进行血液相容性的分析;采用细胞培养MTT法及细胞形态学观察方法,研究其细胞相容性.结果 血管支架的径向顺应性为（4.03±0.56）%/100mmHg,缝合强度为（204.5±72.1）N/cm2,爆破压力约为（102±8）kPa;支架的溶血率为1.75%,小于ISO规定的5%;在体外动态凝血实验中,血管支架的抗凝血性能显著优于对照组载玻片（P＜0.05）,而且扫描电镜观察到血小板黏附较少,显示出该血管支架具有良好抗凝血性能;MTT比色法结果显示其细胞毒性为0～1级;细胞形态学观察显示L929细胞在该血管支架膜片的浸提液中呈梭形或三角形,贴壁良好.结论 该血管支架具有良好的力学性能、血液相容性和细胞相容性,可以满足组织工程血管支架的要求.     

GY—131医用硅橡胶生物学评价

在医用高分子材料及其制品的发展中,硅橡胶应用最广泛,效果最显著。用硅橡胶作为基质材料占半数以上,因此我们对用于口腔、眼面及输卵管等部位的GY—131医用硅橡胶材料及制品进行了生物学考察。根据材料本身的特点及使用要求,参考国内外有关资料及标准,作了系统的生物学评价。主要针对材料局部刺激、热原、细胞毒性、致突变、整体毒性及肌肉埋植等几方面进行研究。结果显示材料溶血率小于2％;未见细胞毒性或热原反应;整体毒性未见动物死亡;未诱发微核率增加效应或突变菌株的突变现象,局部无刺激,植入初期材料周围可见轻度的炎症反应,但随时间的延长而消失。表明该材料具有良好的生理惰性,是一种不被机体降解吸收,无毒无刺激、生物相容性好的医用生物材料。     

异种脱细胞真皮替代睑板材料重建眼睑的可行性

背景：眼睑后层重建是眼睑重建的重点和难点,其中睑板替代物更是研究的焦点。异种脱细胞真皮作为一种新型的组织工程材料,在国内外烧伤整形领域,正得到广泛的研究和应用。 目的：观察异种(猪)脱细胞真皮植入兔眼睑后的组织相容性极其组织病理学变化。 方法：剥取健康小白猪全层皮肤20 cm×20 cm,制备异种(猪)脱细胞真皮基质。同时制备兔睑板全层缺损模型并植入脱细胞真皮基质,观察大体情况,并分别于第1,2,3周取移植交界处眼睑组织光镜下观察组织学的改变。 结果与结论：大体观察未见明显排斥反应及眼睑的变形;光镜下1周时可见局部炎症细胞浸润,2周时炎症细胞减少,3周时正常纤维组织长入,逐渐分割代替植入的胶原纤维,炎症反应消失。提示异种脱细胞真皮免疫原性低,并可引导新生胶原的生长,是一种良好的睑板替代物。     

脱细胞异体真皮基质与人脂肪干细胞的生物相容性

背景：脱细胞异体真皮基质具有优良的生物相容性和组织细胞诱导功能。 目的：评价人脂肪干细胞与脱细胞异体真皮基质的生物相容性。 方法：取健康成年人吸脂术后的脂肪组织分离脂肪干细胞,并行原代与传代培养,传至第3代,将细胞与脱细胞异体真皮基质联合体外培养3,7 d,倒置相差显微镜和扫描电镜观察细胞在支架材料上的黏附、生长及增殖情况,并计算细胞在材料上的黏附率;XTT比色法检测细胞的生长增殖情况。 结果与结论：脂肪干细胞在支架材料上分布均匀,24 h内细胞开始伸展、黏附,二三天完全伸展变形,以梭形为主,呈网状排列;随着培养时间延长,支架上的细胞逐渐增多;人脂肪干细胞细胞与脱细胞异体真皮基质混合培养后平均黏附率为95.03%,并保持正常的生长增殖速度,表明支架对细胞具有良好的黏附性;脱细胞异体真皮基质材料与人脂肪干细胞复合后相容性良好。     

猪脂肪干细胞与脱细胞真皮基质的生物相容性

背景：脱细胞真皮基质已广泛应用于器官或组织缺损的修补。 目的：观察猪脂肪源性干细胞与脱细胞真皮基质的相容性。 方法：酶消化法原代培养猪脂肪源性干细胞,利用流式细胞仪和多向诱导分化方法鉴定脂肪干细胞,将其与脱细胞真皮基质共培养。 结果与结论：实验成功培养出猪脂肪源性干细胞,流式细胞仪检测结果显示阳性表达CD44和CD105,不表达CD34和CD45;在诱导培养基条件下,可向脂肪细胞、成骨细胞分化。苏木精-伊红染色及扫描电镜发现干细胞能在脱细胞真皮基质表面黏附生长。脂肪源性干细胞与脱细胞真皮基质具有良好的相容性,将两者体外复合培养后,有望将复合物植入体内进行缺损组织、器官的修复。     

Promoted dermis healing from full-thickness skin defect by porous silk fibroin scaffolds (PSFSs)

Studies on skin substitutes and dermal scaffolds have been extensively carried out in the past several decades and some commercial products derived from collagen and polymers have been in marketing. Yet little research on silk fibroin based dermal scaffolds and products has been reported so far. In the present study, therefore, porous silk fibroin scaffolds (PSFSs) have been prepared by freeze drying method. The effects of PSFSs on skin recovery from full thickness defect have been examined by histological evaluation with respect to neovascularization, dermal regeneration and infiltration of inflammatory cells. In addition, tissue compatibility between PSFSs and polyvinyl alcohol (PVA) sponges (as control) has been semiquantitatively compared by scoring method. The results showed that at day 18 after implantation, new tissues formed in PSFSs whose structure was almost equal to normal skin structure where proportional distribution of functional blood vessels could be found. Furthermore, infiltration of inflammatory cells in PSFSs disappeared within 7 days. By contrast, a variety of interstices, fibrous encapsulization and moderate infiltration of inflammatory cells could be found in PVA sponges at day 18 after implantation. In summary, PSFSs has significantly promoted the skin recovery from full thickness defect, showing fibroin's outstanding tissue compatibility.     

皮肤组织工程-细胞支架的构筑及其生物相容性评价

皮肤组织工程的发展提供了一种无损伤修复创伤和功能重建的皮肤治疗模式.作为组织工程的三要素之一,细胞支架发挥着重要的作用.为满足组织工程中对细胞支架在力学性能、物理结构及生物相容性等方面的要求,我们首先制备了聚乳酸(PDLLA)、聚乳酸-己内酯(PLACL)多孔支架,并以生物相容性较好的猪的无细胞真皮(acellular dermis matrix,ADM)为参比,分别把三种材料植入大鼠背部肌层,术后定期取大鼠皮下埋藏组织进行组织学检测.结果发现PDLLA与PLACL多孔支架的降解周期、力学性能、孔隙率及其孔径都可以根据皮肤组织工程中的要求进行调控.组织学检查,移植物内无明显炎性细胞,21天后,均完全血管化且分布较均匀.说明PDLLA与PLACL的生物相容性较ADM差,但并未出现明显的异物排斥反应,两者的生物相容性基本上可以满足组织工程中对支架的要求,这为聚乳酸类人工皮肤的进一步研究提供了有意义的实验依据.     

人新型脱细胞真皮制备及性状分析

BACKGROUND: Early vascularization is crucial for wound healing. A high-porosity, macrovoid allogeneic skin leads to the rapid vascularization and cellular infiltration. OBJECTIVE: To obtain a new allogeneic skin product with high porosity, good cell permeability and good histocompatibility using an improved preparation method of human acellular dermal matrix. METHODS: Cell components of healthy human skins were removed by the improved method and the traditional method, respectively. The improved method was to remove the subcutaneous fat, eliminate the epidermis (1 mol/L NaCl solution at 37 ℃ for 24 hours) followed by shaking processing (2% NaOH at 45 ℃ for 4 hours), and then, the solution was neutralized with PBS rinsing, dried and stored at 4 ℃ for standby. We detected the porosity and degradation time in vitro of the acellular dermal matrices prepared by two methods and the cytotoxicity of the material infiltration liquid on the adipose-derived stem cells. Hematoxylin-eosin staining was used for the detection of the cell residual, the integrity of collagen and cell biocompatibility. Scanning electron microscopy was used to detect the pore size. RESULTS AND CONCLUSION: Both the two methods could completely remove the cell components, and maintain the integrity of the collagen scaffold. The porosity of acellular dermal matrix with the improved method was (93.22±0.99)%, which was significantly higher than that with the traditional method [(74.28±2.06)%; P < 0.001]. However, there was no significant difference in in vitro degradation time between the two kinds of acellular dermal matrices (P > 0.05). No obvious cytotoxicity of the acellular dermal matrix prepared with the improved method was detected. At 3-7 days of co-culture, the adipose-derived stem cells cultured on the acellular dermal matrix prepared with the improved method could penetrate the basement membrane to the deep dermis, while there was no obvious cell invasion and growth in the deep dermis prepared by the traditional method. Compared with the traditional method, the improved method is more suitable for cell infiltration and growth with higher porosity and larger pore size.       

Expansion and delivery of human fibroblasts on micronized acellular dermal matrix for skin regeneration

In order to obtain an abundant supply of autologous dermal fibroblasts for the manufacture of engineered autologous dermal substitutes, we fabricated the micronized acellular dermal matrix (MADM) microcarriers and expanded human fibroblasts on them. This novel approach eliminated the need for the repeated trypsinizations that may disrupt cell–extracellular matrix interactions and impair cell viability. This cell expansion protocol simultaneously formed an engineered particulate dermal substitute (EPDS) avoiding cell reseeded on the scaffolds process. We further tested its feasibility and effectiveness in athymic murine subcutaneous injection and full-thickness cutaneous wound model. Our results showed that MADM microcarriers retained the ultrastructure of the acellular dermal matrix, had good biocompatibility, and supported human fibroblast expansion either as a direct culture substrate or through culturing cells in conditioned medium prepared from them. In the animal study, EPDS formed a thick layer of tissue below the subcutaneous muscle tissue at 3 weeks following EPDS injection into subcutaneous tissue. In full-thickness cutaneous wound, the degree of wound healing with EPDS implantation was better than that without EPDS although healing rates were not significantly different between wounds implanted with or without EPDS. This demonstrates the potential utility of MADM not only as a cell culture substrate to expand fibroblasts but also as a cell transplantation vehicle for skin regeneration, with several advantages over current expansion–transplantation protocols for skin regeneration. In addition, EPDS may be used for cosmetic or reconstructive soft tissue augmentation in a minimally invasive fashion.     

Development of an esophagus acellular matrix tissue scaffold

A cell-extraction protocol yielding an esophagus acellular matrix (EAM) scaffold for use in tissue engineering of an esophagus, including hypotonic lysis, multiple detergent cell extraction steps, and nucleic acid digestion, was developed in a rat model. Histological techniques, burst pressure studies, in vitro esophageal epithelial cell seeding, and in vivo implantation were used to assess cell extraction, extracellular matrix (ECM) preservation, and biocompatibility. Microscopy demonstrated that cell extraction protocols using sodium dodecyl sulfate (SDS) (0.5%, wt/vol) as a detergent resulted in cell-free EAM with retained ECM protein collagen, elastin, laminin, and fibronectin. Burst pressure studies indicated a loss of tensile strength in EAMs, but at intraluminal pressures that were unlikely to affect in vivo application. In vitro cell seeding studies exhibited epithelial cell proliferation with stratification similar to native esophagi after 11 days, and subcutaneously implanted EAMs displayed neovascularization and a minimal inflammatory response after 30 days of implantation. This study presents an esophagus acellular matrix tissue scaffold with preserved ECM proteins, biomechanical properties, and the ability to support esophageal cell proliferation to serve as the foundation for a tissue-engineered esophagus.     

灌注法制备全肾脏脱细胞基质支架的体内生物相容性

背景：应用灌注法制备的大鼠全肾脏脱细胞基质支架具有良好的体外细胞相容性,但其体内生物相容性尚不明确。 目的：应用灌注法制备大鼠全肾脏脱细胞基质支架,检测其体内生物相容性。 方法：应用灌注法制备Wistar大鼠全肾脏脱细胞基质支架,进行以下实验：①急性毒性实验：在小鼠腹腔分别注射全肾脏脱细胞基质支架浸提液、生理盐水及苯酚。②溶血实验：将抗凝新西兰兔血分别与全肾脏脱细胞基质支架浸提液、生理盐水及蒸馏水混合。③热源实验：向新西兰兔耳缘静脉注射全肾脏脱细胞基质支架浸提液。④内皮刺激实验：在新西兰兔皮下注射全肾脏脱细胞基质支架浸提液,观察有无皮肤刺激反应。⑤皮下植入实验：将全肾脏脱细胞基质支架埋入新西兰兔背部皮下。 结果与结论：全灌注法制备的Wistar大鼠全肾脏脱细胞基质支架无细胞残留,未引起全身毒性反应、急性溶血反应、热源反应及皮肤刺激反应,植入兔体内具有良好的组织相容性。说明大鼠全肾脏脱细胞基质材料在动物体内具有很好的生物相容性。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

移植异种(猪)去细胞真皮基质修复深度烧伤创面的可行性：12个月随访

BACKGROUND: Skin grafting is crucial for patients with deep burns, but limited source of autologous skin grafts is an existing difficulty. OBJECTIVE: To investigate the effect of xenogeneic (porcine) acellular dermal matrix in the treatment of deep burn wounds and the feasibility of its application. METHODS: Forty-one patients with deep burn were divided into two groups according to the intention of the patients, 21 cases in control group and 20 cases in observational group, followed by autologous split-thickness skin grafting alone or combined with different (porcine) acellular dermal matrix, respectively. After 12 months of follow-up, the graft success rate at postoperative 1, 2, 3, 4 weeks and skin graft contraction rate and wound repair at postoperative 3, 6, 9, 12 months were observed and compared between two groups. Moreover, levels of inflammatory factors were detected and compared between two groups at postoperative 1, 2, 3 months. RESULTS AND CONCLUSION: The skin graft success rates showed no difference between two groups at postoperative 1, 2, 3, 4 weeks (P > 0.05). The skin graft contraction rates also showed no difference between two groups at postoperative 3, 6, 9, 12 months (P > 0.05). After 12 months of follow-up, no serious scar hyperplasia, but soft texture appeared in the control group. In the observational group, three cases presented with local pigmentation at the early stage, but it gradually subsided with time; no obvious scar, but only small, point-like scar, was visible, and the repaired wound exhibited soft touch. No adverse events and death occurred in both two groups. Experimental results show that the treatment of deep burns with autologous split-thickness skin grafting combined with xenogeneic (porcine) acellular dermal matrix is safe and effective, which can improve the quality of wound healing.