聚乙二醇交联去细胞瓣构建组织工程瓣膜复合支架

目的 采用分子量20 kDa 的枝化状丙烯酰化聚乙二醇(PEG)交联巯基化去细胞瓣,构建组织工程瓣膜复合支架,并对支架的结构稳定性、力学性能和细胞相容性进行初步检测.方法 取猪新鲜主动脉瓣叶,进行脱细胞瓣处理,制备去细胞瓣并巯基化.戊二醛固定去细胞瓣,PEG 交联巯基化去细胞瓣.形态学观察去细胞及交联效果,差示扫描量热法(DSC)测定热收缩温度,Ⅰ型胶原酶溶液测定其抗酶性分解能力,拉伸实验测定其抗拉强度和弹性模量,并采用AlamarBlue 法测定支架对MRC-5 人胚肺细胞的细胞毒性.结果 HE 染色和扫描电镜(SEM)显示去细胞完全,细胞外基质保存完整;PEG 交联后瓣叶变薄,纤维增粗呈束.PEG 交联瓣的热收缩温度为(75.16 ±0.69)℃,与去细胞瓣[(67.63 ±1.09)℃]相比有显著提高,其差异有统计学意义(P ＜0.05).PEG 交联瓣6 h 及12 h酶性分解率分别为(17.52 ±1.69)%和(48.83 ±2.67)%,与去细胞瓣[(64.91 ±2.05)%和(98.23 ±1.20)%]相比,分解率显著下降.PEG 交联瓣抗拉强度较去细胞瓣显著为高,并达到天然瓣水平,弹性模量则远高于其他各组.细胞毒性试验显示PEG 组和对照组活力细胞差别为(1.82 ±0.03)%,无统计学意义(P ＞0.05).结论 PEG 交联可显著改善去细胞瓣结构稳定性和力学性能,而无明显细胞毒性,有望用于组织工程瓣膜复合支架的构建.     

牙周细胞在离子化胶原表面的黏附与生长

目的:评价离子化胶原对不同交联剂的作用及交联后对牙周细胞的黏附与生长的影响。方法:实验于2006-10/2007-01在四川大学生物材料工程研究中心实验室完成。①通过胶原蛋白与甲醇在低温反应得到化学修饰离子化胶原,将胶原海绵和离子化胶原海绵分别与戊二醛、碳化二亚胺,京尼平在室温条件下发生交联反应,控制交联剂的终质量分数为0.006,交联时间均为24h。②通过测溶液中羟脯氨酸含量测交联产物的交联度;用胶原酶测交联产物体外降解时间;通过牙周细胞在交联产物上的生长情况(A570)确定交联产物对细胞的作用。nm结果:①吸液率和交联度:离子化胶原与各交联剂交联后吸液率都低于胶原交联后,交联度相应提高,胶原和离子化胶原与戊二醛交联其交联度最高,分别为81%,85%。②降解率:离子化胶原的降解率总的趋势低于胶原样品,离子化胶原、胶原被炭化二亚胺交联后,其降解率分别为0.5%,45.0%。③牙周细胞生长情况:离子化胶原交联产物上牙周细胞A570值高于胶原交联nm产物。结论:离子化胶原在交联过程中显示了较好的抗生物降解性能,离子化胶原交联产物在牙周细胞培养过程中显示出了低的细胞毒性,提示其对牙周细胞黏附与生长具有促进作用,可以代替胶原应用于牙周组织治疗。     

经EX-810处理的食管用修补材料

背景：天然生物组织来源材料能逐渐被吸收,同时能够诱导组织特异性结构重塑,是理想的食管修补材料。目的：观察环氧类交联剂EX-810处理的猪食管组织的性能特点。方法：用4%EX-810处理猪食管组织1～120 h,通过对交联指数的测定观察交联情况;扫描电镜观察组织交联前后的形态学变化;将新鲜的,交联3,7 d的样品进行力学性能测定,观察交联对材料力学性能的影响。结果与结论：EX-810能有效地交联组织,并能通过去除细胞、组织中的自由氨基等抗原性物质而有效降低生物组织的抗原性;同时还能保持生物组织中胶原的结构形态不受大的破坏;力学性能较处理前也有较大改善,并且还可以改变材料的应力应变曲线形状,使其更加符合食管修补材料的力学要求。提示经EX-810处理的食管修补材料既保持了天然食管材料的结构特征,同时又具有低抗原性、易于保存及更适合的力学性能等优点。     

经EX-810处理的食管用修补材料

背景:天然生物组织来源材料能逐渐被吸收,同时能够诱导组织特异性结构重塑,是理想的食管修补材料。目的:观察环氧类交联剂EX-810处理的猪食管组织的性能特点。方法:用4%EX-810处理猪食管组织1～120 h,通过对交联指数的测定观察交联情况;扫描电镜观察组织交联前后的形态学变化;将新鲜的,交联3,7 d的样品进行力学性能测定,观察交联对材料力学性能的影响。结果与结论:EX-810能有效地交联组织,并能通过去除细胞、组织中的自由氨基等抗原性物质而有效降低生物组织的抗原性;同时还能保持生物组织中胶原的结构形态不受大的破坏;力学性能较处理前也有较大改善,并且还可以改变材料的应力应变曲线形状,使其更加符合食管修补材料的力学要求。提示经EX-810处理的食管修补材料既保持了天然食管材料的结构特征,同时又具有低抗原性、易于保存及更适合的力学性能等优点。     

戊二醛交联对壳聚糖／羟基磷灰石-庆大霉素缓释材料性能的影响

背景：交联是骨组织工程材料改性的一种常用方法,但目前仍缺乏交联剂对载药人工骨材料性能影响的相关研究与报道。 目的：研究戊二醛交联对壳聚糖,羟基磷灰石-庆大霉素载药人工骨材料力学性能、降解性能及体外药物缓释行为的影响。 方法：分别制备壳聚糖质量分数为10％,20％,30％的壳聚糖,羟基磷灰石-庆大霉素载药人工骨材料与戊二醛交联壳聚糖,羟基磷灰石-庆大霉素载药人工骨材料,检测各组材料的机械强度、吸水率、降解率及体外药物释放行为。 结果与结论：壳聚糖含量为10％,20％,30％壳聚糖,羟基磷灰石-庆大霉素的抗压强度分别为（10．16±1．17）,（28．40±0．64）,（23．28±1．30）MPa,经戊二醛交联后材料的抗压强度分别增大至（36．30±1-20）,（51．60±2．08）,（36．90±3．22）MPa。壳聚糖含量为10％,20％,30％壳聚糖,羟基磷灰石-庆大霉素交联后的吸水率与降解率均低于交联前。在体外缓释的第1天,30％壳聚糖,羟基磷灰石-庆大霉素的药物释放量为42．2％,材料经戊二醛交联处理后药物释放量降至33．6％,在随后的9d,交联壳聚糖／羟基磷灰石-庆大霉素的总释放量均低于壳聚糖,羟基磷灰石-庆大霉素。表明戊二醛交联赋予了材料更好的生物稳定性,减缓了材料降解速率,显著改善了药物突释现象。     

血管内皮细胞生长因子修饰的聚乙二醇化去细胞瓣构建心脏瓣膜复合支架

目的对去细胞瓣进行聚乙二醇(PEG)化改性和血管内皮细胞生长因子(VEGF)共价修饰,以改善去细胞瓣力学性能和生物学性能,并联合内皮祖细胞构建心脏瓣膜复合支架。方法枝化状PEG末端的丙烯酰基与引入到去细胞瓣上的巯基,通过迈克尔加成反应完成去细胞瓣的PEG化,并作去细胞瓣PEG化前后以及天然主动脉瓣的生物力学测定;通过PEG化去细胞瓣上未饱和的丙烯酰基与VEGF中半胱氨酸残基上的巯基发生另一个迈克尔加成反应,对去细胞瓣进行VEGF共价修饰,免疫荧光测定VEGF修饰效果;在VEGF修饰的PEG化去细胞瓣、去细胞瓣和PEG化去细胞瓣上种植内皮祖细胞(EPCs),培养10d后行瓣膜上DNA含量测定、苏木素-伊红染色和扫描电镜。结果力学测试显示:PEG化去细胞瓣的最大抗张强度较单纯去细胞主动脉瓣明显提高(P<0.05),而与天然主动脉瓣相比无差异(P>0.05);免疫荧光检测显示:VEGF可有效地共价修饰PEG化去细胞瓣;与PEG化去细胞瓣和单纯去细胞瓣相比,VEGF修饰的PEG化去细胞瓣明显促进EPCs的增殖,并且可在瓣膜表面形成一层连续的单细胞层。结论 PEG化可明显改善去细胞瓣的力学性能,并且VEGF修饰PEG化去细胞瓣可促进支架上黏附细胞的增殖,有利于改善组织工程心脏瓣膜的构建。     

精氨酸甘氨酸天冬氨酸多肽表面修饰促进生物支架材料对骨髓来源种子细胞的黏附

背景:精氨酸甘氨酸天冬氨酸多肽具有较强的黏附性和生物支架材料可接枝结合,且不会改变材料的表面理化性质.目的:观察应用精氨酸甘氨酸天冬氨酸多肽表面修饰猪主动脉瓣去细胞支架材料对骨髓干细胞黏附性的影响.方法:采用胰蛋白酶+TritonX-100 法制备猪主动脉瓣去细胞支架材料,用YGRGDSP多肽(酪氨酸-甘氨酸-精氨酸-甘氨酸-天冬氨酸-丝氨酸-脯氨酸)进行处理,按照精氨酸甘氨酸天冬氨酸多肽的质量浓度(0.5,1.0,1.5,2.0 g/L)、反应时间(4,8,12,24 h)、反应pH值(7.0,7.4,8.0)分为不同实验组.结果与结论:茚三酮显示精氨酸甘氨酸天冬氨酸多肽可很好的交联到猪主动脉瓣去细胞支架材料,最佳反应条件为:室温、1.5 g/L精氨酸甘氨酸天冬氨酸、pH 7.4、持续振荡12 h.提示利用YGRGDSP多肽对猪主动脉瓣去细胞支架材料进行表面修饰可显著改善骨髓来源种子细胞的黏附性.     

一种新型潜在骨固定聚合物的合成与弯曲强度

目的:通过熔融缩聚法合成一种新型的不饱和聚酯酰胺(即顺酐、苯酐、丙二醇、新戊二醇、己二胺共聚物)并对其进行表征.方法:为用作可降解骨内固定高分子材料,加入一定量的交联剂及引发-促进剂室温预交联后热处理深度交联,观察热处理条件对交联后的不饱和聚酯酰胺弯曲强度和在模拟体液介质及0.1 mol/L氢氧化钠标准水溶液中降解(水解)性能的影响.结果:提高热处理时间或温度能显著增加热处理后的不饱和聚酯酰胺的弯曲强度,热处理后的不饱和聚酯酰胺(含有50%交联剂)弯曲强度最大可以达到123 MPa.在195 ℃热处理18 h后的不饱和聚酯酰胺(含有50%交联剂)在模拟体液介质中降解3个月后弯曲强度可以保持114.3 MPa.同时热处理条件和交联剂的含量对控制热处理后的不饱和聚酯酰胺水解率也起重要作用,在整个水解过程中,试样表现整体溶蚀行为.结论:实验合成的新型不饱和聚酯酰胺具有骨内固定材料的潜在优势.     

生物交联剂京尼平对静电纺明胶纳米纤维膜改性的影响

背景:生物高分子纳米纤维膜极不稳定,易水解,所以需要进行交联改性.而以往所采用的交联剂具有一定的细胞毒性,降低了材料的生物相容性.目的:用生物交联剂京尼平对静电纺明胶纤维膜进行交联处理,观察交联产物的理化性能和生物相容性.设计、时间及地点:观察性实验,于2008 03/10在东华大学生物材料与组织工程研究实验室完成.材料:将交联剂京尼平按质量比为0.0%,2.5%,5.0%,7.5%,10%加入明胶溶液中共混,通过静电纺制各纳米纤维膜.方法:扫描电镜样品纤表面喷金后在10 kV加速电压下观察纤维表面的形貌.在万能材料测试机测试其拉伸力学性能.采用MTT法测试猪动脉血管内皮细胞在纳米纤维膜上的黏附与增殖情况.主要观察指标:其混静电纺明胶纳米纤维的形态结构,力学性能、生物相容性.结果:通过扫描电镜观测发现京尼平共混交联的明胶纳米纤维尺寸略有增大,当京尼平含量为5.0%时,纤维直径最大,增大了约200 nm;力学测试显示材料的力学性能在添加京尼平之后有了明显提高,当京尼平含量为5.0%时,应力达到了(2.45±0.09)MPa,应变达到了(3.85±0.57)%;生物相容性实验表明猪动脉血管内皮细胞在经京尼平处理过的明胶纳米纤维膜上能有效地黏附与增殖.结论:含有京尼平的明胶纳米纤维膜各项理化性能都有了显著的提高,与猪动脉血管内皮细胞复合具有良好的生物相容性.     

海藻酸钠交联肝素涂层在体外循环及人工心肺支持装置管路中的应用

背景：目前国内体外循环心脏手术使用的非肝素涂层管路和插管对血液破坏大、炎性反应重,影响心脏手术后患者的恢复和生存。目的：采用生物医用高分子材料研制新型体外循环管道肝素涂层技术,并对其稳定性及抗凝血性能进行研究。方法：利用CaCl2将活化医用聚氯乙烯体外循环管道内表面修饰形成Ca2＋膜,并与海藻酸钠和肝素交联;其中Ca2＋与海藻酸钠、肝素钠中的Na＋反应,从而使线型聚合物分子发生交联,形成化学交联海藻酸钠-肝素复合物的网状结构,实现生物型材料肝素化涂层。结果与结论：CaCl2修饰活化医用聚氯乙烯体外循环管道并与海藻酸钠和肝素交联反应,形成生物型高分子材料肝素化涂层管道,试验证明肝素化涂层管道具有良好的血液相容性、稳定性、抗凝血性能,可满足体外循环中短期转流的要求。     

应用猪主动脉脱细胞基质制备新型组织工程血管支架：生物相容性及力学性能评价（英文）

背景：组织工程血管构建的关键依赖于理想的支架。猪血管作为组织工程血管构建材料已有广泛应用,但其较高的免疫原性及较差的力学强度限制了该材料作为组织工程支架的应用。目的：应用猪主动脉脱细胞基质制备一种新的具有良好机械性能及生物相容性的组织工程血管支架。方法：对猪主动脉进行脱细胞处理和热交联改性制备脱细胞血管基质支架,采用苏木精-伊红染色及生物力学分析评估其脱细胞效果及血管基质的力学性能。将人脐静脉血管内皮细胞接种于脱细胞血管基质支架中进行体外培养,评估其生物相容性。结果与结论：用1%的TritonX-100溶液处理猪主动脉84h可完全脱除血管细胞,同时不破坏血管基质结构;经真空下120℃热交联处理12h,脱细胞基质的拉伸断裂强度得到明显提高,达到1.70MPa。在该改性血管基质支架上接种人脐带静脉内皮细胞体外培养7d,扫描电镜显示内皮细胞呈现典型的血管内皮层状结构。表明猪主动脉经过脱细胞处理能够维持血管基质完整,冷冻干燥和真空热交联处理可有效提高其拉伸强度,且对血管内皮细胞具有良好的相容性。     

应用猪主动脉脱细胞基质制备新型组织工程血管支架:生物相容性及力学性能评价

背景:组织工程血管构建的关键依赖于理想的支架.猪血管作为组织工程血管构建材料已有广泛应用,但其较高的免疫原性及较差的力学强度限制了该材料作为组织工程支架的应用.目的:应用猪主动脉脱细胞基质制备一种新的具有良好机械性能及生物相容性的组织工程血管支架.方法:对猪主动脉进行脱细胞处理和热交联改性制备脱细胞血管基质支架,采用苏木精-伊红染色及生物力学分析评估其脱细胞效果及血管基质的力学性能.将人脐静脉血管内皮细胞接种于脱细胞血管基质支架中进行体外培养,评估其生物相容性.结果与结论:用1%的Triton X-100溶液处理猪主动脉84 h可完全脱除血管细胞,同时不破坏血管基质结构;经真空下120 ℃热交联处理12 h,脱细胞基质的拉伸断裂强度得到明显提高,达到1.70 MPa.在该改性血管基质支架上接种人脐带静脉内皮细胞体外培养7 d,扫描电镜显示内皮细胞呈现典型的血管内皮层状结构.表明猪主动脉经过脱细胞处理能够维持血管基质完整,冷冻干燥和真空热交联处理可有效提高其拉伸强度,且对血管内皮细胞具有良好的相容性.     

Biomaterial characterization of off‐the‐shelf decellularized porcine pericardial tissue for use in prosthetic valvular applications

Fixed pericardial tissue is commonly used for commercially available xenograft valve implants, and has proven durability, but lacks the capability to remodel and grow. Decellularized porcine pericardial tissue has the promise to outperform fixed tissue and remodel, but the decellularization process has been shown to damage the collagen structure and reduce mechanical integrity of the tissue. Therefore, a comparison of uniaxial tensile properties was performed on decellularized, decellularized‐sterilized, fixed, and native porcine pericardial tissue versus native valve leaflet cusps. The results of non‐parametric analysis showed statistically significant differences (p < .05) between the stiffness of decellularized versus native pericardium and native cusps as well as fixed tissue, respectively; however, decellularized tissue showed large increases in elastic properties. Porosity testing of the tissues showed no statistical difference between decellularized and decell‐sterilized tissue compared with native cusps (p > .05). Scanning electron microscopy confirmed that valvular endothelial and interstitial cells colonized the decellularized pericardial surface when seeded and grown for 30 days in static culture. Collagen assays and transmission electron microscopy analysis showed limited reductions in collagen with processing; yet glycosaminoglycan assays showed great reductions in the processed pericardium relative to native cusps. Decellularized pericardium had comparatively low mechanical properties among the groups studied; yet the stiffness was comparatively similar to the native cusps and demonstrated a lack of cytotoxicity. Suture retention, accelerated wear, and hydrodynamic testing of prototype decellularized and decell‐sterilized valves showed positive functionality. Sterilized tissue could mimic valvular mechanical environment in vitro, therefore making it a viable potential candidate for off‐the‐shelf tissue‐engineered valvular applications.     

Porcine radial artery decellularization by high hydrostatic pressure

Many types of decellularized tissues have been studied and some have been commercially used in clinics. In this study, small‐diameter vascular grafts were made using HHP to decellularize porcine radial arteries. One decellularization method, high hydrostatic pressure (HHP), has been used to prepare the decellularized porcine tissues. Low‐temperature treatment was effective in preserving collagen and collagen structures in decellularized porcine carotid arteries. The collagen and elastin structures and mechanical properties of HHP‐decellularized radial arteries were similar to those of untreated radial arteries. Xenogeneic transplantation (into rats) was performed using HHP‐decellularized radial arteries and an untreated porcine radial artery. Two weeks after transplantation into rat carotid arteries, the HHP‐decellularized radial arteries were patent and without thrombosis. In addition, the luminal surface of each decellularized artery was covered by recipient endothelial cells and the arterial medium was fully infiltrated with recipient cells. Copyright © 2012 John Wiley & Sons, Ltd.     

改良酶学方法获得异种血管脱细胞支架

背景:扩大脱细胞基质的孔径与孔隙率,保证其作为移植物的力学特性,是目前血管组织工程研究的热点之一。目的:观察改良的酶学方法处理异种血管的力学性能和组织相容性。方法:取猪的颈动脉作为基质,采用传统胰蛋白酶-EDTA加1%TritonX-100和0.1%氨水顺序脱细胞方案,并做了适当的方法改进,延长胰酶处理的时间,分别为胰酶处理4h,5h,6h。结果与结论:经组织学分析,脱细胞基质中无细胞成分,脱细胞的支架结构完整,随着胰酶处理时间的延长,其基质内弹力板破坏明显,孔径和孔隙率逐渐增大,缝合强度与爆破强度相对于自然血管虽略有降低,但差异无显著性意义。组织成分胶原蛋白含量也有所下降,尤其胰酶延长至6h组,其胶原蛋白含量明显低于正常未处理的自然血管(P<0.05)。结果显示延长胰蛋白酶处理时间后得到的脱细胞血管基质,具有良好的孔隙率、生物力学性能和组织相容性。     

A comparison study of different decellularization treatments on bovine articular cartilage

Previous researches have emphasized on suitability of decellularized tissues for regenerative applications. The decellularization of cartilage tissue has always been a challenge as the final product must be balanced in both immunogenic residue and mechanical properties. This study was designed to compare and optimize the efficacy of the most common chemical decellularization treatments on articular cartilage. Freeze/thaw cycles, trypsin, ethylenediaminetetraacetic acid (EDTA), sodium dodecyl sulfate (SDS), and Triton‐X 100 were used at various concentrations and time durations for decellularization of bovine distal femoral joint cartilage samples. Histological staining, scanning electron microscopy, DNA quantification, compressive strength test, and Fourier‐transform infrared spectroscopy were performed for evaluation of the decellularized cartilage samples. Treatment with 0.05% trypsin/EDTA for 1 day followed by 3% SDS for 2 days and 3% Triton X‐100 for another 2 days resulted in significant reduction in DNA content and simultaneous maintenance of mechanical properties. Seeding the human adipose‐derived stem cells onto the decellularized cartilage confirmed its biocompatibility. According to our findings, an optimized physiochemical decellularization method can yield in a nonimmunogenic biomechanically compatible decellularized tissue for cartilage regeneration application.     

Decellularized human corneal stromal cell sheet as a novel matrix for ocular surface reconstruction

The shortage of donor corneas as well as the limitations of tissue substitutes leads to the necessity to develop alternative materials for ocular surface reconstruction. Corneal surface substitutes must fulfill specific requirements such as high transparency, low immunogenicity, and mechanical stability combined with elasticity. This in vitro study evaluates a decellularized matrix secreted from human corneal fibroblasts (HCF) as an alternative material for ocular surface reconstruction. HCF from human donors were cultivated with the supplementation of vitamin C to form a stable and thick matrix. Furthermore, due to enhanced cultivation time, a three‐dimensional like multilayered construct which partly mimics the complex structure of the corneal stroma could be generated. The formed human cell‐based matrices (so‐called cell sheets [CS]) were subsequently decellularized. The complete cell removal, collagen content, ultrastructure, and cell toxicity of the decellularized CS (DCS) as well as biomechanical properties were analyzed. Surgical feasibility was tested on enucleated porcine eyes. After decellularization and sterilization, a transparent, thick, cell free, and sterile tissue substitute resulted, which allowed expansion of limbal epithelial stem cells with no signs of cytotoxicity, and good surgical feasibility. DCS seem to be a promising new corneal tissue substitute derived from human cells without the limitation of donor material; however, future in vivo studies are necessary to further elucidate its potential for ocular surface reconstruction.     

The development and implantation of a biologically derived allograft scaffold

Biologically derived scaffolds are becoming viable treatment options for tissue/organ repair and regeneration. A continuing hurdle is the need for a functional blood supply to and from the implanted scaffold. We have addressed this problem by constructing an acellular ileal scaffold with an attached vascular network suitable for implantation and immediate reperfusion with the host's blood. Using a vascular perfusion approach, a segment of porcine ileum up to 30 cm long, together with its attached vasculature, was decellularized as a single entity. The quality of the decellularized scaffold was assessed histologically and using molecular tools. To establish vascular perfusion potentials of the scaffold, a right‐sided nephrectomy and end‐to‐end anastomosis of the decellularized scaffold's vasculature to a renal artery and vein were performed in a pig of similar size to the donor animal. Lengths of ileal scaffold, together with its attached vasculature, were successfully decellularized, with no evidence of intact cells/nuclear material or collagen degradation. The scaffold's decellularized vascular network demonstrated optimum perfusion at 1, 2 and 24 h post‐implantation and the mesenteric arcade remained patent throughout the assessment. The 1, 2 and 24 h explanted scaffolds demonstrated signs of cellular attachment, with cells positive for CD68 and CD133 on the vascular luminal aspect. It is possible to decellularize clinically relevant lengths of small intestine, together with the associated vasculature, as a single segment. The functional vascular network may represent a route for recellularization for future regeneration of bowel tissue for patients with short bowel syndrome. Copyright © 2013 John Wiley & Sons, Ltd.     

A straightforward method to produce decellularized dermis‐based matrices for tumour cell cultures

Decellularized matrices are steadily gaining popularity to study the biology of cells and tissues, as they represent a biomimetic environment in which cells can recapitulate certain behaviours that share similarities with those observed in vivo. Basically, biochemistry, microstructure and mechanics of the decellularized matrices are the most valuable properties that differentiate these culturing systems from conventional bidimensional models. Several procedures to decellularize tissues have been proposed so far, with the common aim to preserve the tissue chemical/physical properties of the original tissue. However, these processes are complex, time‐consuming and expensive. In this work, we propose a cost‐effective, easy‐to‐produce decellularized dermal matrix, derived from animal skin. The chemical/physical processes to obtain the matrices proved to not alter matrix structure and did not induce cytotoxicity issues. To test the validity of the decellularized matrices as a model to study the behaviour of tumour cells in vitro, we performed microstructural and mechanical investigations as well as cell proliferation assays. In particular, three different tumour cell lines were used, which proliferated and invaded the matrix with no additional treatments. Decellularized skin scaffold, presented in this work, could be a strong competitor for conventional 3D systems like synthetic porous scaffolds or hydrogels. Copyright © 2016 John Wiley & Sons, Ltd.     

改良酶学方法获得异种血管脱细胞支架

背景：扩大脱细胞基质的孔径与孔隙率,保证其作为移植物的力学特性,是目前血管组织工程研究的热点之一。目的：观察改良的酶学方法处理异种血管的力学性能和组织相容性。方法：取猪的颈动脉作为基质,采用传统胰蛋白酶-EDTA加1%TritonX-100和0.1%氨水顺序脱细胞方案,并做了适当的方法改进,延长胰酶处理的时间,分别为胰酶处理4h,5h,6h。结果与结论：经组织学分析,脱细胞基质中无细胞成分,脱细胞的支架结构完整,随着胰酶处理时间的延长,其基质内弹力板破坏明显,孔径和孔隙率逐渐增大,缝合强度与爆破强度相对于自然血管虽略有降低,但差异无显著性意义。组织成分胶原蛋白含量也有所下降,尤其胰酶延长至6h组,其胶原蛋白含量明显低于正常未处理的自然血管（P〈0.05）。结果显示延长胰蛋白酶处理时间后得到的脱细胞血管基质,具有良好的孔隙率、生物力学性能和组织相容性。