MRI体外示踪SPIO标记人脐带间充质干细胞的实验研究

目的:研究超顺磁性纳米铁颗粒(superparamagnetic ironoxide particles,SPIO)体外标记人脐带间充质干细胞(HuCMScs)及MRI成像示踪的可行性.方法:从人胎儿脐带中分离培养、扩增脐带间充质干细胞(HUCMSCs),分别采用0 μg/ml,25μg/ml,50μg/ml浓度的SPIO标记0.5×106,1×106,2×106和10×106 HUCMSCs.普鲁士蓝染色和透射电镜鉴定细胞内铁颗粒情况,并用3.0T MR/离体扫描T1WI,T2WI,GRE/300序列成像,测定细胞群信号.结果:不同数量的HUCMSCs与0 μg/ml,25 μg/ml,50 μg/ml浓度的SP10共同培养18小时,普鲁士蓝染色发现标记的细胞随标记浓度的升高染色程度逐渐加深.透射电镜检查显示细胞内含致密铁颗粒.离体MRI不同序列测定不同浓度SPIO标记相同数量的细胞群,GRE/30°和T2WI测定的各组之间均有统计学差别,501μg/ml与25μg/ml组分别与0μg/ml组之间有显著统计学差别(P<0.05);同一浓度SPIO标记人脐带间充质干细胞,信号强度与细胞数量有关(P<0.05).结论:SPIO可以标记人脐带间充质干细胞,应用MRI可以对其进行体外示踪和监测.     

硅化超顺磁氧化铁纳米颗粒标记人羊膜间充质细胞的效率及其对细胞增殖的影响

旨在探讨一种新型硅化超顺磁性氧化铁纳米颗粒标记人羊膜间充质细胞的最佳方法, 并检测其对细胞增殖的影响. 用不同浓度的Si-SPIO和多聚赖氨酸混合制备PLL-Si-SPIO复合物, 标记体外培养的hAMCs. 利用普鲁士蓝染色和透射电子显微镜等方法对Si-SPIO的标记情况进行分析鉴定. 分析Si-SPIO标记后1~4周铁颗粒在细胞内的维持与稳定. 应用MTS分析法探讨经Si-SPIO标记后hAMCs的增殖活性. Si-SPIO标记后的hAMCs移植到小鼠纹状体内1周, 鉴定Si-SPIO阳性细胞的存活与分布. 观察发现, hAMCs经Si-SPIO标记后细胞内可检测到大量铁颗粒, 铁颗粒能在细胞内维持4周以上. Si-SPIO标记具有浓度依赖性, 最适浓度为20 µg/mL; 较低浓度的Si-SPIO对细胞增殖活力没有显著影响. 移植到小鼠脑内1周后可见Si-SPIO阳性细胞. 结果可知, 浓度为20 µg/mL的Si-SPIO标记hAMCs可获得良好的标记效果, 并且不影响细胞的增殖活力.     

脂肪干细胞与间充质干细胞体外诱导分化为心肌细胞的差别

本文旨在探讨脂肪干细胞(adipose-derived stem cells, ASCs)和骨髓间充质干细胞(mesenchymal stem cells, MSCs)在组织含量、体外培养和诱导分化为心肌细胞方面的差别.ASCs从新西兰白兔皮下脂肪组织提取,MSCs从大鼠四肢长骨骨髓提取,体外培养扩增,免疫细胞学方法鉴定.采用细胞集落形成法检测组织中干细胞的含量.将不同代的干细胞用不同浓度的5-氮胞苷诱导,观察其形态变化,免疫细胞化学方法检测诱导后细胞是否转化为心肌细胞.结果显示,体外培养的ASCs呈短梭形,分布均匀,生长迅速,细胞形态单一、稳定.MSCs原代生长非常缓慢,呈簇生长,细胞纯度偏低,容易混杂其它细胞类型,传代细胞容易分化和老化.脂肪组织中ASCs含量显著高于骨髓中MSCs含量,且前者含量受年龄影响小.5-氮胞苷诱导ASCs分化为心肌细胞的有效浓度为6～9μmol/L,而MSCs在3～15μmol/L 5-氮胞苷诱导下可见心肌细胞形成.ASCs诱导分化的心肌细胞呈球形细胞团,MSCs分化的心肌细胞呈条形或棒状,其心肌细胞分化率低于ASCs.幼年动物MSCs的组织含量和心肌细胞分化率均高于老年动物,而ASCs受动物年龄影响较小.结果表明,ASCs在组织含量、细胞纯度、生长速度和心肌细胞分化率等方面均明显优于骨髓MSCs,在心肌细胞再生方面较MSCs具有更大的优势.     

VEGF靶向分子探针的制备及对卵巢癌细胞SKOV3体外成像的初步研究

目的:制备分子探针VEGF-USPIO和研究其在体外对卵巢癌细胞的靶向成像作用。方法:采用化学偶联法将VEGF抗体与USPIO连接,构建成具有免疫活性的靶向分子探针VEGF-USPIO。用CCK-8法检测该探针对卵巢癌细胞株SKOV3细胞活性的影响;普鲁士蓝染色法检测细胞磁性标记情况,并对经过标记的细胞进行体外磁共振成像,观察其对磁共振信号强度的影响。结果:成功合成了分子靶向探针VEGF-USPIO;当探针浓度在60μg/mL及以下时对细胞活性无影响(P0.05);细胞普鲁士蓝染色结果显示标记了靶向探针VEGF-USPIO的细胞其胞膜及胞浆含铁颗粒沉积较多;细胞体外磁共振成像结果显示经靶向探针标记的细胞,T2WI信号强度较未标记探针的对照组细胞降低(P0.05)。结论:所合成的VEGF-USPIO通过磁共振信号强度的变化实现MRI成像。     

分离与培养人脂肪组织间充质干细胞的研究

目的：探索人脂肪组织源性间充质干细胞（ASCs）的分离、体外培养,为其广泛应用提供实验依据。方法：无菌条件下获取腹部手术病人皮下脂肪组织,酶消化法分离、培养ASCs,观察细胞形态并绘制细胞生长曲线,计算细胞群体倍增时间;对第2代细胞进行免疫组织化学染色,鉴定其表面分子CD44表达;取2—4代细胞用含体积分数为10％胎牛血清、1％青链霉素原液、1μmmol／L地塞米松、10μmmol／L胰岛素、0．5mmmol／LIBMX的高糖DMEM培养基中诱导培养一周,观察细胞形态变化,并用油红“O”染色定性。结果：人脂肪组织中含有大量间充质干细胞,呈成纤维细胞样贴壁生长,细胞群体倍增时间为55h左右;免疫化学染色鉴定CD44阳性;成脂诱导分化一周,可见细胞内有大量脂滴,油红“0”染色可见胞浆内有大量红染颗粒。结论：建立了一种自人体脂肪组织分离,培养ASCs经济简便的方法,为其能够作为组织工程理想的种子细胞及广泛应用于临床提供实验依据。     

荧光磁性纳米粒子标记间充质干细胞靶向胃癌研究

目的:探索一种高效的早期胃癌诊断体系.方法:荧光磁性纳米粒子与间充质干细胞共培养,不同时间点检测干细胞存活率.利用荧光显微镜、普鲁士蓝染色、透射电子显微镜等方法观测干细胞被标记情况;建立裸鼠的胃癌模型,并将标记后的干细胞尾静脉注射入裸鼠体内,14d后用动物成像仪和核磁共振成像仪检测.结果:荧光磁性纳米粒子在低于 100μg/mL浓度下对间充质干细胞没有毒性作用,荧光磁性纳米粒子与干细胞共培养6h后,荧光显微镜、普鲁士蓝染色、透射电子显微镜检测结果显示粒子被内吞后定位于细胞质中,动物实验显示通过检测肿瘤部位的荧光信号和核磁信号,被标记的干细胞能够准确定位到胃癌的发生部位.结论:这种双模式检测体系为胃癌的早期诊断与治疗提供一种新的方法.     

可见光响应的CdTe/TiO2复合纳米颗粒体外PDT灭活HL60细胞的研究

文章采用溶胶凝胶法制备核壳CdTe/TiO_2复合纳米颗粒,探讨了该复合纳米颗粒体外PDT对HL60细胞的灭活作用。通过扫描电镜(TEM)、X射线光电子衍射仪(XPS)对CdTe/TiO_2进行表征。文中,用紫外可见光吸收光谱(UV-vis)测得尺寸为2-5 nm的CdTe QDs吸收峰为460 nm。研究表明,CdTe/TiO_2复合纳米颗粒尺寸在80 nm左右,其吸收光谱相较于TiO_2的光响应区拓展至可见光区。将CdTe/TiO_2与HL60细胞进行共同孵育,采用CCK-8法研究了其在暗室条件下细胞的生长情况和浓度对细胞相对存活率的影响以及在不同浓度的CdTe/TiO_2复合纳米颗粒PDT后的细胞活性。实验结果表明:在共同孵育16 h后CdTe/TiO_2对HL60细胞的毒性最强,10~320μg/mL浓度的CdTe/TiO_2样品对HL60细胞均具有较强的灭活作用。当添加CdTe/TiO_2样品浓度为320μg/mL时,光照1 h后PDT灭活效率达到87.7%。     

Ⅱ型胶原-硫酸软骨素支架的制备及组织工程软骨的构建

研究经乙基-(3-二甲基氨基丙基)碳化二亚胺盐酸盐(EDC)处理的Ⅱ型胶原-硫酸软骨素支架材料的性能特点,并在体外构建组织工程软骨。从鸡软骨中提取Ⅱ型胶原,以不同浓度的EDC为交联剂通过冷冻干燥的方法制备Ⅱ型胶原与硫酸软骨素复合支架并测定其理化性质。将体外培养的新生兔关节软骨细胞接种在Ⅱ型胶原与硫酸软骨素复合支架上,观察软骨细胞在支架上的生长形态并检测支架上软骨细胞分泌的糖胺聚糖含量及Ⅱ型胶原含量。结果表明:采用EDC与硫酸软骨素交联增加了支架的稳定性,最适的交联剂质量浓度为7 mg/mL。软骨细胞在复合支架上增殖分化良好,并保持软骨细胞特异分化的表型,分泌Ⅱ型胶原与蛋白多糖(GAG)。培养14 d后已有软骨样组织形成。     

叶酸修饰增强CdS-TiO_2光动力体外灭活HL60细胞效果探究

本文研究了叶酸修饰硫化镉掺杂二氧化钛(FA-CdS-TiO_2)纳米颗粒体外光动力(PDT)灭活HL60细胞的作用效果,探讨了叶酸修饰增强CdS-TiO_2体外PDT效果的作用机理。采用表面修饰的方法制备FA-CdSTiO_2;通过透射电镜(TEM)、傅里叶红外光谱(FTIR)、荧光激发光谱(FS)方法,对纳米颗粒进行结构和光学性质的表征;采用CCK-8法检测细胞活性;利用荧光探针标记技术分析细胞内活性氧水平和细胞对纳米颗粒的摄取效率。实验结果表明:叶酸修饰后,纳米颗粒粒径大小和光学性质符合光敏剂要求,且不会引起新的细胞毒性;通过叶酸修饰,细胞对纳米颗粒的摄取效率提升,细胞内活性氧产量提高,FA-CdS-TiO_2纳米颗粒的PDT效率明显增强,在FA-CdS-TiO_2浓度为20μg/m L时,暗室细胞存活率约为85%,可见光下对HL60细胞的灭活率达78%,实现了较低暗毒性下的较高PDT灭活效率。     

IL-6通过MAPK/ERK信号通路调节BMSCs软骨定向分化

白细胞介素-6(IL-6)是参与骨髓间充质干细胞(BMSCs)软骨定向分化的重要调节因子.MAPK/ERK信号通路可介导骨关节炎软骨损伤.然而,IL-6调节BMSCs定向分化为软骨细胞的分子机制尚不清楚.IL-6通过激活MAPK/ERK信号途径,抑制BMSCs的成软骨分化.本文发现,BMSCs在体外向软骨细胞分化时,Il-6基因表达水平显著下调,同时分泌到培养基中的IL-6蛋白水平亦明显降低.重组IL-6可抑制BMSCs向软骨细胞分化,软骨分化标志蛋白Runx2和Sox9的诱导表达亦相应下调.IL-6可诱导MAPK/ERK信号通路活化,加入ERK特异性阻断剂后,Runx2和Sox9的诱导表达恢复正常.结果提示,IL-6通过激活MAPK/ERK信号通路抑制BMSCs的软骨细胞分化.炎症因子IL-6对软骨细胞的再生具有不利的影响,该研究为软骨组织工程研究和骨关节炎等软骨疾病的治疗提供有价值的参考.     

Structures, magnetic properties, and XPS of one-dimensional cyanide-bridged Cu-Ni/Pt bimetallic assembly complexes

Synthesis, crystal structures, and spectroscopic and magnetic properties of new one-dimensional cyano-bridged bimetallic complexes, [Cu^II(N-Eten)₂][M^II(CN)₄] (N-Eten = N-ethylethylenediamine; M^II = Ni^II (1) and Pt^II (2)), have been reported. Both complexes consist of one-dimensional alternate chains of Cu^II and M^II moieties. The Pt-C bond distances of 1.997(3) and 2.001(3) Å for 2 are considerably longer than the Ni-C bond lengths of 1.866(3) and 1.872(3) Å for 1. Because of pseudo Jahn-Teller distortion, the axial Cu-N bond distances of 2.554(2) and 2.550(3) Å for 1 and 2 are longer than those of equatorial ones of 2.008(2) and 2.056(2) Å for 1 and 2.010(2) and 2.054(2) Å for 2. In contrast to M^II-C bond distances, the Cu-N ones of 1 are similar to those of 2 regardless of element-substitution. These complexes indicate weak antiferromagnetic interactions with Weiss constants = − 4.68 and −3.95 K for 1 and 2, respectively. The emission spectrum of 2 (λ_ex = 360 nm) exhibits a broad band with peaks at 22 800 and 24 000 cm⁻¹ at 298 K. The Cu 2p_1/2 and 2p_3/2 peaks of XPS spectra are compared systematically to various copper(II) complexes showing different bridging features or distorted coordination geometries as models for excited structures induced by external physical conditions. The spectroscopic properties are discussed from the viewpoint of magneto-optical properties.     

脑囊虫的MRI 表现

目的:探讨脑囊虫不同分期、分型的磁共振表现及应用价值。方法:收集脑囊虫病例127例,行MR轴、矢、冠位扫描及增强扫描。结果:脑囊虫分型:单发小囊型44例,单发大囊型6例,多发小囊型34例,脑实质钙化型18例;脑室型11例;蛛网膜下腔型7例;混合型7例。脑囊虫分期:活动期32例,退变死亡期49例,钙化期18例,混合期28例。结论:磁共振扫描能够清晰显示各种细节,是诊断脑囊虫的有力手段。     

Synthesis and anomalous magnetic properties of heterometal dinuclear Fe(II)-Cr(III) complex bridged by 3,5-bis(pyridin-2-yl)-pyrazolate

A Cr(III)-Fe(II) dinuclear complex [(nta)Cr(μ-bpypz)Fe(dpea)]BF₄, where bpypz=3,5-bis(pyridin-2-yl)-pyazolate, nta=nitrilotriacetate and dpea=(2-aminoethyl)-bis(2-pyridylmethyl)amine, has been synthesized and structurally characterized by ESI-MS and ²H NMR spectra. The proposed structure is mostly more planar in the bridging moiety as compared to that of the previously reported [(nta)Cr(μ-bpypz)Fe(picen)]BF₄(N,N^′-bis-(2-pyridylmethyl)ethylenediamine(picen)). The dpea complex is found to be a high spin state, but to exhibit no spin-crossover by magnetic susceptibility and Mössbauer spectra measurements; showing a large antiferromagnetic interaction, which could make the spin transition temperature lower and/or destroy spin-crossover, suggesting complementarity between spin-crossover and magnetic interaction.     

Synthesis, structure and properties of copper(II) and manganese(II) coordination polymers with 1,5-dinitronaphthalene-3,7-dicarboxylate

Two new metal-organic coordination polymers with 1,5-dinitronaphthalene-3,7-dicarboxylate (NNDC), [Cu₂(NNDC)₂(DMF)_1.8(DMSO)_2.2(H₂O)₂]·H₂O (1) and [Mn₃(NNDC)₃(DMSO)₄]·2DMSO (2) have been synthesized under solvothermal conditions and characterized by single crystal X-ray diffraction and Thermogravimetric Analysis (TGA). The structure of compound 1 consists of one-dimensional chains with copper ions being linked by the dicarboxylate ligands. The coordination chains are associated into ladder-like double chains through O-H?O hydrogen bonds and π-π interactions, and the ladders are packed in a cross fashion through further π-π interactions to give the three-dimensional structure. The Mn(II) compound exhibits a 3D framework with the pcu topology, in which [Mn₃(COO)₆] clusters as octahedral secondary building blocks are linked by the naphthalene spacers. Magnetic analyses were carried out based on both temperature- and field-dependent data, consistently suggesting relatively weak antiferromagnetic interactions within the carboxylate bridged [Mn₃(COO)₆] cluster.     

多核19F磁共振成像研究进展

~(19)氟(~(19)F)的磁共振成像(MRI)研究可追溯到35年以上。在这段时间里,~1H磁共振成像的蓬勃发展使磁共振成为影像医学的支柱,但~(19)F磁共振成像研究的进展却较为缓慢。然而最近几年,~(19)F磁共振成像的研究受到了广泛的关注。在某种程度上,这是由于MR成像中软件与硬件的发展,更因为分子影像学的概念的提出与发展。本文将对~(19)F多核磁共振成像的应用,特别是使用全氟化碳纳米成像探针的19F多核磁共振成像进行综述。     

Iron(III), chromium(III) and cobalt(II) complexes with squarate: Synthesis, crystal structure and magnetic properties

The preparation and variable temperature-magnetic investigation of three squarate-containing complexes of formula [Fe₂(OH)₂(C₄O₄)₂(H₂O)₄]·2H₂O (1) [Cr₂(OH)₂(C₄O₄)₂(H₂O)₄]·2H₂O (2) and [Co(C₄O₄)(H₂O)₄]_n (3) [H₂C₄O₄ = 3.4-dihydroxycyclobutene-1,2-dione (squaric acid)] together with the crystal structures of 1 and 3 are reported. Complex 1 contains discrete centrosymmetric [Fe₂(OH)₂(C₄O₄)₂(H₂O)₄] diiron(II) units where the iron pairs are joined by a di-μ-hydroxo bridge and two squarate ligands acting as bridging groups through adjacent oxygen atoms. Two coordinated water molecules in cis position complete the octahedral environment at each iron atom in 1. The iron-iron distance with the dinuclear unit is 3.0722(6) Å and the angle at the hydroxo bridge is 99.99(7)°, values which compare well with the corresponding ones in the isostructural compound 2 (2.998 Å and 99.47°) whose structure was reported previously. The crystal structure of 3 contains neutral chains of squarato-O¹,O³-bridged cobalt(II) ions where four coordinated water molecules complete the six-coordination at each cobalt atom. The cobalt-cobalt separation across the squarate bridge is 8.0595(4) Å. A relatively important intramolecular antiferromagnetic coupling occurs in 1 whereas it is very weak in 2, the exchange pathway being the same [J = −14.4 (1) and −0.07 cm⁻¹ (2), the spin Hamiltonian being defined as ]. A weak intrachain antiferromagnetic interaction between the high-spin cobalt(II) ions occurs in 3 (J = −0.30 cm⁻¹). The magnitude and nature of these magnetic interactions are discussed in the light of their respective structures and they are compared with those reported for related systems.     

Preparation, structural, magnetic and thermal properties of two heterobimetallic cyano-bridged nickel(II)-copper(II)/platinum(II) coordination polymers

Two unique bimetalic Pt(II) coordination polymers of composition [Ni(hydeten)₂Pt(CN)₄] (Ni-Pt) and [Cu(hydeten)₂Pt(CN)₄] (Cu-Pt) [hydeten = N-(2-hydroxyethyl-ethylenediamine) or 2-(2-aminoethylamino)ethanol] have been synthesized and structurally characterized by various methods in this study. The crystal structure of Cu-Pt was determined by single-crystal X-ray diffraction analysis. The structure of Cu-Pt forms of infinite 2,2-TT type [-Cu(hydeten)₂-NC-Pt(CN)₂-CN-] chains containing paramagnetic copper atoms bridged by tetracyanoplatinate species. In this complex, Cu(II) centers display an axially elongated octahedron with two chelating hydeten molecules in the equatorial positions and N-bonded bridging cyano groups in the axial positions, whereas Pt(II) centers are four coordinate with four cyanide-carbon atoms in a square-planar arrangement. The decrease of the moments of these complexes in temperature range of 50 305 K can assigned to the antiferromagnetic interactions in the structures. The thermal decomposition of Cu-Pt comprise of five distinguished stages, while the thermal decomposition of Ni-Pt take place four different stages.     

急性弥漫性轴索损伤CT与MRI对比的研究

目的:比较急性弥漫性轴索损伤患者CT及MRI表现的不同。方法:选取60例急性弥漫性轴索损伤患者,而CT无异常改变或改变轻微的病例行MRI检查。全部检查均在损伤后6天内完成。GCS评分＜8分44例,9—12分16例。结果:MRI显示具有DAI损伤表现的共41例,阳性率68．3%,其中单独具有DAIⅠ、Ⅱ和Ⅲ型损伤表现的分别为7例、12例和1例,同时具有DAIⅠ和Ⅱ型损伤表现的共12例,同时具有DAJⅠ和Ⅲ型损伤表现的共2例,同时具有DAJⅡ和Ⅲ型损伤表现的共2例,同时具有三型DAI损伤表现的共5例;CT显示具有DAI损伤表现的共3例,阳性率5．0%,其中具有DAIⅠ型损伤表现的2例,具有DAIⅡ型损伤表现的1例,具有DAIⅢ型损伤表现的0例。结论:CT缺乏异常表现或轻微异常表现的急性弥漫性轴索损伤,MRI常常能显示损伤,包括特殊的损伤部位和一些其他类型合并损伤。     

磁共振成像应用于检测胎盘植入介入治疗

为了探讨磁共振成像(magnetic resonance imaging, MRI)在胎盘植入介入治疗中的诊断作用和为临床治疗提供依据,本研究选取30例于2012年6月至2015年12月间在我院进行介入治疗的胎盘植入患者作为研究对象,根据病理诊断标准,分析患者胎盘植入介入治疗前后的MRI检查结果。结果显示,粘连性胎盘的敏感性和特异性分别为77.5%和90.2%,植入性胎盘的敏感性和特异性分别为75.5%和87.7%,穿透性胎盘的敏感性和特异性分别为85%和100%。最好的预测胎盘植入的MRI特征是在T2W磁共振成像(T2W-MRI)序列上存在暗色的胎盘内条带。介入治疗1年后复查时,发现患者子宫恢复为正常大小,宫腔内的胎盘组织基本消失,宫壁与植入胎盘融合、宫腔内膜线和子宫结合带的信号完整。综上结果,说明MRI可作为检测胎盘植入可靠性和可重复性的工具,并且能够显示胎盘植入部位及子宫肌层受侵程度,可用于评价胎盘植入介入治疗的疗效。     

In vivo assessment of nodularin-induced hepatotoxicity in the rat using magnetic resonance techniques (MRI, MRS and EPR oximetry)

Acute nodularin-induced hepatotoxicity was assessed in vivo, in rats using magnetic resonance (MR) techniques, including MR imaging (MRI), MR spectroscopy (MRS), and electron paramagnetic resonance (EPR) oximetry. Nodularin is a cyclic hepatotoxin isolated from the cyanobacterium Nodularia spumigena. Three hours following the intraperitoneal (i.p.) administration of nodularin (LD50), a region of 'damage', characterized by an increase in signal intensity, was observed proximal to the porta hepatis (PH) region in T2-weighted MR images of rat liver. Image analysis of these regions of apparent 'damage' indicated a statistically significant increase in signal intensity around the PH region following nodularin administration, in comparison with controls and regions peripheral to the PH region. An increase in signal intensity was also observed proximal to the PH region in water chemical shift selective images (CSSI) of nodularin-treated rat livers, indicating that the increased signal observed by MRI is an oedematous response to the toxin. Microscopic assessment (histology and electron microscopy) and serum liver enzyme function tests (aminotransferase (ALT) and aspartate ALT (AST)) confirmed the nodularin-induced tissue injury observed by MRI. In vivo and in vitro MRS was used to detect alterations in metabolites, such as lipids, Glu+Gln, and choline, during the hepatotoxic response (2-3 h post-exposure). Biochemical assessment of perchloric acid extracts of nodularin-treated rat livers were used to confirm the MRS results. In vivo EPR oximetry was used to monitor decreasing hepatic pO2 (approximately 2-fold from controls) 2-3 h following nodularin exposure. In vivo MR techniques (MRI, MRS and EPR oximetry) are able to highlight effects that may not have been evident in single end point studies, and are ideal methods to follow tissue injury progression in longitudinally, increasing the power of a study through repeated measures, and decreasing the number of animals to perform a similar study using histological or biochemical techniques.