Histomorphometry of the Testes, Epididymides, and Spermatozoa of Four Indigenous Breeds of Bulls in Ibadan, Nigeria

Forty testicles were used to carry out histomorphometry study on the testes, epididymides and spermatozoa of four indigenous breeds of bull found in Ibadan (Red Bororo (RB), Sokoto Gudali (SG), White Fulani (WF) and Dhali (DL)). The testicles were harvested immediately after the slaughter at the Bodija abattoir and transported to the laboratory in an insulated flask containing warm water at 37°C within 30 minutes. In the laboratory, the samples were dissected and histological sections of the right testis, right and left caudal epididymides taken from two bulls in each group. The tissues were fixed in Bouin’s fluid. They were processed in an automatic tissue processor, embedded in paraffin wax using the embedding system (Leica EG 1160) and sectioned with microtome at 4 microns. The sections were then stained by Haematoxylin and Eosin method and mounted and examined under a microscope. There was no significant difference between breed and seminiferous tubular diameter (STD) and testicular germinal height (TGEH). The mean STD ranged from 223.00 ± 28.35 to 316.00 ± 37.70 μm, while the TGEH ranged from 81.60 ± 10.05 to 89.80 ± 4.83 μm. The mean epididymal tubular diameters (ETD) and epididymal lumen diameters (ELD) had the highest value of 378.00 ± 10.95 and 298.20 ± 26.0489 μm in DL and lowest of 373.80 ± 37.70 and 278.10 ± 27.71 μm in RB, respectively. The mean epididymal germinal epithelia heights (EGEH) were highest (61.20 ± 5.70 μm) in WF and lowest (39.80 ± 0.86 μm) in RB. The mean sperm head length (SHL), sperm midpiece length (SMPL), sperm tail length (STL) and sperm total length (TL) had the highest values of 5.90 ± 0.10, 8.80 ± 0.25, 24.20 ± 2.59 and 38.90 ± 2.94 μm, respectively in SG. SHL and SMPL were lowest in DL bulls (5.00 ± 0.00 and 6.30 ± 0.20 μm), while STL and TL were lowest (18.90 ± 0.24 and 31.30 ± 0.85 μm). The results of this study provide baseline data on the histomorphometry of the testes and epididymides and spermatozoa of some indigenous bulls.

Cytotoxicity Research of Recombinant Human Lactoferrin on Primary Hepatocyte and Nephrocyte Cell

[Objective] The aim was to study whether recombinant Human Lactoferrin has toxic effect on Primary Hepatocyte and Nephrocyte Cell of rat to provide reference for further safety evaluation.[Method] Recombinant Human Lactoferrin and its digested products were taken as tested compound,cow Lactoferrin was used for contrast.Primary Hepatocyte and Nephrocyte Cell of rat were cultured and IC50 values were tested by MTT,and cytotoxic dose-response relationship was tested.[Result]Target toxicity was not found from recombinant Human Lactoferrin on hepatocytes and nephrocytes,in accordance with sub-chronic toxicity test.[Conclusion] This study is of reference value for further safety evaluation of recombinant Human Lactoferrin and safety of evaluation method of GM food.

In Vitro Cytotoxicity of Polyphosphoester as a Novel Injectable Alveolar Replacement Material

The aim of this study was to investigate the in vitro cytotoxicity of polyphosphoester polymer used as a novel injectable alveolar bone substitutes for controlled delivery of tetracycline. Cell culture medium was exposed to the polymer （0.01-10 mg/mL） for 24 h. The L-929 mouse fibro- blasts were then exposed to the treated cell culture medium for 24 h. Finally, cell viability and growth were assessed by using MTT assay and Alamar Blue assay. No significant cytotoxicity of the polyphosphoester against L-929 mouse fibroblasts was observed at a concentration up to 10 mg/mL （P〉0.05）. The two evaluation methods showed no significant differences （P〉0.05）. This study suggests that polyphosphoester does not demonstrate any significant toxic effects to cells in vitro and has the potential to be used both as a medical device and as scaffolds in tissue engineering applications.

In vitro corrosion behavior and cytotoxicity property of magnesium matrix composite with chitosan coating

Mg-6%Zn-10%β-Ca3(PO4)2 composite was prepared through powder metallurgy methods with different chitosan coatings on its surface. The properties of the chitosan coatings on the surface of Mg-6%Zn-10%β-Ca3(PO4)2 composite, such as the adhesion ability, the corrosion behavior and the cytotoxicity properties, were investigated, and the microstructure of the chitosan coating was observed by scanning electron microscope(SEM). The results show that chitosan coating improves the corrosion resistance of the magnesium composite specimens significantly. Mg-6%Zn-10%β-Ca3(PO4)2 composite specimens exhibit good corrosion resistance and low p H values in simulated body fluid(SBF) at 37 °C in the immersion test with 7-layer chitosan coating whose relative molecular mass is 30×104 Da. The cytotoxicity tests indicate that Mg-6%Zn-10%β-Ca3(PO4)2 with chitosan coating is nontoxic with a cytotoxicity grade of zero against L-929 cells, which is better than that of uncoated composites.

Cytotoxicity evaluation of extracts and fractions of five marine sponges from the Persian Gulf and HPLC fingerprint analysis of cytotoxic extracts

Objective: To screen the cytotoxic effects of some marine sponges extracts on HeLa and PC12 cells.Methods: Five marine sponges including Ircinia echinata(I. echinata), Dysidea avara,Axinella sinoxea, Haliclona tubifera and Haliclona violacea were collected from the Persian Gulf(Hengam Island). The cytotoxic effect of these sponges was evaluated by using MTT assay. The metabolic high performance liquid chromatography fingerprint of I. echinata was also carried out at two wavelengths(254 and 280 nm).Results: Among the sponges tested in this study, the extracts of I. echinata and Dysidea avara possessed the cytotoxic effect on HeLa and PC12 cells. The obtained fractions from high performance liquid chromatography were evaluated for their cytotoxic properties against the cell lines. The isolated fractions did not show significant cytotoxic properties.Conclusions: I. echinata could be considered as a potential extract for chemotherapy.Further investigation is needed to determine the accuracy of mechanism.

市政污泥及其热处理渣对CHO-K1细胞的毒性研究

评估市政污泥资源化产物的健康风险对其安全规范利用具有指导意义。分别对某污水处理厂的污泥进行了热解和好氧发酵处理,运用中国仓鼠卵巢细胞(CHO-K1)对市政污泥及其热处理渣开展细胞毒性试验和遗传毒性试验,评估其对典型哺乳动物细胞的危害和潜在风险。结果显示,与原污泥样品相比,污泥热解渣和好氧发酵渣的细胞毒性和遗传毒性均降低。与原污泥样品相比,RTCA细胞增殖试验结果显示,污泥热解渣和好氧发酵渣对CHO-K1细胞的生长抑制性明显减弱;CCK-8细胞毒性试验结果显示,污泥热解渣和好氧发酵渣的细胞相对存活率分别增加了161%和137%;Caspase 3细胞凋亡蛋白酶活力试验结果显示,暴露于污泥热解渣和好氧发酵渣中的CHO-K1细胞的Caspase 3凋亡蛋白酶的活性分别降低了29.3%和20.1%;彗星试验结果显示,污泥热解渣和好氧发酵渣的尾长分别缩短了64.5%和25.9%,尾矩分别减少了78.4%和28.1%。研究结果表明,热处理技术(热解处理和好氧发酵处理)可减少市政污泥的细胞毒性和遗传毒性,降低市政污泥对生态环境和人体健康的潜在风险。

溶剂纯化改善聚苯胺纳米材料生物相容性的研究

目的考察不同有机溶剂纯化对聚苯胺(polyaniline,PANI)纳米材料生物相容性的影响,为开发新型高分子医用材料奠定基础。方法分别利用二甲基甲酰胺(N,N-dimethylformamide,DMF)和二甲基亚砜(dimethyl sulfoxide,DMSO)对化学氧化法合成的PANI纳米线进行纯化处理,比较纯化前后材料的微观形态变化;将纯化前后的PANI材料配制成100%、50%、25%、10%、5%和1%的浸提液,以BV2细胞为研究对象,采用CCK-8法测定各浸提液组的细胞存活率;将纯化前后的PANI材料配制成100、80、64、51和41 mg·mL^(-1)的混悬液,通过灌胃给药考察材料对小鼠口服急性毒性及对肝肾的损伤。结果所得材料为直径约20 nm的PANI纳米线,经过纯化后其形貌未发生明显改变。未经纯化的100%PANI浸提液具有很强的细胞毒性,但是随着浓度的降低,其毒性逐渐减弱。经DMF和DMSO纯化后,细胞在其浸提液中的存活率均高于同浓度下未纯化材料(P均<0.01)。PANI材料用DMSO纯化后给药,各组小鼠的体质量增长率高于未纯化材料(P<0.05或P<0.01),其最大耐受量(MTD)>3000 mg·kg^(-1)。解剖后,各组小鼠肾脏无明显变化,但是肝细胞受到不同程度的损伤,材料经纯化后的损伤效应低于未纯化材料。结论溶剂处理是一种简便可行的PANI纳米材料纯化策略,可在提高材料生物相容性的同时维持其形貌不变。

不同浸提条件对医疗器械体外细胞毒性试验结果的影响

目的探讨不同浸提条件(浸提24、72 h)对医疗器械体外细胞毒性试验中细胞生长与细胞毒性结果的影响。方法采用含血清MEM细胞培养液作为浸提介质,按照GB/T 16886.12-2017推荐的浸提比例,将空白对照品、阴性对照品及5种不同医疗器械常用材料样品在(37±1)℃条件下分别浸提24、72 h,依据GB/T 16886.5-2017中的MTT法进行试验。比较浸提24、72 h对细胞生长与细胞毒性结果的影响。结果与浸提24 h比较,浸提72 h后空白对照组吸光度偏低,相对偏差为-3.0%,差异无统计学意义(P>0.05);被检样品试验结果显示,样品1、样品2有细胞毒性,样品3和样品5无细胞毒性,浸提24、72 h结果一致,相对增殖率(RGR)变化小;样品4有细胞毒性,但浸提72 h后RGR明显降低,降低幅度为93.7%,提示有毒物质溶出量增多。结论结合显微镜观察细胞形态,确认延长浸提时间至72 h对细胞生长的影响可以忽略。对于被检样品组,延长浸提时间至72 h各样品RGR呈降低趋势。对于RGR>80%、<60%的医疗器械样品,浸提24、72 h的评价结果一致,评价风险较小。而对于RGR处于临界值(60%~80%)的医疗器械样品,浸提24、72 h的结果可能不一致,评价风险较大,尤其是长期接触人体的医疗器械,建议采取浸提72 h的条件进行体外细胞毒性试验,以降低评价风险。

硫化铜/氧化石墨烯/壳聚糖/纳米羟基磷灰石复合材料的抗菌及促成骨作用

目的探讨硫化铜(CuS)/氧化石墨烯(GO)/壳聚糖(CS)/纳米羟基磷灰石(nHA)复合材料(CGCHs)的抗菌和促成骨作用及其作用机制。方法采用水热法合成CuS/GO纳米颗粒,通过原位沉淀法合成CS/nHA支架和CGCHs支架,检测材料表征、光热转换性能和生物安全性,评估CGCHs组和近红外光(NIR)照射下CGCHs(CGCHs+NIR)组的细菌抑制效果及其对细菌生物膜相关基因表达的影响,观察CGCHs和CS/nHA不同材料组的促成骨分化和成骨、破骨相关基因表达。结果CGCHs是具有高度孔隙率的三维支架,在CuS/GO浓度为200μg/mL时CGCHs同时兼具良好的红外升温效果和生物安全性。琼脂糖平板涂菌和细菌死活染色结果均表明CGCHs+NIR组抗菌性能最佳,生物膜相关基因qPCR检测证实其具有抑制细菌生物膜相关基因表达的作用。茜素红染色结果表明CGCHs具有良好的体外促成骨性能,体外共培养3、7、14、21和28 d qPCR结果表明CGCHs对成骨早期和晚期相关基因表达均具有促进作用。与破骨细胞共培养结果可观察到CGCHs具有抑制破骨细胞形成的作用,细胞凋亡检测结果进一步验证这一结论,破骨分化相关基因qPCR检测结果表明,CGCHs主要通过抑制抗酒石酸酸性磷酸酶、组织蛋白酶K、CTR、P65和P38在共培养7、14 d的表达来抑制破骨细胞的分化。结论作为纳米复合材料,CGCHs生物安全性好,具有良好的红外光热协同抗菌作用,在促成骨分化的同时抑制破骨细胞分化,有望为感染性骨缺损治疗提供新的思路。

海藻酸钠/壳聚糖复合凝胶的制备与细胞毒性评价

背景:海藻酸钠和壳聚糖分别是聚阳离子和聚阴离子的天然高分子材料,二者可以互为交联剂形成复合凝胶,避免使用普通交联剂产生的细胞毒性。目的:制备海藻酸钠壳聚糖复合凝胶并评价其体外细胞毒性。方法:以0.25 mol/L乙酸溶解壳聚糖,制成质量浓度30 g/L溶液,再以0.1 mol/L的NaOH中和酸性得到壳聚糖絮状沉淀,将壳聚糖絮状沉淀与质量浓度3%海藻酸钠等比例混合,高频振动混匀,使二者形成复合凝胶。使用傅里叶变换红外光谱、扫描电镜分析观察凝胶成分和交联纤维网络结构。分别以海藻酸钠壳聚糖复合凝胶24,72 h浸提液、聚乙烯24,72 h浸提液及苯酚溶液培养L-929细胞,体外检测其细胞毒性。结果与结论:傅里叶变换红外光谱检测到海藻酸钠壳聚糖复合凝胶特征性峰值改变,扫描电镜显示其内部形成丰富间隙的空间网络结构。浸提液法检测海藻酸钠壳聚糖复合凝胶的细胞毒性为合格,表明海藻酸钠/壳聚糖复合凝胶具有成为组织工程支架材料的良好条件。

敌敌畏和氧乐果对小鼠精子畸形的影响

目的探讨敌敌畏(DDV)和氧乐果染毒对雄性动物生殖细胞的毒性作用,为有机磷农药对农副产品污染的综合防治提供科学依据。方法选择雄性昆明种小鼠42只,随机分为7组(6个染毒组和1个对照组)每组6只。染毒组小鼠每天分别以5.0、10.0、20.0mg/kg剂量DDV,2.5、5.0、10.0mg/kg剂量氧乐果经口染毒;对照组给予等容积的生理盐水。每天每只小鼠染毒1次,连续染毒21d。首次染毒后5周,以脱颈臼法处死小鼠,制作精子标本,观察精子畸形率。结果各染毒组精子畸形率(DDV染毒组分别为4.33%,8.40%,13.65%,氧乐果染毒组分别为4.35%,6.63%,7.93%)均高于对照组(1.97%),差异均有统计学意义(P<0.01),且呈剂量-反应关系(P<0.01)。结论DDV和氧乐果均可引起雄性小鼠生殖细胞的遗传损伤,导致生殖毒性效应。

新型医用钛合金生物相容性评价

目的:评价新型医用钛合金Ti12.5Zr2.5Nb2.5Ta(TZNT)的生物相容性,并和3种标准医用钛材料Ti6Al4V、Ti6Al7Nb和TA2进行对比。方法:通过细胞毒性试验观察细胞在TZNT、Ti6Al4V和Ti6Al7Nb合金离子析出培养液中的生长情况,计算在不同时间组的相对细胞增殖率,评价其毒性等级;通过皮下埋植试验观察4种材料植入白鼠体内4周和8周的组织变化情况,计算炎细胞密度和纤维膜厚度的变化情况,评价其引起的组织反应程度。结果:TZNT、Ti6Al4V和Ti6Al7Nb在3个时间组(2、4、6d)的相对细胞增殖率均不低于100%,细胞毒性等级评定为0级。和Ti6Al4V和Ti6Al7Nb相比,TZNT和TA2植入白鼠体内4周和8周后,无论是炎细胞密度还是植入物周围的纤维膜厚度均有所降低,引起的组织反应程度较低。结论:TZNT具有良好的生物相容性,是一种理想的生物医用钛合金。

甲醛对雄性小鼠的生殖遗传毒性

目的:观察甲醛对雄性小鼠精子畸变、生殖能力和子代胎鼠肝细胞微核率的影响。方法:随机将小鼠分为阴性对照组(NC)、甲醛染毒组和阳性对照组(PC),每组12只,采用静式吸入染毒,染毒组吸入甲醛浓度分别是21mg.m-3(1/24LC50)、42mg.m-3(1/12LC50)和84mg.m-3(1/6LC50),每天2h,每周6d,共13周。染毒结束后处死小鼠,检测小鼠精子畸形率,采用显性致死试验方法,检测雄性小鼠生殖能力及子代胎鼠肝细胞微核率。结果:各染毒组雄性小鼠精子头畸形率显著高于阴性对照组(P<0.05),并与甲醛染毒剂量呈明显正相关(r=0.83,P<0.001)。各剂量染毒组孕鼠的胎吸收率较阴性对照组显著升高(P<0.01),1/6LC50组的每窝平均活胎数较阴性对照组显著下降(P<0.05)。结论:甲醛可以造成雄性小鼠精子畸形,降低雄性小鼠生殖能力,对雄性小鼠具有一定的生殖遗传毒性。

纳米羟基磷灰石/胶原复合材料的制备及生物学评价

低温下,水热合成的纳米羟基磷灰石浆液与中性胶原溶胶混合,升温使羟基磷灰石晶体与胶原自组装形成复合凝胶,经洗涤、冻干,获得羟基磷灰石/胶原复合材料。采用XRD,FTIR和TEM对复合材料的特性进行研究,结果表明:羟基磷灰石晶体尺寸为16nm×40nm;复合材料的晶相组成,羟基磷灰石晶体大小、胶原纤维的结构等都与天然骨相似。采用骨髓嗜多染细胞微核实验、致敏实验、以及细胞毒性实验对复合材料进行生物相容性评价。小鼠骨髓微核率为1.6±0.9,与阴性对照组无显著性差异,和阳性对照组有显著性差异。最大剂量法实验表明复合材料无致敏作用。滤膜扩散实验证明复合材料无细胞毒性。复合材料骨内植入的组织学分析表明,4周时,在材料和骨组织的界面处有新骨生成,12周时,界面处有大量的新骨形成。皮下植入44周后,材料降解成碎片,纤维组织长入材料降解区域。本研究制备的纳米羟基磷灰石/胶原复合材料具有体内降解和促进骨形成的能力。不具有致染色体畸变作用、皮肤致敏作用和细胞毒性,是一种生物相容性良好的潜在的骨替换材料。

蝎毒组分对人大肠癌细胞的体外抑杀作用研究

目的观察蝎毒不同组分对人大肠癌细胞的体外抑杀作用。方法 用噻唑蓝(MTT法)、集落形成抑制实验,以丝裂霉素C(MMC)为阳性对照药品,以蝎毒提取物SVC Ⅰ、SVC Ⅱ、SVC Ⅲ,SVC Ⅳ各组分、浓度分别为45μg/ml、60μg/ml、75μg/ml、90μg/ml处理人大肠癌细胞Lovo,观察Lovo的生长情况。结果 SVCⅢ对Lovo细胞的细胞毒性和集落形成的抑制作用略强于MMC组,SVC Ⅰ、SVC Ⅱ、SVC Ⅳ对细胞Lovo的影响在所用药物浓度内与对照组相比未见统计学差异,细胞毒性作用不显著。结论SVC Ⅲ对人大肠癌细胞具有明显的细胞毒性和生长抑制作用。

纤维蛋白止血敷料的生物相容性研究

目的评价纤维蛋白止血敷料的生物相容性。方法对纤维蛋白止血敷料进行如下生物学检测:细胞毒性试验(MTT法)、急性全身毒性试验、皮肤刺激试验、皮内刺激试验和溶血试验,根据标准对实验数据进行分析和评估。结果纤维蛋白止血敷料的细胞毒性分级0—1级;无急性全身毒性作用,无皮肤刺激反应、皮内刺激反应,无溶血性。结论纤维蛋白止血敷料具有良好的生物相容性。

角蒿中的环己乙醇类化学成分研究

利用Sephadex LH-20及硅胶等柱色谱技术从药用民族植物角蒿(Incarvillea dissectifoliolaQ.S.Zhao)的根茎中分离得到了11个环己乙醇类化合物。经理化数据对照和波谱分析,确定它们的结构分别为rengyolone(1),(3aS,6R)-2,3,3a,6,7,7a-hexahydrobenzofuran-3a,6-diol(2),(3aR)-hexahydro-3a-hydroxybenzofuran-6(2H)-one(3),(3aR,6R)-octahydrobenzofuran-3a,6-diol(4),(3aR,6S)-octahydrobenzofuran-3a,6-diol(5),4-hydroxy-4-(2-hydroxyethyl)cyclohexanone(6),(3aS,4S)-hexahydro-3a,4-dihydroxybenzofuran-6(2H)-one(7),(3aR,4S)-hexa-hydro-3a,4-dihydroxy benzofuran-6(2H)-one(8),rengyol(9),isorengyol(10)和rengyoside B(11)。这11个化合物均为首次从角蒿中分离得到。化合物4,5,6,7,8,9,10的细胞毒活性未见报道,对它们的细胞毒活性进行了测定,结果表明它们对HL-60细胞株的IC50分别为102,44.7,24.6,98.0,47.8,25.3,92.3和47.2μmol/L。

中药鹿藿的抑菌实验研究

研究发现中药鹿藿在体外可瞬间抑制人类精子的运动。为了研究中药鹿藿的抑菌作用,本实验对几种女性生殖道常见致病菌和条件致病菌进行了体外抑菌实验,发现鹿藿对金黄色葡萄球菌、淋病奈瑟氏菌和大肠杆菌有抑制作用,并可抑制常见耐药菌株的生长。金黄色葡萄球菌的MIC折合生药为31.25mg/mL,大肠杆菌的MIC250mg/mL,淋球菌的MIC为31.25mg/mL。结果表明中药鹿藿在能有效抑制精子运动的浓度可以抑制某些男、女性生殖道常见细菌的生长。

东亚钳蝎毒抗癌多肽对Eca109 细胞和HeLa 细胞的毒性作用

目的和方法：采用ＭＴＴ 比色法、生长抑制实验、集落形成抑制实验，探讨东亚钳蝎毒抗癌多肽（ａｎｔｉｃａｎｃｅｒｐｏｌｙｐｅｐｔｉｄｅ ｆｒｏｍ Ｂｕｔｈｕｓ Ｍａｒｔｅｎｓｉｉ Ｖｅｎｏｍ ，ＡＰＢＭＶ） 对Ｅｃａ １０９ 细胞和ＨｅＬａ 细胞的细胞毒作用。结果：ＡＰＢＭＶ 对Ｅｃａ１０９ 细胞有明显的毒性作用，且呈显著量效关系，处理细胞２４ ｈ 和７２ ｈ 的ＩＣ５０ 分别为０－０２７μｇ·ｍＬ－１ 和０－０２４ μｇ·ｍＬ－１ 。ＡＰＢＭＶ对Ｅｃａ １０９ 细胞的生长抑制作用呈明显的时效和量效关系。处理Ｅｃａ １０９ 细胞２４ ｈ 、４８ ｈ 、７２ ｈ 的ＩＣ５０ 分别为０－０３２μｇ·ｍＬ－１，０－０２８ μｇ·ｍ Ｌ－１ ，０－０２３ μｇ·ｍＬ－１ 。ＡＰＢＭＶ 还显著抑制Ｅｃａ１０９ 细胞的集落形成，并有明显的量效关系，其ＩＣ５０ 值为０－０２３μｇ·ｍＬ－１ 。但ＡＰＢＭＶ 对ＨｅＬａ 细胞的作用不明显。结论： ＡＰＢＭＶ 对肿瘤细胞的杀伤或生长抑制作用具有一定的肿瘤差异性。

新型可注射性人工髓核的体外细胞毒性测试

目的检测前期制备的新型人工髓核材料的细胞毒性。方法材料分为4组，编号为1～4。样品1组为合成的聚碳酸酯型聚氨酯；样品2组为在室温下由硅氢加成反应制成的缩合硅橡胶材料；样品3组是在2号样品中添加二氧化硅后制成；样品4组为室温下固化的聚醚型聚氨酯。采用细胞培养基作为阴性对照组。采用小鼠成纤维细胞（L929细胞）在材料浸提液中培养的方法检测其细胞毒性，使用3-（4，5-二甲基-2-噻唑）-2，5-二苯基溴化四唑（MTT）比色法比较材料组与对照组间吸光度值和细胞相对增值率。结果材料组的吸光度值和L929细胞相对增殖率相对于对照组没有明显差异（P〉0．05），在细胞毒性分级中处于0～1级，属于合格材料。结论该材料没有细胞毒性，具有作为髓核替代材料的应用潜力。