3D Collagen Gels:A Promising Platform for Dendritic Cell Culture in Biomaterials Research

The three-dimensional(3D)cell culture system has garnered significant attention in recent years as a means of studying cell behavior and tissue development,as opposed to traditional two-dimensional cultures.These systems can induce specific cell reactions,promote specific tissue functions,and serve as valuable tools for research in tissue engineering,regenerative medicine,and drug discovery.This paper discusses current developments in the field of three-dimensional cell culture and the potential applications of 3D type 1 collagen gels to enhance the growth and maturation of dendritic cells.

绝经后盆底组织TGF-β_1和CollagenⅠ、Ⅲ表达与SUI的关系

目的:探讨绝经后压力性尿失禁(SUI)患者盆底支持结构转化生长因子β1(TGF-β1)、胶原蛋白Ⅰ、Ⅲ(CollagenⅠ、Ⅲ)与SUI的关系。方法:采用免疫组化方法,测定30例压力性尿失禁患者(SUI组)、28例盆底器官膨出(POP)患者(POP组)阴道前壁组织中TGF-β1及Collagen阳性率,并选择同期30例非卵巢功能性肿瘤和宫颈病变患者作为对照(对照组)。结果:SUI组、POP组及对照组患者阴道前壁组织中,TGF-β1表达阳性率分别为20.00%、14.30%、93.33%,SUI组与对照组比较,差异有极显著性(P<0.01);SUI组与POP组比较,差异无显著性(P>0.05)。3组患者阴道前壁组织中胶原Ⅰ含量的表达以辉度值为标准,在SUI组、POP组及对照组分别为98.62、98.15和100.03。SUI组与对照组比较,差异有极显著性(P<0.01);POP组与对照组比较,差异有极显著性(P<0.01);SUI组与POP组比较,差异无显著性(P>0.05)。3组患者阴道前壁组织中胶原Ⅲ含量的表达以辉度值为标准,在SUI组、POP组及对照组分别为98.46、98.23和100.12。SUI组与对照组比较,差异有极显著性(P<0.01);POP组与对照组比较,差异有极显著性(P<0.01);SUI组与POP组比较,差异无显著性(P>0.05)。结论:绝经后SUI患者盆底支持结构的退行性病变与组织中的转化生长因子β1减少及胶原蛋白含量水平低下有关。

瘦素对成骨细胞增殖及CollagenⅠ和Cbfa1 mRNA表达的影响

目的研究瘦素在体外对原代培养乳鼠颅盖骨成骨细胞增殖及对CollagenⅠ(Ⅰ型胶原蛋白)和Cbfa1 mRNA表达的影响。方法培养乳鼠成骨细胞,以乳鼠成骨细胞为体外实验模型,采用MTT法检测不同浓度瘦素(0、40、80和160ng/ml)作用于成骨细胞后的增殖情况;通过RT-PCR方法检测不同浓度瘦素对CollagenⅠ和Cbfa1 mRNA的表达的影响。结果瘦素作用于成骨细胞后,可促进其增值,OD值显著增加(P<0.01);瘦素增加CollagenⅠ和Cbfa1 mRNA的表达,呈剂量效应关系。结论瘦素促进成骨细胞增殖,同时通过调节CollagenⅠ和Cbfa1的表达,促进成骨细胞分化,进而促进骨形成。

氧化苦参碱对皮肤创面愈合中PCNA、α-SMA及Type Ⅰ collagen的影响

目的探讨氧化苦参碱对小鼠皮肤创面愈合中血清增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)及细胞Ⅰ型胶原蛋白(Type Ⅰ collagen)的影响。方法昆明小鼠背部手术制备1.5 cm×1.5 cm全层皮肤缺损创面模型。自第1 d始,除对照组给予等量生理盐水外,余各组分别按20、40、80 mg/kg给予氧化苦参碱。Van Gieson纤维胶原染色法观察小鼠背部皮肤创面组织胶原纤维的表达;免疫组学法评价创面组织中PCNA、α-SMA及Type Ⅰ collagen的表达。结果Van Gieson纤维胶原染色显示,氧化苦参碱可促进新生肉芽组织、毛细血管及新生纤维胶原生长。免疫组学研究显示,在第7 d时,氧化苦参碱20、40、80 mg/kg使小鼠皮肤创面组织中PCNA表达明显升高(P<0.05);在第9 d、11 d时,氧化苦参碱20 mg/kg使小鼠皮肤创面组织中α-SMA显著增加;氧化苦参碱20 mg/kg在第3 d、11 d,氧化苦参碱40 mg/kg在第3 d,氧化苦参碱80 mg/kg在第3 d、7 d引起Type Ⅰ collagen的表达显著升高(P<0.05)。结论氧化苦参碱能增加皮肤创面愈合中PCNA、α-SMA及Type Ⅰ collagen的表达。

白细胞介素1β与机械牵拉共同作用角膜成纤维细胞CollagenⅠ及Lumican的表达

背景:继发性圆锥角膜是角膜屈光手术后的并发症之一,但其发生机制尚不清楚。目的:探讨白细胞介素1β与机械牵拉对角膜成纤维细胞CollagenⅠ和Lumican表达的影响。方法:体外培养角膜成纤维细胞,对其施加不同幅度(5%,10%,15%)的机械牵拉和不同质量浓度的白细胞介素1β,共同作用12,24,36 h,通过实时荧光定量PCR检测CollagenⅠ和Lumican m RNA的表达变化。结果与结论:(1)白细胞介素1β单独作用12 h使CollagenⅠα1的表达降低(P<0.05),作用24 h使Lumican的表达升高(P<0.05);(2)5%牵拉单独作用24,36 h使CollagenⅠα1表达升高(P<0.05);10%和15%牵拉单独作用12,24,36 h使CollagenⅠα1和CollagenⅠα2的表达降低(P<0.05);(3)5%牵拉单独作用12 h使Lumican表达升高(P<0.05),而10%和15%牵拉单独作用12 h使Lumican表达降低(P<0.05);5%,10%,15%牵拉24 h和36 h时使Lumican表达升高(P<0.05);(4)白细胞介素1β和机械牵拉共同作用24,36 h使CollagenⅠα1和CollagenⅠα2表达降低,Lumican表达升高;(5)结果提示,白细胞介素1β能抑制CollagenⅠ的合成,而促进Lumican的表达。低幅度机械牵拉能促进角膜成纤维细胞胶原的合成,中高幅度机械牵拉抑制其胶原的合成。两者共同作用能抑制角膜成纤维细胞胶原合成且促进Lumican的表达。

TGF-β/Smad信号转导通路对紫外线致人工皮肤光损伤中MMP-1、pro-collagen Ⅰ mRNA表达影响的初步研究

目的探讨人工皮肤光损伤中TGF-β/Smad信号转导通路与基质金属蛋白酶1及Ⅰ型前胶原蛋白的相互作用。方法以紫外线照射人工皮肤后,通过Real-Time RT-PCR法(荧光染料掺入法)检测其中基质金属蛋白酶1、Ⅰ型前胶原蛋白、TGFβRⅠ及TGFβRⅡmRNA的表达量。结果紫外线使人工皮肤中基质金属蛋白酶1(MMP-1)的mRNA表达量明显增加,而Ⅰ型前胶原蛋白mRNA的表达下调(P<0.05),TGFβRⅡmRNA表达显著下调(P<0.01);加入TGF-β1后MMP-1表达显著下调(P<0.01),Ⅰ型前胶原蛋白及TGFβRⅡmRNA表达显著增高(P<0.01);而UV照射后8小时加入TGF-β1,MMP-1表达量较对照组高(P<0.05),Ⅰ型前胶原蛋白及TGFβRⅡmRNA表达量均较对照组低(P<0.05)。结论 UV可以抑制人工皮肤中Ⅰ型前胶原mRNA的表达、上调MMP-1的表达,而这种调控作用可能与TGF-βRⅡ的表达受抑制,TGF-β/Smad信号传导通路被削弱有关。

IGF-Ⅰ对体外培养的胎鼠肺成纤维细胞collagenⅠ合成的作用及其细胞内信号转导机制

目的探讨IGF-Ⅰ对体外培养的胎鼠肺成纤维细胞collagenⅠ合成的作用及其细胞内的信号转导机制。方法体外培养胎鼠肺成纤维细胞,给予不同剂量的rhIGF-Ⅰ,作用不同的时间,研究rhIGF-Ⅰ对胎鼠肺成纤维细胞collagenⅠ合成的量效和时效作用,并通过用MEK1途径的特异性阻断剂PD98059或PI3K途径的特异性阻断剂LY294002预处理胎鼠肺成纤维细胞,研究rhIGF-Ⅰ刺激胎鼠肺成纤维细胞collagenⅠ合成的细胞内信号转导机制。结果 rhIGF-Ⅰ在10～200 ng/mL剂量作用下能剂量依赖性地刺激胎鼠肺成纤维细胞collagenⅠ mRNA的合成;rhIGF-Ⅰ在200 ng/mL时分别作用于胎鼠肺成纤维细胞0.5～24 h能时间依赖性地刺激collagenⅠ mRNA的合成;LY294002能部分抑制rhIGF-Ⅰ的促胎鼠肺成纤维细胞collagenⅠ mRNA合成的作用,PD98059对rhIGF-I促胎鼠肺成纤维细胞collagenⅠ mRNA合成的作用无影响。结论 IGF-Ⅰ是影响胎鼠肺成纤维细胞collagenⅠ合成的生长因子之一,rhIGF-Ⅰ促胎鼠肺成纤维细胞collagenⅠ合成的作用是通过PI3K-Akt途径实现的。

葡甘露聚糖凝胶对宫颈炎大鼠CollagenⅠ、CollagenⅢ表达的影响

目的:探讨葡甘露聚糖凝胶对宫颈炎大鼠宫颈组织中Ⅰ型胶原蛋白(CollagenⅠ)、Ⅲ型胶原蛋白(CollagenⅢ)的调节作用。方法:采用苯酚胶浆进行阴道内注射建立宫颈炎模型,造模结束后进行局部给药治疗,将18只雌性SD大鼠造模后分为模型组、葡甘露聚糖凝胶高、中、低剂量组(1.2 g·kg-1、0.6 g·kg-1、0.3 g·kg-1)、复方甲硝唑阴道栓组(0.4 g·kg-1)、凝胶基质组各3只,另外3只作为空白组。阴道内灌注给药7天后,RT-PCR和Western blot法检测大鼠宫颈组织中CollagenⅠ、CollagenⅢmRNA及蛋白的表达。结果:RT-PCR法结果显示与空白组比较,模型组大鼠宫颈组织中CollagenⅠ、CollagenⅢmRNA的表达均明显降低(P﹤0.01);与模型组比较,葡甘露聚糖凝胶给药组大鼠宫颈组织中CollagenⅠ、CollagenⅢ的表达有不同程度的提高(P﹤0.01或P﹤0.05)。Western blotting法检测宫颈组织中CollagenⅠ、CollagenⅢ蛋白的表达:与空白组比较,模型组大鼠宫颈组织中CollagenⅠ、CollagenⅢ蛋白的表达均有所降低(P﹤0.01);与模型组比较,葡甘露聚糖凝胶给药组大鼠宫颈组织中CollagenⅠ、CollagenⅢ蛋白的表达有不同程度的提高(P﹤0.01或P﹤0.05)。结论:葡甘露聚糖凝胶对宫颈炎大鼠宫颈组织中CollagenⅠ、CollagenⅢ表达具有促进作用,从而促进宫颈黏膜的组织修复。

Growth and differentiation of neural stem cells in a three-dimensional collagen gel scaffold

Collagen protein is an ideal scaffold material for the transplantation of neural stem cells. In this study rat neural stern cells were seeded into a three-dimensional collagen gel scaffold, with suspension cultured neural stem cells being used as a control group. Neural stem cells, which were cultured in medium containing epidermal growth factor and basic fibroblast growth factor, actively expanded and formed neurospheres in both culture groups. In serum-free medium conditions, the processes extended from neurospheres in the collagen gel group were much longer than those in the suspension culture group. Immunofluorescence staining showed that neurespheres cultured in collagen gels were stained positive for nestin and differentiated cells were stained positive for the neuronal marker βIII-tubulin, the astrocytic marker glial fibrillary acidic protein and the oligodendrocytic marker 2＇,3＇-cyclic nucleotide 3＇-phosphodiesterase. Compared with neurospheres cultured in suspension, the differentiation potential of neural stem cells cultured in collagen gels increased, with the formation of neurons at an early stage. Our results show that the three-dimensional collagen gel culture system is superior to suspension culture in the proliferation, differentiation and process outgrowth of neural stem cells.

Effects of gentiana scabra bage on expression of hepatic type Ⅰ,Ⅲ collagen proteins in Paragonimus skrjabini rats with liver fibrosis

Objective:To exploit- the effects of gentiana scabra bage on the expression of hepatic collagen proteins in Paragonimus skrjabini rats with liver fibrosis.Methods:Immunohistochemical technique was used to observe the changes of content of hepatic type Ⅰ,Ⅲ collagen proteins in Paragonimus skrjabini rats with Liver fibrosis before and after the gentiana scabra bage treatmeat.Results:Comparing with the model group,changes of hepatic tvpe Ⅰ and type Ⅲ collagen proteins in gentiana scabra bage treated group were significantly weakened.Conclusions:Gentiana scabra bage treatment can reduce the content of hepatic type Ⅲ and type Ⅰ collagen protein significantly in Paragonimus skrjabini rats with liver fibrosis,thereby,playing the role against hepatic fibrosis.

温经汤加味对EM肾虚血瘀证大鼠局部微环境MMP-9、TNF-α、Collagen Ⅰ、Collagen Ⅲ的影响及其意义

目的探讨子宫内膜异位症(Endometosis,EM)肾虚血瘀证大鼠异位病灶局部MMP-9,TNF-α,CollagenⅠ、CollagenⅢ表达水平,以及温经汤加味对其干预作用及机制。方法SPF级雌性未孕健康SD大鼠设为空白组;构建肾虚血瘀证大鼠模型,造模成功后,设为假手术组(仅开腹);其余大鼠以自体内膜移植法构建EM肾虚血瘀证模型,造模成功后的大鼠分为模型组(不用药物干预)、阳性药(达那唑)组、温经汤加味组(低、中、高3种剂量)。阳性药对照组给予达那唑63 mg·kg^(-1)连续4周灌胃,中药低、中、高剂量组分别以5 g·kg^(-1)、10 g·kg^(-1)、20 g·kg^(-1)连续4周灌胃。取各组大鼠子宫内膜组织观察组织病理变化;免疫组化法(IHC)检测各组大鼠子宫内膜组织MMP-9,TNF-α的蛋白表达;蛋白免疫印迹法(WB)、实时荧光聚合酶链式反应法(RT-PCR)检测各组大鼠子宫内膜组织MMP-9、TNF-α、CollagenⅠ、CollagenⅢ基因表达。结果与空白组、假手术组比较,EM肾虚血瘀证模型组大鼠在位、异位内膜组织中MMP-9、TNF-α、CollagenⅠ、CollagenⅢ蛋白及其基因表达在模型组子宫内膜组织升高(P<0.01);与模型组比较,阳性药对照组、温经汤加味治疗组大鼠MMP-9、TNF-α、CollagenⅠ、CollagenⅢ蛋白及其基因表达水平均降低(P<0.01或P<0.05)。结论局部微环境免疫抑制微血管新生导致异位内膜侵袭是EM发生、发展过程中的重要病理表现,MMP-9、TNF-α、CollagenⅠ、CollagenⅢ过度表达可能促进了这一过程的发生、发展;温经汤加味通过"逆转"MMP-9、TNF-α、CollagenⅠ、CollagenⅢ等细胞因子异常升高,起到了解除局部微环境免疫抑制、阻断微血管新生的作用,在治疗EM肾虚血瘀证过程中表现出确切的疗效。

Effects of Icariin on Expression of OPN mRNA and Type ⅠCollagen in Rat Osteoblasts in Vitro

To study the effects of Icariin on expression of osteopontin （OPN） mRNA and type Ⅰ collagen in rat osteoblasts in vitro and to explore its possible mechanisms in preventing osteoporosis. OB was isolated from calvaria of new-born new-born fetal Sprague-Dawley （SD） rats by means of modified sequential collagenase digestion and incubated in MEM medium and the cell morphology was observed under inverted phase contrast microscope, OB was identified by alkaline phosphatase （ALP） staining. Different concentration （0.1μg/mL, 1.0 μg/mL, 10 μ/mL） of Icariin was added to the OB and incubated. The effect of Icariin on the proliferation and osteogenesis of OB was monitored by MTT analysis. The expression of type l collagen was estimated with immunohistochemistry techniques. The expression levels of mRNA of OPN in the cells in every group were examined by reverse-transcriptase ploymerase chain reaction （RT-PCR）. The expression of OPN mRNA and type Ⅰ collagen was strengthened gradually with the increase of Icariin concentration and peaked with 10 μg/mL Icariin on the 5th day. Icariin could significantly promote the expression of OPN mRNA and type Ⅰ collagen in rat osteoblasts in vitro. The levels of expression of OPN mRNA and type Ⅰ collagen were changed with different concentration of Icariin. Icariin could effectively prevent and treat osteoporosis and promote the bone formation.

硅橡胶硬度通过自噬调控成纤维细胞增殖及α- SMA、CollagenⅠ的表达

目的探讨不同硬度硅橡胶对人真皮成纤维细胞增殖及其表达α-SMA和CollagenⅠ的影响,并研究自噬在其中所起的作用。方法将A、B组分液体硅胶按不同比例混合固化制成一系列不同硬度的硅橡胶片,测定其邵氏硬度、断裂伸长率、水接触角、表面形貌等机械性能。利用CCK-8检测成纤维细胞在不同硬度硅橡胶表面的增殖情况,而后选取细胞增殖存在显著差异的两种硬度硅橡胶进一步实验,采用Western blot和ELISA法分别检测正常条件下、施加自噬抑制剂3-MA和诱导剂西罗莫司后成纤维细胞自噬标志蛋白和α-SMA、CollagenⅠ的表达情况。结果成纤维细胞增殖能力随硅橡胶硬度的增加而增加,差异有统计学意义(P<0.05)。高硬度硅橡胶上的成纤维细胞具有更高的自噬活性,且α-SMA、Collagen I的表达量比低硬度硅橡胶更低,差异有统计学意义(P<0.01)。结论硅橡胶硬度可通过自噬调控成纤维细胞的增殖,高硬度硅橡胶上调成纤维细胞自噬活性从而促进成纤维细胞增殖并抑制其表达α-SMA和CollagenⅠ。

Expression and role of specificity protein 1 and collagenⅠin recurrent pterygial tissues

AIM:To investigate the expression profiles of the transcription factor specificity protein 1(Sp1)and collagenⅠin recurrent pterygial tissues.What is more,to compare the changes of Sp1 and collagen I among primary pterygial tissue,recurrent pterygial tissue and conjunctival tissue.METHODS:In the prospective study,we collected the pterygial tissues of 40 patients who underwent resection of primary pterygial tissue and recurrent pterygial tissue,and the conjunctival tissues of 10 patients with enucleation due to trauma.The relative expression levels of Sp1 and collagen I were analyzed by reverse transcription quantitative-polymerase chain reaction and Western blot.Paired t-test was performed to compare the Sp1 and collagen I of recurrent pterygial tissues,as well as the primary pterygial tissues and conjunctival tissues.In further,Pearson’s hypothesis testing of correlation coefficients was used to compare the correlations of Sp1 and Collagen I.RESULTS:The content of Sp1 and collagen I m RNA and protein was significantly greater in recurrent pterygial tissue than that was in primary and conjunctival tissue(P<0.05).There was a positive correlation between the m RNA and protein levels of Sp1 and collagen I in recurrent pterygial tissues(protein:r=0.913,P<0.05;m RNA:r=0.945,P<0.05).CONCLUSION:Sp1 and collagen I are expressed in normal conjunctival,primary,and recurrent pterygial tissues,but expression is significantly greater in the latter.Sp1 and collagen I may be involved in the regulation of the development of recurrent pterygium.

CaMKⅡ抑制剂抑制AngⅡ或电场刺激诱导的心肌成纤维细胞TNF-α、TGF-β_1及collagenⅠ、Ⅲ的表达

目的:观察钙-钙调素依赖性蛋白激酶(CaMK)Ⅱ在血管紧张素Ⅱ(AngⅡ)或电场刺激(EFS)诱导的大鼠心肌成纤维细胞增殖、分泌细胞因子及胶原酶表达中的作用及其机制。方法:培养新生1-3 d乳鼠心肌成纤维细胞(3代),分为正常对照组(control)、0.1μmol/L AngⅡ组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂KN92组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂KN93组、0.1μmol/L AngⅡ+0.5μmol/L CaMKⅡ抑制剂AIP组;10 V1.0 Hz EFS组、10 V 1.0 Hz EFS+0.5μmol/L KN92组、10 V 1.0 Hz EFS+0.5μmol/L KN93组、10 V1.0 Hz EFS+0.5μmol/L AIP组、10 V1.0 Hz EFS+0.1μmol/L AngⅡ组。MTT法测定心肌成纤维细胞增殖;ELISA法测定细胞因子(TGF-β1,TNF-α)分泌;RT-PCR检测TGF-β1、TNF-α、collagenⅠ、ⅢmRNA水平。结果:CaMKⅡ抑制剂(0.5μmol/L KN93,0.5μmol/L AIP)能预防AngⅡ或EFS诱导的心肌成纤维细胞增殖;CaMKⅡ抑制剂(0.5μmol/L KN93,0.5μmol/L AIP)可预防AngⅡ或EFS引起的细胞培养上清液TGF-β1、TNF-α含量及相应mRNA表达增加。CaMKⅡ抑制剂(0.5μmol/L AIP,1.0μmol/L AIP)预防0.1μmol/L AngⅡ引起的collagenⅠ、Ⅲ表达增加。结论:抑制CaMKⅡ对AngⅡ或EFS诱导的心肌成纤维细胞增殖具有预防作用,其机制可能与CaMKⅡ抑制剂抑制TGF-β1、TNF-α以及胶原的表达有关。

Novel electrospun poly(ε-caprolactone)/type Ⅰ collagen nanofiber conduits for repair of peripheral nerve injury

Recent studies have shown the potential of artificially synthesized conduits in the repair of peripheral nerve injury. Natural biopolymers have received much attention because of their biocompatibility. To investigate the effects of novel electrospun absorbable poly(ε-caprolactone)/type Ⅰ collagen nanofiber conduits(biopolymer nanofiber conduits) on the repair of peripheral nerve injury, we bridged 10-mm-long sciatic nerve defects with electrospun absorbable biopolymer nanofiber conduits, poly(ε-caprolactone) or silicone conduits in Sprague-Dawley rats. Rat neurologica1 function was weekly evaluated using sciatic function index within8 weeks after repair. Eight weeks after repair, sciatic nerve myelin sheaths and axon morphology were observed by osmium tetroxide staining, hematoxylin-eosin staining, and transmission electron microscopy.S-100(Schwann cell marker) and CD4(inflammatory marker) immunoreactivities in sciatic nerve were detected by immunohistochemistry. In rats subjected to repair with electrospun absorbable biopolymer nanofiber conduits, no serious inflammatory reactions were observed in rat hind limbs, the morphology of myelin sheaths in the injured sciatic nerve was close to normal. CD4 immunoreactivity was obviously weaker in rats subjected to repair with electrospun absorbable biopolymer nanofiber conduits than in those subjected to repair with poly(ε-caprolactone) or silicone. Rats subjected to repair with electrospun absorbable biopolymer nanofiber conduits tended to have greater sciatic nerve function recovery than those receiving poly(ε-caprolactone) or silicone repair. These results suggest that electrospun absorbable poly(ε-caprolactone)/type Ⅰ collagen nanofiber conduits have the potential of repairing sciatic nerve defects and exhibit good biocompatibility. All experimental procedures were approved by Institutional Animal Care and Use Committee of Taichung Veteran General Hospital, Taiwan, China(La-1031218) on October 2, 2014.

Neurotrophins differentially stimulate the growth of cochlear neurites on collagen surfaces and in gels

The electrodes of a cochlear implant are located far from the surviving neurons of the spiral ganglion, which results in decreased precision of neural activation compared to the normal ear. If the neurons could be induced to extend neurites toward the implant, it might be possible to stimulate more discrete subpopulations of neurons, and to increase the resolution of the device. However, a major barrier to neurite growth toward a cochlear implant is the fluid filling the scala tympani, which separates the neurons from the electrodes. The goal of this study was to evaluate the growth of cochlear neurites in three-dimensional extracellular matrix molecule gels, and to increase biocompatibility by using fibroblasts stably transfected to produce neurotrophin-3 and brain-derived neurotrophic factor. Spiral ganglion explants from neonatal rats were evaluated in cultures. They were exposed to soluble neurotrophins, cells transfected to secrete neurotrophins, and/or collagen gels. We found that cochlear neurites grew readily on collagen surfaces and in three-dimensional collagen gels. Co-culture with cells producing neurotrophin-3 resulted in increased numbers of neurites, and neurites that were longer than when explants were cultured with control fibroblasts stably transfected with green fluorescent protein. Brain-derived neurotrophic factor-producing cells resulted in a more dramatic increase in the number of neurites, but there was no significant effect on neurite length. It is suggested that extracellular matrix molecule gels and cells transfected to produce neurotrophins offer an opportunity to attract spiral ganglion neurites toward a cochlear implant.

Significance of elevated serum concentration of type Ⅰ collagen metabolites and its correlation with serum interleukin 6 activities in multiple myeloma

In this study, serum concentrations of carboxyterminal propeptide of type Ⅰ collagen(PICP) and carboxyterminal cross-linked telopeptide of type Ⅰ collagen (ICTP), which represent the rates of synthesis and degradation of type Ⅰ collagen, were determined by radioimmunoassay in 56 patients with multiple myeloma (MM) and 22 healthy controls. It was discovered that serum concentrations of both PICP and ICTP were higher in MM than those in healthy controls (P<0. 01 ). With the disease progressing and the number of bone lesions increasing,serum concentration of ICTP elevated while serum concentration of PICP showed no significant change. Neither serum PICP nor ICTP concentration was related to M-component classes. Our results indicated that serum ICTP concentration was a good serological marker to reflect severity of bone lesions in MM and elevated serum PICP concentration might be due to compensatory increase in type Ⅰ collagen synthesis. Moreover, we also found that serum ICTP concentrations in MM correlated with serum interleukin-6 (IL-6) activities (r= 0. 610, P< 0. 01),which confirms the effectiveness of IL-6 as an osteoclast activating factor.

Inhibition of α_1(Ⅰ) collagen gene in vitro transcription by antisense oligodeoxynucleotides

Objective and Methods: Excessive accumulation of collagen typeⅠ and type Ⅲ causes the formation of keloids and hypertrophic scars. To understand the mechanism by which antisense oligodeoxynucleotide (Oligo) acts on in vitro transcrption α1 (I) collagen gene, isotopes (α-32pGTP) was incorporated into 2 SP6 in vitro transcription systems. Results and Conclu- sion: Oligo 2 (at the transcription start region) could effectively inhibit in vitro transcription of pGEM3-Col13 and the control (random oligodeoxynucleotides) showed no inhibition. However, oligo 1 (at the transcription start region) obviously inhibited the in vitro transcription of pGEM3-Col14, while Oligo 2, which targeted at the down stream region (about 200 bp) of the promoter showed no significant inhibition effect.

透明质酸联合注射血小板血浆及A型肉毒毒素对猪真皮Ⅰ型胶原蛋白表达的影响

目的探讨透明质酸联合A型肉毒毒素(botulinum toxin type-A,Botox A)和富含血小板血浆(platelet rich plasma,PRP)进行猪真皮内注射,检测真皮层成纤维细胞数目及Ⅰ型胶原蛋白mRNA分泌代谢的变化。方法采用巴马猪建立动物模型,使用交联透明质酸(cross-linked hyaluronic acid,HA)和非交联透明质酸(non-cross-linked hyaluronic acid,N-HA)分别与Botox A、PRP联合作为实验组(HA组、HA+Botox A组、HA+PRP组、N-HA组、N-HA+Botox A组、N-HA+PRP组),并与生理盐水组和正常组比较。于注射第1和4周后,通过组织切片染色、实时荧光定量聚合酶链反应(PCR)等方法,定性定量观察、比较真皮层成纤维细胞数量变化,真皮层Ⅰ型胶原蛋白mRNA分泌表达变化。结果HA组的成纤维细胞数量、Ⅰ型胶原蛋白mRNA表达量在第1周和第4周时比N-HA组高,并且HA+PRP组的成纤维细胞数量、Ⅰ型胶原蛋白mRNA表达水平显著高于同期HA+Botox A组、生理盐水组和正常组(P<0.05)。结论应用HA与PRP联合注射更易促进成纤维细胞的增殖和Ⅰ型胶原蛋白mRNA的表达,其主要机制可能是通过改变细胞外基质的理化性质进而促进真皮层细胞因子的合成,从而影响胶原合成。