钛表面粗糙度对牙周韧带细胞早期附着影响的荧光观察

背景:种植体表面粗糙程度对组织细胞的增殖、分化及基因表达有直接影响.目的:观察牙周韧带细胞在不同纯钛表面粗糙度的早期附状态,以及材料表面性能对组织细胞分化的影响.设计、时间及地点:随机对照观察,多样本比较实验,于2005-01/2006-07在赣南医学院科研中心完成.材料:采用机床切割机将商业纯钛棒制备成直径为10mm厚2mm的钛片,共24个,分为4组:机械处理组,硝酸处理组,喷砂处理组,综合处理组,每组6个样品.方法:采用TR240便携式表面粗糙度仪测定表面粗糙度.机械处理组:进行单纯的机械处理.硝酸处理组:单纯的机械处理+65%的硝酸(100℃,1 h)处理.喷砂处理组:单纯的机械处理+100 μ m的Al2O3喷砂处理.综合处理组:单纯的机械处理+100 μ m的Al2O3喷砂处理后+65%的硝酸(100℃,1 h)处理.主要观察指标:样品在DMEM中预置30 m后接种第3代的细胞,分别于30,60,120,240 min,1,3,7 d取出钛片,于荧光显微镜下观察各组纯钛表面的粗糙度和牙周韧带细胞在不同表面粗糙度的纯钛表面早期附着情况.结果:①定量分析:4组粗糙度分别为:机械处理组(599.5±8.3)nm、硝酸处理组(406.5±4.6)nm、喷砂处理组(358.8±11.8)nm、综合处理组(8.7±2.0)nm,机械处理组高于硝酸处理组、喷砂处理组和综合处理组(P<0.01),差异有显著性意义(P<0.01).②定性分析:牙周韧带细胞在纯钛表面早期附着的荧光观察显示,随着时间的推移,各组早期附着在纯钛表面的牙周韧带细胞增多,细胞增殖良好.但粗糙度大的钛表面,细胞附着较少,粗糙度小的钛表面,细胞附着较致密些.结论:钛表面粗糙度越小,牙周韧带细胞早期附着越多,有利于细胞黏附和增殖.     

加速角膜胶原交联术与离子导入跨上皮角膜胶原交联术治疗圆锥角膜的效果观察

目的探讨加速角膜胶原交联术与离子导入跨上皮角膜胶原交联术治疗圆锥角膜的效果。方法本研究为随机对照研究, 选取2020年6月至2022年1月郑州市中心医院眼科和郑州市第一人民医院眼科收治的17例(26只眼)临床诊断为圆锥角膜并接受治疗的患者, 男11例(17只眼), 女6例(9只眼), 年龄(25.7±4.5)岁, 年龄范围为16～37岁。根据手术方法的不同将患者分为加速交联组与离子导入组, 每组13只眼睛(如患者需双眼治疗, 则双眼归入同组)。加速交联组患者行加速角膜胶原交联术, 离子导入组患者行离子导入跨上皮角膜胶原交联术, 比较两组最佳矫正远视力、角膜前表面曲率、交联线增强信号情况及角膜内皮细胞的情况。结果术后6个月加速交联组矫正远视力[(0.31±0.13)log MAR]高于离子导入组[(0.17±0.05)log MAR];加速交联组角膜前表面曲率(48.59±3.99)高于离子导入组(47.96±6.14), 差异均有统计学意义(P<0.05)。术前、术后两组角膜内皮密度、六边形角膜内皮细胞比及细胞面积变异系数比较, 差异无统计学意义(P>0.05)。结论加速交...     

老年牙周致病菌生长与血糖变化关系研究

目的探讨老年牙周致病菌与血糖之间的关系。方法选取2013年1月至2014年12月在该院确诊的糖尿病(DM)患者50例、糖耐量异常患者50例及在门诊随机选取50例血糖正常的患者,检测其糖化血红蛋白、三酰甘油(TG)、总胆固醇(TC)、口服糖耐量实验2h的血糖浓度(2hPG)、口腔中各种菌群分布(球菌、杆菌、丝状菌、梭状菌、弯曲菌、螺旋菌)、口腔中各种致病菌的分布情况及碱性磷酸酶(ALP)的活性。结果三组受试者口腔内菌群分布结果显示,口腔中的球菌、杆菌、梭状菌和丝状菌的比较中,DM组和糖耐量异常组的值明显高于对照组,组间差异有统计学意义(P0.05)。而弯曲菌和螺旋体的比较中,各组间差异无统计学意义(P0.05)。对于口腔中的伴放线杆菌(Aa)、类杆菌(Bg)、卟啉单胞菌(Pg)、福塞类杆菌(Bf)值,DM组明显高于对照组(P0.05),糖耐量异常组的Aa、Pg、Bg、Bf值明显高于对照组(P0.05),DM组的值也明显高于糖耐量异常组(P0.05)。结论血糖可以影响到牙周致病菌分布的异常,并促进致病菌的生长繁殖,且与血糖值呈正相关,所以临床上高血糖且口腔存在牙周炎的患者要严格控制血糖值。     

牙周牙髓联合病变与牙周感染的相关性分析

目的探讨牙周牙髓联合病变与牙周感染的相关性。方法牙周牙髓联合病变患者62例（81颗牙）为观察组,同期行牙齿矫正或行其他治疗的牙病患者60例（79颗牙）为对照组,收集2组患牙根管内残留组织提取DNA,应用PCR技术对可能引起牙周牙髓部位感染的致病菌进行检测,并进行2组间比较。结果对照组均未检出致病菌,而观察组32例（37颗牙）检出致病菌,阳性率为45．68％（37／81）。结论牙周感染是牙周牙髓联合病变主要症状之一,可作为牙周牙髓联合病变的主要诊断依据。     

细胞因子的相互作用对角膜基质细胞胶原代谢的影响

目的观察细胞因子IL-1α与TGF-β对兔角膜基质细胞胶原降解的作用,并探讨其相互作用对角膜基质细胞胶原分解代谢的影响。方法角膜基质细胞表面添加含纤溶酶原及不同浓度的IL-1α和/或TGF-β的MEM液,培养24小时,测定培养液中的羟脯氨酸含量作为胶原降解活性。结果IL-1α促进角膜基质细胞胶原降解;TGF-β1抑制角膜基质细胞的胶原降解,IL-1α与TGF-β同时存在,在IL-1α低浓度时,抑制角膜基质细胞胶原降解,高浓度时,TGF-β1不能抑制角膜基质细胞胶原降解。结论IL-1α与TGF-β的相互作用调控角膜基质细胞胶原代谢,在角膜基质创伤修复过程中起着重要作用。     

胶原海绵和温敏型Ⅰ型胶原细胞相容性比较

背景:因心脏结构高度复杂、心肌组织有很高的机械顺应性,心肌组织工程对材料的要求高.以往研究表明Ⅰ型胶原和胶原海绵材料均适宜心肌细胞生长及分化.目的:拟进一步对比观察胶原海绵和温敏型Ⅰ型胶原与骨髓间充质干细胞的相容性,为心肌组织工程选择更理想的载体材料.设计、时间及地点:在解放军第四军医大学西京医院心血管内科实验室2007-03/10完成对比观察设计的实验.材料:胶原海绵由上海其胜制剂有限公司提供;温敏型Ⅰ型胶原由第四军医大学组织工程中心提供.方法:将体外培养的大鼠骨髓间充质干细胞分别复合于胶原海绵和温敏型l型胶原材料,共同培养7 d,分别于培养第1,3,5和7天取材.主要观察指标:扫描电镜和苏木精.伊红染色观察骨髓间充质干细胞细胞形态及附着情况;MTT检测骨髓间充质干细胞细胞增殖情况.结果:①扫描电镜检测显示温敏型Ⅰ型胶原材料中骨髓间充质干细胞细胞黏附优于胶原海绵材料,细胞在Ⅰ型胶原材料中生长连接成片,表面有大量分泌颗粒.②MTT检测表明在培养第1,3,5,7天温敏型Ⅰ型胶原材料中骨髓间充质干细胞细胞活性(吸光度值)及细胞分裂增殖速度均高于胶原海绵材料(P＜0.01).结论:温敏型Ⅰ型胶原体外与骨髓间充质干细胞的相容性优于胶原海绵.     

原子力显微镜对不同类型胶原蛋白与细胞黏附情况的研究

目的研究不同类型胶原蛋白与成纤维细胞及角质形成细胞的黏附情况,并用原子力显微镜观察其相互作用。方法分别用牛肌腱胶原、鼠尾胶原和标准Ⅳ型胶原包被培养板,接种相同数量的原代角质细胞,在不同时相用活细胞电子计数仪测定未黏附细胞的数量,观察不同类型胶原所未黏附的细胞数量的多少,并用原子力显微镜观察胶原蛋白与细胞的黏附情况。结果接种后5min,三种胶原（鼠尾胶原、牛肌腱胶原、标准Ⅳ型胶原）中未贴壁的细胞数差别较大,Ⅳ型胶原中未贴壁的细胞最少;但培养至1h,三种胶原中未贴壁的细胞数无明显差别,三者的贴壁率几乎相同;培养至第6小时三种胶原中未贴壁的细胞数仍然无明显差别,三者的贴壁率仍然基本相近。原子力显微镜可以清晰观察到胶原及胶原与细胞间黏附的微观结构。结论牛肌腱胶原和鼠尾胶原与Ⅳ型胶原对角质形成细胞的黏附作用无明显差别,原子力显微镜可以用于观察胶原及胶原与细胞间黏附的微观结构。     

纯钛表面微弧氧化结合Ⅰ型胶原对成骨细胞黏附增殖的影响

目的 探讨微弧氧化(microarc oxidation,MAO)结合应用于纯钛种植体表面处理的可行性.方法 根据对纯钛钛片处理的不同将实验分为对照组(A组,不作处理)、MAO组(B组,纯钛片上进行MAO处理)及MAO加Ⅰ型胶原组(C组,纯钛片上MAO处理后吸附Ⅰ型胶原).将成骨细胞培养于各组钛片上,通过扫描电镜、MT...     

交联对胶原生物学和物理性能的影响

背景:水溶性1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺和N-羟基琥珀酰亚胺是无毒、生物相容性良好的交联剂,可促发胶原发生交联,形成酰胺交联键.目的:分析交联对胶原生物学和物理性能有何影响.设计、时间及地点:胶原的物理化学性质观察实验,于2008-09/11在福建工程学院材料科学与工程系实验室完成胶原交联的实验.材料:I型生胶原溶液由广州创尔公司提供.1-乙基-(3-3-二甲基氨丙基)-碳化-二亚胺和N-羟基琥珀酰亚胺为交联剂分别由上海吉尔生化公司和上海伯奥生物科技有限公司提供.以N-吗啡林和乙撑磺酸为缓冲剂,由上海源聚生物科技有限公司提供.方法:将胶原溶液置于冷冻机中,在-80℃下冷冻12 h.之后,转入冷冻干燥机中在真空状态下冷冻干燥24 h至完全脱水,得到胶原网络基质.再将此干燥胶原基质浸入分别浸入不同浓度的1-乙基-3-3-二甲基氨丙基)·碳化二亚胺溶液,将N-羟基琥珀酰哑胺按质量比为1-乙基-3-3-二甲基氨丙基)-碳化二亚胺:N-羟基琥珀酰亚胺=4:1的量加入,以乙撑磺酸为缓冲剂调pH至5.5.经去离予水清洗后在真空状态下冷冻干燥至完全脱水.主要观察指标:胶原交联前后显微结构、缩水温度、膨胀动力学特征以及抵抗降解能力.结果:对胶原交联前后性能的研究表明,交联后胶原分子中自南-NH2官能团转换成N-H官能团,胶原分子内及分子间的氨基链接形成了;当1-乙基-3-(3-二甲基氨丙基)-碳化二业胺浓度达到2 g/L时,胶原的交联程度达到最大;交联后,胶原的显微结构从交联前的无序状态变成紧密有序的结构,热稳定性、形态稳定性增强,抵抗降解的能力显著增加.结论:采片1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺-和N-羟基琥珀酰亚胺-作为交联剂对胶原进行交联处理,可有效改善胶原作为骨组织工程材料的生物学和物理性能.     

骨髓基质细胞在胞外基质涂层钛表面的黏附和铺展

背景:为模拟胞外基质对生物行为的调控功能,通常采用基质中的活性成分对钛表面进行改性,但这些单一的活性成分与天然基质仍有差距.目的:在体外观察成骨细胞自身分泌的胞外基质修饰的钛表面对骨髓基质干细胞生物学行为的影响,为进一步指导钛种植体表面的仿生构建提供实验依据.设计、时间及地点:细胞-材料学体外对照观察,于2008-12/2009-02在安徽医科大学医学生物工程实验室完成.材料:SPF级新生1～3 d龄SD乳鼠2只,SD大鼠2只,由安徽省实验动物中心提供.TA2纯钛棒为宝鸡金埠钛设备有限公司产品.方法:取SD乳鼠,采用改良组织块法培养成骨细胞.取直径12 mm的钛棒,加工成厚1 mm纯钛片,消毒后放入24孔培养板中,用DMEM培养液将成骨细胞浓度调整为3×10.L-1,取1 mL/孔接种于钛片上,经过反复冻融脱玄细胞留下基质,即为基质化钛片.取SD大鼠,采用全血贴壁法培养骨髓摹质干细胞,取传至第3代的细胞,分为基质化钛片组、纯钛片组,用DMEM培养液将其浓度调整为1×108L-1,分别接种于基质化钛片和纯铁片上,1 mL/孔.主要观察指标:通过荧光免疫组化、扫描电镜观察和MTT检测,评价两组钛片上骨髓基质细胞的早期黏附及铺展情况.结果:接种4 h时与纯钛片组比较,基质化钛片组细胞黏附率显著升高(P＜0.05).接种4 h后,骨髓基质干细胞在基质化钛片表面已呈现出一定的附着形态:24 h后细胞呈良好的铺展形态,紧贴附于基质化钛片表面,出现伪足突起,细胞间以这种突起建立相互联系;72 h后细胞充分铺展,胞体平铺面积迅速扩大.结论:胞外基质的存在能够促进骨髓基质细胞的早期黏附,并且有利于细胞的进一步铺展.     

红细胞膜表面黏附分子与红细胞寿命的相关性研究

目的 探讨红细胞寿命与其膜表面蛋白黏附分子的关系,以期建立1种检测红细胞贮存时间的方法 .方法 采集10人(份)健康无偿献血者的新鲜红细胞标本10 ml/份,应用Percoll密度梯度离心法将红细胞分成5个年龄(层),流式检测术检测各分层红细胞膜表面黏附分子CD47、CD44、CD147的表达量;应用SPSS统计软件对...     

Effect of surface energy and roughness on cell adhesion and growth – facile surface modification for enhanced cell culture

In vitro, cellular processing on polymeric surfaces is fundamental to the development of biosensors, scaffolds for tissue engineering and transplantation. However, the effect of surface energy and roughness on the cell–surface interaction remains inconclusive, indicating a lack of complete understanding of the phenomenon. Here, we study the effect of surface energy (E_s) and roughness ratio (r) of a polydimethylsiloxane (PDMS) substrate on cell attachment, growth, and proliferation. We considered two different cell lines, HeLa and MDA MB 231, and rough PDMS surfaces of different surface energy in the range E_s = 21–100 mJ m⁻², corresponding to WCA 161°–1°, and roughness ratio in the range r = 1.05–3, corresponding to roughness 5–150 nm. We find that the cell attachment process proceeds through three different stages marked by an increase in the number of attached cells with time (stage I), flattening of cells (stage II), and elongation of cells (III) on the surface. Our study reveals that moderate surface energy (E_s ≈ 70 mJ m⁻²) and intermediate roughness ratio (r ≈ 2) constitute the most favourable conditions for efficient cell adhesion, growth, and proliferation. A theoretical model based on the minimization of the total free energy of the cell–substrate system is presented and is used to predict the spread length of cells that compares well with the corresponding experimental data within 10%. The performance and reusability of the rough PDMS surface of moderate energy and roughness prepared via facile surface modification are compared with standard T-25 cell culture plates for cell growth and proliferation, which shows that the proposed surface is an attractive choice for efficient cell culture.

In vitro, cellular processing on polymeric surfaces is fundamental to the development of biosensors, scaffolds for tissue engineering and transplantation.     

Effects of molecular weight and surface functionalization on surface composition and cell adhesion to Hyaluronan coated titanium.

This paper describes the effect of surface functionalization on surface composition and cell adhesion to titanium samples by high and low molecular weight Hyaluronan (HA). HA was covalently linked to aminated Ti surfaces obtained by two different surface functionalization techniques, that is polyethyleneimine (PEI) adsorption and deposition from allylamine plasma. The two approaches yield very different surface densities of available amino groups, affecting this way the number and frequency of surface-HA bonds and the configurational freedom of the latter. Results of cell adhesion test are dependent on the surface functionalization approach adopted, low molecular weight HA coupled to PEI functionalized Ti does not yield the same degree of resistance to cell adhesion found on other samples. These results indicate that the details of the surface functionalization step are crucial for surface engineering of implant devices by biological molecules.     

Scanning and non-scanning surface plasmon microscopy to observe cell adhesion sites

We observe adhesion sites of a cell on a substrate with high resolution. Since this observation requires interfacial measurements between the cell and the substrate, we employ scanning localized surface plasmon microscopy. We experimentally show that focal adhesion sites of a mouse muscle cell can be observed without fluorescent labeling. We also show that a non-scanning surface plasmon microscope combined with the scanning localized surface plasmon microscope contributes to observing an entire cell adhesion site and identify regions of interest.     

Novel gene-transferring scaffolds having a cell adhesion molecule-DNA-apatite nanocomposite surface

A low efficiency has long been the most critical problem of conventional gene-transferring systems using calcium phosphates, and this was successfully improved on by using a laminin-DNA-apatite composite (LD-Ap) layer. The gene-transferring efficiency of the LD-Ap surface was 1-2 orders of magnitude higher than that of a DNA-calcium phosphate composite surface. This is because laminin enhances cell adhesion and spreading, and this provides regions of high DNA concentration between a cell and the LD-Ap surface. The efficiency of gene transfer of the LD-Ap surface was equivalent to, or even higher than that mediated using a commercial lipid-based transfection reagent applied using the manufacturer's recommended optimum conditions. In addition, the gene-transferring efficiency of our system could be controlled by changing the laminin and DNA content in the LD-Ap layer. Moreover, our system is composed of highly safe reagents: apatite, DNA and laminin, all of which are present in the human body. Hence, the LD-Ap surface, which enhances cell attachment on its surface, and mediates a safe, highly efficient and controllable gene transfer, is highly applicable to tissue engineering and gene therapy applications.     

肺炎链球菌细胞壁表面粘附相关蛋白的初步鉴定

通过Ｗｅｓｔｅｒｎｂｌｏｔ将肺炎链球菌细胞壁表面蛋白质转移到ＰＶＤＦ膜上，然后与生物素标记的巨噬细胞在膜上进行粘附实验，初步鉴定出六条细胞壁表面粘附相关蛋白，它们的分子量分别是２０，３０，３７，５９，６６，８５ｋＤ其生物学特性，功能有待进一步研究确定，这为进一步研究肺炎链球菌致病机理，发展新一代多价疫苗和开发抗菌药物奠定了一定的基础。     

Effects of interferon-gamma on expression of cell surface receptors for collagen and deposition of newly synthesized collagen by cultured human lung fibroblasts.

We used cultured human diploid lung fibroblasts as a model system to examine the effects of recombinant IFN-gamma on synthesis of collagen, matrix deposition of newly synthesized collagen, and the expression of cell surface receptors for collagen. Using [3H]proline-labeled cells we found that IFN-gamma resulted in dose-dependent inhibition of fibroblast collagen synthesis. Pulse-chase experiments to analyze compartmentalization of newly synthesized collagen showed that the decrease in collagen synthesis was confined to the soluble pool of procollagen in the medium, while extracellular matrix associated collagen was not changed, indicating that a larger proportion of newly synthesized collagen was deposited into the matrix in IFN-gamma exposed fibroblasts (34.2 vs. 25.3%). This increase in the efficiency of collagen matrix deposition was associated with enhanced expression of a cell surface receptor for collagen as detected by indirect immunofluorescence labeling and analysis by flow cytometry. Fibroblasts (IMR-90) cultured in the presence of IFN-gamma (1,000 U/ml) exhibited a twofold increase in mean linear fluorescence intensity compared with cells cultured under control conditions. The distribution of log fluorescence intensity in both control and IFN-gamma exposed cells was normally distributed about the mean, indicating that discrete subpopulations with respect to receptor expression were not present. Increased fluorescence intensity and log normal distribution of fluorescence intensity also were identified in IFN-gamma-treated lung fibroblasts from a normal adult individual and two strains obtained from patients with pulmonary fibrosis. These results indicate that IFN-gamma modulates fibroblast collagen matrix deposition as well as collagen synthesis. The associated increase in collagen receptors suggests that cytokine-mediated modulation of the cell surface maybe a contributing factor in regulation of fibroblast collagen accumulation in the extracellular matrix or in cellular interaction with collagen-containing matrix. Such an effect could modulate the interaction of fibroblasts with extracellular matrix at sites of inflammation and play an important role in the remodeling of matrix during repair from tissue injury.     

磷酸果糖抑制肌腱胶原产生和趾屈指肌腱吻合后的粘连

背景：有实验证实外源性6-磷酸果糖能降低肌腱细胞Ⅰ型胶原的产生量,减轻肌腱术后粘连的形成。目的：观察6-磷酸果糖对肌腱术后粘连形成的影响。方法：取72只兔行中趾屈指肌腱切断吻合,随机分成实验组与对照组（n=36）,分别于腱鞘内注入6-磷酸果糖与生理盐水,术后4,8周后行肌腱粘连检测、生物力学测定、组织学观察和扫描电镜观察;术后1,2,4,8周采用原位杂交方法测定肌腱转化生长因子β1和Ⅰ型胶原mRNA的表达。结果与结论：术后4,8周,与对照组比较,实验组肌腱缝合处光滑,屈趾肌腱滑动距离较长,肌腱滑动受限较轻（P〈0．05）,但两组最大抗断裂载荷差异无显著性意义（P〉0．05）;扫描电镜和组织学观察结果显示,对照组胶原纤维排列紊乱,实验组胶原纤维排列整齐。实验组转化生长因子β1和Ⅰ型胶原mRNA表达水平均低于对照组（P〈0．05）。证实6-磷酸果糖能有效抑制转化生长因子β1在肌腱损伤修复中的作用,减轻粘连形成。     

Role of adhesion molecules in the pathogenesis of collagen diseases

Adhesion molecules play important roles and involve in many aspects of cell-cell or cell-extracellular matrix interactions in collagen diseases. In the present article, we describe the role of adhesion molecules in the pathogenesis of collagen diseases. Adhesion molecules on endothelial cells participate in leukocyte recruitment in collagen diseases. Adhesion molecules on RA synoviocytes which interact with MNCs and destroy chondrocytes by pannus formation. In PSS, adhesion molecules on fibroblasts play important roles to form fibrosis by interacting with MNCs or collagen fibrils. We further mention on adhesion molecules in interaction between MNCs and muscle fibers in myositis, those in vasculitis in SLE and other diseases, and autoantibodies inducing adhesion molecules on endothelial cells in PSS and Wegener's granulomatosis.