碱性成纤维细胞生长因子对瘢痕成纤维细胞生物学行为的影响

目的探讨碱性成纤维细胞生长因子（bFGF）对瘢痕成纤维细胞生物学特性的影响。方法采用细胞培养、MTT、ELISA法、氯胺T和RT—PCR法检测bFGF在不同作用剂量下对瘢痕来源的成纤维细胞生长增殖和细胞外基质（ECM）合成的影响。结果（1）MTT检测bFGF对瘢痕成纤维细胞有明确的促增殖作用，并具有剂量相关性；（2）氯胺T法显示实验组和对照组HPr含量无显著性差异，RT—PCR法显示各组Ⅰ、Ⅲ型胶原蛋白mRNA表达水平无明显变化；说明bFGF对瘢痕成纤维细胞胶原蛋白的合成无促进作用；（3）ELISA法表明随着bFGF作用浓度的升高，FN的表达表现为增高趋势，且以100ng／ml浓度下作用最显著。结论bFGF可以促进创面愈合，但不引起瘢痕的过度形成。     

成纤维细胞的生物学特性及其成骨作用的研究进展

骨缺损（尤其是大型骨缺损）的治疗,由局部伤情复杂和缺乏理想的修复材料,一直是困扰临床医生和基础医学工作者的一大难题,而寻找一种尽可能达到或接近自体骨移植效果的理想的骨替代材料更是无数学者热切探索、孜孜以求的目标。近年来日趋活跃的骨组织工程（ｂｏｎｅｔｉｓｓｕｅｅｎｇｉｎｅｅｒｉｎｇ）技术为这一课题的研究带来了新的亮点和希望。目前动物实验已能从骨膜、骨髓等定向性骨祖细胞（ｄｅｔｅｒｍｉｎｅｄｏｓｔｅｏｇｅｎｉｃｐｒｅｃｕｒｓｏｒｃｅｌｌｓ,ＤＯＰＣ）密集处分离培养出成骨细胞,经体外扩增并与载体结合…     

止痒软疤擦剂对人成纤维细胞生物学行为的影响及其机制研究

目的 研究止痒软疤擦剂对人成纤维细胞生物学行为的影响，探讨其对瘢痕形成的抑制作用及其可能机制。方法采用梯度浓度的止痒软疤擦剂对体外培养的人皮肤成纤维细胞进行干预，观察干预后成纤维细胞的组织学改变，并以MTF法检测细胞增殖状态，ELISA法检测TGF-β1的蛋白表达水平，RT—PCR法检测干预后成纤维细胞的Ⅰ、Ⅲ型前胶原mRNA表达。结果与正常组比较，1：800至1：200各稀释度干预条件下人成纤维细胞的增殖均被显著抑制，1：800稀释度干预后成纤维细胞Ⅲ型前胶原mRNA表达显著降低，TGF-β1的蛋白表达明显升高。结论止痒软疤擦剂可通过抑制人成纤维细胞增殖，并在转录水平下调Ⅲ型前胶原mRNA表达，减少细胞外胶原的沉积，从而减少和／或减轻瘢痕增生的发生。     

虎杖苷对成纤维细胞生物学特性的影响

背景：虎杖多年来一直是烧伤、创伤创面愈合治疗方剂中的一味主药,因成分复杂,具体药理作用难以进一步研究,对于虎杖苷促愈合作用目前未见文献报道。 目的：分析不同浓度虎杖苷对成纤维细胞生物学特性的影响。 方法：取烧伤后行瘢痕切除植皮4例患者剩余小中厚皮片,原代培养人成纤维细胞。用含有10^-6,10^-5,10^-4,10^-3,10^-2 mol/L不同浓度虎杖苷培养液作用第2代人成纤维细胞,未加虎杖苷的培养液作为对照。MTT法检测细胞增殖情况;流式细胞仪检测细胞周期及凋亡;ELISA法检测上清中纤维结合蛋白、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白的表达情况。 结果与结论：10^-5、10^-4 mol/L组促进成纤维细胞增殖最明显,10^-2 mol/L组吸光度值显著下降,细胞生长受抑制。10^-3 mol/L组G1期细胞大幅度下降,细胞有S期阻滞现象。10^-2 mol/L组有明确的促凋亡作用。10^-2 mol/L组上清中Ⅰ、Ⅲ型胶原蛋白较其他各组显著增高(P < 0.05);纤维结合蛋白较对照组显著增高(P < 0.05),较其他各组有所下降(P < 0.05)。说明低浓度虎杖苷有促进成纤维细胞增殖、保护细胞免于凋亡及促进纤维结合蛋白的表达与合成分泌的作用,促进成纤维细胞增殖的最适浓度应为10^-5~10^-4 mol/L。     

成纤维细胞超微结构的研究

本文用实验组织学方法,系统观察了成纤维细胞6种基本细胞形态的超微结构。发现成胶原细胞有前、中、后期三个功能阶段,中期细胞又分为合成期和分泌期;破纤维细胞可区分出吞噬型和溶酶体型2种亚型;纤维细胞存在静止型和衰老型2种亚型。成胶原细胞和破纤维细胞分别是胶原合成和胶原降解的主要细胞。本文从超微结构方面,证实成纤维细胞来源于毛细血管旁未分化间充质细胞,少分化成纤维细胞是其他形态的成纤维细胞的来源。成纤维细胞各种成熟的细胞形态可以互相转化,在功能上互相协调。本文提出成纤维细胞系统的概念,认为该系统包括成纤维细胞的6种基本细胞形态,其主要机能是合成胶原蛋白和其他细胞间质,降解胶原,使结缔组织(包括瘢痕组织)收缩。     

肿瘤相关成纤维细胞影响肿瘤微环境的研究进展

肿瘤相关成纤维细胞(CAF)是从肿瘤组织中分离到的一种活化的成纤维细胞,是肿瘤生长和进展的重要因素。近年来发现CAF不仅可以通过旁分泌的方式促进肿瘤细胞的生长和转移,而且通过与其他细胞的相互作用调节血管的形成和肿瘤免疫。     

人胚胎成纤维细胞与小鼠胚胎成纤维细胞生物学特性比较

探讨人胚胎成纤维细胞用于人胚胎干细咆体外长期培养的可行性，以含10％胎牛血清的DMEM(低糖)溶液为培养基，对人胚胎成纤维细胞的生长形态、对胰酶的敏感性、生长曲线及细胞周期进行研究，并与小鼠胚胎成纤维细胞作比较。结果显示，人胚胎成纤维细胞和小鼠胚胎成纤维细胞在体外均为贴壁生长型细胞，与小鼠胚胎成纤维细胞相比，人胚胎成纤维细胞生长更旺盛，且细胞寿命更长；在室温条件下，对0．25％胰酶更敏感，消化时间不宜超过3min。提示人胚胎成纤维细胞不仅在生长状况上与小鼠胚胎成纤维细胞类似，而且作为人胚胎干细胞体外长期培养的饲养层，在使用期限上优于后者，值得进一步研究。     

飞秒激光处理促进人牙周成纤维细胞在纯钛材料表面的增殖和分化

背景：目前对种植体表面优化处理促进人工牙种植体骨整合的形成是近年来这一领域的研究热点,而通过飞秒激光这种表面处理钛金属表面改性促进成纤维细胞的增殖和分化的相关性研究报道较少。目的：观察飞秒激光处理对于人牙周膜成纤维细胞在钛金属表面增殖和分化的影响。方法：将人牙周膜成纤维细胞接种在分别经过飞秒激光处理、TiO₂颗粒喷砂和砂纸顺向抛光的纯钛片表面(飞秒激光组,TiO₂颗粒喷砂组和对照组),观察细胞增殖和分化情况。结果与结论：与对照组相比,飞秒激光组和TiO₂颗粒喷砂组钛片表面粗糙度明显增加,表面细胞密度明显增加,RUNX2和OSX mRNA表达水平增加;且飞秒激光组上述指标高于TiO₂颗粒喷砂组。提示飞秒激光处理的纯钛表面更加粗糙,更有利于人牙周膜成纤维细胞的增殖和分化。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

微弧氧化技术在骨科钛植入体生物功能改性中的应用进展

微弧氧化是用于增强钛植入体的生物相容性和抗菌性的有效表面处理技术。在钛植入体表面制备生物活性元素组成的多孔涂层,是微弧氧化技术最具吸引力的特征。本研究主要介绍了微弧氧化的基本原理,阐述了该方法的技术优势,并总结了几种微弧氧化涂层的国内外研究进展;对含钙磷、银、铜、锌、硅的微弧氧化涂层,重点关注了对该类型涂层的骨整合性、抗菌性以及毒性的研究报道,旨在为研究者提供较为全面的视角,评估微弧氧化在骨科钛植入体上的应用进展,为后续临床研究提供参考。     

Effect of the surface geometry of smooth and porous-coated titanium alloy on the orientation of fibroblasts in vitro

The migration and orientation of human gingival fibroblasts in relation to the rim of smooth-surfaced and porous-coated titanium discs placed on multilayers in vitro was investigated. Samples were examined after 6 h, 24 h, 3 days, and 7 days of culture using phase-contrast and scanning electron microscopy. The cells migrated from the multilayer onto the smooth-surfaced discs forming bridges between them, and orientated along parallel circumferential grooves in the rim of the discs. This resulted in the cellular bridges orientating at an acute angle to the rim of the disc, and adjacent cells in the multilayer orientating parallel to the rim. Cellular bridges were also formed between the porous-coated discs and the multilayer but, because the cells that migrated onto, and between, the spheres of the porous-coat showed no preferred orientation, the bridges retained their orientation at right angles to the surface of the rim. This in turn resulted in the cells of the adjacent multilayer becoming similarly orientated. These observations suggest that the geometrical configuration of the surface of implants could influence whether a capsule or an orientated fibrous attachment is developed in relation to implants in vivo.     

冠状病毒的分子生物学研究进展

冠状病毒是所有RNA病毒中基因组最大的病毒，具有胃肠道、呼吸道或神经系统嗜性，常引起人普通感冒或动物胃肠道、呼吸系统疾病，而SARS-CoV则引起人的严重急性呼吸道综合征。研究冠状病毒分子生物学特征对阐明其病毒粒子的发生、感染和致病机制有重要意义。本文就冠状病毒的分子生物学特征包括冠状病毒基因组特征，编码蛋白的结构及功能等方面的研究进展作一综述。     

In vitro biological effects of titanium rough surface obtained by calcium phosphate grid blasting

Surface roughness modulates the osseointegration of orthopaedic and dental titanium implants. High surface roughness are currently obtained by blasting of titanium implants with silica or aluminium oxide abrasive particles. This process may cause the release of cytotoxic silicium or aluminium ions in the peri-implant tissue. To generate a biocompatible roughened titanium surface, we currently develop an innovative grid-blasting process using biphasic calcium phosphate (BCP) particles. Titanium alloy (Ti6Al4V) discs were either polished, BCP grid-blasted or left as-machined. BCP grid-blasting created an average surface roughness of 1.57 +/- 0.07 microm compared to the original machined surface of 0.58 +/- 0.05 microm. X-ray photoelectron spectroscopy indicated traces of calcium and phosphorus and relatively less aluminium on the BCP grid-blasted surface than on the initial titanium specimen. Scanning electronic microscopy observations and measurement of mitochondrial activity (MTS assay) showed that osteoblastic MC3T3-E1 cells were viable in contact with the BCP grid-blasted titanium surface. In addition, our results indicate that MC3T3-E1 cells expressed ALP activity and conserved their responsiveness to bone morphogenetic protein BMP-2. The overall results clearly indicate that this calcium phosphate grid-blasting technique increases the roughness of titanium implants and provides a non-cytotoxic surface with regard to mouse osteoblasts.     

钛表面纳米仿生磷灰石涂层对成骨样细胞活性的生物增强效应

目的：观察钛表面纳米仿生磷灰石涂层对成骨样细胞行为的影响，为骨科常用钛植入体的表面改性及其生物效应提供实验依据。方法: 商业用纯钛经过物理、化学和生物处理，表面生成均匀薄层仿生的纳米磷灰石涂层，将仿生涂层的钛金属板与成骨样细胞复合培养，以纯钛和只经磨砂、酸蚀处理的钛板作为对照，采用MTT法检测细胞活力和增殖变化、扫描电镜和激光共聚焦荧光显微镜观察细胞形态、RT-PCR检测碱性磷酸酶基因表达。结果: 纳米仿生磷灰石涂层比非涂层钛金属表面细胞的增殖数量明显增高，细胞的形态和分布也优于对照组；培养12 d，涂层对细胞ALP基因表达的量明显高于对照组。结论: 钛金属表面纳米仿生磷灰石涂层可以增强细胞的生物效应，提高钛植入体的骨界面早期结合，具有很好的应用前景。     

Preparation of graded porous titanium coatings on titanium implant materials by plasma spraying

Graded porous titanium coatings have been deposited on titanium substrates for dental implants by plasma spraying in an argon atmosphere. X-ray diffraction (XRD), scanning electron microscopy (SEM), surface roughness measurement, and tensile strength tests were performed on graded porous coatings. The results showed that Ti(3)O(5) was formed in the outermost surface of the porous coatings due to oxidation. The graded porous coatings consisted of three layers. The outer layer was full of macropores with a surface roughness of approximately 100 microm. The diameter of many macropores reached and even surpassed 150 microm, which could be beneficial for tissue to grow into the coating. The middle layer consisted of a mixture of micropores and macropores. The inner layer was a very dense and tight interface layer that included mechanical, physical, and metallurgical bonding. In tensile strength tests, testing bars peeled off the coatings, because the adhesive agent fractured, but the coatings remained intact.     

Composition of surface oxide film of titanium with culturing murine fibroblasts L929

Changes in the composition of surface oxide film on titanium specimens in the presence of amino acids, serum proteins, and cells were characterized using X-ray photoelectron spectroscopy. The surface oxide film on titanium formed in the air is so protective that the further oxidation of titanium is prevented in various circumstances. During immersion of the specimen in Hanks' solution, Eagle's minimum essential medium (MEM), and MEM with the addition of fetal bovine serum (MEM+FBS), calcium phosphate precipitated, causing the increase in thickness of the surface oxide film. Calcium phosphate was also precipitated with culturing murine fibroblast L929, but the amount of the calcium phosphate was smaller than those in Hanks' solution, MEM, and MEM+FBS. The relative concentration ratio of calcium to phosphorous, [Ca]/[P], increased with proteins charging negatively, while the ratio decreased with the cells whose extracellular matrix charging positively. In addition, sulfur precipitated as S(0) and/or S(2-) only with culturing the cells. Sulfate ions in the MEM+FBS are reduced at the interface between titanium and the solution with the existence of cells.     

Effect of surface processing on the attachment, orientation, and proliferation of human gingival fibroblasts on titanium.

The adhesion, orientation, and proliferation of human gingival fibroblasts was studied on electropolished (elpTi), etched (etchTi), and sandblasted (sblTi) titanium surfaces. The texture, chemical state, and composition of the titanium surfaces were analyzed using a surface tracing instrument and electron spectroscopy for chemical analysis. Considerable differences were evident in the surface texture and chemical composition of the differently treated titanium plates. Electropolishing produced the smoothest and cleanest surface. Human gingival fibroblasts attached, spread, and proliferated on all titanium surfaces. However, cells on elpTi exhibited an extremely flat morphology and seemed to form cellular bridges with adjacent cells, whereas the etchTi and sblTi surfaces harbored both round and flat cells with many long processes. Cells on elpTi appeared to grow in thick layers with no specific orientation, whereas on etchTi surfaces they were migrating along the parallel, irregular minor grooves caused by mechanical polishing, and on sblTi surfaces they seemed to grow in clusters. Stress-fiber type actin bundles and vinculin-containing focal adhesions were present in cells spreading on elpTi and etchTi surfaces but not in cells spreading on sblTi surfaces. Cell shape, orientation, and proliferation appear to depend on the texture of the titanium surface and probably also on the properties of the oxide layer and adjacent bulk material. Our findings suggest that smooth or finely grooved titanium surfaces could be optimal in implants adjacent to soft tissues as they support the attachment and growth of human gingival fibroblasts.