外源性胶原对人工活性肌腱影响的体外形态学观察

目的探讨外源性胶原在人工活性肌腱的构建中对细胞功能的影响. 方法将人发、碳纤维(carbon fiber, CF)及聚羟基乙酸(polyglycolic acid,PGA)3种材料复合外源性胶原(实验组)及不复合外源性胶原(对照组),与密度为3×106/mm3的转化人胚腱细胞进行体外联合培养,于培养后2小时、3天和5天3个时间点行倒置显微镜观察,以及培养5天行扫描电镜观察细胞数量及形态. 结果有胶原复合的实验组人工活性肌腱培养2小时后,细胞向材料周围聚集,附着于材料的部分细胞呈圆形;3天时附着的细胞增多,细胞与材料相容性好,细胞密度大.而无胶原复合的对照组3种材料上细胞附着较少.5天后实验组细胞形态由圆形→椭圆形→梭形演变,对照组细胞形态大部分仍为圆形及椭圆形,细胞数量仍少于实验组. 结论外源性胶原有助于细胞对材料的附着以及细胞自身的增殖.     

增生性瘢痕成纤维细胞与正常皮肤成纤维细胞生长差异比较

【目的】探讨增生性瘢痕成纤维细胞与正常皮肤成纤维细胞基因表现型的差异以及EGF、PDGF和bFGF对其的影响。【方法】测定细胞生长曲线;应用液闪计数测定成纤维细胞胶原蛋白的合成。【结果】增生性瘢痕成纤维细胞增殖速度比正常皮肤成纤维细胞慢;胶原蛋白的合成高于正常皮肤成纤维细胞。EGF、PDGF和bFGF均对两种成纤维细胞的增殖具有强烈的刺激作用。EGF和bFGF可减少增生性瘢痕成纤维细胞的胶原蛋白合成,PDGF可以使两种成纤维细胞的胶原蛋白合成增加。【结论】增生性瘢痕成纤维细胞是一种特殊基因表现型的成纤维细胞,它具有自己特殊的生物学特性。EGF、PDGF和bFGF在增生性瘢痕的形成过程中可能具有重要作用。     

成纤维细胞移植促进人工真皮内血管新生的研究

目的观察成纤维细胞与人工真皮共同移植对移植床及真皮海绵内微血管形成、胶原沉积的影响,促进人工真皮海绵内类真皮组织形成,缩短"二期植皮"的间隔时间.方法 Wistar大鼠18只,背部两侧各制作2.5 cm×2.5 cm全层皮肤缺损创面,在移植人工真皮之前分为两组,即成纤维细胞移植组(A组):向创面喷洒混入密度为1.0×105/cm2的同种真皮成纤维细胞的纤维蛋白胶0.5 ml;对照组(B组):只喷洒纤维蛋白胶0.5 ml.于移植后5和10天分别切取移植的真皮及周围组织,进行HE染色、Masson染色、VEGF免疫抗体染色和墨汁灌注染色,观察移植床及真皮海绵内微血管新生及胶原沉积状态;于移植后5天行伊文蓝组织灌注,分光光度计测定真皮海绵内染料含量,定量检测真皮海绵内循环渗透情况.结果移植后5天,新生血管主要集中在真皮下移植床,A组新生血管较B组明显增多,A组(9.64±2.36)个/高倍视野,B组(3.88±1.62)个/高倍视野,差异有统计学意义(P＜0.05);VEGF阳性细胞亦明显增多.移植后10天,移植床及真皮内均有微血管形成,A组血管数量和管径均较B组增加,胶原沉积量也明显增加.A组(46.04±8.90)个/高倍视野,B组(30.08±7.76)个/高倍视野,有统计学意义(P＜0.05).结论同种成纤维细胞移植可明显加快人工真皮内及移植床的血管新生和胶原沉积,加快真皮海绵内类真皮组织的形成.     

可注射性人胎盘胶原豚鼠皮内、皮下注射实验研究

目的 研究可注射性人胎盘胶原的理化性质及临床应用特点.方法 将实验豚鼠随机分为两组,并将浓度为8mg/ml及30mg/ml的两种胶原分别注于两组豚鼠的皮下和皮内,以生理盐水作对照.实验鼠分别在注射后1、3、5、7、14、28、49、91、196天作大体观察和组织切片观察.结果 显示可注射性人胎盘胶原不引起明显的过敏反应及炎症反应.注入宿主皮内后,能较长期稳定留存,并逐步被宿主的自身胶原替代,共同进入宿主的生命代谢过程.它还能使真皮的胶原纤维增加,真皮增厚,延缓了真皮的衰老.结论 可注射性人胎盘胶原在整形美容外科中作为改善皮肤衰老中出现的皱褶及修复皮肤浅表凹陷性缺损具有实用价值.     

PLGA/Ⅰ型胶原复合支架用于组织工程化骨再造的研究

目的:采用PLGA/Ⅰ型胶原复合改良生物支架,构建组织工程化骨组织.方法:采用Ⅰ型胶原和聚乳酸-聚乙醇酸共聚物(PLGA)复合,制作改良的生物支架,将原代培养的成骨细胞接种于复合支架上,培养1周,扫描电镜观察成骨细胞在支架上的生长及黏附情况;同时,将细胞-支架复合体自体异位植入,并取材观察其成骨情况.结果:经鉴定,原代培养的细胞符合成骨细胞的特征;扫描电镜:在生物支架上大量成骨细胞呈簇状生长,并形成多个细胞突起;大体标本:4个月时可见骨块形成;自体异位植入后1个月可见新生骨组织形成,周围有多个活性成骨细胞和骨母细胞,至4个月时骨组织渐趋成熟.结论:Ⅰ型胶原和PLGA复合支架是一种理想的生物可降解支架,可用于组织工程化骨再造的研究.     

Evaluation of bone turnover in type I osteoporosis using biochemical markers specific for both bone formation and bone resorption

The aims of the study were to evaluate the use of bone-specific biochemical markers of turnover in type I osteoporosis, to test for evidence of heterogeneity of bone turnover in this condition, and to attempt to devise an uncoupling index by using the relationship between bone-specific biochemical markers of bone formation and bone resorption. In women with type I osteoporosis (mean age 64 years, SD 5;n=63) the mean level of serum osteocalcin, a specific biochemical marker of bone formation, was 9.9 ng/ml (SD 2.0), which was higher than the level in normal postmenopausal women (mean age 65 years, SD 6;n=8.9 ng/ml (SD 2.0;p<0.01). The variance of serum osteocalcin levels in the two groups was similar. Compared with this 11% increase in the biochemical marker for bone formation, the markers of bone resorption, total urinary deoxypyridinoline (bone-specific), pyridinoline and hydroxyproline were increased by 40% (p<0.0001), 61% (p<0.0001) and 25% (p<0.01), respectively. Furthermore, these biochemical markers of bone resorption had greater variance in women in type I osteoporosis than in the normal postmenopausal women (p<0.001). The urinary excretion of the free crosslinks deoxypyridinoline, pyridinoline and glycosylated pyridinoline were increased by 26% (p<0.001), 17% (p<0.01) and 13% (NS) respectively. An uncoupling index was calculated for the difference between urinary deoxypyridinoline and serum osteocalcin using the results from the normal women and expressed asz-scores. We conclude that the pyridinium crosslinks of collagen enable better discrimination between normal and osteoporotic women than does hydroxyproline. In osteoporosis there appears to be heterogeneity of bone resorption. Finally, an uncoupling index indicated that in osteoporosis bone resorption was increased to a greater extent than bone formation as compared with normal postmenopausal women.     

重组人白细胞介素—1受体拮抗剂抗实验性肝纤维化作用

目的：观察ｒｈＩＬ－１ｒａ抗大鼠实验性肝纤维化的作用。方法：以Ｗｉｓｔａｒ大鼠为实验观察对象,采用４０５四氯化碳皮下注射,按每１００ｇ体重０．３ｍｌ（首剂为０．５ｍｌ／１００ｇ体重）皮下注射,每周２次,共１６次,诱导肝纤维化模型。ｒｈＩＬ－１ｒａ理组于实验第１天开始注射四氯化碳时按每１００ｇ体重０．５ｍｇ肌肉注射ｒｈＩＬ－１ｒａ,每日１次。于实验１７、２９、４３、５７日每组分别处死动物６ １０只,     

Evaluation of a cross-linked versus non-cross-linked collagen matrix in full-thickness skin defects

Autologous skin transplantation is the gold standard for treatment of full-thickness skin defects such as deep burn injuries, but has the disadvantages of limited donor sites and donor site morbidities. Alternative skin replacement products, such as xenografts and allografts, are not a permanent solution. Numerous manufactured skin substitutes already show promising approaches, but have limited efficacy. Therefore, wound dressings adaptable to the physiology of wound healing are still needed.In a randomized controlled in vivo study, a newly designed biocompatible collagen nonwoven matrix was compared to the Integra® bilayer dermal substitute and untreated controls in 48 full-thickness skin defects in a swine model.The take of all templates was complete, and all the tissue-engineered products accelerated dermal wound healing compared to the untreated controls, as identified by planimetric measurements. The higher collagen dose treatments and Integra®-covered wounds developed the thickest, cell-rich neoepidermal tissue in histological examination.The innovative biocompatible collagen matrix is flexibly applicable and modifiable, and offers potential as a carrier membrane for therapeutic supplemental products such as growth factors to further develop effective wound dressings.     

人胶原和自体脂肪声带注射的实验研究

目的探讨国产人胶原狗声带注射的组织学转归及其在矫治声门闭合不全中的应用价值和前景。方法实验动物分为４组，Ａ组２只分别注射人胶原和自体脂肪矫正左侧声带麻痹引起的声门闭合不全，并对治疗效果进行主、客观观察，术后５个月取材，透射电镜观察。Ｂ、Ｃ、Ｄ组９只狗（每组３只）于右侧声带注入人胶原、左侧声带注入自体脂肪各０．３ｍｌ，分别于术后１、３、５个月取材并观察其组织学变化和体积变化。结果自体脂肪能在声带组织中存活，但吸收率很高，１、３、５个月体积平均减少速度为３８．５１％，疗效不能持久。人胶原易于注射，组织相容性好，并能被宿主成纤维细胞和血管移生，同时伴有宿主胶原的再生，显示出最终被宿主组织完全取代的趋势，１～５个月体积平均减少速度为０．６５％。矫治效果长期稳定。结论自体脂肪应用有局限性，只能作为短期的声带植入材料；人胶原可用于各种原因所致声门闭合不全的长期矫治材料，具有良好的应用前景。     

血府逐瘀汤对心肌成纤维细胞增殖和胶原合成的影响

目的:探讨血府逐瘀汤对心肌成纤维细胞增殖与胶原合成的影响,为血府逐瘀汤防治多种由于心肌成纤维细胞增殖而导致的心脏疾病提供理论依据。方法:采用直接加药和血清药理学方法观察血府逐瘀汤对心肌成纤维细胞增殖与胶原合成的影响。结果:1:80稀释的1 g/ml血府逐瘀汤与含10％血府逐瘀汤鼠血清已经对心肌成纤维细胞增殖有抑制作用,同对照组比较具有显著性差异(P<0.05);1:80稀释的1 g/ml血府逐瘀汤能显著抑制心肌成纤维细胞分泌胶原,达17.9％。结论:血府逐瘀汤不仅能够抑制心肌成纤维细胞增殖,而且具有抑制心肌成纤维细胞分泌胶原的作用,血府逐瘀汤具有抗心肌纤维化的作用。