微等离子体（Plasma）治疗不同时期增生性瘢痕的效果研究

发布时间：2018-05-01 02:34

本文选题：微等离子体 + 增生性瘢痕　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:组织损伤后会进行自我修复,在修复过程中组织生长过度和异常增生,形成瘢痕,是创伤后愈合最常见的并发症,是外科修复的热点、难点话题。分析影响临床预后的原因,可能是由于Ⅰ型胶原、Ⅲ型胶原构成比发生了变化。本实验采用Plasma对不同时期的兔耳增生性瘢痕进行照射,经过观察各实验组与对照组中Ⅰ型胶原、Ⅲ型胶原构成比的变化情况及瘢痕组织中胶原纤维的分布特征,探讨Plasma治疗增生性瘢痕不同时期的区别,为临床确定Plasma治疗增生性瘢痕的最佳时期提供理论依据。方法:选取体重在2.5±0.18kg的雄性新西兰大白兔6只,用圆形环钻沿兔耳耳腹长轴各做6个直径为8mm,深及软骨表面的圆形全层皮肤创面缺损,各创面间隔1.0cm,6只兔子共形成72个圆形创面。随机将72个瘢痕创面分成6组,分别行Plasma治疗和对照(不做任何治疗):1增生期治疗组(n=12):造模后1个月(增生期)行Plasma照射治疗。Plasma参数:定点TIP,直径12毫米,像束点间距1毫米,能量设定为80瓦,无套管治疗头(热效应弱,剥脱效应强)重复扫描4次。Plasma治疗术后21天取材。2消退期治疗组(n=12):造模后2个月(消退期)行Plasma照射治疗。治疗参数、取材时间同1组。3成熟期治疗组(n=12):造模后3个月(成熟期)行Plasma照射治疗。治疗参数、取材时间同1组。4增生期对照组(n=12):作为增生期治疗组的空白对照,不做治疗,与1组同时取材。5消退期对照组(n=12):作为消退期治疗组的空白对照,不做治疗,与2组同时取材。6成熟期对照组(n=12):作为成熟期治疗组的空白对照,不做治疗,与3组同时取材。经脱水、浸蜡、石蜡包埋等步骤后,将标本做成厚度约为5.0μm组织切片。苏木精-伊红染色(hematoxylin-eosinstaining,he)、masson和天狼猩红染色后分别观察每个实验组与对照组中兔耳增生性瘢痕胶原纤维的分布特征;用苦味酸-天狼猩红偏振光法以及图像分析技术对各实验组及对照组兔耳增生性瘢痕组织中Ⅰ型胶原纤维、Ⅲ型胶原纤维的构成比进行测定,以反映plasma治疗增生性瘢痕的成效。将各实验组及对照组兔耳增生性瘢痕组织中Ⅲ型胶原占的比例进行统计,数据用均数±标准差表示。选择spss13.0软件进行数据分析,进行完全随机分组两因素析因设计与方差分析,设定p0.01时差异有统计学意义。结果:1组织形态学变化对照组增生期:局部肿胀充血,逐渐增厚,高出周围正常皮肤,凸出表面,外形不规则,表面呈红色,质实韧,未超出原术野范围;光镜下见排布紊乱的胶原纤维大量增生,可见旋涡结构,微血管增生明显。消退期:瘢痕组织厚度有所减低,硬度也开始向软转化,颜色由红色向浅红转变、轻度萎缩;光镜下可见到外形粗大、较增生期更密集、排列进一步紊乱、胶原结节少见胶原纤维及减少的微血管。成熟期:瘢痕组织厚度明显减低,但仍然高出于皮肤表面,质地较正常皮肤硬,中央部位色白,明显萎缩;光镜下显示胶原纤维外形变细、更加密集、排列明显紊乱且无胶原结节,同时微血管进一步减少。治疗组增生期:瘢痕组织质地较软、触之有弹性,颜色变浅,光镜下可见到胶原纤维变细,颜色由深变浅,胶原排列有序。消退期:瘢痕组进一步变软、弹性增加,颜色变淡,光镜下可见到纤维变细,颜色明显变浅,胶原排列疏散、规则的瘢痕组织胶原纤维。成熟期:瘢痕组织变得更加平软,弹性和颜色更与周围皮肤更加接近,光镜下可见真皮层变薄,成纤维细胞减少;与消退期相比,胶原纤维变得细而长,排列更加有序。2偏光显微镜观察与图象分析对照组增生期:Ⅰ型胶原纤维约占57%,束状分布、黄色、粗大且排列无序;Ⅲ型胶原纤维约占43%,网疏状分布、绿色、细丝状,分布于Ⅰ型胶原周围。消退期:Ⅰ型胶原纤维增加,约占全视野的83%,外形粗大,排列明显紊乱,相互聚集形成团块,相比之下Ⅲ型胶原纤维少,所占比约17%,分布于Ⅰ型胶原周围。成熟期:Ⅰ型胶原纤维大量增加,约占整个视野的92%以上,排列更加紊乱,呈竹节状,Ⅲ型胶原少见,所占比约为8%。治疗组增生期:胶原纤维排列较增生期对照组有序,Ⅰ型胶原纤维为53%左右,低于对照组,而Ⅲ型胶原在47%左右,比例提高。消退期:可见排列较对照组规则、疏散的胶原纤维,Ⅰ型胶原纤维所占比例为69%,且型胶原纤维所占比明显高于消退期对照组,约占31%。成熟期:可见排列疏散并趋于一致的胶原纤维,更接近于正常组织,Ⅰ型胶原纤维所占比例为75%,Ⅲ型胶原纤维所占比显著增高,约占25%。3统计学分析分别统计各实验组及对照组兔耳增生性瘢痕组织中Ⅰ型胶原、Ⅲ型胶原所占的比例;应用spss13.0对各组标本Ⅲ型胶原所占的比例行两因素多水平的析因分析,以p0.01作为判断差异显著性的标准。结果表明:在增生性瘢痕的三个不同阶段,Ⅲ型胶原所占的比例不同且具有统计学差异(f=307.038,p=0.0000.01),plasma治疗后,Ⅲ型胶原在瘢痕组织中所占的比例明显增加(f=142.806,p=0.0000.01)。分析表明,plasma对增生性瘢痕中Ⅲ型胶原的增加程度在不同时期的亦有显著差别(f=16.214,p=0.0000.01)。与对照组相比分别为:成熟期增加17%;消退期增加14%;增生期增加4%。因而,跟据统计学结果认为:plasma治疗兔耳增生性瘢痕在成熟期效果最好,其次为消退期,最后为增生期。结论:1plasma治疗后,瘢痕组织质地改善明显,由质地实韧变得柔软而有弹性。将正常皮肤与瘢痕组织中的胶原纤维形态进行比较后发现,治疗组瘢痕组织中Ⅰ、Ⅲ型胶原比值降低,更接近于正常皮肤中比例,胶原纤维形态也趋向于正常皮肤胶原纤维,这可能是微等离子体治疗增生性瘢痕的机理之一。2 Plasma对不同时期增生性瘢痕的治疗效果截然不同。在增生期微等离子体治疗增生性瘢痕的效果不明显;在消退期微等离子体治疗增生性瘢痕的效果较增生期有所提高;在成熟期微等离子体治疗增生性瘢痕的效果是最好、最明显的。
[Abstract]:Objective: after tissue injury, self repair will be carried out. Hypergrowth and abnormal hyperplasia and scar formation during the repair process are the most common complications of post-traumatic healing. It is a hot and difficult topic for surgical repair. The analysis of the causes of clinical prognosis may be due to the change in the ratio of type I collagen and type III collagen. Plasma was used to irradiate the hypertrophic scars of rabbit ears at different times. After observing the change of collagen type I, the ratio of collagen type III and the distribution characteristics of collagen fibers in the scar tissue, the difference of Plasma in the treatment of hypertrophic scars in different periods was observed, and the best treatment for the treatment of hypertrophic scar in the clinic was to determine the most clinical value of Plasma in the treatment of hypertrophic scar. The theoretical basis was provided at the good time. Methods: 6 male New Zealand white rabbits with a weight of 2.5 + 0.18kg were selected, with round ring drilling along the long axis of the ears of the rabbit ear, 6 diameters of 8mm, deep and cartilaginous surface of the full layer skin wound defect, each wound interval 1.0cm, 6 rabbits were formed 72 circular wounds, and 72 scar wounds were randomly divided into 6 groups. Plasma treatment and control (no treatment): 1 hyperplastic period treatment group (n=12):.Plasma parameters were treated by Plasma irradiation after 1 months of modeling: designated TIP, 12 mm in diameter, 1 mm like beam point spacing, 80 watts of energy, and 21 days after 4.Plasma treatments without cannula therapy head (heat effect weak, exfoliation effect) .2 treatment group (n=12): 2 months after the model (regression period) treated with Plasma irradiation. Treatment parameters, time and 1 groups of.3 mature period treatment group (n=12): 3 months after the model (mature stage) Plasma irradiation treatment. Treatment parameters, time and 1 groups of.4 proliferation period control group (n=12): as a blank control group of hyperplastic treatment group, no treatment, no treatment, The control group (n=12) was taken from the 1 group at the same time as the control group (.5). As a blank control group in the treatment group, the control group was not treated with the 2 group, and the.6 mature control group (n=12) was obtained simultaneously. As a blank control group of the mature period treatment group, no treatment was done, and the 3 groups were taken simultaneously. After dehydration, wax impregnation and paraffin embedding, the specimens were made into a thickness of about 5 mu m tissue. The distribution characteristics of collagen fibers in rabbit ear hypertrophic scar were observed by hematoxylin-eosinstaining (he), Masson and Sirius red staining in each experimental group and the control group, respectively. In the experimental group and the control group, the experimental group and the control group were treated with the image analysis technique. The composition ratio of type I collagen fiber and type III collagen fiber was measured to reflect the effect of plasma in the treatment of hypertrophic scar. The proportion of type III collagen in the hypertrophic scar tissue of the experimental group and the control group was counted and the data were expressed with the mean standard deviation of mean number. The SPSS13.0 software was selected to carry out the data analysis and carry out the complete random grouping. Two factor analysis of factorial design and variance analysis, the difference was statistically significant when setting P0.01. Results: 1 the hyperplastic period in the control group of the control group: local swelling and hyperemia, gradually thickening, high out of the normal skin, convex surface, irregular shape, red surface, solid and tough, not beyond the scope of the original field; under the light microscope, the collagen fibrous fiber arranging disorder was seen in the light microscope. A large number of hyperplasia, visible vortex structure, microvascular hyperplasia obvious. Decline period: the thickness of scar tissue decreased, the hardness began to change to soft, color from red to light red, light atrophy, light microscope can be seen thicker, more dense than the proliferation stage, a step disorder, collagenoid nodules rare collagen fibers and reduced microvessels. Mature period: the thickness of scar tissue decreased obviously, but still high out of the skin surface, the texture of the skin was more hard, the central part was white and atrophy, and the collagen fibers were thinner and denser in the light microscope, the arrangement of the collagen fibers was obviously disorderly and no collagen nodules, and the microvessels were reduced in one step. The hyperplastic period of the treatment group was softer and touch. It is elastic, light color, light microscope can see collagen fiber thinner, color from deep to shallow, collagen arrangement orderly. Decline period: scar group further soften, increase elasticity, color fade, light microscope can see fiber thinner, the collagen arrangement is evacuate, regular scar tissue collagen fiber. Mature period: scar tissue becomes even more Soft, elastic and color more close to the surrounding skin, the light microscope showed that the dermis thinned and fibroblasts decreased. Compared with the fading period, the collagen fibers became thinner and longer, arranged more orderly.2 polarizing microscope observation and image analysis control group proliferation period: type I collagen fibers accounted for about 57%, fascicular distribution, yellow, coarse and disordered. Type III collagen fibers accounted for about 43%, mesh sparse distribution, green, filamentous, distributed around type I collagen. Phase I collagen fibers increased, accounting for about 83% of the whole field of vision, the shape was large, the arrangement was obviously disorderly and formed a group, compared with type III collagen fibers, about 17%, distributed around type I collagen. Collagen fibers increased in a large amount, accounting for more than 92% of the whole field of vision, and the arrangement was more disorder. The collagen fibers were more disorder. The type III collagen was rare. The proportion of collagen fiber was about 8%. treatment group. The collagen fiber arrangement was more orderly than the control group, and the type I collagen fiber was about 53%, lower than the control group, and the proportion of type III collagen was about 47%. The proportion of collagen fiber in evacuation was 69%, and the proportion of type I collagen fiber was 69%, and the proportion of type collagen fibers was significantly higher than that in the control group, which accounted for the mature period of the collagen fibers, which were arranged to be identical to the normal tissue, and the proportion of type I collagen fibers was 75%, and the ratio of type III collagen fibers was the ratio of collagen fiber. The proportion of type I collagen and type III collagen in the rabbit ear hypertrophic scar tissue was statistically analyzed by 25%.3 statistical analysis, and the proportion of type III collagen in the rabbit ear hypertrophic scar tissue was measured by SPSS13.0, and two factors were used to analyze the proportion of type III collagen in each group, and P0.01 was used as the criterion to judge the difference significance. The results showed that: In the three different stages of hypertrophic scar, the proportion of type III collagen is different and has statistical difference (f=307.038, p=0.0000.01). After plasma treatment, the proportion of type III collagen in scar tissue is significantly increased (f=142.806, p=0.0000.01). The analysis shows that the increase of type III collagen in hypertrophic scar is different at the same time. There were also significant differences in the period (f=16.214, p=0.0000.01). Compared with the control group, the maturation period increased by 17%, the stage of extinction increased by 14%, and the proliferation period increased by 4%., and according to the statistical results, the effect of plasma in the treatment of hypertrophic scar in the rabbit ears was the best in the mature stage, followed by the elimination period, and the final hyperplastic period. Conclusion: after 1plasma treatment, scar tissue was treated. The texture improvement is obvious, and the texture is soft and flexible. After comparing the collagen fibers in the normal skin and the scar tissue, it is found that the ratio of type I and type III collagen in the scar tissue of the treatment group is lower, which is closer to the normal skin ratio, and the collagen fiber form tends to the normal skin collagen fiber, which may be the micro separation. The therapeutic effect of.2 Plasma on hypertrophic scar in different periods is very different. The effect of microplasma treatment on hypertrophic scar is not obvious in the stage of proliferation. The effect of microplasma treatment on hyperplastic scar in the stage of regression is higher than that in the proliferative stage. The effect of sexual scarring is the best and most obvious.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R622

【参考文献】