微等离子束对豚鼠皮肤超微结构分析和羟脯氨酸测定

Effects of radio-frequency micro-plasma on the ultrastructure of dermal collagen and hydroxyproline content in ginea pigs

目的观察微等离子束对豚鼠皮肤胶原组织作用效应的组织学和超微结构变化及羟脯氨酸含量测定,探讨微等离子束的作用机理。方法选择15只豚鼠,每只豚鼠背部划分为实验侧和空白对照侧2个等分区域,给予60W/10 kJ微等离子束照射,于作用后即刻、1周后和1月后分别切取实验侧及空白对照部位皮肤行组织病理维多利亚-立春红染色,透射电镜分析和羟脯氨酸检测试剂盒进行含量测定。结果 60 W/10 kJ即刻表现为表皮局灶性出现点阵状改变,部分表皮出现汽化缺失或者坏死变性,真皮浅层胶原组织出现点阵化表现和明显均质化;特殊染色显示微等离子束主要影响真皮胶原纤维,形成局灶性胶原纤维凝集和变性。1周后皮肤浅层胶原组织结构逐渐致密,排列有序,有少量组织细胞。1月后皮肤浅层胶原组织明显增厚,胶原纤维增粗并排列致密,弹力纤维呈局灶性增粗。透射电镜显示微等离子束作用后表皮细胞较完整,细胞间结构正常,但真皮胶原丧失正常结构,细胞结构消失,大量细胞凋亡明显,1月后仍可见少量细胞凋亡的表现但胶原结构逐渐恢复,浅层胶原纤维排列明显致密。羟脯氨酸测定显示微等离子束作用1周后羟脯氨酸含量要高于作用之前,但是差异性不具有统计学意义(P>0.05);1月后羟脯氨酸含量要明显高于作用前,差异性具有统计学意义(P<0.05)。结论微等离子束对豚鼠皮肤胶原组织作用有明显的刺激效应,其主要靶组织为真皮胶原组织,可以明显促进皮肤新生胶原的增生。

Objective To investigate the effect of micro-plasma radio-frequency on the ultrastructure of skin collagen and the level of hydroxyproline in ginea pigs.Methods Fifteen guinea pigs were enrolled in this study.One side of their dorsal skin was exposed to 60 W/10 kJ radiofrequency plasma treatment and the other side was taken as control.Samples were taken immediately,one week and one month after the micro-plasma treatment and examined by light and transmission electron microscopy（TEM）,and the level of hydroxyproline content by hydroxyproline assay kit.Results Immediately after the 60 W/10 kJ plasma treatment,the epidermis showed focal lesions,obviously homogenization and complete vaporization loss or degeneration and necrosis of the superficial and middle-level layer derma,and a large area of collagen homogenization.Special staining demonstrated that the micro-plasma beam mainly affected the dermal collagen fibers,forming focal collagen fiber agglutination and degeneration.One week after the micro-plasma treatment,the superficial dermal collagen fibers gradually became into an orderly dense arrangement.One month after the micro-plasma treatment,the superficial collagen fibers showed markedly compact and dense arrangement.TEM showed that the epidermal cells were relatively complete,with normal intercellular structures.But the dermal collagen fibers lost their original normal structure and dermal cells showed disintegration and numerous apoptosis.A small amount of apoptosis remained but collagen structure gradually restored after one month.The level of hydroxyproline after one week was higher than that of before plasma treatment,but the difference was not significant（P〉0.05）.The level of hydroxyproline after one month was significantly higher than that before plasma treatment,with a statistically significant difference（P〈0.05）.Conclusions The novel micro-plasma radio-frequency technique has obvious stimulating effect on skin collagen.Its main tissue target is dermal collagen tissue.It can stimulate neo-collagen hyperplasia in the skin to a certain degree.