肉芽组织下注射微粒皮浆对大鼠创伤性慢性创面愈合的影响

目的研究肉芽组织下注射微粒皮浆对大鼠创伤性慢性创面愈合的作用。 方法选取60只SD雄性大鼠，于背部制作大小为3.0 cm×3.0 cm的创面，并将钢丝圈缝于创面内缘。按照随机数字表法将大鼠分为3组，分别为一般创面组、慢性创面组和微粒皮浆组，每组各20只。一般创面组背部造成创面后给予抗感染治疗并常规换药；慢性创面组背部形成创面后给予抗感染治疗、常规换药，并连续7 d肌内注射氢化可的松干预形成慢性创面；微粒皮浆组背部形成创面后给予抗感染治疗、常规换药，连续7 d肌内注射氢化可的松干预形成慢性创面，取大鼠右大腿外侧皮肤制备微粒皮浆注射于肉芽组织下。造模完成次日开始观察创面情况，定为观察第1天。观察第7、14、21、28天各组创面愈合情况，并计算创面愈合率；留取观察第14天的肉芽组织进行苏木精-伊红染色以及CD31免疫组织化学染色，观察苏木精-伊红染色下创面新生毛细血管分布情况及CD31免疫组织化学染色下CD31表达情况与微血管密度。对数据行单因素方差分析和LSD-t检验。 结果观察第14天，一般创面组创面明显缩小，慢性创面组皮肤爬伸不明显，微粒皮浆组创面大部分愈合；观察第28天，一般创面组剩余部分残留创面，慢性创面组创面愈合不明显，微粒皮浆组创面基本愈合。观察第14、21、28天，一般创面组愈合率分别为(51.09±0.94)%、(75.43±0.92)%、(86.51±0.57)%，慢性创面组创面愈合率分别为(20.30±0.95)%、(35.59±1.18)%、(45.82±1.35)%，微粒皮浆组创面愈合率分别为(39.00±0.86)%、(64.62±0.15)%、(91.25±0.87)%，比较差异均有统计学意义(F＝1 993.60、6 475.02、9 984.47，P值均小于0.05)；观察第14天，慢性创面组创面愈合率分别与一般创面组、微粒皮浆组比较，差异均有统计学意义(t＝89.90、50.93，P值均小于0.05)；观察第21天，慢性创面组创面愈合率分别与一般创面组、微粒皮浆组比较，差异均有统计学意义(t＝117.90、116.10，P值均小于0.05)；观察第28天，慢性创面组创面愈合率分别与一般创面组、微粒皮浆组比较，差异均有统计学意义(t＝86.43、94.29，P值均小于0.05)。观察第14天，创面苏木精-伊红染色观察，一般创面组可见少许新生毛细血管，慢性创面组未见明显新生毛细血管，微粒皮浆组其间有大量新生毛细血管。观察第14天，创面CD31免疫组织化学染色观察(CD31阳性表达呈棕黄色)，一般创面组棕黄色的颗粒散在分布，慢性创面组棕黄色颗粒稀疏分布，微粒皮浆组可见大量棕黄色的颗粒分布。观察第14天，创面微血管密度比较，一般创面组、慢性创面组、微粒皮浆组微血管密度分别为(49.20±17.96)、(37.32±9.57)、(64.93±20.29)个/视野，比较差异有统计学意义(F＝34.09，P<0.05)；慢性创面组创面微血管密度分别与一般创面组、微粒皮浆组比较，差异有统计学意义(t＝3.23、11.50，P值均小于0.05)。 结论微粒皮浆肉芽组织下注射可促进大鼠创伤性慢性创面血管增生，其创面愈合率明显升高，创面愈合时间缩短。

Effect of granulation tissue injection of micropar skin pulp on the healing of traumatic chronic wound in rats

ObjectiveTo study the effect of granulation tissue injection of micropar skin pulp on the healing of traumatic chronic wound in rats. MethodsSixty SD male rats were selected to make wounds of 3.0 cm×3.0 cm in the back and sewed on the inner edge of the wound with wire coils. The rats were divided into 3 groups according to the random number table method, which were general wound group, chronic wound group and micropar skin pulp group. There were 20 rats in each group. The general wound group was treated with anti-infective therapy and the routine dressing change after the wound was caused by the back. The chronic wound group was treated with anti-infection treatment and routine dressing change after the formation of the back wound. Chronic wound formation was completed by intramuscular injection of hydrocortisone for 7 days. After the formation of the wound on the back of the micropar skin pulp group, anti-infection treatment and the routine dressing change were given, intramuscular injection of hydrocortisone was performed for 7 days in a row to interfere with the formation of the chronic wound, and the microne was prepared from the skin on the lateral side of the right thigh of the rats and injected under granulation tissue. The observation of wound surface was started next day after the completion of modeling, which was set as the observation day. The wound healing of each group was observed on the 7th, 14th, 21st, 28th day of observation and the wound healing rate was calculated. Granulation tissues on 14th day of observation were retained and observed for hematoxylin-eosin staining and CD31 immunohistochemical staining. The distribution of new capillaries was observed under hematoxylin -eosin staining and the expression of CD31, microvessel density of the wound were observed by CD31 immunohistochemical staining. Data were analyzed with one-way analysis of variance and LSD-t test. ResultsOn the 14th day of observation, the wound of the general wound group was significantly reduced, the skin climbing of the chronic wound group was not obvious, and most of the wound was healed in the micropar skin pulp group. On the 28th day of observation, there was remaining wound in the general wound group, the wound healing in the chronic wound group were not obvious, most of wounds in the micropar skin pulp group healed. On the 14th, 21st, 28th day of observation, the healing rates of the general wound group were (51.09±0.94)%, (75.43±0.92)%, (86.51±0.57)%, and healing rates of the chronic wound group were (20.30±0.95)%, (35.59±1.18)%, (45.82±1.35)%, and the healing rates of the micropar skin pulp group were (39.00±0.86)%, (64.62±0.15)%, (91.25±0.87)%, the comparative differences were statistically significant (F=1 993.60, 6 475.02, 9 984.47, with P values below 0.05). Comparing the chronic wound group with the general wound group and the micropar skin pulp group respectively on the 14th day of observation, the differences were statistically significant (t=89.90, 50.93, with P values below 0.05). Comparing the chronic wound group with the general wound group and the micropar skin pulp group respectively on the 21st day of observation, the differences were statistically significant (t=117.90, 116.10, with P values below 0.05). Comparing the chronic wound group with the general wound group and the micropar skin pulp group respectively on the 28th day of observation, the differences were statistically significant (t=86.43, 94.29, with P values below 0.05). On the 14th day of observation, hematoxylin-eosin staining was observed on the wound surface, a little new capillaries were observed in the general wound group, no obvious new capillaries were observed in chronic wound group, a large number of new capillaries were observed in the micropar skin pulp group.On the 14th day of observation, the immunohistochemical comparison of CD31 (The positive expression of CD31 was brown-yellow) showed that brown-yellow particles were scattered in the general wound group, while brown-yellow particles were sparsely distributed in the chronic wound group, and a large number of brown-yellow particles could be seen in the micropar skin pulp group.On the 14th day of observation, the comparison of microvascular density was observed, the comparison of microvascular density in the general wound group, the chronic wound group, the micropar skin pulp group were (49.20±17.96), (37.32±9.57), (64.93±20.29) (individual/field of view), the comparative differences were statistically significant (F=34.09, P<0.05), and the chronic wound group was compared with the general wound group, micropar skin pulp group, the differences were statistically significant (t=3.23, 11.50, with P values below 0.05). ConclusionGranulation tissue injection of micropan skin pulp can promote vascular proliferation in traumatic chronic wounds in rats, and the wound healing rate is significantly higher, and the wound healing time is shorter.