目的 评估VSD联合含氧液冲洗对下肢慢性静脉性溃疡(CVLU)患者创面肉芽组织生长及巨噬细胞活化的影响. 方法 2010年12月-2014年7月,将笔者单位收治的34例CVLU患者,按随机数字表法分为单纯VSD组(简称A组)11例、VSD+生理盐水组(简称B组)11例、VSD+含氧液组(简称C组)12例.入院后清创,术中留取创面中心肉芽组织标本,术后A组仅行VSD治疗(负压值为-30～-25 kPa,下同),B组采用VSD联合生理盐水冲洗,C组采用VSD联合含氧生理盐水(含氧液)冲洗(氧流量1 L/min).治疗7d,去除VSD装置.清创前及治疗7d行创面大体观察.治疗7d,计算创面肉芽组织覆盖率,并取创面中心肉芽组织标本以HE染色、Masson染色后行组织病理学观察.清创后VSD治疗前(以下简称治疗前)及治疗7d,用经皮氧分压测定仪检测创周皮肤局部氧分压.治疗7d,取创面肉芽组织用免疫组织化学染色法检测血管内皮生长因子(VEGF)表达.治疗前及治疗7d,取创面肉芽组织行免疫荧光染色观察CD68与诱导型一氧化氮合酶双阳性(Ⅰ型巨噬细胞)、CD68与精氨酸酶1双阳性(Ⅱ型巨噬细胞)细胞并计数.对数据行Fisher确切概率法检验、单因素方差分析、协方差分析、配对t检验、LSD检验. 结果 (1)大体观察示,清创前3组患者创面均存在坏死组织,肉芽组织较少.治疗7d,3组患者创面均出现新生肉芽组织,且C组创面新生肉芽组织最多.(2)治疗7d,C组患者创面肉芽组织覆盖率高于A、B组(P＜0.05或P＜0.01).(3)治疗7d,HE染色示C组患者创面新生毛细血管和Fb较A、B组更丰富;Masson染色示C组患者创面新生胶原较A、B组更丰富,且分布均匀.(4)治疗7d,C组患者创周皮肤局部氧分压为(40.7±4.1)mmHg(1 mmHg=0.133 kPa),高于A组的(35.0±3.1)mmHg和B组的(35.4±2.7)mmHg(P值均小于0.01),且3组均较治疗前升高(t值为10.38 ～ 22.52,P值均小于0.01).(5)治疗7d,C组患者创面肉芽组织的VEGF表达高于A、B组(P＜0.05或P＜0.01).(6)治疗7d,A、B、C组患者创面肉芽组织Ⅰ型巨噬细胞计数分别为每400倍视野下(14.3±2.3)、(11.5±3.0)、(10.7±2.3)个(F=25.14,P＜0.01),其中C组低于A、B组(P＜0.05或P＜0.01),且3组均较治疗前降低(t值为14.76～23.73,P值均小于0.01).治疗7d,A、B、C组患者创面肉芽组织Ⅱ型巨噬细胞计数分别为每400倍视野下(32.7±3.2)、(35.1±3.3)、(41.3±3.2)个(F=81.10,P＜0.01),其中C组高于A、B组(P值均小于0.01),且3组均较治疗前升高(t值为-69.34 ～-47.95,P值均小于0.01). 结论 VSD联合含氧液冲洗可有效提高CVLU患者创面局部氧分压,促进创面肉芽组织中巨噬细胞从以Ⅰ型为主转向以Ⅱ型为主,利于肉芽组织生长,从而改善创基条件.
目的 評估VSD聯閤含氧液遲洗對下肢慢性靜脈性潰瘍(CVLU)患者創麵肉芽組織生長及巨噬細胞活化的影響. 方法 2010年12月-2014年7月,將筆者單位收治的34例CVLU患者,按隨機數字錶法分為單純VSD組(簡稱A組)11例、VSD+生理鹽水組(簡稱B組)11例、VSD+含氧液組(簡稱C組)12例.入院後清創,術中留取創麵中心肉芽組織標本,術後A組僅行VSD治療(負壓值為-30～-25 kPa,下同),B組採用VSD聯閤生理鹽水遲洗,C組採用VSD聯閤含氧生理鹽水(含氧液)遲洗(氧流量1 L/min).治療7d,去除VSD裝置.清創前及治療7d行創麵大體觀察.治療7d,計算創麵肉芽組織覆蓋率,併取創麵中心肉芽組織標本以HE染色、Masson染色後行組織病理學觀察.清創後VSD治療前(以下簡稱治療前)及治療7d,用經皮氧分壓測定儀檢測創週皮膚跼部氧分壓.治療7d,取創麵肉芽組織用免疫組織化學染色法檢測血管內皮生長因子(VEGF)錶達.治療前及治療7d,取創麵肉芽組織行免疫熒光染色觀察CD68與誘導型一氧化氮閤酶雙暘性(Ⅰ型巨噬細胞)、CD68與精氨痠酶1雙暘性(Ⅱ型巨噬細胞)細胞併計數.對數據行Fisher確切概率法檢驗、單因素方差分析、協方差分析、配對t檢驗、LSD檢驗. 結果 (1)大體觀察示,清創前3組患者創麵均存在壞死組織,肉芽組織較少.治療7d,3組患者創麵均齣現新生肉芽組織,且C組創麵新生肉芽組織最多.(2)治療7d,C組患者創麵肉芽組織覆蓋率高于A、B組(P＜0.05或P＜0.01).(3)治療7d,HE染色示C組患者創麵新生毛細血管和Fb較A、B組更豐富;Masson染色示C組患者創麵新生膠原較A、B組更豐富,且分佈均勻.(4)治療7d,C組患者創週皮膚跼部氧分壓為(40.7±4.1)mmHg(1 mmHg=0.133 kPa),高于A組的(35.0±3.1)mmHg和B組的(35.4±2.7)mmHg(P值均小于0.01),且3組均較治療前升高(t值為10.38 ～ 22.52,P值均小于0.01).(5)治療7d,C組患者創麵肉芽組織的VEGF錶達高于A、B組(P＜0.05或P＜0.01).(6)治療7d,A、B、C組患者創麵肉芽組織Ⅰ型巨噬細胞計數分彆為每400倍視野下(14.3±2.3)、(11.5±3.0)、(10.7±2.3)箇(F=25.14,P＜0.01),其中C組低于A、B組(P＜0.05或P＜0.01),且3組均較治療前降低(t值為14.76～23.73,P值均小于0.01).治療7d,A、B、C組患者創麵肉芽組織Ⅱ型巨噬細胞計數分彆為每400倍視野下(32.7±3.2)、(35.1±3.3)、(41.3±3.2)箇(F=81.10,P＜0.01),其中C組高于A、B組(P值均小于0.01),且3組均較治療前升高(t值為-69.34 ～-47.95,P值均小于0.01). 結論 VSD聯閤含氧液遲洗可有效提高CVLU患者創麵跼部氧分壓,促進創麵肉芽組織中巨噬細胞從以Ⅰ型為主轉嚮以Ⅱ型為主,利于肉芽組織生長,從而改善創基條件.
목적 평고VSD연합함양액충세대하지만성정맥성궤양(CVLU)환자창면육아조직생장급거서세포활화적영향. 방법 2010년12월-2014년7월,장필자단위수치적34례CVLU환자,안수궤수자표법분위단순VSD조(간칭A조)11례、VSD+생리염수조(간칭B조)11례、VSD+함양액조(간칭C조)12례.입원후청창,술중류취창면중심육아조직표본,술후A조부행VSD치료(부압치위-30～-25 kPa,하동),B조채용VSD연합생리염수충세,C조채용VSD연합함양생리염수(함양액)충세(양류량1 L/min).치료7d,거제VSD장치.청창전급치료7d행창면대체관찰.치료7d,계산창면육아조직복개솔,병취창면중심육아조직표본이HE염색、Masson염색후행조직병이학관찰.청창후VSD치료전(이하간칭치료전)급치료7d,용경피양분압측정의검측창주피부국부양분압.치료7d,취창면육아조직용면역조직화학염색법검측혈관내피생장인자(VEGF)표체.치료전급치료7d,취창면육아조직행면역형광염색관찰CD68여유도형일양화담합매쌍양성(Ⅰ형거서세포)、CD68여정안산매1쌍양성(Ⅱ형거서세포)세포병계수.대수거행Fisher학절개솔법검험、단인소방차분석、협방차분석、배대t검험、LSD검험. 결과 (1)대체관찰시,청창전3조환자창면균존재배사조직,육아조직교소.치료7d,3조환자창면균출현신생육아조직,차C조창면신생육아조직최다.(2)치료7d,C조환자창면육아조직복개솔고우A、B조(P＜0.05혹P＜0.01).(3)치료7d,HE염색시C조환자창면신생모세혈관화Fb교A、B조경봉부;Masson염색시C조환자창면신생효원교A、B조경봉부,차분포균균.(4)치료7d,C조환자창주피부국부양분압위(40.7±4.1)mmHg(1 mmHg=0.133 kPa),고우A조적(35.0±3.1)mmHg화B조적(35.4±2.7)mmHg(P치균소우0.01),차3조균교치료전승고(t치위10.38 ～ 22.52,P치균소우0.01).(5)치료7d,C조환자창면육아조직적VEGF표체고우A、B조(P＜0.05혹P＜0.01).(6)치료7d,A、B、C조환자창면육아조직Ⅰ형거서세포계수분별위매400배시야하(14.3±2.3)、(11.5±3.0)、(10.7±2.3)개(F=25.14,P＜0.01),기중C조저우A、B조(P＜0.05혹P＜0.01),차3조균교치료전강저(t치위14.76～23.73,P치균소우0.01).치료7d,A、B、C조환자창면육아조직Ⅱ형거서세포계수분별위매400배시야하(32.7±3.2)、(35.1±3.3)、(41.3±3.2)개(F=81.10,P＜0.01),기중C조고우A、B조(P치균소우0.01),차3조균교치료전승고(t치위-69.34 ～-47.95,P치균소우0.01). 결론 VSD연합함양액충세가유효제고CVLU환자창면국부양분압,촉진창면육아조직중거서세포종이Ⅰ형위주전향이Ⅱ형위주,리우육아조직생장,종이개선창기조건.
Objective To evaluate the therapeutic effects of VSD combined with irrigation of oxygen loaded fluid on the growth of granulation tissue and macrophage polarization in chronic venous leg ulcers.Methods Thiry-four patients with chronic venous leg ulcers hospitalized in our department from December 2010 to July 2014 were divided into VSD group (A,n =11),VSD+irrigation group (B,n =11),and VSD + oxygen loaded fluid irrigation group (C,n =12) according to the random number table.After admission,debridement was performed,and granulation tissue in the center of the wound was harvested during the operation.After debridement,the patients in group A were treated with VSD only (negative pressure from -30 to-25 kPa,the same below) ; the patients in group B were treated with VSD combining irrigation of normal saline; the patients in group C were treated with VSD combining normal saline loaded with oxygen irrigation (flow of 1 L/min).On post treatment day (PTD) 7,the VSD devices were removed.Gross observation was conducted before debridement and on PTD 7.Ou PTD 7,the granulation tissue in the center of the wound was harvested for histopathological observation with HE staining and Masson staining,following calculation of granulation tissue coverage rate.After debridement but before the negative pressure therapy (hereinafter referred to as before treatment) and on PTD 7,partial pressure of oxygen of the skin around the wound was measured by transcutaneous tissue oxygen tension survey meter.On PTD 7,expression of vascular endothelial growth factor (VEGF) was determined with immunohistochemistry.Before treatment and on PTD 7,cells with double positive expressions of induced nitric oxide synthase plus CD68 (type Ⅰ macrophage) and arginase 1 plus CD68 (type Ⅱ macrophage) were observed with immunofluorescence staining and quantified.Data were processed with Fisher's exact test,one-way analysis of variance,covariance analysis,paired t test,and LSD test.Results (1) The gross observation showed that before debridement there was a certain amount of necrotic tissue and little granulation tissue in the wounds of patients in all the 3 groups.On PTD 7,new granulation tissue was found in the wounds of patients in all the 3 groups,and in group C its amount was the largest.(2) On PTD 7,the granulation tissue coverage rate of wounds in patients of groupC was higher than that of group A or B (P ＜0.05 orP ＜0.01).(3) On PTD7,HE staining showed that there appeared more abundant new born microvessels and fibroblasts in the wounds of patients in group C than those in groups A and B; Masson staining showed that there was more abundant fresh collagen distributed orderly in the wounds of patients in group C compared with group A or B.(4) On PTD 7,it was found that partial pressure of oxygen of the skin around the wounds in patients of group C [(40.7 ±4.1) mmHg,1 mmHg =0.133 kPa] was higher than that of group A [(35.0 ±3.1)mmHg] or B [(35.4 ± 2.7) mmHg,with P values below 0.01] ; the partial pressure of oxygen of the skin around the wounds of patients in all the 3 groups was increased significantly compared with that before treatment (with t values from 10.38 to 22.52,P values below 0.01).(5) On PTD 7,the expression of VEGF in the wounds of patients in group C was higher than that in group A or B (P ＜0.05 orP ＜0.01).(6) On PTD7,the number of type Ⅰ macrophages in granulation tissue of patients was respectively 14.3 ± 2.3,11.5 ± 3.0,and 10.7 ±2.3 per 400 times vision field in groups A,B,and C (F =25.14,P ＜0.01),while the number in groupC was less than that in group A or B (P ＜0.05 orP ＜0.01).Compared with that before treatment,the number of type Ⅰ macrophages was significantly decreased on PTD 7 in all the 3 groups (with t values from 14.76 to 23.73,P values below 0.01).On PTD 7,the number of type Ⅱ macrophages in granulation tissue of patients was respectively 32.7 ± 3.2,35.1 ± 3.3,and 41.3 ± 3.2 per 400 times vision field in groups A,B,and C (F =81.10,P ＜0.01),and the number in group C was lager than that in group A or B (with P values below 0.01).Compared with that before treatment,the number of type Ⅱ macrophages in all the 3 groups was significantly increased (with t values from-69.34 to-47.95,P values below 0.01).Conclusions VSD combined with irrigation of oxygen loaded fluid can raise the partial pressure of oxygen of the skin around the wounds effectively,promoting the transition of macrophages from type Ⅰ to type Ⅱ,thus it may promote the growth of granulation tissue,resulting in a better recipient for skin grafting or epithelization.
