益腎活血股密保方治療早中期股骨頭壞死的臨床研究及其含藥血清對體外破骨細胞影響的實驗研究
發(fā)布時間：2018-08-06 22:05
【摘要】：目的:1.通過收集臨床病例進行隨機對照研究,觀察益腎活血股密保中藥方治療早中期股骨頭壞死(Osteonecrosis of the femoral head,ONFH)的臨床療效。2.通過實驗研究,檢測股密保含藥血清的毒性;利用體外誘導(dǎo)骨髓單核細胞的方式獲取足夠數(shù)量、性狀穩(wěn)定的滿足實驗要求的破骨細胞;觀測股密保含藥血清對骨髓單核細胞誘導(dǎo)分化為破骨細胞的影響;觀測股密保含藥血清對破骨細胞骨吸收功能的影響。方法:1.按照西醫(yī)納入標準及排除標準的范圍,對2016年1月至2017年2月在蘇州市中醫(yī)醫(yī)院骨傷科門診進行治療并完成隨訪的0NFH患者,共43例(51髖),采用隨機法將病人分為兩組,對照組口服阿侖膦酸鈉(福善美)70mgqw8周;治療組口服股密保方,4周為一個療程,共2個療程。治療前后采用Harris評分、疼痛直觀模擬量表(VAS表)和影像學(xué)檢查結(jié)果等進行詳細的記錄,最后統(tǒng)計數(shù)據(jù)進行分析。2.用體外誘導(dǎo)骨髓單核細胞的方式,在培養(yǎng)及傳代完成的骨髓單核細胞中加入含25ng/ml的M-CSF、100ng/ml的RANKL、1%PS、5%FBS的α-MEN溶液進行破骨細胞的誘導(dǎo);并進行破骨細胞的觀察及鑒定;采用CCK-8法檢測含藥血清毒性;骨髓單核細胞誘導(dǎo)后經(jīng)多聚甲醛固定后行TRAP染色計算其中陽性細胞數(shù)目;對骨片進行甲苯胺藍染色,然后觀察骨片上面骨吸收陷窩形成的個數(shù)及面積;最后進行掃描電鏡觀察。結(jié)果:1、治療后兩組患者的Harris評分及VAS評分均較治療前有明顯改善,(p0.05)但兩組臨床療效之間無顯著統(tǒng)計學(xué)差異。2、兩組比較治療前后壞死指數(shù)評分差值研究顯示,在緩解股骨頭壞死進程方面,治療組優(yōu)于對照組。3、兩組比較治療前后療效結(jié)果顯示,在緩解骨髓水腫及關(guān)節(jié)積液方面,股密保有一定療效,與阿侖膦酸鈉療效相似。4.通過骨髓單核細胞體外誘導(dǎo)的方式,可以獲取滿足實驗要求的破骨細胞。5.股密保高、中、低濃度含藥血清組與生理鹽水對照組相比,在12h,24h,3d及5d四個觀察期內(nèi)毒性差異均無統(tǒng)計學(xué)意義(P0.05)。6.通過觀察TRAP陽性細胞數(shù),在抑制骨髓單核細胞分化為破骨細胞的作用方面,股密保高、中、低濃度含藥血清組與生理鹽水對照組相比有顯著差異(P0.05),但股密保各濃度組之間療效無顯著差異(P0.05)。7.在限制骨吸收陷窩計數(shù)作用方面,通過數(shù)據(jù)分析,高濃度股密保含藥血清組與生理鹽水對照組療效之間存在顯著性差異(P0.05);其余各組之間無顯著差異。8.在限制骨吸收陷窩面積作用方面,通過數(shù)據(jù)分析,高濃度股密保含藥血清組與生理鹽水對照組療效之間存在顯著性差異(P0.05);低濃度及中濃度含藥血清組與高濃度含藥血清組療效之間存在顯著差異(P0.05);其余各組之間無顯著差異。結(jié)論:1.對于早中期股骨頭壞死患者,股密保方有一定的療效,可緩解疼痛,改善髖關(guān)節(jié)功能,減輕骨髓水腫及關(guān)節(jié)積液,延緩股骨頭壞死塌陷進程,療效與阿侖膦酸鈉相似。2.股密保含藥血清沒有毒性,可以抑制骨髓單核細胞誘導(dǎo)分化為破骨細胞;可以抑制破骨細胞的骨吸收功能。
[Abstract]:Objective: 1. through the collection of clinical cases, a randomized controlled study was carried out to observe the clinical efficacy of Yishen Huoxue shares in the treatment of Osteonecrosis of the femoral head (ONFH) in the treatment of early and mid-term osteonecrosis of the femoral head (head, ONFH) through experimental study to detect the toxicity of the blood serum of the femoral head containing drug, and to obtain sufficient numbers of bone marrow mononuclear cells in vitro. Quantity, stable character of the osteoclasts that meet the requirements of the experiment; the effect of the observation on the induced differentiation of bone marrow mononuclear cells into osteoclasts of the bone marrow mononuclear cells; the effect of the observation of the serum on the bone resorption of osteoclasts. Method: 1. according to the standard and exclusion criteria of Western medicine, from January 2016 to February 2017. A total of 43 patients (51 hips) were treated and followed up in the orthopedics clinic of the State Medical Hospital of Chinese medicine hospital. The patients were divided into two groups by random method. The control group was taken orally alendronate (fuxamy) for 70mgqw8 weeks, and the treatment group was treated with 43 courses for 4 weeks, with a total of 2 courses. The Harris score and the pain visual analogue scale were used before and after treatment (VA The S table) and the results of the imaging examination were recorded in detail. Finally, the statistical data were used to analyze the way that.2. was used to induce bone marrow mononuclear cells in vitro. In the culture and generation of bone marrow mononuclear cells, M-CSF, 100ng/ml RANKL, 1%PS, 5%FBS's alpha -MEN solution was used to induce osteoclasts, and osteoclasts were carried out. Observation and identification; CCK-8 method was used to detect the toxicity of serum containing drugs. The number of positive cells was calculated by TRAP staining after bone marrow mononuclear cells were fixed and fixed with POM. The bone slices were stained with toluidine blue, then the number and area of bone resorption lacunae formed on the bone were observed. Finally, the scanning electron microscope was observed. Results: 1, after treatment. The Harris score and the VAS score of the two groups were significantly better than those before the treatment, (P0.05), but there was no significant difference between the two groups of clinical effects. The difference between the two groups before and after the treatment of the necrosis index showed that the treatment group was better than the control group.3 in alleviating the necrosis of the femoral head, and the two groups were compared before and after the treatment. In alleviating bone marrow edema and joint effusion, it has a certain effect, which is similar to the effect of alendronate in the form of.4. induced by bone marrow mononuclear cells in vitro. It is possible to obtain the.5. strands of osteoclast to meet the requirements of the experiment. The middle, low concentration serum containing group and the saline control group are compared with the saline control group, four observations in 12h, 24h, 3D and 5D. There was no statistical significance (P0.05) in the period of endotoxicity (.6.), by observing the number of TRAP positive cells and the effect of inhibiting the differentiation of bone marrow mononuclear cells into osteoclast, there was a significant difference between the group and the normal saline group (P0.05), but there was no significant difference (P0.05).7 between the group and the concentration group (P0.05).7. There was a significant difference in the effect of limiting the counting of bone resorption lacunae by data analysis. There was a significant difference between the high concentration of the blood serum group and the saline control group (P0.05). There was no significant difference between the other groups and the effect of.8. on the area of bone resorption lacunae. There was significant difference between the curative effect of the saline control group (P0.05), and there was a significant difference between the low concentration and medium concentration serum group and the high concentration serum group (P0.05). There was no significant difference between the other groups. Conclusion: 1. for the early and middle stage of the femoral head necrosis, the femoral head has a certain effect, relieving pain and improving the hip. Joint function, reducing bone marrow edema and joint effusion, postponing the process of necrosis and collapse of the femoral head, the curative effect is not toxic with the.2. share containing alendronate, which can inhibit the differentiation of bone marrow mononuclear cells into osteoclast, and inhibit the bone resorption function of osteoclast.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R274.9
[Abstract]:Objective: 1. through the collection of clinical cases, a randomized controlled study was carried out to observe the clinical efficacy of Yishen Huoxue shares in the treatment of Osteonecrosis of the femoral head (ONFH) in the treatment of early and mid-term osteonecrosis of the femoral head (head, ONFH) through experimental study to detect the toxicity of the blood serum of the femoral head containing drug, and to obtain sufficient numbers of bone marrow mononuclear cells in vitro. Quantity, stable character of the osteoclasts that meet the requirements of the experiment; the effect of the observation on the induced differentiation of bone marrow mononuclear cells into osteoclasts of the bone marrow mononuclear cells; the effect of the observation of the serum on the bone resorption of osteoclasts. Method: 1. according to the standard and exclusion criteria of Western medicine, from January 2016 to February 2017. A total of 43 patients (51 hips) were treated and followed up in the orthopedics clinic of the State Medical Hospital of Chinese medicine hospital. The patients were divided into two groups by random method. The control group was taken orally alendronate (fuxamy) for 70mgqw8 weeks, and the treatment group was treated with 43 courses for 4 weeks, with a total of 2 courses. The Harris score and the pain visual analogue scale were used before and after treatment (VA The S table) and the results of the imaging examination were recorded in detail. Finally, the statistical data were used to analyze the way that.2. was used to induce bone marrow mononuclear cells in vitro. In the culture and generation of bone marrow mononuclear cells, M-CSF, 100ng/ml RANKL, 1%PS, 5%FBS's alpha -MEN solution was used to induce osteoclasts, and osteoclasts were carried out. Observation and identification; CCK-8 method was used to detect the toxicity of serum containing drugs. The number of positive cells was calculated by TRAP staining after bone marrow mononuclear cells were fixed and fixed with POM. The bone slices were stained with toluidine blue, then the number and area of bone resorption lacunae formed on the bone were observed. Finally, the scanning electron microscope was observed. Results: 1, after treatment. The Harris score and the VAS score of the two groups were significantly better than those before the treatment, (P0.05), but there was no significant difference between the two groups of clinical effects. The difference between the two groups before and after the treatment of the necrosis index showed that the treatment group was better than the control group.3 in alleviating the necrosis of the femoral head, and the two groups were compared before and after the treatment. In alleviating bone marrow edema and joint effusion, it has a certain effect, which is similar to the effect of alendronate in the form of.4. induced by bone marrow mononuclear cells in vitro. It is possible to obtain the.5. strands of osteoclast to meet the requirements of the experiment. The middle, low concentration serum containing group and the saline control group are compared with the saline control group, four observations in 12h, 24h, 3D and 5D. There was no statistical significance (P0.05) in the period of endotoxicity (.6.), by observing the number of TRAP positive cells and the effect of inhibiting the differentiation of bone marrow mononuclear cells into osteoclast, there was a significant difference between the group and the normal saline group (P0.05), but there was no significant difference (P0.05).7 between the group and the concentration group (P0.05).7. There was a significant difference in the effect of limiting the counting of bone resorption lacunae by data analysis. There was a significant difference between the high concentration of the blood serum group and the saline control group (P0.05). There was no significant difference between the other groups and the effect of.8. on the area of bone resorption lacunae. There was significant difference between the curative effect of the saline control group (P0.05), and there was a significant difference between the low concentration and medium concentration serum group and the high concentration serum group (P0.05). There was no significant difference between the other groups. Conclusion: 1. for the early and middle stage of the femoral head necrosis, the femoral head has a certain effect, relieving pain and improving the hip. Joint function, reducing bone marrow edema and joint effusion, postponing the process of necrosis and collapse of the femoral head, the curative effect is not toxic with the.2. share containing alendronate, which can inhibit the differentiation of bone marrow mononuclear cells into osteoclast, and inhibit the bone resorption function of osteoclast.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R274.9
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相關(guān)期刊論文 前10條
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3 李s，
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