【摘要】 探討血管內皮細胞生長因子（VEGF）對肌肉骨膜瓣、松質骨顆粒和骨髓聯合移植修復骨缺損中的 影響 .［ 方法 ］製作家兔橈骨骨缺損模型，在缺損區用肌肉骨膜瓣、松質骨顆粒及骨髓進行聯合移植，於術後第1、2、4、6、8周時利用光學顯微鏡和免疫組織化學等方法檢測VEGF在骨缺損修復不同階段中對各種細胞內的表達情況.［結果］VEGF在骨缺損修復過程中的不同階段的表達程度不同，在術後1～6周時(即軟骨內化骨階段)，新生的骨細胞、成骨細胞及軟骨細胞內均可見大量的VEGF，其高峰出現在第4周.［結論］VEGF是骨缺損修復過程中重要的生長因子，聯合移植有利於骨缺損的快速修復.
【關鍵詞】 血管內皮生長因子類 骨缺損 骨移植
Effects of vascular endothelial growth factor for repairing bone defect
ABSTRACT:OBJECTIVETo study the expression and effects of vascular endothelial growth factor (VEGF) for combined transplanting the musculoperiosteal flap, bone marrow and spongy bone granules to repairing bone defect.METHODSSegmental bone defect was created in the mid upper of radial shaft of adult rabbits, and combination of musculoperiosteal flap, bone marrow and spongy bone granules was transplanted to the bone defect. VEGF expression in the different phase of repairing bone defect was defected by immunohistochemistry at postoperative of 1, 2, 4, 6, 8 weeks.RESULTSExpression of VEGF was different in the different phase of repairing bone defect. VEGF was expressed in neoosteocytes, osteoblasts and chondrocytes in the bone defects sites from one to six weeks (phase of os endochondrale) after operation, and the high expression of VEGF was detected at 4 weeks.CONCLUSIONVEGF is key factor in healing bone defects, and combined transplantation makes for repairing the bone defect.
Key words:vascular endothelial growth factors;bone defect;bone transplantation
目前 ，車禍造成的開放性骨折、骨壞死及骨缺損日益增多，而骨折不癒合及骨缺損是 治療 難點.雖然多種方法可用於修復骨缺損，包括自體骨移植、同種異體骨移植及人工骨移植等，但均有弊端.本實驗通過肌肉骨膜瓣、松質骨顆粒及骨髓聯合移植修復了家兔橈骨骨缺損，並利用免疫組織化學方法觀察了修復效果，旨在為臨床治療骨缺損提供依據.
1 材料與方法
取健康家兔30隻，體重為2.5～3.0kg，雌雄不限，由延邊大學醫學部實驗動物科提供，分籠飼養.實驗前術區備皮，常規消毒，用5g/L利多卡因行局部浸潤麻醉，取雙前肢前內側縱行切口，經肌間隙進入，顯露橈骨幹，截除橈骨中段15mm，製作骨缺損動物模型，選擇右前肢為實驗肢，左前肢為對照肢.取髂後上棘切口，長度約為20mm，取出15mm×10mm大小的髂骨，刮取其松質骨顆粒備用；於脛骨上段前內側切開皮膚約10mm,抽取骨髓2.0mL, 自然 狀態下凝固備用.將實驗組旋前圓肌置入骨缺損模型家兔實驗肢骨缺損區，並將松質骨顆粒及骨髓植入旋前圓肌組織內，用橈骨骨膜包裹，用50無損傷線仔細縫合骨膜，再逐層縫合切口；而對照肢則未給予任何處理，直接縫合.分別於術後1、2、4、6、8周時隨機選擇模型家兔5隻處死，分別於實驗肢及對照肢橈骨骨缺損處取材，置於100g/L甲醛溶液固定24h，EDTA脫鈣4周，每周換液1次，梯度酒精脫水，石蠟包埋，5μm厚組織切片，行HE及免疫組織化學染色.方法為組織切片脫臘，水化，EDTA熱修復，過氧化氫封閉非特異性抗原，PBS沖洗3次，滴加第一抗體，4℃過夜；沖洗3次，滴加第二抗體，室溫孵育10min；PBS沖洗3次，滴加辣根酶標記鏈霉卵白素，室溫孵育10min；PBS沖洗3次，DAB顯色，梯度酒精脫水，二甲苯透明，封片.
【關鍵詞】 血管內皮生長因子類 骨缺損 骨移植
Effects of vascular endothelial growth factor for repairing bone defect
ABSTRACT:OBJECTIVETo study the expression and effects of vascular endothelial growth factor (VEGF) for combined transplanting the musculoperiosteal flap, bone marrow and spongy bone granules to repairing bone defect.METHODSSegmental bone defect was created in the mid upper of radial shaft of adult rabbits, and combination of musculoperiosteal flap, bone marrow and spongy bone granules was transplanted to the bone defect. VEGF expression in the different phase of repairing bone defect was defected by immunohistochemistry at postoperative of 1, 2, 4, 6, 8 weeks.RESULTSExpression of VEGF was different in the different phase of repairing bone defect. VEGF was expressed in neoosteocytes, osteoblasts and chondrocytes in the bone defects sites from one to six weeks (phase of os endochondrale) after operation, and the high expression of VEGF was detected at 4 weeks.CONCLUSIONVEGF is key factor in healing bone defects, and combined transplantation makes for repairing the bone defect.
Key words:vascular endothelial growth factors;bone defect;bone transplantation
目前 ，車禍造成的開放性骨折、骨壞死及骨缺損日益增多，而骨折不癒合及骨缺損是 治療 難點.雖然多種方法可用於修復骨缺損，包括自體骨移植、同種異體骨移植及人工骨移植等，但均有弊端.本實驗通過肌肉骨膜瓣、松質骨顆粒及骨髓聯合移植修復了家兔橈骨骨缺損，並利用免疫組織化學方法觀察了修復效果，旨在為臨床治療骨缺損提供依據.
1 材料與方法
取健康家兔30隻，體重為2.5～3.0kg，雌雄不限，由延邊大學醫學部實驗動物科提供，分籠飼養.實驗前術區備皮，常規消毒，用5g/L利多卡因行局部浸潤麻醉，取雙前肢前內側縱行切口，經肌間隙進入，顯露橈骨幹，截除橈骨中段15mm，製作骨缺損動物模型，選擇右前肢為實驗肢，左前肢為對照肢.取髂後上棘切口，長度約為20mm，取出15mm×10mm大小的髂骨，刮取其松質骨顆粒備用；於脛骨上段前內側切開皮膚約10mm,抽取骨髓2.0mL, 自然 狀態下凝固備用.將實驗組旋前圓肌置入骨缺損模型家兔實驗肢骨缺損區，並將松質骨顆粒及骨髓植入旋前圓肌組織內，用橈骨骨膜包裹，用50無損傷線仔細縫合骨膜，再逐層縫合切口；而對照肢則未給予任何處理，直接縫合.分別於術後1、2、4、6、8周時隨機選擇模型家兔5隻處死，分別於實驗肢及對照肢橈骨骨缺損處取材，置於100g/L甲醛溶液固定24h，EDTA脫鈣4周，每周換液1次，梯度酒精脫水，石蠟包埋，5μm厚組織切片，行HE及免疫組織化學染色.方法為組織切片脫臘，水化，EDTA熱修復，過氧化氫封閉非特異性抗原，PBS沖洗3次，滴加第一抗體，4℃過夜；沖洗3次，滴加第二抗體，室溫孵育10min；PBS沖洗3次，滴加辣根酶標記鏈霉卵白素，室溫孵育10min；PBS沖洗3次，DAB顯色，梯度酒精脫水，二甲苯透明，封片.
收藏