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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:

第24卷

期數(shù):

2012年06期

頁碼:

13-17

欄目:

基礎(chǔ)研究

出版日期:

2012-06-30

文章信息/Info

Title:

Mechanism of action of spongy bone autografting combined with BMSCs transplantation on avascular necrosis of femoral head in rabbits

作者:

馮建書¹; 胡永成²; 王保芝³; 王江泳¹; 劉長安⁴; 曹沛宏²

1.河北醫(yī)科大學(xué)附屬石家莊市第三醫(yī)院，河北石家莊050011；
2.天津醫(yī)院， 天津300211；
3.河北醫(yī)科大學(xué)，河北石家莊050017；
4.白求恩國際和平醫(yī)院，河北石家莊050010

Author(s):

FENG Jian-shu*; HU Yong-cheng; WANG Bao-zhi; WANG Jiang-yong; LIU Chang-an; CAO Pei-hong. *

The Third Hospital of Shijiazhuang affiliated to Hebei Medical University,Shijiazhuang 050011,Hebei,China

關(guān)鍵詞:

股骨頭壞死　間質(zhì)干細胞移植　骨移植　動物實驗　兔

Keywords:

Femur head necrosis; Mesenchymal stem cell transplantation; Bone transplantation; Animal experimentation; Rabbits

摘要:

目的：探討自體松質(zhì)骨和骨髓間充質(zhì)干細胞聯(lián)合移植治療股骨頭缺血性壞死的可能機制，為該方法應(yīng)用于臨床提供依據(jù)。 方法：兔齡4～5個月的新西蘭大耳白兔24只，雌雄不限，隨機分為2組，每組12只。采用液氮冷凍股骨頭負重區(qū)的方法造成股骨 頭缺血壞死模型后，A組進行自體松質(zhì)骨移植，B組進行自體松質(zhì)骨和骨髓間充質(zhì)干細胞聯(lián)合移植。每組術(shù)后2、4、6、8周分別 處死3只動物，制作股骨頭標本，進行X線檢查和組織學(xué)觀察，并對術(shù)后8周的股骨頭標本進行掃描電鏡觀察。結(jié)果：①X線觀察 結(jié)果。A組術(shù)后2周時，有2個標本鉆孔區(qū)密度不均勻，鉆孔邊緣清晰可見；4周時，有3個標本鉆孔區(qū)密度增高不顯著，鉆孔邊緣 出現(xiàn)硬化線；6周時有4個標本鉆孔區(qū)顯示有骨吸收現(xiàn)象，出現(xiàn)股骨頭關(guān)節(jié)面塌陷；8周時有3個標本骨吸收現(xiàn)象明顯，而骨小梁 結(jié)構(gòu)不清晰。B組術(shù)后2周，股骨頭形態(tài)均正常，鉆孔區(qū)有密度增高影，質(zhì)地均勻，鉆孔邊緣尚可分辨；4周時股骨頭形態(tài)均正常 ，鉆孔區(qū)密度進一步升高，質(zhì)地均勻，鉆孔邊緣變得模糊；6周時股骨頭形態(tài)仍正常，鉆孔區(qū)密度高而均勻，鉆孔邊緣出現(xiàn)放射 狀骨小梁接合現(xiàn)象；8周時鉆孔區(qū)密度接近周圍骨質(zhì)，股骨頭內(nèi)出現(xiàn)張力性和壓力性骨小梁，鉆孔邊緣交界區(qū)骨小梁接合良好。 ②組織學(xué)觀察結(jié)果。A組術(shù)后2周時，移植骨發(fā)生壞死，4個標本出現(xiàn)再血管化，移植區(qū)出現(xiàn)破骨和成骨細胞，成骨細胞的數(shù)目較 少，移植骨周圍附加新生骨較薄，有少量連接現(xiàn)象；另2個標本局部新生血管較少，成骨細胞募集不良，新生骨組織的形成較少 ，無連接現(xiàn)象。術(shù)后4周時，3個標本新生骨增加，連接現(xiàn)象增多，出現(xiàn)骨塑形；另3個標本移植骨無明顯修復(fù)現(xiàn)象。術(shù)后6周時 ，有2個標本新生骨量增加，有骨塑形現(xiàn)象；另4個標本移植骨無明顯修復(fù)表現(xiàn)，出現(xiàn)骨吸收現(xiàn)象。 術(shù)后8周時，3個標本可見到 塑形的板層骨小梁和成熟的骨髓組織；另3個標本出現(xiàn)較明顯的骨吸收現(xiàn)象，局部仍有壞死骨組織存在，無明顯的骨組織修復(fù)現(xiàn) 象。B組術(shù)后2周時，植入的松質(zhì)骨發(fā)生壞死，出現(xiàn)局部再血管化，破骨細胞和大量的成骨細胞出現(xiàn)在移植區(qū)，植入的松質(zhì)骨被 邊緣生成的1層較厚的新生骨連接在一起；4周時骨小梁增粗變大，連接進一步加強，并出現(xiàn)骨髓組織；6周時骨塑形現(xiàn)象活躍； 8周時骨塑形現(xiàn)象仍然存在，可以見到塑形的板層骨小梁和成熟的骨髓組織。③掃描電鏡觀察結(jié)果。術(shù)后8周時，A組有3個標本 在掃描電鏡下顯示移植骨保持原有骨小梁結(jié)構(gòu)，部分骨小梁出現(xiàn)蟲蝕樣破壞。B組標本在掃描電鏡下顯示，骨小梁密集，骨髓腔 間隙較小，力學(xué)結(jié)構(gòu)欠完善，骨膠原纖維排列不均勻、不整齊。結(jié)論：自體松質(zhì)骨和骨髓間充質(zhì)干細胞聯(lián)合移植治療股骨頭缺 血性壞死，細胞移植為股骨頭的骨質(zhì)修復(fù)提供了充足的骨髓間充質(zhì)干細胞，骨修復(fù)現(xiàn)象活躍，可提高骨移植的成功率，促進股 骨頭骨組織修復(fù)。

Abstract:

Objective:To investigate the mechanism of action of spongy bone autografting combined with bone marrow mesenchymal stem cells(BMSCs)transplantation on the experimental avascular necrosis of femoral head （ANFH）for the purpose of application in clinic.Methods:Twenty-four New Zealand white rabbits,4 to 5 months old,were divided into 2 groups randomly,12 cases in each group.After the rabbit model of ANFH were built by freezing the weight loading region of femoral head with liquid nitrogen,rabbits in group A were administrated with spongy bone autografting,while rabbits in group B were administrated with spongy bone autografting combined with BMSCs transplantation.Three rabbits selected from each group were sacrificed 2,4,6,8 weeks after the transplantation for making specimens of femoral head,which were observed with radioscopy,histological examination.Moreover,those specimens obtained 8 weeks after the transplantation were observed with scanning electron microscope.Results:Radiological observations:In group A,two specimens presented uneven density in drilled area and clear images of drilled holes 2 weeks after operation,sclerosis line was showed at the margin of drilled area in 3 specimens and increase in density were not significant in drilled area 4 weeks after operation,bone resorption of drilled area and femoral head collapse appeared in 4 specimens 6 weeks after operation,three specimens presented obvious bone resorption and disordered structure of trabeculae of bone 8 weeks after operation.In group B,the shape of femoral head was normal and the bone density increased uniformly in the drilled area while the margin of drilled area could be distinguished 2 weeks after operation,the shape of femoral head was normal and the bone density further increased uniformly in the drilled area and the margin of drilled area became obscured 4 weeks after operation,the shape of femoral head was still normal and the bone density was high and uniform in the drilled area and radial conjunction of the trabeculae was presented at the margin of drilled area 6 weeks after operation,the bone density in the drilled area approached that of the surrounding substantia ossea and tensile trabeculae and compression trabeculae formed in the femoral head and good conjunctions of the trabeculae were presented in the junctional zone of the margin of drilled area 8 weeks after operation.Histological observations:In group A,two weeks after operation,osteonecrosis of bone graft appeared,revascularization appeared in 4 specimens,osteoclasts and fewer osteoblasts appeared in transplantation zone,the new bone around bone graft was slight and a few conjunctions were presented,there were no conjunction and few neovascularization were presented in the other 2 specimens,osteoblasts recruitment was poor.Four weeks after operation,new bone and conjunction increased and bone modeling appeared in 3 specimens,while no remodeling of bone graft was presented in the other 3 specimens.Six weeks after operation,new bone mass increased and bone remodeling occured in 2 specimens,there were no obvious repair of bone graft phenomenon and bone resorption occurred in the other 4 specimens.Eight weeks after operation,moulded lamellar bone trabeculae and mature myeloid tissue could be observed in 3 specimens.In the other 3 specimens obvious bone resorption phenomenon were observed while no obvious bone repair phenomenon appeared,and bone necrosis could be seen in partial area.In group B,two weeks after operation,necrosis of bone graft and partial revascularization appeared,osteoblasts and osteoclasts appeared in the transplant area, grafted spongy bone were connected by new bone.Bone trabecula augmented and conjunction enhanced and myeloid tissue appeared 4 weeks after operation.Bone remodeling was active 6 weeks after operation and it still existed 8 weeks after operation,meanwhile,the lamellar bone trabecula and matured myeloid tissue could be observed.Scanning electron microscope observations:Eight weeks after operation,trabecular structure was maintained and moth-eaten bony destruction was found in part bone trabecula in 3 specimens of group A.Grafted bone trabeculae in group B were different from normal biomechanics structure,they arranged closely and had a narrow cavitas medullaris,whlie,osteocollagenous fibers arranged unevenly and irregularly.Conclusion:In the treatment of ANFH,cellular transplant provides sufficient BMSCs and the result is active repairing of bone tissue of the femoral head.Spongy bone autografting combined with BMSCs transplantation can obviously promote the repairing of bone tissues of the femoral head and raise the achievement rate of bone transplantation.

備注/Memo

備注/Memo:

2012-02-28收稿2012-05-11修回
基金項目：河北省科技廳科技攻關(guān)項目（2007276171）
通訊作者：王江泳E-mail：[email protected]〖HJ2.4mm〗

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更新日期/Last Update: 2012-06-30