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[1]趙守軍,熊文化,許柯.細(xì)胞因子基因轉(zhuǎn)染的骨骺干細(xì)胞復(fù)合明膠-硫酸軟骨素-透明質(zhì)酸鈉支架修復(fù)大鼠骨骺生長(zhǎng)板缺損的實(shí)驗(yàn)研究[J].中醫(yī)正骨,2018,30(02):10-15.
　ZHAO Shoujun,XIONG Wenhua,XU Ke.A combination of epiphysis stem cell transfected with cytokine genes and bio-scaffold consists of gelatin,chondroitin sulfate and sodium hyaluronate for repairing epiphyseal growth plate defect in rats:an experimental study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2018,30(02):10-15.
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細(xì)胞因子基因轉(zhuǎn)染的骨骺干細(xì)胞復(fù)合明膠-硫酸軟骨素-透明質(zhì)酸鈉支架修復(fù)大鼠骨骺生長(zhǎng)板缺損的實(shí)驗(yàn)研究()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第30卷
期數(shù):
2018年02期
頁(yè)碼:
10-15
欄目:
基礎(chǔ)研究
出版日期:
2018-02-20

文章信息/Info

Title:
A combination of epiphysis stem cell transfected with cytokine genes and bio-scaffold consists of gelatin,chondroitin sulfate and sodium hyaluronate for repairing epiphyseal growth plate defect in rats:an experimental study
作者:
趙守軍; 熊文化; 許柯
浙江省寧波市第二醫(yī)院,浙江 寧波 315010
Author(s):
ZHAO Shoujun; XIONG Wenhua; XU Ke
The Second Hospital of Ningbo City,Ningbo 315010,Zhejiang,China
關(guān)鍵詞:
骨骺;  組織工程;  缺氧誘導(dǎo)因子1; α亞基;  甲狀旁腺激素相關(guān)肽;  干細(xì)胞移植;  組織支架;  大鼠; Sprague-Dawley;  動(dòng)物實(shí)驗(yàn)
Keywords:
Keywords epiphyses;  tissue engineering;  hypoxia-inducible factor 1; alpha subunit;  parathyroid hormone-related peptide;  stem cell transplantation;  tissue scaffolds;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:觀(guān)察細(xì)胞因子基因轉(zhuǎn)染的骨骺干細(xì)胞復(fù)合明膠-硫酸軟骨素-透明質(zhì)酸鈉支架修復(fù)大鼠骨骺生長(zhǎng)板缺損的效果。方法:取SD大鼠肋軟骨透明帶細(xì)胞,用Percoll密度梯度離心法獲取骨骺干細(xì)胞。從低氧誘導(dǎo)因子(hypoxia-inducible factor,HIF)-1α、甲狀旁腺激素相關(guān)肽(parathyroid hormone-related peptide,PTHrP)、重組人源核因子-κB(recombinant human nuclear factor-κB,rhNF-κB)、HIF-1α+PTHrP、rhNF-κB+PTHrP、HIF-1α+rhNF-κB、rhNF-κB+HIF-1α+PTHrP共7種細(xì)胞因子及細(xì)胞因子組合中篩選出對(duì)骨骺干細(xì)胞增殖作用最佳的細(xì)胞因子或細(xì)胞因子組合。利用篩選出的細(xì)胞因子或細(xì)胞因子組合,構(gòu)建能穩(wěn)定表達(dá)重組細(xì)胞因子的骨骺干細(xì)胞。取20只SD大鼠,制作右脛骨上端骨骺生長(zhǎng)板缺損模型。造模后將大鼠隨機(jī)分為2組,每組10只,分別采用從自體肋軟骨透明帶細(xì)胞獲取的骨骺干細(xì)胞移植(自體移植組)和重組骨骺干細(xì)胞植入明膠-硫酸軟骨素-透明質(zhì)酸鈉載體(組織工程組)兩種方法修復(fù)骨骺生長(zhǎng)板缺損。分別于修復(fù)術(shù)后4周、8周時(shí),拍攝2組大鼠雙下肢X線(xiàn)片,計(jì)算雙側(cè)脛骨長(zhǎng)度差值和雙側(cè)脛骨角差值。X線(xiàn)檢查結(jié)束后從每組隨機(jī)選取5只大鼠,取肋軟骨獲取骨骺干細(xì)胞,以Western Blot法測(cè)定CollagenⅡ及CollagenⅩ蛋白水平; 同時(shí)切取大鼠右脛骨上端,制作切片,HE染色后光鏡下觀(guān)察骨骺生長(zhǎng)板損傷修復(fù)情況。結(jié)果:最終篩選出的細(xì)胞因子組合為HIF-1α+PTHrP,將其構(gòu)建入pIRES2-EGFP真核表達(dá)載體,轉(zhuǎn)染獲取的骨骺干細(xì)胞,獲得穩(wěn)定表達(dá)重組細(xì)胞因子的骨骺干細(xì)胞。骨骺生長(zhǎng)板缺損修復(fù)術(shù)后4周、8周時(shí),組織工程組大鼠的雙側(cè)脛骨長(zhǎng)度差值、雙側(cè)脛骨角差值均小于自體移植組[(0.07±0.01)cm,(0.35±0.05)cm,t=17.360,P=0.000;(0.83±0.45)cm,(1.51±0.13)cm,t=4.590,P=0.001; 1.50°±0.69°,2.55°±0.40°,t=4.160,P=0.001; 14.26°±1.87°,33.98°±2.17°,t=21.770,P=0.000],組織工程組大鼠的CollagenⅡ、CollagenⅩ蛋白水平均高于自體移植組[0.21±0.02,0.10±0.01,t=15.560,P=0.000; 0.37±0.03,0.14±0.01,t=16.260,P=0.000; 0.22±0.03,0.08±0.01,t=14.010,P=0.000; 0.33±0.02,0.11±0.01,t=21.000,P=0.000]。HE染色結(jié)果顯示,修復(fù)術(shù)后4周時(shí),組織工程組脛骨上端骨骺生長(zhǎng)板缺損由新生軟骨部分填充,自體移植組尚未形成骨骺生長(zhǎng)板組織結(jié)構(gòu); 修復(fù)術(shù)后8周時(shí),組織工程組脛骨上端骨骺生長(zhǎng)板部分接近閉合,呈薄層柱狀結(jié)構(gòu),自體移植組有新骨骺形成。結(jié)論:以骨骺干細(xì)胞作為種子細(xì)胞,以HIF-1α+PTHrP為細(xì)胞因子,以明膠-硫酸軟骨素-透明質(zhì)酸鈉載體作為生物支架,能有效修復(fù)大鼠骨骺生長(zhǎng)板缺損。
Abstract:
ABSTRACT Objective:To observe the role of a combination of epiphysis stem cell transfected with cytokine genes and bio-scaffold consists of gelatin,chondroitin sulfate and sodium hyaluronate in repairing epiphyseal growth plate defect in rats.Methods:The zona-pellucida cells were fetched out from costicartilage of SD rats and the epiphysis stem cells were obtained by using Percoll density gradient centrifugation method.Cytokine or cytokine combinations which had the best promoting effect on proliferation of epiphysis stem cells were selected out of seven kinds of cytokines and cytokine combinations,including hypoxia-inducible factor(HIF)-1α,

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備注/Memo

備注/Memo:
基金項(xiàng)目:寧波市科技局社會(huì)發(fā)展資助項(xiàng)目(2009C50022) 通訊作者:熊文化 E-mail:[email protected]
更新日期/Last Update: 2018-07-02