重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2015年
23期
3176-3179
,共4页
谭伟%吕海%曹纬%张朋%杨刘柱%周初松
譚偉%呂海%曹緯%張朋%楊劉柱%週初鬆
담위%려해%조위%장붕%양류주%주초송
组织工程%支架%髓核%去细胞%骨髓间充质干细胞
組織工程%支架%髓覈%去細胞%骨髓間充質榦細胞
조직공정%지가%수핵%거세포%골수간충질간세포
tissue engineering%scaffold%nucleus pulposus%acellular matrix%bone marrow mesenchymal stem cells
目的:体外培养兔骨髓间充质干细胞(rBMSCs)-髓核去细胞基质支架(NPAMS)复合体(rBMSCs-NPAMS)构建组织工程髓核。方法制备若干 NPAMS,接种 BMSCs 至 NPAMS 体外培养分为 NPAMS 组、实验组和正常对照组(正常髓核)。肉眼及显微镜观察复合体形态变化,并行扫描电镜(SEM)、苏木素-伊红(HE)染色、免疫组织化学、实时定量 PCR(qRT-PCR)、支架的生物力学等检测。结果肉眼下 rBMSCs-NPAMS 形态接近正常髓核;SEM 显示细胞在支架表面大量黏附、并向深部迁移,表面细胞密度比横截面细胞密度大;HE 染色表明随时间延长,rBMSCs-NPAMS 内细胞量递增,分布更广泛;免疫组织化学显示细胞外基质2型胶原(CollagenⅡ)分泌量随时间递增,且实验组 CollagenⅡ表达量大于 NPAMS 组,小于正常对照组;qRT-PCR结果:NPAMS 组未提取到 mRNA,实验组 CollagenⅡ、聚集蛋白聚糖(Aggrecan)mRNA 相对表达量呈时间依赖性递增,但均小于正常对照组(P <0.01),支架与正常髓核在相同位移下的压缩载荷差异无统计学意义(P >0.05)。结论采用髓核体外 NPAMS复合 rBMSCs 可成功构建组织工程髓核。
目的:體外培養兔骨髓間充質榦細胞(rBMSCs)-髓覈去細胞基質支架(NPAMS)複閤體(rBMSCs-NPAMS)構建組織工程髓覈。方法製備若榦 NPAMS,接種 BMSCs 至 NPAMS 體外培養分為 NPAMS 組、實驗組和正常對照組(正常髓覈)。肉眼及顯微鏡觀察複閤體形態變化,併行掃描電鏡(SEM)、囌木素-伊紅(HE)染色、免疫組織化學、實時定量 PCR(qRT-PCR)、支架的生物力學等檢測。結果肉眼下 rBMSCs-NPAMS 形態接近正常髓覈;SEM 顯示細胞在支架錶麵大量黏附、併嚮深部遷移,錶麵細胞密度比橫截麵細胞密度大;HE 染色錶明隨時間延長,rBMSCs-NPAMS 內細胞量遞增,分佈更廣汎;免疫組織化學顯示細胞外基質2型膠原(CollagenⅡ)分泌量隨時間遞增,且實驗組 CollagenⅡ錶達量大于 NPAMS 組,小于正常對照組;qRT-PCR結果:NPAMS 組未提取到 mRNA,實驗組 CollagenⅡ、聚集蛋白聚糖(Aggrecan)mRNA 相對錶達量呈時間依賴性遞增,但均小于正常對照組(P <0.01),支架與正常髓覈在相同位移下的壓縮載荷差異無統計學意義(P >0.05)。結論採用髓覈體外 NPAMS複閤 rBMSCs 可成功構建組織工程髓覈。
목적:체외배양토골수간충질간세포(rBMSCs)-수핵거세포기질지가(NPAMS)복합체(rBMSCs-NPAMS)구건조직공정수핵。방법제비약간 NPAMS,접충 BMSCs 지 NPAMS 체외배양분위 NPAMS 조、실험조화정상대조조(정상수핵)。육안급현미경관찰복합체형태변화,병행소묘전경(SEM)、소목소-이홍(HE)염색、면역조직화학、실시정량 PCR(qRT-PCR)、지가적생물역학등검측。결과육안하 rBMSCs-NPAMS 형태접근정상수핵;SEM 현시세포재지가표면대량점부、병향심부천이,표면세포밀도비횡절면세포밀도대;HE 염색표명수시간연장,rBMSCs-NPAMS 내세포량체증,분포경엄범;면역조직화학현시세포외기질2형효원(CollagenⅡ)분비량수시간체증,차실험조 CollagenⅡ표체량대우 NPAMS 조,소우정상대조조;qRT-PCR결과:NPAMS 조미제취도 mRNA,실험조 CollagenⅡ、취집단백취당(Aggrecan)mRNA 상대표체량정시간의뢰성체증,단균소우정상대조조(P <0.01),지가여정상수핵재상동위이하적압축재하차이무통계학의의(P >0.05)。결론채용수핵체외 NPAMS복합 rBMSCs 가성공구건조직공정수핵。
Objective To construct tissue engineering nucleus pulposus by culture of rabbit bone marrow mesenchymal stem cells (rBMSCs)-nucleus pulposus acellular matrix scaffold (NPAMS)complexes (rBMSCs-NPAMS).Methods Several NPAMS were prepared,and rBMSCs was seeded into NPAMS.The scaffolds and complex were detected by general observation,HE stai-ning,immunohistochemical,qRT-PCR,scanning electron microscopy.Results The scanning electron microscopy showed the seed cell in nucleus pulposus ECM-derived scaffold could adhesion and growth.The cell attachment and proliferation were observed by HE staining.Immunohistochemical examination with typeⅡ collagen showed positive results.qRT-PCR revealed the time-depend-ent of the mRNA expression of collagen Ⅱ,and which was smaller than positive control(P<0.01).The relative expression of Ag-grecan mRNA grew in a time dependent fashion,which was still lower than positive control(P<0.01).Scaffolds and normal nucle-us pulposus had no statistical significance of the compression load under the same displacement of the compression(P>0.05).Con-clusion Natural nucleus pulposus acellular matrix scaffold composite allogeneic bone marrow mesenchymal stem cells can be suc-cessfully built into tissue engineering nucleus pulposus.