Insufficient donor dermis and the shortage of three-dimensional vascular networks are the main limitations in the tissue-engineered dermis(TED).To solve these problems,we initially constructed pre-vascularized bone ma...Insufficient donor dermis and the shortage of three-dimensional vascular networks are the main limitations in the tissue-engineered dermis(TED).To solve these problems,we initially constructed pre-vascularized bone marrow mesenchymal stem cell sheet(PBMCS)and pre-vascularized fibroblasts cell sheet(PFCS)by cell sheet technology,and then superimposed or folded them together to construct a pre-vascularized TED(PTED),aiming to mimic the real dermis structure.The constructed PTED was implanted in nude mice dorsal dermis-defect wound and the wound-healing effect was quantified at Days 1,7 and 14 via the methods of histochemistry and immunohistochemistry.The results showed that PTED could rapidly promote the wound closure,especially at Day 14,and the wound-healing rate of three-layer PTED could reach 97.2%(P<0.01),which was faster than the blank control group(89.1%),PBMCS(92.4%),PFCS(93.8%)and six-layer PTED(92.3%).In addition,the vessel density in the PTED group was higher than the other groups on the 14th day.Taken together,it is proved that the PTED,especially three-layer PTED,is more conducive to the fullthickness dermis-defect repair and the construction of the three-dimensional vascular networks,indicating its potential application in dermis-defect repair.展开更多
目的:构建根尖牙乳头干细胞(stem cells from apical papilla,SCAP)细胞膜片,并对其组织学形态和成骨/成牙本质分化性能进行研究,为SCAP细胞膜片应用于年轻恒牙牙髓再生提供实验基础。方法:分离年轻恒牙第三磨牙牙乳头,进行SCAP原代培...目的:构建根尖牙乳头干细胞(stem cells from apical papilla,SCAP)细胞膜片,并对其组织学形态和成骨/成牙本质分化性能进行研究,为SCAP细胞膜片应用于年轻恒牙牙髓再生提供实验基础。方法:分离年轻恒牙第三磨牙牙乳头,进行SCAP原代培养。采用流式细胞术检测SCAP表面标志物,茜素红S染色检测其成骨/成牙本质分化。取第3代SCAP,膜片诱导液连续培养14d,形成细胞膜片。对SCAP细胞膜片进行H-E染色,组织学观察;流式细胞术检测SCAP细胞膜片的细胞周期变化;实时定量PCR(qRT-PCR)检测细胞膜片中Runx2、ALP、Col-I基因表达;Western印迹法检测细胞膜片Runx2、ALP蛋白表达水平。采用SPSS17.0软件包对实验结果进行统计学分析,两样本细胞周期检测和q RT-PCR基因检测均数的比较采用t检验。结果:组织学H-E染色显示,SCAP细胞膜片由5~6层细胞组成,细胞量大,细胞之间排列紧密,且富含细胞外基质。SCAP细胞膜片细胞周期G2+S期所占比例较低,而G1期比例较高,提示SCAP细胞膜片增殖性能受到显著抑制(P<0.05)。同时,qRT-PCR和Western印迹法结果显示,与SCAP相比,SCAP细胞膜片成骨/成牙本质分化能力显著升高(P<0.05)。结论:SCAP细胞膜片富含细胞外基质,且成骨/成牙本质分化能力高于单纯SCAP,SCAP细胞膜片应用于牙髓再生更具有优势。展开更多
基金supported by The Natural Science Foundation of China(81571829)The Fundamental Research Funds for the Central Universities(lzujbky-2020-it29)the open project of State Key Laboratory of Solid Lubrication,Lanzhou Institute of Chemical Physics,Chinese Academy of Sciences(LSL-1907).
文摘Insufficient donor dermis and the shortage of three-dimensional vascular networks are the main limitations in the tissue-engineered dermis(TED).To solve these problems,we initially constructed pre-vascularized bone marrow mesenchymal stem cell sheet(PBMCS)and pre-vascularized fibroblasts cell sheet(PFCS)by cell sheet technology,and then superimposed or folded them together to construct a pre-vascularized TED(PTED),aiming to mimic the real dermis structure.The constructed PTED was implanted in nude mice dorsal dermis-defect wound and the wound-healing effect was quantified at Days 1,7 and 14 via the methods of histochemistry and immunohistochemistry.The results showed that PTED could rapidly promote the wound closure,especially at Day 14,and the wound-healing rate of three-layer PTED could reach 97.2%(P<0.01),which was faster than the blank control group(89.1%),PBMCS(92.4%),PFCS(93.8%)and six-layer PTED(92.3%).In addition,the vessel density in the PTED group was higher than the other groups on the 14th day.Taken together,it is proved that the PTED,especially three-layer PTED,is more conducive to the fullthickness dermis-defect repair and the construction of the three-dimensional vascular networks,indicating its potential application in dermis-defect repair.
