Periosteum is a thin fibrous layer that covers most bones. It resides in a dynamic mechanically loaded environment and provides a niche for pluripotent cells and a source for molecular factors that modulate cell behav...Periosteum is a thin fibrous layer that covers most bones. It resides in a dynamic mechanically loaded environment and provides a niche for pluripotent cells and a source for molecular factors that modulate cell behaviour. Elucidating periosteum regenerative poten-tial has become a hot topic in orthopaedics. This review discusses the state of the art of osteochondral tissue engineering rested on periosteum derived progenitor cells(PDPCs) and suggests upcoming research direc-tions. Periosteal cells isolation, characterization and migration in the site of injury, as well as their differen-tiation, are analysed. Moreover, the role of cell mecha-nosensing and its contribution to matrix organization, bone microarchitecture and bone stenght is examined. In this regard the role of periostin and its upregulation under mechanical stress in order to preserve PDPC sur-vival and bone tissue integrity is contemplated. The re-view also summarized the role of the periosteum in the field of dentistry and maxillofacial reconstruction. The involvement of microRNAs in osteoblast differentiation and in endogenous tissue repair is explored as well. Fi-nally the novel concept of a guided bone regenerationbased on the use of periosteum itself as a smart mate-rial and the realization of constructs able to mimic the extracellular matrix features is talked out. Additionally, since periosteum can differentiate into insulin produc-ing cells it could be a suitable source in allogenic trans-plantations. That innovative applications would takeadvantage from investigations aimed to assess PDPCimmune privilege.展开更多
The use of periosteum-derived progenitor cells (PCs) combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of ...The use of periosteum-derived progenitor cells (PCs) combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional (3D) poly-lactic-co-glycolic acid (PLGA) fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic me- dium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type I collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reac- tion (RT-PCR). The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules of neo-bone tissue formation and the presence of matrix mineralization were confirmed by positive von Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.展开更多
目的研究向胫骨骨膜下单次注射转化生长因子(TGF)β1和β2后成年大白兔骨膜生发层细胞组织学的变化,探讨生长因子促进骨膜生发层细胞增殖的效果。方法 4月龄新西兰大白兔22只,分为3组,分别向两侧胫骨近端内侧骨膜下注射Tris溶液(对照组...目的研究向胫骨骨膜下单次注射转化生长因子(TGF)β1和β2后成年大白兔骨膜生发层细胞组织学的变化,探讨生长因子促进骨膜生发层细胞增殖的效果。方法 4月龄新西兰大白兔22只,分为3组,分别向两侧胫骨近端内侧骨膜下注射Tris溶液(对照组)(6只)、40 ng TGFβ1(8只)、40 ng TGFβ2(8只),2个实验组均于2个相距5 mm的部位分别注射20 ng。于术后第3、5、7及10天取材。标本均进行HE染色,对生发层厚度以及细胞密度进行观察测量及统计学分析。结果各组不同时间点生发层厚度比较,对照组注射后各时间点差异无显著性(P>0.05);注射后第5天,TGFβ1组生发层厚度(73±10)μm和TGFβ2组生发层厚度(112±13)μm较第3天明显增加;两实验组最大厚度(117±26)μm、(130±19)μm均出现在注射后第7天(P<0.05),注射后第10天与第3天差异无显著性(P>0.05)。同时间点3组间生发层厚度比较,注射后第3天、第10天差异无显著性(P>0.05);注射后第5天、第7天TGFβ1和TGFβ2组均明显高于对照组,且注射后第5天TGFβ2组厚度(112±13)μm明显大于TGFβ1组(73±10)μm(P=0.017)。细胞密度比较,3组注射后不同时间点以及3组间差异均无显著性(P>0.05)。结论单次骨膜下注射TGFβ1和TGFβ2均能一过性促进成年新西兰大白兔骨膜生发层细胞增殖。相比于TGFβ1,TGFβ2促进成年兔骨膜生发层细胞增殖效应较强。展开更多
目的探讨外源性降钙素基因相关肽(calcitonin gene related peptide,CGRP)对糖尿病大鼠骨膜微血管病变和膜性成骨的影响。方法建立糖尿病大鼠模型,外源性CGRP静脉注射,随机分为对照组(CON)、糖尿病组(DM)、CGRP干预组(CGRP),分别在5w、...目的探讨外源性降钙素基因相关肽(calcitonin gene related peptide,CGRP)对糖尿病大鼠骨膜微血管病变和膜性成骨的影响。方法建立糖尿病大鼠模型,外源性CGRP静脉注射,随机分为对照组(CON)、糖尿病组(DM)、CGRP干预组(CGRP),分别在5w、10w后观察各组大鼠骨膜微组织结构及组织计量学测定;墨汁灌注观测骨膜微血管单位面积。结果DM1骨祖细胞数较CON均增大(P<0.01),DM2骨膜厚度等均明显小于CON组(P<0.01);微血管单位面积增大,但渗透性大。CGRP骨膜厚度较DM1增多(P<0.01)。CGRP2较DM2骨膜厚度、骨祖细胞数均增大(P<0.01),微血管连续性好。结论外源性CGRP可改善糖尿病大鼠骨膜的微循环损伤,促进膜性成骨对糖尿病大鼠骨质疏松症起到修复作用。展开更多
基金Supported by Italian FIRB and PRIN project grants,No.2010J8RYS7 and No.RBAP10MLK7
文摘Periosteum is a thin fibrous layer that covers most bones. It resides in a dynamic mechanically loaded environment and provides a niche for pluripotent cells and a source for molecular factors that modulate cell behaviour. Elucidating periosteum regenerative poten-tial has become a hot topic in orthopaedics. This review discusses the state of the art of osteochondral tissue engineering rested on periosteum derived progenitor cells(PDPCs) and suggests upcoming research direc-tions. Periosteal cells isolation, characterization and migration in the site of injury, as well as their differen-tiation, are analysed. Moreover, the role of cell mecha-nosensing and its contribution to matrix organization, bone microarchitecture and bone stenght is examined. In this regard the role of periostin and its upregulation under mechanical stress in order to preserve PDPC sur-vival and bone tissue integrity is contemplated. The re-view also summarized the role of the periosteum in the field of dentistry and maxillofacial reconstruction. The involvement of microRNAs in osteoblast differentiation and in endogenous tissue repair is explored as well. Fi-nally the novel concept of a guided bone regenerationbased on the use of periosteum itself as a smart mate-rial and the realization of constructs able to mimic the extracellular matrix features is talked out. Additionally, since periosteum can differentiate into insulin produc-ing cells it could be a suitable source in allogenic trans-plantations. That innovative applications would takeadvantage from investigations aimed to assess PDPCimmune privilege.
基金Project supported by the Investitionsbank Berlin (IBB), Germany (No. 10020666) and the Science and Technology Bureau of ZhejiangProvince, China (No. 991110052)
文摘The use of periosteum-derived progenitor cells (PCs) combined with bioresorbable materials is an attractive approach for tissue engineering. The aim of this study was to characterize the osteogenic differentiation of PC in 3-dimensional (3D) poly-lactic-co-glycolic acid (PLGA) fleeces cultured in medium containing allogeneic human serum. PCs were isolated and expanded in monolayer culture. Expanded cells of passage 3 were seeded into PLGA constructs and cultured in osteogenic me- dium for a maximum period of 28 d. Morphological, histological and cell viability analyses of three-dimensionally cultured PCs were performed to elucidate osseous synthesis and deposition of a calcified matrix. Furthermore, the mRNA expression of type I collagen, osteocalcin and osteonectin was semi-quantitively evaluated by real-time reverse transcriptase-polymerase chain reac- tion (RT-PCR). The fibrin gel immobilization technique provided homogeneous PCs distribution in 3D PLGA constructs. Live-dead staining indicated a high viability rate of PCs inside the PLGA scaffolds. Secreted nodules of neo-bone tissue formation and the presence of matrix mineralization were confirmed by positive von Kossa staining. The osteogenic differentiation of PCs was further demonstrated by the detection of type I collagen, osteocalcin and osteonectin gene expression. The results of this study support the concept that this tissue engineering method presents a promising method for creation of new bone in vivo.
文摘目的研究向胫骨骨膜下单次注射转化生长因子(TGF)β1和β2后成年大白兔骨膜生发层细胞组织学的变化,探讨生长因子促进骨膜生发层细胞增殖的效果。方法 4月龄新西兰大白兔22只,分为3组,分别向两侧胫骨近端内侧骨膜下注射Tris溶液(对照组)(6只)、40 ng TGFβ1(8只)、40 ng TGFβ2(8只),2个实验组均于2个相距5 mm的部位分别注射20 ng。于术后第3、5、7及10天取材。标本均进行HE染色,对生发层厚度以及细胞密度进行观察测量及统计学分析。结果各组不同时间点生发层厚度比较,对照组注射后各时间点差异无显著性(P>0.05);注射后第5天,TGFβ1组生发层厚度(73±10)μm和TGFβ2组生发层厚度(112±13)μm较第3天明显增加;两实验组最大厚度(117±26)μm、(130±19)μm均出现在注射后第7天(P<0.05),注射后第10天与第3天差异无显著性(P>0.05)。同时间点3组间生发层厚度比较,注射后第3天、第10天差异无显著性(P>0.05);注射后第5天、第7天TGFβ1和TGFβ2组均明显高于对照组,且注射后第5天TGFβ2组厚度(112±13)μm明显大于TGFβ1组(73±10)μm(P=0.017)。细胞密度比较,3组注射后不同时间点以及3组间差异均无显著性(P>0.05)。结论单次骨膜下注射TGFβ1和TGFβ2均能一过性促进成年新西兰大白兔骨膜生发层细胞增殖。相比于TGFβ1,TGFβ2促进成年兔骨膜生发层细胞增殖效应较强。
文摘目的探讨外源性降钙素基因相关肽(calcitonin gene related peptide,CGRP)对糖尿病大鼠骨膜微血管病变和膜性成骨的影响。方法建立糖尿病大鼠模型,外源性CGRP静脉注射,随机分为对照组(CON)、糖尿病组(DM)、CGRP干预组(CGRP),分别在5w、10w后观察各组大鼠骨膜微组织结构及组织计量学测定;墨汁灌注观测骨膜微血管单位面积。结果DM1骨祖细胞数较CON均增大(P<0.01),DM2骨膜厚度等均明显小于CON组(P<0.01);微血管单位面积增大,但渗透性大。CGRP骨膜厚度较DM1增多(P<0.01)。CGRP2较DM2骨膜厚度、骨祖细胞数均增大(P<0.01),微血管连续性好。结论外源性CGRP可改善糖尿病大鼠骨膜的微循环损伤,促进膜性成骨对糖尿病大鼠骨质疏松症起到修复作用。
