These tools will no longer be maintained as of December 31, 2024. Archived website can be found here. PubMed4Hh GitHub repository can be found here. Contact NLM Customer Service if you have questions.


Pubmed for Handhelds

PUBMED FOR HANDHELDS

Search MEDLINE/PubMed

  • Title: [EFFECT OF CHANGE OF TISSUE INTERFACE STIFFNESS ON OSTEOGENIC DIFFERENTIATION OF RAT BONE MARROW MESENCHYMAL STEM CELLS].
    Author: Chen L, Xu M, Liu Y, Duan H, Hua W, He F.
    Journal: Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi; 2016 Dec 08; 30(12):1524-1531. PubMed ID: 29786346.
    Abstract:
    OBJECTIVE: To investigate the effect of tissue interface stiffness change on the spreading, proliferation, and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and to find the suitable stiffness range for stem cell differentiation. METHODS: Bone marrow of male Sprague Dawley rats (4 weeks old) were selected to isolate and culture BMSCs by whole bone marrow cell adherent method. The third generation BMSCs (1×105 cells/mL) were inoculated into the ordinary culture dishes covered with polyacrylamide hydrophilic gel (PA) which elastic modulus was 1, 4, 10, 40, and 80 kPa (cells seeded on PA), and ordinary culture dish (75 MPa extreme high elastic modulus) as control. Spreading of cells in different stiffness of PA was observed under light microscope. The elastic modulus values of 4, 10, and 40 kPa PA were selected as groups A, B, and C respectively; the ordinary culture dish (75 MPa extreme high elastic modulus) was used as control group (group D). Cell counts was used to detect the growth conditions of BMSCs, alkaline phosphatase (ALP) kit to detect the concentration of ALP, alizarin red staining technique to detect calcium deposition status, and real-time quatitative PCR technique to detect the expressions of bone gla protein (BGP), Runx2, and collagen type I mRNA. RESULTS: With increased PA stiffness, BMSCs spreading area gradually increased, especially in 10 kPa and 40 kPa. At 1 and 2 days after culture, the growth rate showed no significant difference between groups (P>0.05); at 3-5 days, the growth rate of groups B and C was significantly faster than that of groups A and D (P<0.05), but difference was not statistically significant between groups A and D (P<0.05); at 5 days, the proliferation of group C was significantly higher than that of group B (P<0.05). ALP concentrations were (53.69±0.89), (97.30±1.57), (126.60±14.54), and (12.93±0.58) U/gprot in groups A, B, C, and D respectively; groups A, B, and C were significantly higher than group D, and group C was significantly higher than groups A and B (P<0.05). Alizarin red staining showed that the percentages of calcium nodules was 20.07%±4.24% in group C; group C was significantly higher than groups A, B, and D (P<0.05). The expression levels of BGP and collagen type I mRNA were significantly higher in groups A, B, and C than group D, and in group C than groups A and B (P<0.05). The expression level of Runx2 mRNA was significantly higher in groups B and C than group D, and in group C than group B (P<0.05), but no significant difference was found between groups A and D (P>0.05). CONCLUSIONS: PA elastic modulus of 10-40 kPa can promote the proliferation and osteogenic differentiation of BMSCs, and the higher the stiffness, the stronger the promoting effect. 目的: 探讨组织界面刚度变化对大鼠BMSCs铺展、增殖和成骨分化的影响，寻找适合其成骨分化的刚度范围。. 方法: 取4周龄SPF级雄性SD大鼠骨髓，采用全骨髓细胞贴壁法分离培养BMSCs并鉴定。取第3代BMSCs，按1×105个/mL密度接种于覆盖弹性模量分别为1、4、10、40、80 kPa聚丙烯酰胺水凝胶（polyacrylamide hydrophilic gel，PA）的普通培养皿（细胞接种于PA基底上），普通培养皿（弹性模量75 MPa）作为对照，光镜下观察细胞在不同刚度PA上的铺展状态。选取4、10和40 kPa PA覆盖作为研究对象（分别为A、B、C组），以普通培养皿为对照组（D组），细胞计数检测BMSCs生长情况，ALP试剂盒检测BMSCs的ALP浓度，茜素红染色法检测BMSCs的钙沉积状况，实时荧光定量PCR检测BMSCs的骨钙素（bone gla protein，BGP）、Runx2、Ⅰ型胶原mRNA表达。. 结果: 随着PA刚度的提升，BMSCs铺展面积增加，尤其在10、40 kPa PA上铺展较好。生长曲线示，培养1、2 d各组间细胞增殖情况差异无统计学意义（P＞0.05），3～5 d B、C组增殖情况明显优于A、D组（P＜0.05），5 d时C组显著高于B组（P＜0.05）。A、B、C、D组ALP浓度分别为（53.69±0.89）、（97.30±1.57）、（126.60±14.54）、（12.93±0.58）U/gprot，A、B、C组显著高于D组，C组高于A、B组，差异均有统计学意义（P＜0.05）。茜素红染色示，C组钙结节百分比为20.07%±4.24%，显著高于A、B、D组，差异有统计学意义（P＜0.05）。实时荧光定量PCR检测示，A、B、C组BGP、Ⅰ型胶原mRNA相对表达量均高于D组，C组高于A、B组，差异均有统计学意义（P＜0.05）。B、C组Runx2 mRNA相对表达量高于D组，C组高于B组，差异均有统计学意义（P＜0.05）；A、D组间差异无统计学意义（P＞0.05）。. 结论: 10～40 kPa范围的PA能促进大鼠BMSCs增殖及成骨分化，刚度越高，作用越强。.
    [Abstract] [Full Text] [Related] [New Search]