Effect of platelet-rich fibrin released transforming growth factor beta 1 on the proliferation of bone marrow mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2014.45.004

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells are the ideal cells for tissue repair. Whether the ability of in vitro proliferation can be enhanced is a key factor to promote tissue repair.
OBJECTIVE: To observe the effect of transforming growth factor β1 on the proliferation of bone marrow mesenchymal stem cells in vitro.
METHODS: Blood samples were taken from the central artery of rabbits to prepare platelet-rich fibrin by centrifugation method which was then placed into fresh DMEM at 37 ℃ for 7, 14, 21, 28 days to collect exudates. The mass concentrations of transforming growth factor-β1 in the exudates of platelet-rich fibrin were detected. Rabbit bone marrow mesenchymal stem cells were collected and cultured in the conditioned medium made by the exudates of platelet-rich fibrin, and the proliferation of cells was observed.
RESULTS AND CONCLUSION: Concentration of transforming growth factor β1 was increased with time increasing, increased fastest at 21-28 days, and peaked at 28 days. Under the same stimulus concentration, the proliferation of bone marrow mesenchymal stem cells was reduced at 0-1 day, increased obviously at 1-2 days, and entered into a steady phase at 2-3 days. Under 150 ng/L transforming growth factor β1, bone marrow mesenchymal stem cells proliferated fastest. Experimental findings indicate that with the increase of time, the concentration of transforming growth factor β1 in the exudates of platelet-rich fibrin increase gradually, and the conditioned media containing different concentrations of transforming growth factor β1 play different roles in promoting the proliferation of bone marrow mesenchymal stem cells. Bone marrow mesenchymal stem cells cultured in the conditioned medium containing 150 ng/L transforming growth factor β1 for 2-3 days can proliferate fastest.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: bone marrow, mesenchymal stem cells, transforming growth factor beta1

CLC Number:

R394.2

Chen Cheng, Li Shu-hui, Zhang Wen-li, Li Yi-ming, Zhou Jing, Wu Pei-ling. Effect of platelet-rich fibrin released transforming growth factor beta 1 on the proliferation of bone marrow mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(45): 7233-7238.

Figures/Tables 5

References

[1]王悦,朱喆,成荣杰,等. 富血小板血浆对人真皮细胞增殖及PDGF、TGF-β和VEGF表达的影响[J].实用口腔医学杂志, 2012,28(1):64-69.
[2]Demir B, Sengün D, Berbero?lu A.Clinical evaluation of platelet-rich plasma and bioactive glass in the treatment of intra-bony defects.J Clin Periodontol. 2007;34(8):709-715.
[3]胡图强,何俐,李祖兵,等.纳米羟基磷灰石/PRP修复牙槽突裂的实验研究[J].口腔医学研究,2008,24(3):262-265.
[4]李艳秋,周延民,孙晓琳,等.富血小板纤维蛋白体外释放TGF-β和PDGF-AB影响因素的探讨[J].现代口腔医学杂志,2012, 26(6): 404-407.
[5]王宇,李琦.Choukroun富血小板纤维蛋白机制及应用的研究进展[J].口腔医学研究,2012,28(4):388-390.
[6]孙英华,王稚英.富血小板纤维蛋白凝胶和膜显微与超微结构研究[J].中国医学工程,2011,19(7):65-67.
[7]罗晓丁,李丹,张剑明.富血小板纤维蛋白促进组织愈合机制的探讨[J].中国口腔种植学杂志,2011,16(4):198-200.
[8]孙洁,张剑明,李彦秋.富血小板纤维蛋白超微结构的观察与探讨[J].口腔医学研究,2010,26(1):98-101.
[9]杨世茂,王明国,李静,等.富血小板纤维蛋白与富血小板血浆体外释放生长因子的比较及其对脂肪干细胞增殖分化的影响[J].华西口腔医学杂志,2012,30(6):641-644,649.
[10]Lee HS, Huang GT, Chiang H,et al.Multipotential mesenchymal stem cells from femoral bone marrow near the site of osteonecrosis.Stem Cells. 2003;21(2):190-199.
[11]Fisher M, Hyzy S, Guldberg RE,et al.Regeneration of bone marrow after tibial ablation in immunocompromised rats is age dependent.Bone. 2010;46(2):396-401.
[12]Yamaguchi S, Kuroda S, Kobayashi H,et al.The effects of neuronal induction on gene expression profile in bone marrow stromal cells (BMSC)--a preliminary study using microarray analysis.Brain Res. 2006;1087(1):15-27.
[13]王劲,张勇,罗成基,等.骨髓间充质干细胞成骨分化中转化生长因子受体的表达[J].中国组织工程研究与临床康复,2009,13(6): 1049-1052.
[14]鲁玲玲,赵焕英,赵春礼,等.骨髓间充质干细胞基因工程改造方法的比较[J].中国组织工程研究与临床康复,2007,11(3):471-474.
[15]Lee RH, Hsu SC, Munoz J,et al.A subset of human rapidly self-renewing marrow stromal cells preferentially engraft in mice.Blood. 2006;107(5):2153-2161.
[16]Pan HC, Yang DY, Chiu YT,et al.Enhanced regeneration in injured sciatic nerve by human amniotic mesenchymal stem cell.J Clin Neurosci. 2006;13(5):570-575.
[17]王鲲,茯小平,杨志华,等.骨髓间充质干细胞移植修复高原大鼠股骨缺损[J].中国组织工程研究,2014,18(14):2185-2190.
[18]王琰,陈希哲.口腔颌面部骨组织缺损修复材料研究进展[J].中国实用口腔科杂志,2011,4(3):175-177.
[19]杨全全,何家才,杨瑞,等. 富血小板纤维蛋白修复牙种植体周围骨缺损的研究[J].安徽医科大学学报,2012,47(5):581-584.
[20]杨全全,何家才.富血小板纤维蛋白在口腔种植中应用的研究进展[J].安徽医科大学学报,2011,46(10):1066-1068.
[21]崔丹,王佳卉,曹羽茜,等.Choukroun富血小板纤维蛋白复合Bio-oss修复兔颅骨缺损[J].微量元素与健康研究,2014, 7(4): 7-9.
[22]中华人民共和国科学技术部. 关于善待实验动物的指导性意见. 2006-09-30.
[23]冯玉华,董静,卢蕾,等.富血小板纤维蛋白诱导骨髓间充质干细胞向许旺细胞的分化[J].中国组织工程研究,2012,16(49): 9157-9161.
[24]宋明艳,李娜,姜彦,等.全骨髓贴壁法体外分离培养兔骨髓间充质干细胞[J].山东医药,2014,54(27):31-33.
[25]王宇,周延民,车彦海,等.Choukroun富血小板纤维蛋白对兔拔牙窝愈合修复的影响[J].口腔医学研究,2011,27(6):456-459.
[26]牛玉梅,陈野,李艳萍,等.犬自体骨髓间充质干细胞介导HA-TCP修复牙槽骨缺损的实验研究[J].口腔医学研究,2012,28(1):9-11.
[27]杜刚,李林,张波,等.富血小板血浆联合骨髓间充质干细胞不同配比浓度对兔软骨缺损的影响[J].中国实验方剂学杂志,2013, 19(19): 258-261.
[28]肖仕辉,韦庆军,赵劲民,等.全骨髓贴壁法培养兔骨髓间充质干细胞体外定向成骨诱导分化及鉴定[J].中国组织工程研究,2013, 17(6):1069-1074.
[29]Aldahmash A, Zaher W, Al-Nbaheen M, et al.Human stromal (mesenchymal) stem cells: basic biology and current clinical use for tissue regeneration.Ann Saudi Med. 2012;32(1): 68-77.
[30]Gu Y, Wang Y, Dai H,et al.Chondrogenic differentiation of canine myoblasts induced by cartilage-derived morphogenetic protein-2 and transforming growth factor-β1 in vitro.Mol Med Rep. 2012;5(3):767-772.
[31]陈松,符培亮,丛锐军,等. TGF-β3、BMP-2及地塞米松诱导兔滑膜MSCs成软骨分化的研究[J].中国修复重建外科杂志,2014, 28(1):92-99.
[32]王善正,王宸,芮云峰.自体激活富血小板血浆干预兔骨髓间充质干细胞体外成软骨分化的研究[J].中国组织工程研究,2013, 17(1):1-8.
[33]江华,肖增明.骨髓间充质干细胞在骨科疾病修复中的应用[J].中国临床康复,2006,10(45):118-120.
[34]郭璇,霍然,吕仁荣,等.兔骨髓间充质干细胞培养及向成软骨细胞的诱导分化[J].中国组织工程研究与临床康复,2011,15(6): 963-966.
[35]黄涛,孟志斌,贾丙申,等.全骨髓贴壁法分离培养rBMSCs及成骨诱导探讨[J].广东医学,2010,31(9):1089-1091.
[36]钱文慧,徐艳,孙颖.富血小板血浆与富血小板纤维蛋白在牙周组织再生中的应用[J].口腔生物医学,2011,2(2):96-99.
[37]于佳,郝永明,陆家瑜,等.犬PRF复合BMSCs修复拔牙窝颊侧骨壁缺损的实验研究[J].口腔颌面外科杂志,2014,24(4):266-271.
[38]许丰伟,柳忠豪.Choukroun富血小板纤维蛋白在口腔种植骨缺损中的研究与进展[J].中国组织工程研究,2012,16(4):741-744.
[39]许丰伟,柳忠豪,董凯.富血小板纤维蛋白膜(PRF)引导成骨能力的比较研究[J].中国口腔种植学杂志,2012,17(1):10-13.
[40]Dohan Ehrenfest DM, de Peppo GM, Doglioli P,et al.Slow release of growth factors and thrombospondin-1 in Choukroun's platelet-rich fibrin (PRF): a gold standard to achieve for all surgical platelet concentrates technologies. Growth Factors. 2009;27(1):63-69.
[41]崔冬,张腾,刁建升,等.富血小板纤维蛋白对人脂肪干细胞增殖和成脂分化的影响[J].中华医学美学美容杂志,2013,19(3):203-206.
[42]董凯,柳忠豪,张晓洁,等.富血小板纤维蛋白提取液对成骨细胞影响的实验研究[J].实用口腔医学杂志,2013,29(4):472-476.
[43]陈润良,刘磊,裴瑛波.骨髓间充质干细胞在多学科领域的临床应用[J].中国组织工程研究与临床康复,2009,13(6):1143-1146.
[44]张卫兵,洪光祥,康皓. 富血小板血浆对免骨髓间充质干细胞增殖的作用[J].中国组织工程研究与临床康复,2008,12(34): 6639-6642.
[45]王艾丽,陈玲玲,胡剑峰,等.TGF-β1对骨髓间充质干细胞在无氧无血清条件下凋亡的影响[J].武汉大学学报:医学版,2007, 28(6):729-732.
[46]曾晖,肖德明,陶可,等.转化生长因子-β1对骨髓干细胞分化过程中Wnt信号通路相关基因表达的影响[J].中华实验外科杂志, 2013,30(7):1347-1350.