锶纳米纤维能促进骨组织再生
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摘要
为了探究锶纳米纤维在骨组织再生中的作用机制,本研究通过静电纺丝来制备聚合物纳米复合材料工程支架,促进骨组织再生。将碳酸锶纳米粒子(nSrCO3)以10%和15%重量比添加到聚己酸内酯(PCL)中,做成纤维直径在300～500 nm范围内的纳米复合材料纤维支架(PCL+10%SrCO3和PCL+15%SrCO3),掺入nSrCO3后降低了纤维支架的结晶度和弹性模量,复合PCL+15%SrCO3支架可在4 d内释放出高达58 ppm的Sr2+离子。细胞研究证实,体外使用的含有15%n Sr CO3的复合支架增强了人间充质干细胞的增殖。在PCL+15%SrCO3中,最小沉积量显著增加达约4倍,促进了骨形成。在PCL+15%SrCO3纤维中,成骨标志物如BMP-2和Runx2的mRNA和蛋白的高表达也可以证明锶纳米纤维能促进骨形成。本研究为研究锶纳米纤维在促进骨组织再生中的运用提供了一定的参考价值。
To explore the mechanism of strontium nanofibers in bone tissue regeneration, in this study, polymer nanocomposite engineering scaffolds were prepared by electrospinning to promote bone tissue regeneration. Strontium carbonate nanoparticles(nSrCO3) was added to polycaprolactone(PCL) at a ratio of 10% and 15% by weight to form a nanocomposite fiber scaffold with a fiber diameter in the range of 300～500 nm(PCL+10% SrCO3 and PCL+15% SrCO3). The crystallinity and elastic modulus of the fiber scaffold were reduced after incorporation of nSrCO3.The composite PCL+15% SrCO3 scaffold can release up to 58 ppm of Sr2+ions within 4 days. Cell studies confirmed that in vitro using of a composite scaffold containing 15% n Sr CO3 enhanced the proliferation of human mesenchymal stem cells. In PCL+15% SrCO3, the minimum deposition amount was significantly increased by about 4 times, and promoted bone formation. The high expression of osteogenic markers such as BMP-2 and Runx2 m RNA and protein in PCL+15% SrCO3 fibers also demonstrated that nanofibers promoted bone formation. This study provides some reference value for studying the application of strontium nanofibers in bone tissue regeneration.
To explore the mechanism of strontium nanofibers in bone tissue regeneration, in this study, polymer nanocomposite engineering scaffolds were prepared by electrospinning to promote bone tissue regeneration. Strontium carbonate nanoparticles(nSrCO3) was added to polycaprolactone(PCL) at a ratio of 10% and 15% by weight to form a nanocomposite fiber scaffold with a fiber diameter in the range of 300～500 nm(PCL+10% SrCO3 and PCL+15% SrCO3). The crystallinity and elastic modulus of the fiber scaffold were reduced after incorporation of nSrCO3.The composite PCL+15% SrCO3 scaffold can release up to 58 ppm of Sr2+ions within 4 days. Cell studies confirmed that in vitro using of a composite scaffold containing 15% n Sr CO3 enhanced the proliferation of human mesenchymal stem cells. In PCL+15% SrCO3, the minimum deposition amount was significantly increased by about 4 times, and promoted bone formation. The high expression of osteogenic markers such as BMP-2 and Runx2 m RNA and protein in PCL+15% SrCO3 fibers also demonstrated that nanofibers promoted bone formation. This study provides some reference value for studying the application of strontium nanofibers in bone tissue regeneration.
引文
Atkins G.J.,Welldon K.J.,Halbout P.,and Findlay D.M.,2007,Strontium ranelate treatment of human primary osteoblasts promotes an osteocyte-like phenotype while eliciting an osteoprotegerin response,Osteoporosis International,20(4):653-664
Brennan T.C.,Rybchyn M.S.,Green W.,Atwa S.,Conigrave A.D.,and Mason R.S.,2009,Osteoblasts play key roles in the mechanisms of action of strontium ranelate,Br.J.Pharmacol.,157(7):1291-1300
Choudhary S.,Halbout P.,Alander C.,Raisz L.,and Pilbeam C.,2007,Strontium ranelate promotes osteoblastic differentiation and mineralization of murine bone marrow stromal cells:involvement of prostaglandins,J.Bone Miner.Res.,22(7):1002-1010
Cui C.B.,Wang J.,Zhao Y.X.,Zhang J.W.,Zhao Y.Q.,and Yang Y.T.,2016,Effect of micellar polymer PEG-PLL-OPG on osteoporosis and arteriosclerosis in mice,Shiyong Yixue Zazhi(The Journal of Practical Medicine),32(16):2598-2602(崔春便,王敬,赵迎新,张继伟,赵玉倩,杨艳同,2016,纳米胶束复合物PEG-PLL-OPG对小鼠骨质疏松及动脉硬化的影响,实用医学杂志,32(16):2598-2602)
Hao J.,Acharya A.,Chen K.,Chou J.,Kasugai S.,and Lang N.P.,2015,Novel bioresorbable strontium hydroxyapatite mem brane for guided bone regeneration,Clin.Oral Implants Res.,26(1):1-7
Im Y.M.,Oh T.H.,Jin W.C.,Seo Y.H.,and Hwang J.S.,2014,Preparation of poly(vinyl alcohol)/ZrO2,composite nanofibers via co-axial electrospinning with higher Zr O2,particle content,Fibers&Polymers,15(10):2066-2071
Javed A.,Bae J.F.,Gutierrez S.,Pratap J.,Zaidi S.K.,Lou Y.,van Wijnen A.J.,Stein J.L.,Stein G.S.,and Lian J.B.,2008,Structural coupling of Smad and Runx2 for execution of the BMP2 osteogenic signal,J.Biol.Chem.,283(13):8412-8422
Kumar G.,Tison C.K.,Chatterjee K.,Pine P.S.,McDaniel J.H.,Salit M.L.,Young M.F.,and Simon C.G.,2011,The determination of stem cell fate by 3D scaffold structures through the control of cell shape,Biomaterials,32(35):9188-9196
Kumar S.,Bose S.,and Chatterjee K.,2014,Amine-functionalized multiwall carbon nanotubes impart osteoinductive and bactericidal properties in poly(ε-caprolactone)composites,Rsc.Advances,4(37):19086-19098
Kumar S.,Raj S.,Kolanthai E.,Sood A.K.,Sampath S.,and Chatterjee K.,2015,Chemical functionalization of graphene to augment stem cell osteogenesis and inhibit biofilm formation on polymer composites for orthopedic applications,Acs Appl Mater Interfaces,7(5):3237
Lee H.Y.,Lie D.,Lim K.S.,Thirumoorthy T.,and Pang S.M.,2009,Strontium ranelate-induced toxic epidermal necrolysis in a patient with post,Osteoporosis Int.,20(1):161-162
Liu X.W.,Zhong J.,Peng X.T.,Wei D.X.,Zhou J.,Ye Y.,and Sun G.,2014,Performance testing of biodegradable meshlike microporous balloon combined with calcium phosphate cement for vertebroplasty,Zhongguo Zuzhi Gongcheng Yanjiu(Journal of Clinical Rehabilitative Tissue Engineering Research),18(12):1817-1823(刘训伟,钟建,彭湘涛,魏岱旭,周涓,叶勇,孙钢,2014,联合钙盐骨水泥及可降解网状微孔球囊椎体成形的体外实验,中国组织工程研究,18(12):1817-1823)
Livak K.J.,and Schmittgen T.D.,2001,Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method,Methods,25(4):402-408
Ma J.Q.,Yu M.,and Zhang H.S.,2015,Progress of strontium on bone mineral metabolism,Yixue Yanjiu Yu Jiaoyu(Journal of Hebei Medical College for Continuing Education),32(2):82-86(马家晴,于萌,张海松,2015,锶对骨矿代谢的研究进展,医学研究与教育,32(2):82-86)
Meunier P.J.,Roux C.,Seeman E.,Ortolani S.,Badurski J.E.,Spector T.D.,Cannata J.,Balogh A.,Lemmel E.M.,Pors-N-ielsen S.,Rizzoli R.,Genant H.K.,and Reginster J.Y.,2004,The effects of strontium ranelate on the risk of vertebral fracture in women with postmenopausal osteoporosis.,N.Engl.J.Med.,350(5):459-468
Morohashi T.,Sano T.,Harai K.,and Yamada S.,1995,Effects of strontium on calcium metabolism in rats.Ⅱ.Strontium prevents the increased rate of bone turnover in ovariectomized rats,Jpn.J.Pharmacol.,68(2):153-159
Poh P.S.,Hutmacher D.W.,Stevens M.M.,and Woodruff M.A.,2013,Fabrication and in vitro characterization of bioactive glass composite scaffolds for bone regeneration,Biofabrication,5(4):045005
Pors N.S.,2004,The biological role of strontium,Bone,35(3):583-588
Reginster J.Y.,Seeman E.,De Vernejoul M.C.,Adami.S,Compston J.,Phenekos C.,Devogelaer J.P.,Curiel M.D.,Sawicki A.,Goemaere S.,Sorensen O.H.,Felsenberg D.,and Meunier P.J.,2005,Strontium ranelate reduces the risk of nonvertebral fractures in postmenopausal women with osteoporosis:Treatment of Peripheral Osteoporosis(TROPOS)study,J.Clin.Endocrinol.Metab.,90(5):2816-2822
Ren J.,Blackwood K.A.,Doustgani A.,Poh P.P.,Steck R.,Stevens M.M.,and Woodruff M.A.,2014,Melt-electrospun polycaprolactone strontium-substituted bioactive glass scaffolds for bone regeneration,J.Biomed.Mater.Res.A,102(9):3140-3153
Schumacher M.,Lode A.,Helth A.,and Gelinsky M.,2013,Anovel strontium(Ⅱ)-modified calcium phosphate bone ce ment stimulates human-bone-marrow-derived mesenchymal stem cell proliferation and osteogenic differentiation in vitro,Acta Biomaterialia,9(12):9547-9557
Tu Q.,Valverde P.,and Chen J.,2006,Osterix enhances proliferation and osteogenic potential of bone marrow stromal cells,Biochem.Biophys.Res.Commun.,341(4):1257-1265
Wu M.J.,2012,The relationship between strontium and human health,Weiliang Yuansu Yu Jiankang Yanjiu(Studies of Trace Elements and Health),29(5):66-67(吴茂江,2012,锶与人体健康,微量元素与健康研究,29(5):66-67)
Yang F.,Yang D.,Tu J.,Zheng Q.,Cai L.,and Wang L.,2011,Strontium enhances osteogenic differentiation of mesenchymal stem cells and in vivo bone formation by activating Wnt/catenin signaling,Stem Cells,29(6):981-991
Brennan T.C.,Rybchyn M.S.,Green W.,Atwa S.,Conigrave A.D.,and Mason R.S.,2009,Osteoblasts play key roles in the mechanisms of action of strontium ranelate,Br.J.Pharmacol.,157(7):1291-1300
Choudhary S.,Halbout P.,Alander C.,Raisz L.,and Pilbeam C.,2007,Strontium ranelate promotes osteoblastic differentiation and mineralization of murine bone marrow stromal cells:involvement of prostaglandins,J.Bone Miner.Res.,22(7):1002-1010
Cui C.B.,Wang J.,Zhao Y.X.,Zhang J.W.,Zhao Y.Q.,and Yang Y.T.,2016,Effect of micellar polymer PEG-PLL-OPG on osteoporosis and arteriosclerosis in mice,Shiyong Yixue Zazhi(The Journal of Practical Medicine),32(16):2598-2602(崔春便,王敬,赵迎新,张继伟,赵玉倩,杨艳同,2016,纳米胶束复合物PEG-PLL-OPG对小鼠骨质疏松及动脉硬化的影响,实用医学杂志,32(16):2598-2602)
Hao J.,Acharya A.,Chen K.,Chou J.,Kasugai S.,and Lang N.P.,2015,Novel bioresorbable strontium hydroxyapatite mem brane for guided bone regeneration,Clin.Oral Implants Res.,26(1):1-7
Im Y.M.,Oh T.H.,Jin W.C.,Seo Y.H.,and Hwang J.S.,2014,Preparation of poly(vinyl alcohol)/ZrO2,composite nanofibers via co-axial electrospinning with higher Zr O2,particle content,Fibers&Polymers,15(10):2066-2071
Javed A.,Bae J.F.,Gutierrez S.,Pratap J.,Zaidi S.K.,Lou Y.,van Wijnen A.J.,Stein J.L.,Stein G.S.,and Lian J.B.,2008,Structural coupling of Smad and Runx2 for execution of the BMP2 osteogenic signal,J.Biol.Chem.,283(13):8412-8422
Kumar G.,Tison C.K.,Chatterjee K.,Pine P.S.,McDaniel J.H.,Salit M.L.,Young M.F.,and Simon C.G.,2011,The determination of stem cell fate by 3D scaffold structures through the control of cell shape,Biomaterials,32(35):9188-9196
Kumar S.,Bose S.,and Chatterjee K.,2014,Amine-functionalized multiwall carbon nanotubes impart osteoinductive and bactericidal properties in poly(ε-caprolactone)composites,Rsc.Advances,4(37):19086-19098
Kumar S.,Raj S.,Kolanthai E.,Sood A.K.,Sampath S.,and Chatterjee K.,2015,Chemical functionalization of graphene to augment stem cell osteogenesis and inhibit biofilm formation on polymer composites for orthopedic applications,Acs Appl Mater Interfaces,7(5):3237
Lee H.Y.,Lie D.,Lim K.S.,Thirumoorthy T.,and Pang S.M.,2009,Strontium ranelate-induced toxic epidermal necrolysis in a patient with post,Osteoporosis Int.,20(1):161-162
Liu X.W.,Zhong J.,Peng X.T.,Wei D.X.,Zhou J.,Ye Y.,and Sun G.,2014,Performance testing of biodegradable meshlike microporous balloon combined with calcium phosphate cement for vertebroplasty,Zhongguo Zuzhi Gongcheng Yanjiu(Journal of Clinical Rehabilitative Tissue Engineering Research),18(12):1817-1823(刘训伟,钟建,彭湘涛,魏岱旭,周涓,叶勇,孙钢,2014,联合钙盐骨水泥及可降解网状微孔球囊椎体成形的体外实验,中国组织工程研究,18(12):1817-1823)
Livak K.J.,and Schmittgen T.D.,2001,Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method,Methods,25(4):402-408
Ma J.Q.,Yu M.,and Zhang H.S.,2015,Progress of strontium on bone mineral metabolism,Yixue Yanjiu Yu Jiaoyu(Journal of Hebei Medical College for Continuing Education),32(2):82-86(马家晴,于萌,张海松,2015,锶对骨矿代谢的研究进展,医学研究与教育,32(2):82-86)
Meunier P.J.,Roux C.,Seeman E.,Ortolani S.,Badurski J.E.,Spector T.D.,Cannata J.,Balogh A.,Lemmel E.M.,Pors-N-ielsen S.,Rizzoli R.,Genant H.K.,and Reginster J.Y.,2004,The effects of strontium ranelate on the risk of vertebral fracture in women with postmenopausal osteoporosis.,N.Engl.J.Med.,350(5):459-468
Morohashi T.,Sano T.,Harai K.,and Yamada S.,1995,Effects of strontium on calcium metabolism in rats.Ⅱ.Strontium prevents the increased rate of bone turnover in ovariectomized rats,Jpn.J.Pharmacol.,68(2):153-159
Poh P.S.,Hutmacher D.W.,Stevens M.M.,and Woodruff M.A.,2013,Fabrication and in vitro characterization of bioactive glass composite scaffolds for bone regeneration,Biofabrication,5(4):045005
Pors N.S.,2004,The biological role of strontium,Bone,35(3):583-588
Reginster J.Y.,Seeman E.,De Vernejoul M.C.,Adami.S,Compston J.,Phenekos C.,Devogelaer J.P.,Curiel M.D.,Sawicki A.,Goemaere S.,Sorensen O.H.,Felsenberg D.,and Meunier P.J.,2005,Strontium ranelate reduces the risk of nonvertebral fractures in postmenopausal women with osteoporosis:Treatment of Peripheral Osteoporosis(TROPOS)study,J.Clin.Endocrinol.Metab.,90(5):2816-2822
Ren J.,Blackwood K.A.,Doustgani A.,Poh P.P.,Steck R.,Stevens M.M.,and Woodruff M.A.,2014,Melt-electrospun polycaprolactone strontium-substituted bioactive glass scaffolds for bone regeneration,J.Biomed.Mater.Res.A,102(9):3140-3153
Schumacher M.,Lode A.,Helth A.,and Gelinsky M.,2013,Anovel strontium(Ⅱ)-modified calcium phosphate bone ce ment stimulates human-bone-marrow-derived mesenchymal stem cell proliferation and osteogenic differentiation in vitro,Acta Biomaterialia,9(12):9547-9557
Tu Q.,Valverde P.,and Chen J.,2006,Osterix enhances proliferation and osteogenic potential of bone marrow stromal cells,Biochem.Biophys.Res.Commun.,341(4):1257-1265
Wu M.J.,2012,The relationship between strontium and human health,Weiliang Yuansu Yu Jiankang Yanjiu(Studies of Trace Elements and Health),29(5):66-67(吴茂江,2012,锶与人体健康,微量元素与健康研究,29(5):66-67)
Yang F.,Yang D.,Tu J.,Zheng Q.,Cai L.,and Wang L.,2011,Strontium enhances osteogenic differentiation of mesenchymal stem cells and in vivo bone formation by activating Wnt/catenin signaling,Stem Cells,29(6):981-991