微囊化软骨细胞诱导间充质干细胞定向分化治疗兔椎间盘退变的实验研究
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摘要
目的探讨经微囊化软骨细胞诱导分化的间充质干细胞对兔椎间盘退变的治疗效果。方法将微囊化软骨细胞与间充质干细胞进行共培养,干细胞诱导分化。观察分化后干细胞的免疫学特征、增殖曲线、蛋白多糖及Ⅱ型胶原蛋白合成能力。建立兔椎间盘退变模型,并观察分化后的间充质干细胞对兔椎间盘退变的修复能力。结果经微囊化软骨细胞分化诱导7d后的间充质干细胞,增殖能力未受到明显影响,同时具备了软骨细胞的部分表面特征,能够合成蛋白多糖及Ⅱ型胶原蛋白,且能力优于Transwell对照组。将干细胞植入兔椎间盘退变模型中,在术后16周内,椎间盘的力学性能得到修复,形态及结构也得到了明显改善。结论与传统的Transwell诱导体系相比,微囊诱导体系具有更高的诱导效率;经微囊化软骨细胞诱导的MSCs,植入椎间盘退变模型动物体内后,能更好地维持椎间盘的形态、结构及力学特征。
Objective To investigate the curative effect of mesenchymal stem cells(MSCs)induced by microencapsuled chondrocytes on the intervertebral disc degeneration(IDD)in rabbits.Methods MSCs were induced by co-culture with microencapsuled chondrocytes.The surface antigens,proliferation rate,synthesis of proteoglycan and typeⅡcollagen of MSCs were tested.Induced MSCs were transplanted into the rabbits to establish IDD models and evaluate the repairing effects.Results After induction by microencapsuled chondrocytes for 7d,the MSCs possessed certain surface characteristics of chondrocytes and could produce more proteoglycan and typeⅡcollagen than Transwell system,while the proliferation of MSCs was not affected.After transplanting the MSCs induced by microencapsuled chondrocytes,the form,structure,and mechanical properties were maintained 16 weeks after operations.Conclusion Compared with the Transwell system,the induction efficiency of microcapsule system is higher.Transplanting MSCs induced by microencapsuled chondrocytes into rabbit IDD models can repair degenerated discs effectively.
Objective To investigate the curative effect of mesenchymal stem cells(MSCs)induced by microencapsuled chondrocytes on the intervertebral disc degeneration(IDD)in rabbits.Methods MSCs were induced by co-culture with microencapsuled chondrocytes.The surface antigens,proliferation rate,synthesis of proteoglycan and typeⅡcollagen of MSCs were tested.Induced MSCs were transplanted into the rabbits to establish IDD models and evaluate the repairing effects.Results After induction by microencapsuled chondrocytes for 7d,the MSCs possessed certain surface characteristics of chondrocytes and could produce more proteoglycan and typeⅡcollagen than Transwell system,while the proliferation of MSCs was not affected.After transplanting the MSCs induced by microencapsuled chondrocytes,the form,structure,and mechanical properties were maintained 16 weeks after operations.Conclusion Compared with the Transwell system,the induction efficiency of microcapsule system is higher.Transplanting MSCs induced by microencapsuled chondrocytes into rabbit IDD models can repair degenerated discs effectively.
引文
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[14]SCHNEIDER S,KLEIN HH.Long-term graft function of cryostored alginate encapsulated rat islets[J].Eur J Med Res,2011,16(9):396-400.
[15]KERBY A,BOHMAN S,WESTBERG H,et al.Immunoisolation of islets in high guluronic acid barium-alginate microcapsules does not improve graft outcome at the subcutaneous site[J].Artif Organs,2012,36(6):564-570.
[2]PATRICK N,EMANSKI E,KNAUB MA.Acute and chronic low back pain[J].Med Clin North Am,2016,100(1):169-181.
[3]BALIGA S,TREON K,CRAIG NJ,Low back pain:Current surgical approaches[J].Asian Spine J,2015,9(4):645-657.
[4]BOOS N,WEISSBACH S,ROHRBACH H,et al.Classification of age-related changes in lumbar intervertebral discs:2002Volvo Award in Basic Science[J].Spine,2002,27(23):2631-2644.
[5]CHEUNG KM,KARPPINEN J,CHAN D,et al.Prevalence and pattern of lumbar magnetic resonance imaging changes in a population study of one thousand forty-three individuals[J].Spine,2009,34(9):934-940.
[6]ZHANG C,BERVEN SH,FORTIN M,et al.Adjacent segment degeneration versus disease after lumbar spine fusion for degenerative pathology:A systematic review with meta-analysis of the literature[J].Clin Spine Surg,2016,29(1):21-29.
[7]YANG F,LEUNG VY,LUK KD,et al.Mesenchymal stem cells arrest intervertebral disc degeneration through chondrocytic differentiation and stimulation of endogenous cells[J].Mol Ther,2009,17(11):1959-1966.
[8]RICHARDSON SM,HOYLAND JA,MOBASHERI R,et al.Mesenchymal stem cells in regenerative medicine:opportunities and challenges for articular cartilage and intervertebral disc tissue engineering[J].J Cell Physiol,2010,222(1):23-32.
[9]HUANG YC,LEUNG VY,LU WW,et al.The effects of microenvironment in mesenchymal stem cell-based regeneration of intervertebral disc[J].Spine J,2013,13(3):352-362.
[10]RICHARDSON SM,WALKER RV,PARKER S,et al.Intervertebral disc cell-mediated mesenchymal stem cell differentiation[J].Stem Cells,2006,24(3):707-716.
[11]HASSE C,SCHREZENMEIR J,STINNER B,et al.Successful allotransplantation of microencapsulated parathyroids in rats[J].World J Surg,1994,18(4):630-634.
[12]HANG H,SHI X,GU GX,et al.In vitro analysis of cryopreserved alginate-poly-L-lysine-alginate-microencapsulated human hepatocytes[J].Liver Int,2010,30(4):611-622.
[13]MISTRY H,CONNOCK M,PINK J,et al.Autologous chondrocyte implantation in the knee:systematic review and economic evaluation[J].Health Technol Assess,2017,21(6):1-294.
[14]SCHNEIDER S,KLEIN HH.Long-term graft function of cryostored alginate encapsulated rat islets[J].Eur J Med Res,2011,16(9):396-400.
[15]KERBY A,BOHMAN S,WESTBERG H,et al.Immunoisolation of islets in high guluronic acid barium-alginate microcapsules does not improve graft outcome at the subcutaneous site[J].Artif Organs,2012,36(6):564-570.