可注射水凝胶负载脂肪来源干细胞外泌体促进骨再生
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摘要
目的探究来自人类脂肪来源干细胞(adipose-derived stem cells,ADSCs)的外泌体(exosomes)在水凝胶(hydrogel的成骨能力表达情况。方法制备可注射温敏型含壳聚糖、纳米羟基磷灰石、胶原和β-甘油磷酸钠(CS/nHAC/β-GP)的水凝胶。通过SEM观察,运用Western blot方法,及ALP等指标检测ADSCs诱导产生的外泌体在水凝胶上的成骨能力表达情况。结果⑴在体外实验中,与对照组相比,外泌体组的水凝胶ALP表达显著升高。⑵外泌体对ADSCs在CS/nHAC/β-GP水凝胶内的增殖及其对成骨分化均有明显促进作用。结论由ADSCs产生的外泌体和CS/nHAC/β-GP水凝胶组成的新型材料为骨组织工程提供了一种新的治疗方案,在修复骨缺损方面显示出良好的潜力。
Objective To investigate the expression of the osteogenic ability of exomes from human adipose-derived stem cells(ADSCs) in hydrogels. Methods Injectable thermosensitive hydrogels containing chitosan, nano-hydroxyapatite, gelatin and sodiumβ-glycerophosphate(CS/n HAC/β-GP) were prepared. The expression of the osteogenic capacity of exosomes induced by ADSCs on hydrogels was observed by SEM and examined by Western Blot and ALP detection. Results(1) In vitro, the expression of ALP in hydrogels was significantly increased in the exosome group compared with the control group;(2) The exosomes had obvious promotion effect on proliferation and osteogenic differentiation of ADSCs in CS/n HAC/β-GP hydrogels. Conclusion The new material consisting of exosome derived from ADSCs and CS/n HAC/β-GP hydrogel provides a new treatment strategy for bone tissue engineering and shows good potential in the repair of bone defects.
Objective To investigate the expression of the osteogenic ability of exomes from human adipose-derived stem cells(ADSCs) in hydrogels. Methods Injectable thermosensitive hydrogels containing chitosan, nano-hydroxyapatite, gelatin and sodiumβ-glycerophosphate(CS/n HAC/β-GP) were prepared. The expression of the osteogenic capacity of exosomes induced by ADSCs on hydrogels was observed by SEM and examined by Western Blot and ALP detection. Results(1) In vitro, the expression of ALP in hydrogels was significantly increased in the exosome group compared with the control group;(2) The exosomes had obvious promotion effect on proliferation and osteogenic differentiation of ADSCs in CS/n HAC/β-GP hydrogels. Conclusion The new material consisting of exosome derived from ADSCs and CS/n HAC/β-GP hydrogel provides a new treatment strategy for bone tissue engineering and shows good potential in the repair of bone defects.
引文
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[13] JIANG T, KUMBAR S G, NAIR L S, et al. Biologically active chitosan systems for tissue engineering and regenerative medicine[J].Curr Top Med Chem,2008,8(4):354-364.
[14] MAZZONI E, D′AGOSTINO A, MANFRINI M, et al. Human adipose stem cells induced to osteogenic differentiation by an innovative collagen/hydroxylapatite hybrid scaffold[J]. FASEB J,2017,31(10):4555-4565.
[15] DHIVYA S, SARAVANAN S, SASTRY T P, et al. Nanohydroxyapatite-reinforced chitosan composite hydrogel for bone tissue repair in vitro and in vivo[J]. J Nanobiotechnology, 2015,13:40.
[16] KOSAKA N, IGUCHI H, HAGIWARA K, et al. Neutral sphingomyelinase 2(nSMase2)-dependent exosomal transfer of angiogenic micro RNAs regulate cancer cell metastasis[J]. J Biol Chem, 2013,288(15):10849-10859.
[17] SOLDEVILLA B, RODRGUEZ M, SAN MILLN C, et al.Tumor-derived exosomes are enriched inΔNp73, which promotes oncogenic potential in acceptor cells and correlates with patient survival[J]. Hum Mol Genet,2014,23(2):467-478.
[18] SABER-SAMANDARI S, SABER-SAMANDARI S. Biocompatible nanocomposite scaffolds based on copolymer-grafted chitosan for bone tissue engineering with drug delivery capability[J]. Mater Sci Eng C Mater Biol Appl, 2017,75:721-732.
[19] GARDIN C, FERRONI L, FAVERO L, et al. Nanostructured biomaterials for tissue engineered bone tissue reconstruction[J]. Int J Mol Sci, 2012,13(1):737-757.
[2] TOOI M, KOMAKI M, MORIOKA C, et al. Placenta mesenchymal stem cell derived exosomes confer plasticity on fibroblasts[J]. J Cell Biochem, 2016,117(7):1658-1670.
[3] GEBRAAD A, KORNILOV R, KAUR S, et al. Monocyte-derived extracellular vesicles stimulate cytokine secretion and gene expressionofmatrixmetalloproteinasesbymesenchymalstem/stromal cells[J]. FEBS J, 2018,285(12):2337-2359.
[4] LI W, LIU Y, ZHANG P, et al. Tissue-engineered bone immobilized with human adipose stem cells-derived exosomes promotes bone regeneration[J]. ACS Appl Mater Interfaces, 2018,10(6):5240-5254.
[5] DRURY J L, MOONEY D J. Hydrogels for tissue engineering:scaffold design variables and applications[J]. Biomaterials, 2003,24:4337-4351.
[6] JIANG T, KUMBAR S G, NAIR L S, et al. Biologically active chitosan systems for tissue engineering and regenerative medicine[J]. Curr Top Med Chem, 2008,8(4):354-364.
[7] VAN DER POL E, HOEKSTRA A G, STURK A, et al. Optical and non-optical methods for detection and characterization of microparticles and exosomes[J]. J Thromb Haemost, 2010,8(12):2596-2607.
[8] VAN DER POL E, COUMANS F A, GROOTEMAAT A E, et al.Particle size distribution of exosomes and microvesicles determined by transmission electron microscopy, flow cytometry, nanoparticle tracking analysis, and resistive pulse sensing[J]. J Thromb Haemost, 2014,12(7):1182-1192.
[9] RATAJCZAK M Z, KUCIA M, JADCZYK T, et al. Pivotal role of paracrine effects in stem cell therapies in regenerative medicine:can we translate stem cell-secreted paracrine factors and microvesicles into better therapeutic strategies?[J]. J Ratajczak Leukemia, 2011,26(6):1166-1173.
[10] LIANG X, DING Y, ZHANG Y, et al. Paracrine mechanisms of mesenchymal stem cell-based therapy:current status and perspectives[J]. Cell Transplant, 2014,23(9):1045-1099.
[11] FORERO J C, ROA E, REYES J G, et al. Development of Useful Biomaterial for Bone Tissue Engineering by Incorporating NanoCopper-Zinc Alloy(nCu Zn)in Chitosan/Gelatin/Nano-Hydroxyapatite(Ch/G/n HAp)Scaffold[J]. Materials(Basel), 2017,10(10).pii:E1177.
[12] KAUR S, SINGH S P, ELKAHLOUN A G, et al. CD47-dependent immunomodulatory and angiogenic activities of extracellular vesicles produced by T cells[J]. Matrix Biol, 2014,37:49-59.
[13] JIANG T, KUMBAR S G, NAIR L S, et al. Biologically active chitosan systems for tissue engineering and regenerative medicine[J].Curr Top Med Chem,2008,8(4):354-364.
[14] MAZZONI E, D′AGOSTINO A, MANFRINI M, et al. Human adipose stem cells induced to osteogenic differentiation by an innovative collagen/hydroxylapatite hybrid scaffold[J]. FASEB J,2017,31(10):4555-4565.
[15] DHIVYA S, SARAVANAN S, SASTRY T P, et al. Nanohydroxyapatite-reinforced chitosan composite hydrogel for bone tissue repair in vitro and in vivo[J]. J Nanobiotechnology, 2015,13:40.
[16] KOSAKA N, IGUCHI H, HAGIWARA K, et al. Neutral sphingomyelinase 2(nSMase2)-dependent exosomal transfer of angiogenic micro RNAs regulate cancer cell metastasis[J]. J Biol Chem, 2013,288(15):10849-10859.
[17] SOLDEVILLA B, RODRGUEZ M, SAN MILLN C, et al.Tumor-derived exosomes are enriched inΔNp73, which promotes oncogenic potential in acceptor cells and correlates with patient survival[J]. Hum Mol Genet,2014,23(2):467-478.
[18] SABER-SAMANDARI S, SABER-SAMANDARI S. Biocompatible nanocomposite scaffolds based on copolymer-grafted chitosan for bone tissue engineering with drug delivery capability[J]. Mater Sci Eng C Mater Biol Appl, 2017,75:721-732.
[19] GARDIN C, FERRONI L, FAVERO L, et al. Nanostructured biomaterials for tissue engineered bone tissue reconstruction[J]. Int J Mol Sci, 2012,13(1):737-757.