Effects of Plasmid Fibroblast Growth Factor-2 Magnetic Chitosan Gelatin Microspheres on Proliferation and Differentiation of Mesenchymal Stem Cells_Journal of Biomedical Engineering

Authors：

DINGXingpo ^1,3 , LIMing ² ,  CAOYujiang ² , YANGQiong ^1,2 , HETongchuan ¹ , LUOCong ² , LIHaibing ² , BIYang ¹

1. Stem Cell Biology and Therapy Laboratory, The Children's Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Key Laboratory of Pediatrics in Chongqing, Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Chongqing 400014, China;
2. Department of Orthopedics, The Children's Hospital of Chongqing Medical University, Chongqing 400014, China;
3. Department of Pediatric Orthopedics, Luoyang Orthopedic-Traumatological Hospital, Luoyang 471002, China;

Corresponding author：

CAOYujiang, Email: caoyujiang@126.com

Keywords：

FGF-2 gene; gelatin microsphere; mesenchymal stem cell; proliferation; differentiation

DOI：

10.7507/1001-5515.20150192

Video：

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Abstract Full text Figures/Tables Video References Cited by

The purpose of this study is to investigate the effect of superparamagnetic chitosan FGF-2 gelatin microspheres (SPCFGM) on the proliferation and differentiation of mouse mesenchymal stem cells. The superparamagnetic iron oxide chitosan nanoparticles (SPIOCNs) were synthesized by means of chemical co-precipitation, combined with FGF-2. Then The SPCFGM and superparamagnetic chitosan gelatin microspheres (SPCGM) were prepared by means of crosslinking-emulsion. The properties of SPCFGM and SPIONs were measured by laser diffraction particle size analyser and transmisson electron microscopy. The SPCFGM were measured for drug loading capacity, encapsulation efficiency and release pharmaceutical properties in vitro. The C3H10 cells were grouped according to the different ingredients being added to the culture medium: SPCFGM group, SPCGM group and DMEM as control group. Cell apoptosis was analyzed by DAPI staining. The protein expression level of FGF-2 was determined by Western blot. The proliferation activity and cell cycle phase of C3H10 were examined by CCK8 and flow cytometry. The results demonstrated that both of the SPIOCNs and SPCFGM were exhibited structure of spherical crystallization with a diameter of (25±9) nm and (140±12) μm, respectively. There were no apoptosis cells in the three group cells. Both the protein expression level of FGF-2 and cell proliferation activity increased significantly in the SPCFGM group cells(P<0.05). The SPCFGM is successfully constructed and it can controlled-release FGF-2, remained the biological activity of FGF-2, which can promote proliferation activity of C3H10 cells, and are non-toxic to the cell.

Citation： DINGXingpo, LIMing, CAOYujiang, YANGQiong, HETongchuan, LUOCong, LIHaibing, BIYang. Effects of Plasmid Fibroblast Growth Factor-2 Magnetic Chitosan Gelatin Microspheres on Proliferation and Differentiation of Mesenchymal Stem Cells. Journal of Biomedical Engineering, 2015, 32(5): 1083-1089. doi: 10.7507/1001-5515.20150192 Copy

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14.	金旭红,杨柳,张寿,等.超顺磁性氧化铁标记对骨髓间充质干细胞多向分化诱导的影响[J].生物医学工程学杂志,2012,29(1):125-128, 133.
15.	JAIN T K, REDDY M K, MORALES M A, et al. Biodistribution, clearance, and biocompatibility of Iron oxide magnetic nanoparticles in rats[J]. Mol Pharm, 2008, 5(2):316-327.
16.	方楚玲.超顺磁性氧化铁纳米粒在骨髓间充质干细胞的应用[J].中国矫形外科杂志,2013,21(16):1629-1633.
17.	DETANTE O, VALABLE S, DE FRAIPONT F, et al. Magnetic resonance imaging and fluorescence labeling of clinical-grade mesenchymal stem cells without impacting their phenotype:study in a rat model of stroke[J]. Stem Cells Transl Med, 2012, 1(4):333-341.
18.	WANG A H, CHEN X G, LIU C S, et al. Preparation and characteristics of chitosan microspheres in different acetylation as drug carrier system[J]. J Microencapsul, 2009, 26(7):593-602.
19.	RAFTERY R. O'BRIEN F J, CRYAN S A. chitosan for gene delivery and orthopedic tissue engineering applications[J]. Molecules, 2013, 18(5):5611-5647.
20.	JO J I, TABATA Y. Non-viral gene transfection technologies for genetic engineering of stem cells[J]. Eur J Pharm Biopharm, 2008, 68(1):90-104.
21.	关林波,但卫华,曾睿,等.明胶及其在生物材料中的应用[J].材料导报,2006,20(z2):380-383.
22.	落全祥,高莺,李波宇,等.类胰岛素样生长因子-1与明胶微球复合物对兔下颌骨缺损愈合影响的实验研究[J].中国药物与临床,2013,13(12):1545-1547.
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1. KHAN I M, FRANCIS L, THEOBALD P S, et al. In vitro growth factor-induced bio engineering of mature articular cartilage[J]. Biomaterials, 2013, 34(5):1478-1487.
2. MOW V C, RATCLIFFE A, POOLE A R. Cartilage and diarthrodial joints as paradigms for hierarchical materials and structures[J]. Biomaterials, 1992, 13(2):67-97.
3. KHAN I M, GILBERT S J, SINGHRAO S K, et al. Cartilage integration:evaluation of the reasons for failure of integration during cartilage repair. A review[J]. Eur Cell Mater, 2008, 16(3):26-39.
4. 潘育松,丁国新,王静.组织工程化软骨修复关节软骨损伤和缺损[J].生物医学工程学杂志,2013(2):432-437.
5. VINCENT T, HERMANSSON M, BOLTON M, et al. Basic FGF mediates an immediate response of articular cartilage to mechanical injury[J]. Proc Natl Acad Sci U S A, 2002, 99(12):8259-8264.
6. HANDORF A M, LI W J. Fibroblast growth factor-2 primes human mesenchymal stem cells for enhanced chondrogenesis[J]. PLoS One, 2011, 6(7):e22887.
7. SIPOS P, BERKESI O, TOMBÁCZ E, et al. Formation of spherical Iron(III) oxyhydroxide nanoparticles sterically stabilized by chitosan in aqueous solutions[J]. J Inorg Biochem, 2003, 95(1):55-63.
8. 刘天禄.利用Gerhardt凯氏定氮仪测定凯氏氮[J].石化技术与应用,2000,18(3):180-181.
9. LIU Y, DENG X. Influences of preparation conditions on particle size and DNA-loading efficiency for poly(DL-lactic acid-polyethylene glycol) microspheres entrapping free DNA[J]. J Control Release, 2002, 83(1):147-155.
10. FUKUNAKA Y, IWANAGA K, MORIMOTO K, et al. Controlled release of plasmid DNA from cationized gelatin hydrogels based on hydrogel degradation[J]. J Control Release, 2002, 80(1/3):333-343.
11. COROT C, ROBERT P, IDEE J M, et al. Recent advances in Iron oxide nanocrystal technology for medical imaging[J]. Adv Drug Deliv Rev, 2006, 58(14):1471-1504.
12. TIETZE R, LYER S, DUERR S, et al. Nanoparticles for cancer therapy using magnetic forces[J]. Nanomedicine (Lond), 2012, 7(3):447-457.
13. LIU W, DAHNKE H, RAHMER J, et al. Ultrashort T2^* relaxometry for quantitation of highly concentrated superparamagnetic Iron oxide (SPIO) nanoparticle labeled cells[J]. Magn Reson Med, 2009, 61(4):761-766.
14. 金旭红,杨柳,张寿,等.超顺磁性氧化铁标记对骨髓间充质干细胞多向分化诱导的影响[J].生物医学工程学杂志,2012,29(1):125-128, 133.
15. JAIN T K, REDDY M K, MORALES M A, et al. Biodistribution, clearance, and biocompatibility of Iron oxide magnetic nanoparticles in rats[J]. Mol Pharm, 2008, 5(2):316-327.
16. 方楚玲.超顺磁性氧化铁纳米粒在骨髓间充质干细胞的应用[J].中国矫形外科杂志,2013,21(16):1629-1633.
17. DETANTE O, VALABLE S, DE FRAIPONT F, et al. Magnetic resonance imaging and fluorescence labeling of clinical-grade mesenchymal stem cells without impacting their phenotype:study in a rat model of stroke[J]. Stem Cells Transl Med, 2012, 1(4):333-341.
18. WANG A H, CHEN X G, LIU C S, et al. Preparation and characteristics of chitosan microspheres in different acetylation as drug carrier system[J]. J Microencapsul, 2009, 26(7):593-602.
19. RAFTERY R. O'BRIEN F J, CRYAN S A. chitosan for gene delivery and orthopedic tissue engineering applications[J]. Molecules, 2013, 18(5):5611-5647.
20. JO J I, TABATA Y. Non-viral gene transfection technologies for genetic engineering of stem cells[J]. Eur J Pharm Biopharm, 2008, 68(1):90-104.
21. 关林波,但卫华,曾睿,等.明胶及其在生物材料中的应用[J].材料导报,2006,20(z2):380-383.
22. 落全祥,高莺,李波宇,等.类胰岛素样生长因子-1与明胶微球复合物对兔下颌骨缺损愈合影响的实验研究[J].中国药物与临床,2013,13(12):1545-1547.
23. MASTROGIACOMO M, CANCEDDA R, QUARTO R. Effect of different growth factors on the chondrogenic potential of human bone marrow stromal cells[J]. Osteoarthritis Cartilage, 2001, 9(Suppl A):S36-S40.

Journal of Biomedical Engineering

Effects of Plasmid Fibroblast Growth Factor-2 Magnetic Chitosan Gelatin Microspheres on Proliferation and Differentiation of Mesenchymal Stem Cells

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