可载药的磁性聚己内酯/明胶微球支架的制备及其体外成骨性能的研究

doi:10.13591/j.cnki.kqyx.2022.04.001

Abstract

Abstract: Objective To prepare drug-loaded magnetic granular scaffolds and investigate their osteogenesis behavior in vitro. Methods The polycaprolactone (PCL)/gelatin microsphere (PGM) scaffolds were constructed via a double-emulsion method. The phase contrast microscope and field emission scanning electron microscope (FESEM) were applied for morphology observation. Thermal gravity analysis (TGA) and in vitro degradation test were used to figure out the proportion of ingredients and the trend of mass loss. Cell counting kit-8 (CCK-8) assay was conducted to optimize the concentration of nano oil-acid coated Fe₃O₄ particles (OA@Fe₃O₄), and the final Fe-PGM were prepared which were subsequently loaded with recombinant human bone morphogenetic protein-2 (rhBMP-2). The adhesion of rat bone mesenchymal stem cells (rBMSC) on PGM, Fe-PGM and Fe-PGM+BMP-2 was evaluated by inverted fluorescent microscope (IFM). Osteogenesis-related gene expression of rBMSC on PGM, Fe-PGM and Fe-PGM+BMP-2 was detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) 7 and 14 days after osteogenic induction. Results Photos of phase contrast microscope and FESEM demonstrated that PGM was composed of porous PCL skeleton and inner gelatin hydrogel. TGA and in vitro degradation test indicated that PCL and gelation was about 50% (wt.) of PGM respectively. Curve of mass loss in vitro was smooth. Results of CCK-8 showed that no inhibition on the proliferation of rBMSC was found in 0.4% Fe-PGM group compared to that in PGM group (P＜0.05). Acceptable cell adhesion was observed via IFM in PGM, Fe-PGM and Fe-PGM+BMP-2 group. Additionally, involvement of BMP-2 bettered cell sprawl in Fe-PGM+BMP-2 group. qRT-PCR analysis indicated a higher expression level of osteogenesis-related gene in Fe-PGM and Fe-PGM+BMP-2 group than that in PGM group owing to OA@Fe₃O₄, while application of BMP-2 further broadened the gap between Fe-PGM and Fe-PGM+BMP-2 group (P＜0.05). Conclusion The magnetic PGM scaffold has favorable cytological characteristics, and its drug-loading property could further improve adhesion and osteogenic differentiation of rBMSC on it.

Key words: granular scaffold, nano oil acid coated Fe₃O₄ particles, drug-loading, bone tissue engineering

CLC Number:

R783.1

ZHOU Fang, LIU Jun, HU Shuying, SHI Fan, YAN Jia, ZHANG Feimin. Study on drug-loaded magnetic polycaprolactone/gelatin microsphere scaffolds and their osteogenesis behavior in vitro[J]. Stomatology, 2022, 42(4): 289-295.

References

[1] Elgali I, Omar O, Dahlin C, et al. Guided bone regeneration:Materials and biological mechanisms revisited[J]. Eur J Oral Sci,2017,125(5):315-337.
[2] Kim HD,Amirthalingam S,Kim SL,et al. Biomimetic materials and fabrication approaches for bone tissue engineering[J/OL]. Adv Healthc Mater,2017[2021-12-30]. doi:10.1002/adhm.201700612.
[3] Roseti L,Parisi V,Petretta M,et al. Scaffolds for Bone Tissue Engineering:State of the art and new perspectives[J]. Mater Sci Eng C Mater Biol Appl,2017,78:1246-1262.
[4] Melancon MP,Appleton Figueira T,Fuentes DT,et al. Develop-ment of an electroporation and nanoparticle-based therapeutic platform for bone metastases[J]. Radiology,2018,286(1):149-157.
[5] 竺鑫晨,刘俊,严佳,等. Ⅰ型胶原修饰微球对骨髓间充质干细胞粘附增殖及成骨行为的影响[J]. 口腔医学,2020,40(4):299-303.
[6] Vieira S,Vial S,Reis RL,et al. Nanoparticles for bone tissue engineering[J]. Biotechnol Prog,2017,33(3):590-611.
[7] Xia Y,Sun JF,Zhao L,et al. Magnetic field and nano-scaffolds with stem cells to enhance bone regeneration[J]. Biomaterials,2018,183:151-170.
[8] Li Y,Ye DW,Li MX,et al. Adaptive materials based on iron oxide nanoparticles for bone regeneration[J]. Chemphyschem,2018,19(16):1965-1979.
[9] Hughes S,El Haj AJ,Dobson J. Magnetic micro- and nanoparticle mediated activation of mechanosensitive ion channels[J]. Med Eng Phys,2005,27(9):754-762.
[10] Russo T,Peluso V,Gloria A,et al. Combination design of time-dependent magnetic field and magnetic nanocomposites to guide cell behavior[J]. Nanomaterials (Basel),2020,10(3):577.
[11] Han L,Guo Y,Jia L,et al. 3D magnetic nanocomposite scaffolds enhanced the osteogenic capacities of rat bone mesenchymal stem cells in vitro and in a rat calvarial bone defect model by promoting cell adhesion[J]. J Biomed Mater Res A,2021,109(9):1670-1680.
[12] Atashi F,Modarressi A,Pepper MS. The role of reactive oxygen species in mesenchymal stem cell adipogenic and osteogenic differentiation:A review[J]. Stem Cells Dev, 2015, 24(10):1150-1163.
[13] Wu HH,Yin JJ,Wamer WG,et al. Reactive oxygen species-related activities of nano-iron metal and nano-iron oxides[J]. J Food Drug Anal,2014,22(1):86-94.
[14] Hao LL,Li LL,Wang P,et al. Synergistic osteogenesis promoted by magnetically actuated nano-mechanical stimuli[J]. Nanoscale,2019,11(48):23423-23437.
[15] Garg P,Mazur MM,Buck AC,et al. Prospective review of mesenchymal stem cells differentiation into osteoblasts[J]. Orthop Surg,2017,9(1):13-19.
[16] Zhou N,Li Q,Lin X,et al. BMP2 induces chondrogenic differentiation,osteogenic differentiation and endochondral ossification in stem cells[J]. Cell Tissue Res,2016,366(1):101-111.
[17] Salazar VS,Gamer LW,Rosen V. BMP signalling in skeletal development,disease and repair[J]. Nat Rev Endocrinol,2016,12(4):203-221.
[18] Bessa PC,Casal M,Reis RL. Bone morphogenetic proteins in tissue engineering:The road from laboratory to clinic,part Ⅱ (BMP delivery)[J]. J Tissue Eng Regen Med,2008,2(2/3):81-96.
[19] Annamalai RT,Turner PA,Carson WF 4th,et al. Harnessing macrophage-mediated degradation of gelatin microspheres for spatiotemporal control of BMP2 release[J]. Biomaterials,2018,161:216-227.
[20] Thakur G,Mitra A,Rousseau D,et al. Crosslinking of gelatin-based drug carriers by genipin induces changes in drug kinetic profiles in vitro[J]. J Mater Sci Mater Med,2011,22(1):115-123.
[21] Wang ZH,Liu H,Luo WB,et al. Regeneration of skeletal system with genipin crosslinked biomaterials[J]. J Tissue Eng,2020,11:2041731420974861.
[22] Solorio L,Zwolinski C,Lund AW,et al. Gelatin microspheres crosslinked with genipin for local delivery of growth factors[J]. J Tissue Eng Regen Med, 2010, 4(7):514-523.
[23] Denu RA,Hematti P. Effects of oxidative stress on mesenchymal stem cell biology[J]. Oxid Med Cell Longev,2016,2016:2989076.
[24] Sun YS,Lin Y,Huang HH. Using genipin to immobilize bone morphogenetic protein-2 on zirconia surface for enhancing cell adhesion and mineralization in dental implant applications[J]. Polymers,2020,12(11):2639.
[25] Sajesh KM, Ashokan A, Gowd GS, et al. Magnetic 3D scaffold: A theranostic tool for tissue regeneration and non-invasive imaging in vivo[J]. Nanomedicine: Nanotechnology, Biology and Medicine, 2019, 18:179-188.
[26] Alcorta-Sevillano N,Macías I,Infante A,et al. Deciphering the relevance of bone ECM signaling[J]. Cells,2020,9(12):2630.
[27] Chen Q,Shou P,Zheng C,et al. Fate decision of mesenchymal stem cells:Adipocytes or osteoblasts?[J]. Cell Death Differ,2016,23(7):1128-1139.
[28] Alcorta-Sevillano N,Macías I,Infante A,et al. Deciphering the relevance of bone ECM signaling[J]. Cells,2020,9(12):2630.
[29] Henstock JR,Rotherham M,Rashidi H,et al. Remotely activated mechanotransduction via magnetic nanoparticles promotes mineralization synergistically with bone morphogenetic protein 2:Applications for injectable cell therapy[J]. Stem Cells Transl Med,2014,3(11):1363-1374.
[30] Zhang QC,Tan K,Zhang Y,et al. In situ controlled release of rhBMP-2 in gelatin-coated 3D porous poly(ε-caprolactone) scaffolds for homogeneous bone tissue formation[J]. Biomacromolecules,2014,15(1):84-94.

Study on drug-loaded magnetic polycaprolactone/gelatin microsphere scaffolds and their osteogenesis behavior in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 8

Recommended Articles

Metrics

Comments

[1]	ZHANG Qian, WANG Chang, LIANG Chen, QU Xingyuan, LIU Yue, YAN Baojun, WANG Lei. Progress of research on application of chondroitin sulfate in osteogenic repair materials [J]. Stomatology, 2023, 43(1): 88-91.
[2]	WANG Ying, LI Mingming, ZHAO Xiao, LENG Diya, WU Daming. In vitro biological activity study on the material composited with nanosilver and nanozinc incorporated mespoporous calcium-silicate and PCL [J]. Stomatology, 2022, 42(2): 110-116.
[3]	Yang LIU. The chitosan drug delivery system and its application in bone tissue engineering [J]. , 2019, 39(4): 354-359.
[4]	. Application of poly(lactic-co-glycolic acid) in bone tissue engineering scaffold materials [J]. , 2019, 39(2): 167-170.
[5]	. Applications of composite hydrogel scaffold in bone tissue engineering [J]. , 2018, 38(6): 563-568.
[6]	. Research progress on design and modification of chitosan scaffolds in bone tissue engineering [J]. , 2017, 37(2): 184-187.
[7]	. [J]. , 2014, 34(4): 301-301.
[8]	. Preliminary experimental study of repairing bone defects around dental implants with tissue engineering bone established with Bio-oss collagen scaffoldbone establish by Bio-oss collagen as scaffold [J]. , 2013, 33(2): 95-99.