In this study, we evaluated the use of graphene oxide (GO) mixed with methyl methacrylate gelatin (GelMA) for the construction of a microenvironmental implant to repair bone defects in orthopedic surgery. A scaffold containing a GelMA/GO composite with mesenchymal stem cells (MSCs) was constructed using three-dimensional bioprinting. The survival and osteogenic capacity of MSCs in the composite bioink were evaluated using cell viability and proliferation assays, osteogenesis-related gene expression analysis, and implantation under the skin of nude mice. The printing process had little effect on cell viability. We found that GO enhanced cell proliferation but had no significant effect on cell viability. In vitro experiments suggested that GO promoted material–cell interactions and the expression of osteogenesis-related genes. In vivo experiments showed that GO decreased the degradation time of the material and increased calcium nodule deposition. In contrast to pure GelMA, the addition of GO created a suitable microenvironment to promote the differentiation of loaded exogenous MSCs in vitro and in vivo, providing a basis for the repair of bone defects.
JiangYYangBYangF, et al.One-half wedge osteotomy genioplasty for correction of chin deviation based on three-dimensional computed tomography measurements and simulation. J Craniofac Surg2021; 32(4): 1496–1499.
2.
SongJLiLFangL, et al.Advanced strategies of scaffolds design for bone regeneration. BMEMat2023; 1(4): e12046.
3.
SainiGSegaranNMayerJL, et al.Applications of 3D bioprinting in tissue engineering and regenerative medicine. J Clin Med2021; 10(21): 4966.
4.
VijayavenkataramanSYanWCLuWF, et al.3D bioprinting of tissues and organs for regenerative medicine. Adv Drug Deliv Rev2018; 132: 296–332.
5.
RastinHOrmsbyRTAtkinsGJ, et al.3D bioprinting of methylcellulose/gelatin-methacryloyl (MC/GelMA) bioink with high shape integrity. ACS Appl Bio Mater2020; 3(3): 1815–1826.
6.
SuvarnapathakiSWuXLantiguaD, et al.Hydroxyapatite-incorporated composite gels improve mechanical properties and bioactivity of bone scaffolds. Macromol Biosci2020; 20(10): e2000176.
7.
JiCQiuMRuanH, et al.Transcriptome analysis revealed the symbiosis niche of 3D scaffolds to accelerate bone defect healing. Adv Sci2022; 9(8): e2105194.
8.
HanJJYangHJHwangSJ. Enhanced bone regeneration by bone morphogenetic protein-2 after pretreatment with low-intensity pulsed ultrasound in distraction osteogenesis. Tissue Eng Regen Med2022; 19(4): 871–886.
9.
ParkJLeeSJJungTG, et al.Surface modification of a three-dimensional polycaprolactone scaffold by polydopamine, biomineralization, and BMP-2 immobilization for potential bone tissue applications. Colloids Surf B Biointer2021; 199: 111528.
10.
HuHDongLBuZ, et al.miR-23a-3p-abundant small extracellular vesicles released from gelma/nanoclay hydrogel for cartilage regeneration. J Extracell Vesicles2020; 9(1): 1778883.
11.
HerfordASTandonRStevensTW, et al.Immediate distraction osteogenesis: the sandwich technique in combination with rhBMP-2 for anterior maxillary and mandibular defects. J Craniofac Surg2013; 24(4): 1383–1387.
12.
TexterJ. Graphene oxide and graphene flakes as stabilizers and dispersing aids. COCIS2015; 20: 454–464.
13.
LiuYChenTDuF, et al.Single-layer graphene enhances the osteogenic differentiation of human mesenchymal stem cells in vitro and in vivo. J Biomed Nanotechnol2016; 12(6): 1270–1284.
14.
KolanthaiEBoseSBhagyashreeKS, et al.Graphene scavenges free radicals to synergistically enhance structural properties in a gamma-irradiated polyethylene composite through enhanced interfacial interactions. Phys Chem Chem Phys2015; 17(35): 22900–22910.
15.
LeeWCLimCHShiH, et al.Origin of enhanced stem cell growth and differentiation on graphene and graphene oxide. ACS Nano2011; 5(9): 7334–7341.
16.
DalgicADAlshemaryAZTezcanerA, et al.Silicate-doped nano-hydroxyapatite/graphene oxide composite reinforced fibrous scaffolds for bone tissue engineering. J Biomater Appl2018; 32(10): 1392–1405.
17.
DuranMLuzoACMde SouzaJG, et al.Graphene oxide as scaffolds for stem cells: an overview. Curr Mol Med2017; 17(9): 619–626.
SwansonWBOmiMZhangZ, et al.Macropore design of tissue engineering scaffolds regulates mesenchymal stem cell differentiation fate. Biomaterials2021; 272: 120769.
20.
AhnWBLeeYBJiYH, et al.Decellularized human adipose tissue as an alternative graft material for bone regeneration. Tissue Eng Regen Med2022; 19(5): 1089–1098.
21.
AlexandreXMMoraes SilvaSO’ConnellCD, et al.Enhanced electroactivity, mechanical properties, and printability through the addition of graphene oxide to photo-cross-linkable gelatin methacryloyl hydrogel. ACS Biomater Sci Eng2021; 7(6): 2279–2295.
22.
FengXChenLGuoW, et al.Graphene oxide induces p62/SQSTM-dependent apoptosis through the impairment of autophagic flux and lysosomal dysfunction in PC12 cells. Acta Biomater2018; 81: 278–292.
23.
KangYLiuJWuJ, et al.Graphene oxide and reduced graphene oxide induced neural pheochromocytoma-derived PC12 cell lines apoptosis and cell cycle alterations via the ERK signaling pathways. Int J Nanomed2017; 12: 5501–5510.
24.
ShinSRAghaei-Ghareh-BolaghBDangTT, et al.Cell-laden microengineered and mechanically tunable hybrid hydrogels of gelatin and graphene oxide. Adv Mater2013; 25(44): 6385–6391.
25.
PaulAHasanAKindiHA, et al.Injectable graphene oxide/hydrogel-based angiogenic gene delivery system for vasculogenesis and cardiac repair. ACS Nano2014; 8(8): 8050–8062.
26.
SubhaNRNooeaidPArkudasA, et al.Adipose- and bone marrow-derived mesenchymal stem cells display different osteogenic differentiation patterns in 3D bioactive glass-based scaffolds. J Tissue Eng Regen Med2016; 10(10): E497–E509.
27.
DaveJRChandekarSSBeheraS, et al.Human gingival mesenchymal stem cells retain their growth and immunomodulatory characteristics independent of donor age. Sci Adv2022; 8(25): eabm6504.
28.
KratochvílováAVeseláBLedvinaV, et al.Osteogenic impact of pro-apoptotic caspase inhibitors in MC3T3-E1 cells. Sci Rep2020; 10(1): 7489.
29.
LiuYWangYSunX, et al.RUNX2 mutation reduces osteogenic differentiation of dental follicle cells in cleidocranial dysplasia. Mutagenesis2018; 33(3): 203–214.
30.
MaruyamaZYoshidaCAFuruichiT, et al.Runx2 determines bone maturity and turnover rate in postnatal bone development and is involved in bone loss in estrogen deficiency. Dev Dynam2007; 236(7): 1876–1890.
31.
WeaverCLLaRosaJMLuoXL, et al.Electrically controlled drug delivery from graphene oxide nanocomposite films. ACS Nano2014; 8: 1834–1843.
32.
ChoeGOhSSeokJM, et al.Graphene oxide/alginate composites as novel bioinks for three-dimensional mesenchymal stem cell printing and bone regeneration applications. Nanoscale2019; 11(48): 23275–23285.
33.
YangJWHsiehKYKumarPV, et al.Enhanced osteogenic differentiation of stem cells on phase-engineered graphene oxide. ACS Appl Mater Interfaces2018; 10(15): 12497–12503.
34.
BhadraDSannigrahiJChaudhuriBK, et al.Enhancement of the transport and dielectric properties of graphite oxide nanoplatelets-polyvinyl alcohol composite showing low percolation threshold. Polym Compos2012; 33(3): 436–442.
35.
LiZXiangSLinZ, et al.Graphene oxide-functionalized nanocomposites promote osteogenesis of human mesenchymal stem cells via enhancement of BMP-SMAD1/5 signaling pathway. Biomaterials2021; 277: 121082.
36.
TsimbouriPMMcMurrayRJBurgessKV, et al.Using nanotopography and metabolomics to identify biochemical effectors of multipotency. ACS Nano2012; 6(11): 10239–10249.
37.
QianWGongLCuiX, et al.Nanotopographic regulation of human mesenchymal stem cell osteogenesis. ACS Appl Mater Interfaces2017; 9(48): 41794–41806.
38.
CardosoLFrittonSPGailaniG, et al.Advances in assessment of bone porosity, permeability and interstitial fluid flow. J Biomech2013; 46(2): 253–265.
39.
NagatomiJArulanandamBPMetzgerDW, et al.Frequency- and duration-dependent effects of cyclic pressure on select bone cell functions. Tissue Eng2001; 7(6): 717–728.
40.
WangCWangSKangDD, et al.Biomaterials for in situ cell therapy. BMEMat2023; 1(3): e12039.
41.
QuartoRMastrogiacomoMCanceddaR, et al.Repair of large bone defects with the use of autologous bone marrow stromal cells. N Engl J Med2001; 344(5): 385–386.
42.
MorishitaTHonokiKOhgushiH, et al.Tissue engineering approach to the treatment of bone tumors: three cases of cultured bone grafts derived from patients’ mesenchymal stem cells. Artif Organs2006; 30(2): 115–118.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
