Performance of 3D-printed polylactic acid-nano-hydroxyapatite/chitosan/doxycycline antibacterial scaffold

doi:10.12307/2024.529

Abstract

Abstract: BACKGROUND: Polylactic acid has good biocompatibility and biodegradability, and has become a new orthopedic fixation material. However, the lack of cell recognition signal of this material is not conducive to cell adhesion and osteogenic differentiation, which limits its application in biomaterials.
OBJECTIVE: 3D-printed polylactic acid-nano-hydroxyapatite (nHA)/chitosan (CS) scaffold to evaluate its drug sustained-release and biological properties.
METHODS: The porous polylactic acid scaffold (recorded as PLA scaffold) with interporous pores was printed by fused deposition modeling technique, and the scaffold was soaked in dopamine solution to prepare polylactic acid-dopamine scaffold (recorded as PLA-DA scaffold). Nano-hydroxyapatite was immersed in chitosan solution, and then the PLA-DA scaffold was immersed in it to prepare polylactic acid-nano-hydroxyapatite/chitosan scaffold (recorded as PLA-nHA/CS scaffold). The micro-morphology, porosity, water contact angle, and compressive strength of the three scaffolds were characterized. PLA-nHA/CS scaffold loaded with doxycycline (recorded as PLA-nHA/CS-DOX scaffold) was prepared by freeze-drying method, and its drug release was characterized. PLA, PLA-DA, PLA-nHA/CS, and PLA-nHA/CS-DOX scaffolds were co-cultured with MC3T3-E1 cells, separately, to detect cell proliferation and osteogenic differentiation. Staphylococcus aureus suspensions of different concentrations were co-cultured with four groups of scaffolds. The antibacterial performance of scaffolds was detected by inhibition zone test.
RESULTS AND CONCLUSION: (1) Under scanning electron microscopy, the surfaces of PLA and PLA-DA scaffolders were dense and smooth, and nHA particles were observed on PLA-nHA/CS scaffolders. The porosity of PLA, PLA-DA and PLA-nHA/CS scaffolds decreased gradually, and the compressive strength increased gradually. The elastic modulus of PLA-nHA/CS scaffolds met the requirements of cancelous bone. The water contact angle of PLA-DA and PLA-nHA /CS brackets was smaller than that of PLA scaffolds. The PLA-nHA/CS scaffold sustainably released drugs in vitro for 8 days. (2) CCK-8 assay showed that the proliferation of MC3T3-E1 cells was not significantly affected by the four groups of scaffolds. The activity of alkaline phosphatase in PLA-DA group, PLA-nHA /CS group, and PLA-nHA/CS-DOX group was higher than that in PLA group. Alizarin red staining showed that compared with PLA group, the cells in PLA-nHA/CS group and PLA-nHA/CS-DOX group showed higher mineralized water level. (3) Inhibition zone test exhibited that PLA and PLA-DA scaffolds had no antibacterial properties. PLA-nHA/CS scaffolds had certain antibacterial properties. PLA-nHA/CS-DOX scaffolds had super antibacterial properties. (4) The results showed that the PLA-nHA/CS-DOX scaffold had good drug release performance, cell compatibility, osteogenic properties, and antibacterial properties.

Key words: 3D printing, polylactic acid scaffold, nano-hydroxyapatite, dopamine, chitosan, antibacterial coating, drug sustained release, bone regeneration

CLC Number:

Liu Yan, Zheng Xuexin. Performance of 3D-printed polylactic acid-nano-hydroxyapatite/chitosan/doxycycline antibacterial scaffold[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(22): 3532-3538.

Figures/Tables 9

References

[1] WUBNEH A, TSEKOURA EK, AYRANCI C, et al. Current state of fabrication technologies and materials for bone tissue engineering. Acta Biomater. 2018;80:1-30.
[2] KOONS GL, DIBA M, MIKOS AG. Materials design for bone-tissue engineering. Nat Rev Mater. 2020;5(8):584-603.
[3] TETIK H, FENG D, OXANDALE SW, et al. Bioinspired manufacturing of aerogels with precisely manipulated surface microstructure through controlled local temperature gradients. ACS Appl Mater Interfaces. 2021;13(1):924-931.
[4] ALDEMIR DIKICI BL, REILLY GC, CLAEYSSENS F. Boosting the osteogenic and angiogenic performance of multiscale porous polycaprolactone scaffolds by in vitro generated extracellular matrix decoration. ACS Appl Mater Interfaces. 2020;12(11):12510-12524.
[5] XING H, ZOU B, LIU X, et al. Fabrication strategy of complicated Al2O3-Si3N4 functionally graded materials by stereolithography 3D printing. J Eur Ceram Soc. 2020;40(15):5797-5809.
[6] BOULAALA M, ELMESSAOUDI D, BUJ-CORRAL I, et al. Towards design of mechanical part and electronic control of multi-material/multicolor fused deposition modeling 3D printing. Int J Adv Manuf Tech. 2020; 110:45-55.
[7] ZHANG W, FENG C, YANG G, et al. 3D-printed scaffolds with synergistic effect of hollow-pipe structure and bioactive ions for vascularized bone regeneration. Biomaterials. 2017;135:85-95.
[8] LIM SH, CHIA SMY, KANG L, et al. Three-Dimensional Printing of Carbamazepine Sustained-Release Scaffold. J Pharm Sci. 2016;105(7): 2155-2163.
[9] YANG Y, QIAO X, HUANG R, et al. E-jet 3D printed drug delivery implants to inhibit growth and metastasis of orthotopic breast cancer. Biomaterials. 2020;230:119618.
[10] GAO C, PENG S, FENG P, et al. Bone biomaterials and interactions with stem cells. Bone Res. 2017;5(1):1-33.
[11] BAE EB, PARK KH, SHIM JH, et al. Efficacy of rhBMP-2 loaded PCL/β-TCP/bdECM scaffold fabricated by 3D printing technology on bone regeneration. Biomed Res Int. 2018;2018:1-12.
[12] TERTULIANO OA, GREER JR. The nanocomposite nature of bone drives its strength and damage resistance. Nat Mater. 2016;15(11):1195-1202.
[13] CROSS LM, THAKUR A, JALILI NA, et al. Nanoengineered biomaterials for repair and regeneration of orthopedic tissue interfaces. Acta Biomater. 2016;42:2-17.
[14] QUAN H, HE Y, SUN J, et al. Chemical self-assembly of multifunctional hydroxyapatite with a coral-like nanostructure for osteoporotic bone reconstruction. ACS Appl Mater Interfaces. 2018;10(30):25547-25560.
[15] REZWAN K, CHEN Q, BLAKER JJ, et al. Biodegradable and bioactive porous polymer/inorganic composite scaffolds for bone tissue engineering. Biomaterials. 2006;27(18):3413-3431.
[16] GHORBANI F, ZAMANIAN A, BEHNAMGHADER A, et al. A facile method to synthesize mussel-inspired polydopamine nanospheres as an active template for in situ formation of biomimetic hydroxyapatite. Mat Sci Eng C Mater. 2019;94:729-739.
[17] LÓPEZ-IGLESIAS C, BARROS J, ARDAO I, et al. Vancomycin-loaded chitosan aerogel particles for chronic wound applications. Carbohydr Polym. 2019;204:223-231.
[18] FERREIRA M, RZHEPISHEVSKA O, GRENHO L, et al. Levofloxacin-loaded bone cement delivery system: Highly effective against intracellular bacteria and Staphylococcus aureus biofilms. Int J Pharm. 2017;532(1):241-248.
[19] BAUMANN B, JUNGST T, STICHLER S, et al. Control of nanoparticle release kinetics from 3D printed hydrogel scaffolds. Angew Chem Int Ed. 2017;56(16):4623-4628.
[20] MARTIN V, RIBEIRO IA, ALVES MM, et al. Engineering a multifunctional 3D-printed PLA-collagen-minocycline-nanoHydroxyapatite scaffold with combined antimicrobial and osteogenic effects for bone regeneration. Mat Sci Eng C Mater. 2019;101:15-26.
[21] HUANG Y, ZHAO X, ZHANG Z, et al. Degradable gelatin-based IPN cryogel hemostat for rapidly stopping deep noncompressible hemorrhage and simultaneously improving wound healing. Chem Mater. 2020;32(15):6595-6610.
[22] XIE C, LI P, LIU Y, et al. Preparation of TiO2 nanotubes/mesoporous calcium silicate composites with controllable drug release. Mat Sci Eng C-Mater. 2016;67:433-439.
[23] GOUDARZI A, SADRNEZHAAD SK, JOHARI N. The prominent role of fully-controlled surface co-modification procedure using titanium nanotubes and silk fibroin nanofibers in the performance enhancement of Ti6Al4V implants - ScienceDirect. Surf Coat Technol. 2021;412:127001.
[24] MATSUMOTO T, TASHIRO Y, KOMASA S, et al. Effects of surface modification on adsorption behavior of cell and protein on titanium surface by using quartz crystal microbalance system. Materials. 2020; 14(1):97.
[25] JIA Z, SHI Y, XIONG P, et al. From solution to biointerface: graphene self-assemblies of varying lateral sizes and surface properties for biofilm control and osteodifferentiation. ACS Appl Mater Interfaces. 2016;8(27):17151-17165.
[26] GITTENS RA, SCHEIDELER L, RUPP F, et al. A review on the wettability of dental implant surfaces II: Biological and clinical aspects. Acta Biomater. 2014;10(7):2907-2918.
[27] GOMES FILHO MS, OLIVEIRA FA, BARBOSA MAA. A statistical mechanical model for drug release: Investigations on size and porosity dependence. Physica A. 2016;460:29-37.
[28] WANG Y, SHI X, REN L, et al. Poly (lactide‐co‐glycolide)/titania composite microsphere‐sintered scaffolds for bone tissue engineering applications. J Biomed Mater Res B. 2010;93(1):84-92.
[29] SUN H, ZHANG C, ZHANG B, et al. 3D printed calcium phosphate scaffolds with controlled release of osteogenic drugs for bone regeneration. Chem Eng J. 2022;427:130961.
[30] OLIVEIRA JE, MATTOSO LH, ORTS WJ, et al. Structural and morphological characterization of micro and nanofibers produced by electrospinning and solution blow spinning: a comparative study. Adv Mater Sci Eng. 2013;2013:1-14.
[31] NAZEER MA, YILGOR E, YAGCI MB, et al. Effect of reaction solvent on hydroxyapatite synthesis in sol–gel process. R Soc Open Sci. 2017; 4(12):171098.
[32] WU C, ZHANG Y, ZHOU Y, et al. A comparative study of mesoporous glass/silk and non-mesoporous glass/silk scaffolds: physiochemistry and in vivo osteogenesis. Acta Biomater. 2011;7(5):2229-2236.
[33] VIMALRAJ S. Alkaline phosphatase: Structure, expression and its function in bone mineralization. Gene. 2020;754:144855.
[34] SHI X, LI L, OSTROVIDOV S, et al. Stretchable and micropatterned membrane for osteogenic differentation of stem cells. ACS Appl Mater Interfaces. 2014;6(15):11915-11923.
[35] YU Y, LI X, LI J, et al. Dopamine-assisted co-deposition of hydroxyapatite-functionalised nanoparticles of polydopamine on implant surfaces to promote osteogenesis in environments with high ROS levels. Mat Sci Eng C Mater. 2021;131:112473.
[36] FITZPATRICK V, MARTíN-MOLDES Z, DECK A, et al. Functionalized 3D-printed silk-hydroxyapatite scaffolds for enhanced bone regeneration with innervation and vascularization. Biomaterials. 2021;276:120995.
[37] CAMPANA V, MILANO G, PAGANO E, et al. Bone substitutes in orthopaedic surgery: from basic science to clinical practice. J Mater Sci Mater Med. 2014;25:2445-2461.
[38] BAUER T, MUSCHLER G. Bone graft materials. An overview of the basic science. Clin Oryhop Relat R. 2000;371:10-27.
[39] INZANA JA, OLVERA D, FULLER SM, et al. 3D printing of composite calcium phosphate and collagen scaffolds for bone regeneration. Biomaterials. 2014;35(13):4026-4034.
[40] GAO C, PENG S, FENG P, et al. Bone biomaterials and interactions with stem cells. Bone Res. 2017;5(4):17059.
[41] KRASOWSKA A, SIGLER K. How microorganisms use hydrophobicity and what does this mean for human needs? Front Cell Infect Microbiol. 2014;4:112.
[42] QUAN H, HE Y, SUN J, et al. Chemical Self-Assembly of Multifunctional Hydroxyapatite with a Coral-like Nanostructure for Osteoporotic Bone Reconstruction. ACS Appl Mater Interfaces. 2018;10(30):25547-25560.
[43] ZHANG B, WANG L, SONG P, et al. 3D printed bone tissue regenerative PLA/HA scaffolds with comprehensive performance optimizations. Mater Des. 2021;201:109490.
[44] CAMPOCCIA D, MONTANARO L, ARCIOLA CR. The significance of infection related to orthopedic devices and issues of antibiotic resistance. Biomaterials. 2006;27(11):2331-2339.
[45] VISSCHER L, DANG HP, KNACKSTEDT M, et al. 3D printed Polycaprolactone scaffolds with dual macro-microporosity for applications in local delivery of antibiotics. Mat Sci Eng C Mater. 2018; 87:78-89.