Abstract
Recently, the 3D spheroid cell culture application has been extensively used in the treatment of bone defects. A wide variety of methodologies have been used, which has made the comparison of results complex. Therefore, this systematic review has two aims: (i) to perform an analysis focused on the role of 3D spheroid cell culture in bone regeneration strategies; and (ii) address the main challenges in clinical application. A search of the following keywords "3D cell culture", "spheroid", and "bone regeneration" was carried out in the PubMed, Scopus, and ScienceDirect databases and limited to the years 2010-2020. Studies were included if their primary objective was the behavior of cell aggregates to formed spheroids structures by different 3D cell culture techniques focused on the regeneration of bone tissue. To address the risk of bias for in vitro studies, the United States national toxicology program tool was applied, and descriptive statistics of the data were performed, with the SPSS V.22 program. A total of 16 studies were included, which met the established criteria corresponding to in vitro and in vitro/in vivo studies; most of these studies used stem cells for the 3D cell spheroids. The most often methods used for the 3D formation were low adherence surface and rotational methods, moreover, mesenchymal stem cells were the cell line most frequently used because of their regenerative potential in the field of bone tissue engineering. Although the advances in research on the potential use of 3D spheroids in bone regeneration have made great strides, the constant innovation in cell spheroid formation methodologies means that clinical application remains in the future as strategy for 3D tissue bioprinting.
References
Sui B.D., Hu C.H., Liu A.Q., Zheng C.X., Xuan K., Jin Y. Stem cell-based bone regeneration in diseased microenvironments: challenges and solutions. Biomaterials. 2019; 196:18-30.
Kloss F.R., Offermanns V., Kloss-Brandstätter A. Comparison of allogeneic and autogenous bone grafts for augmentation of alveolar ridge defects -a 12-month retrospective radiographic evaluation. Clin Oral Implants Res. 2018; 29 (11): 1163-1175.
Saba I., Jakubowska W., Bolduc S., Chabaud S. Engineering tissues without the use of a synthetic scaffold: a twenty-year history of the self-assembly method. Biomed Res Int. 2018; 2018: 1-13.
Koledova Z. 3D cell culture: an introduction. Methods Mol Biol. 2017; 1612: 1-11.
Sano K., Usui M., Moritani Y., Nakazawa K., Hanatani T., Kondo H., Nakatomi M., Onizuka S., Iwata T., Sato T., Togari A., Ariyoshi W., Nishihara T., Nakashima K. Co-cultured spheroids of human periodontal ligament mesenchymal stem cells and vascular endothelial cells enhance periodontal tissue regeneration. Regen Ther. 2020; 14: 59-71.
Lin Y.J., Lee Y.W., Chang C.W., Huang C.C. 3D Spheroids of Umbilical Cord Blood MSC-Derived Schwann Cells Promote Peripheral Nerve Regeneration. Front Cell Dev Biol. 2020; 8: 604946.
Rooney, A. Extending a Risk-of-Bias Approach to Address In Vitro Studies. in National Toxicology Program Office of Health Assessment and Translation (USA: Environmental Protection Agency (EPA)). 2015.
He D., Wang R.X., Mao J.P., Xiao B., Chen D.F., Tian W. Three-dimensional spheroid culture promotes the stemness maintenance of cranial stem cells by activating PI3K/AKT and suppressing NF-κB pathways. Biochem Biophys Res Commun. 2017; 488 (3): 528-533.
Moritani Y., Usui M., Sano K., Nakazawa K., Hanatani T., Nakatomi M., Iwata T., Sato T., Ariyoshi W., Nishihara T., Nakashima K. Spheroid culture enhances osteogenic potential of periodontal ligament mesenchymal stem cells. J Periodontal Res. 2018; 53 (5): 870-882.
Suenaga H., Furukawa K.S., Suzuki Y., Takato T., Ushida T. Bone regeneration in calvarial defects in a rat model by implantation of human bone marrow-derived mesenchymal stromal cell spheroids. J Mater Sci Mater Med. 2015; 26 (11): 254.
Yamaguchi Y., Ohno J., Sato A., Kido H., Fukushima T. Mesenchymal stem cell spheroids exhibit enhanced in-vitro and in-vivo osteoregenerative potential. BMC Biotechnol. 2014; 14:105.
Imamura A., Kajiya H., Fujisaki S., Maeshiba M., Yanagi T., Kojima H., Ohno J. Corrigendum to "Three-dimensional spheroids of mesenchymal stem/stromal cells promote osteogenesis by activating stemness and Wnt/β-catenin" [Biochem Biophys Res
Commun. 2020 523 (2) 458-464]. Biochem Biophys Res Commun. 2020; 525 (3): 819-820.
Walser R., Metzger W., Görg A., Pohlemann T., Menger M.D., Laschke M.W. Generation of co-culture spheroids as vascularization units for bone tissue engineering. Eur Cells Mater. 2013; 26: 222-233.
Tiaden A.N., Breiden M., Mirsaidi A., Weber F.A., Bahrenberg G., Glanz S., Cinelli P., Ehrmann M., Richards P.J. Human serine protease HTRA1 positively regulates osteogenesis of human bone marrow-derived mesenchymal stem cells and mineralization of differentiating bone-forming cells through the modulation of extracellular matrix protein. Stem Cells. 2012; 30 (10): 2271-2282.
Rumiński S., Kalaszczyńska I., Lewandowska-Szumieł M. Effect of cAMP signaling regulation in osteogenic differentiation of adipose-derived mesenchymal stem cells. Cells. 2020; 9 (7): 1587.
Yamamoto M., Kawashima N., Takashino N., Koizumi Y., Takimoto K., Suzuki N., Saito M., Suda H. Three-dimensional spheroid culture promotes odonto/osteoblastic differentiation of dental pulp cells. Arch Oral Biol. 2014; 59 (3): 310-317.
Kamoya T., Anada T., Shiwaku Y., Takano-Yamamoto T., Suzuki O. An oxygen-permeable spheroid culture chip (Oxy chip) promotes osteoblastic differentiation of mesenchymal stem cells. Sens. Actuators B Chem. 2016; 232: 75-83.
Saiz A.M. Jr., Gionet-Gonzales M.A., Lee M.A., Leach J.K. Conditioning of myoblast secretome using mesenchymal stem/stromal cell spheroids improves bone repair. Bone. 2019; 125: 151-159.
Fu C., Luo D., Yu M., Jiang N., Liu D., He D., Fu Y., Zhang T., Qiao Y., Zhou Y., Liu Y. Embryonic-like mineralized extracellular matrix/stem cell microspheroids as a bone graft substitute. Adv Healthc Mater. 2018; 7 (19): 1800705.
Murphy K.C., Hoch A.I., Harvestine J.N., Zhou D., Leach J.K. Mesenchymal stem cell spheroids retain osteogenic phenotype through α2β1 signaling. Stem Cells Transl Med. 2016; 5 (9): 1229-1237.
Baraniak P.R., McDevitt T.C. Scaffold-free culture of mesenchymal stem cell spheroids in suspension preserves multilineage potential. Cell Tissue Res. 2012; 347 (3): 701-711.
Gurumurthy B., Bierdeman P.C., Janorkar A.V. Spheroid model for functional osteogenic evaluation of human adipose derived stem cells. J Biomed Mater Res - Part A. 2017; 105 (4): 1230-1236.
Rumiński S., Kalaszczyńska I., Długosz A., Lewandowska-Szumieł M. Osteogenic differentiation of human adipose-derived stem cells in 3D conditions - comparison of spheroids and polystyrene scaffolds. Eur Cell Mater. 2019; 37: 382-401.
Ravi M., Paramesh V., Kaviya S.R., Anuradha E., Paul Solomon F.D. 3D cell culture systems: Advantages and applications. J Cell Physiol. 2015; 230 (1): 16-26.
Nath S.C., Horie M., Nagamori E., Kino-Oka M. Size- and time-dependent growth properties of human induced pluripotent stem cells in the culture of single aggregate. J Biosci Bioeng. 2017; 124 (4): 469-475.
Langan L.M., Dodd N.J.F., Owen S.F., Purcell W.M., Jackson S.K., Jha A.N. Correction: Direct measurements of oxygen gradients in spheroid culture system using electron paramagnetic resonance oximetry. PLoS One. 2016; 11 (8): e0149492.
Yanagihara K., Uchida S., Ohba S., Kataoka K., Itaka K. Treatment of bone defects by transplantation of genetically modified mesenchymal stem cell spheroids. Mol Ther Methods Clin Dev. 2018; 9: 358-366.
Chitturi Suryaprakash R.T., Kujan O., Shearston K., Farah C.S. Three-dimensional cell culture models to investigate oral carcinogenesis: a scoping review. Int J Mol Sci. 2020; 21 (24): 9520.
Vinci M., Box C., Eccles S.A. Three-dimensional (3D) tumor spheroid invasion assay. J Vis Exp. 2015; (99): e52686.
Lee S.J., Lee H.A. Trends in the development of human stem cell-based non-animal drug testing models. Korean J Physiol Pharmacol. 2020; 24 (6): 441-452.
Hamilton G., Rath B.. Applicability of tumor spheroids for in vitro chemosensitivity assays. Expert Opin Drug Metab Toxicol. 2019; 15(1): 15-23.
Ong C.S., Fukunishi T., Zhang H., Huang C.Y., Nashed A., Blazeski A., DiSilvestre D., Vricella L., Conte J., Tung L., Tomaselli G.F., Hibino N. Biomaterial-free three-dimensional bioprinting of cardiac tissue using human induced pluripotent stem cell derived cardiomyocytes. Sci Rep. 2017; 7 (1): 4566.
Zubillaga V., Alonso-Varona A., Fernandes S.C.M., Salaberria A.M., Palomares T. Adipose-derived mesenchymal stem cell chondrospheroids cultured in hypoxia and a 3D porous chitosan/chitin nanocrystal scaffold as a platform for cartilage tissue engineering. Int J Mol Sci 2020; 21(3): 1004.
Trohatou O., Roubelakis M.G. Mesenchymal stem/stromal cells in regenerative medicine: past, present, and future. Cell Reprogram. 2017; 19 (4): 217-224.
Melton D. Stemness: definitions, criteria, and standards. In: Essentials of Stem Cell Biology: Third Edition. Elsevier Inc.; 2014.
Lastra M.L., Gómez Ribelles J.L., Cortizo A.M. Design and characterization of microspheres for a 3D mesenchymal stem cell culture. Colloids Surf B Biointerfaces. 2020; 196: 111322.
Xu P., Jiang F., Zhang H., Yin R., Cen L., Zhang W. Calcium carbonate/gelatin methacrylate microspheres for 3D cell culture in bone tissue engineering. Tissue Eng Part C Methods. 2020; 26 (8): 418-432.