A 3D-printed molybdenum-containing scaffold exerts dual pro-osteogenic and anti-osteoclastogenic effects to facilitate alveolar bone repair

Abstract

The positive regulation of bone-forming osteoblast activity and the negative feedback regulation of osteoclastic activity are equally important in strategies to achieve successful alveolar bone regeneration. Here, a molybdenum (Mo)-containing bioactive glass ceramic scaffold with solid-strut-packed structures (Mo-scaffold) was printed, and its ability to regulate pro-osteogenic and anti-osteoclastogenic cellular responses was evaluated in vitro and in vivo. We found that extracts derived from Mo-scaffold (Mo-extracts) strongly stimulated osteogenic differentiation of bone marrow mesenchymal stem cells and inhibited differentiation of osteoclast progenitors. The identified comodulatory effect was further demonstrated to arise from Mo ions in the Mo-extract, wherein Mo ions suppressed osteoclastic differentiation by scavenging reactive oxygen species (ROS) and inhibiting mitochondrial biogenesis in osteoclasts. Consistent with the in vitro findings, the Mo-scaffold was found to significantly promote osteoblast-mediated bone formation and inhibit osteoclast-mediated bone resorption throughout the bone healing process, leading to enhanced bone regeneration. In combination with our previous finding that Mo ions participate in material-mediated immunomodulation, this study offers the new insight that Mo ions facilitate bone repair by comodulating the balance between bone formation and resorption. Our findings suggest that Mo ions are multifunctional cellular modulators that can potentially be used in biomaterial design and bone tissue engineering.

References

[1] Pedrero SG, Llamas-Sillero P, Serrano-López J. A multidisciplinary journey towards bone tissue engineering. Mater. (Basel), 2021, 14: 4896.

[2] Roohani I, Yeo GC, Mithieux SM, Weiss AS. Emerging concepts in bone repair and the premise of soft materials. Curr. Opin. Biotechnol., 2022, 74: 220-229.

[3] Herbert BA, Novince CM, Kirkwood KL. Aggregatibacter actinomycetemcomitans, a potent immunoregulator of the periodontal host defense system and alveolar bone homeostasis. Mol. Oral. Microbiol., 2016, 31: 207-227.

[4] Weng Y, . Trem2 mediated Syk-dependent ROS amplification is essential for osteoclastogenesis in periodontitis microenvironment. Redox Biol., 2021, 40: 101849.

[5] Zhou J, . Biomaterials and nanomedicine for bone regeneration: progress and future prospects. Exploration, 2021, 1: 20210011.

[6] Zhang Z, . Emerging biomimetic nanotechnology in orthopedic diseases: progress, challenges, and opportunities. Trends Chem., 2022, 4: 420-436.

[7] Bi CS, . The relationship between T-helper cell polarization and the RANKL/OPG ratio in gingival tissues from chronic periodontitis patients. Clin. Exp. Dent. Res., 2019, 5: 377-388.

[8] Bi CS, . Calcitriol inhibits osteoclastogenesis in an inflammatory environment by changing the proportion and function of T helper cell subsets (Th2/Th17). Cell Prolif., 2020, 53: e12827.

[9] Zhou Y, Wu C, Chang J. Bioceramics to regulate stem cells and their microenvironment for tissue regeneration. Mater. Today, 2019, 24: 41-56.

[10] He XT, . The effects of conditioned media generated by polarized macrophages on the cellular behaviours of bone marrow mesenchymal stem cells. J. Cell. Mol. Med., 2018, 22: 1302-1315.

[11] He XT, . Macrophage involvement affects matrix stiffness-related influences on cell osteogenesis under three-dimensional culture conditions. Acta Biomater., 2018, 71: 132-147.

[12] Woo HN, Cho YJ, Tarafder S, Lee CH. The recent advances in scaffolds for integrated periodontal regeneration. Bioact. Mater., 2021, 6: 3328-3342.

[13] Vaquette C, . Tissue engineered constructs for periodontal regeneration: current status and future perspectives. Adv. Healthc. Mater., 2018, 7: e1800457.

[14] Chen X, . Enhanced bone regeneration via spatiotemporal and controlled delivery of a genetically engineered BMP-2 in a composite Hydrogel. Biomaterials, 2021, 277: 121117.

[15] Chen F, . Synergy effects of Asperosaponin VI and bioactive factor BMP-2 on osteogenesis and anti-osteoclastogenesis. Bioact. Mater., 2021, 10: 335-344.

[16] Liu S, Wang YN, Yu L, Li J, Ge S. Development of a thermosensitive hydrogel loaded with DTT and SDF-1 facilitating in situ periodontal bone regeneration. Chem. Eng. J., 2022, 432: 134308.

[17] Chen FM, An Y, Zhang R, Zhang M. New insights into and novel applications of release technology for periodontal reconstructive therapies. J. Control. Release, 2011, 149: 92-110.

[18] Chu M, . Bi-directional regulation functions of lanthanum-substituted layered double hydroxide nanohybrid scaffolds via activating osteogenesis and inhibiting osteoclastogenesis for osteoporotic bone regeneration. Theranostics, 2021, 11: 6717-6734.

[19] Zhu H, Zheng K, Boccaccini AR. Multi-functional silica-based mesoporous materials for simultaneous delivery of biologically active ions and therapeutic biomolecules. Acta Biomater., 2021, 129: 1-17.

[20] Lee NH, . Dual actions of osteoclastic-inhibition and osteogenic-stimulation through strontium-releasing bioactive nanoscale cement imply biomaterial-enabled osteoporosis therapy. Biomaterials, 2021, 276: 121025.

[21] Li J, . Mn-containing bioceramics inhibit osteoclastogenesis and promote osteoporotic bone regeneration via scavenging ROS. Bioact. Mater., 2021, 6: 3839-3850.

[22] Li Z, . Ca(2+)-supplying black phosphorus-based scaffolds fabricated with microfluidic technology for osteogenesis. Bioact. Mater., 2021, 6: 4053-4064.

[23] Schwarz G, Mendel RR, Ribbe MW. Molybdenum cofactors, enzymes and pathways. Nature, 2009, 460: 839-847.

[24] Schwarz G. Molybdenum cofactor biosynthesis and deficiency. Cell. Mol. Life Sci., 2005, 62: 2792-2810.

[25] Schwarz G, Belaidi AA. Molybdenum in human health and disease. Met. Ions Life Sci., 2013, 13: 415-450.

[26] Kyomoto M, . High lubricious surface of cobalt-chromium-molybdenum alloy prepared by grafting poly(2-methacryloyloxyethyl phosphorylcholine). Biomaterials, 2007, 28: 3121-3130.

[27] Qin L, Zeng Q, Wang W, Zhang Y, Dong G. Response of MC3T3-E1 osteoblast cells to the microenvironment produced on Co–Cr–Mo alloy using laser surface texturing. J. Mater. Sci., 2014, 49: 2662-2671.

[28] Dang W, . 3D printing of Mo-containing scaffolds with activated anabolic responses and bi-lineage bioactivities. Theranostics, 2018, 8: 4372-4392.

[29] He XT, . Role of molybdenum in material immunomodulation and periodontal wound healing: targeting immunometabolism and mitochondrial function for macrophage modulation. Biomaterials, 2022, 283: 121439.

[30] Ishii K-A, . Coordination of PGC-1β and iron uptake in mitochondrial biogenesis and osteoclast activation. Nat. Med., 2009, 15: 259-266.

[31] Zheng ZG, . Dual targeting of SREBP2 and ERRα by carnosic acid suppresses RANKL-mediated osteoclastogenesis and prevents ovariectomy-induced bone loss. Cell Death Differ., 2020, 27: 2048-2065.

[32] Tonetti MS, Jepsen S, Jin L, Otomo-Corgel J. Impact of the global burden of periodontal diseases on health, nutrition and wellbeing of mankind: a call for global action. J. Clin. Periodontol., 2017, 44: 456-462.

[33] Genco RJ, Sanz M. Clinical and public health implications of periodontal and systemic diseases: an overview. Periodontol 2000, 2020, 83: 7-13.

[34] Helal O, . Predictors for tooth loss in periodontitis patients: systematic review and meta-analysis. J. Clin. Periodontol., 2019, 46: 699-712.

[35] Ravida A, . The influence of the interaction between staging, grading and extent on tooth loss due to periodontitis. J. Clin. Periodontol., 2021, 48: 648-658.

[36] Xu XY, . Concise review: periodontal tissue regeneration using stem cells: strategies and translational considerations. Stem Cells Transl. Med, 2019, 8: 392-403.

[37] Mo Y, . Local delivery of naringin in beta-cyclodextrin modified mesoporous bioactive glass promotes bone regeneration: from anti-inflammatory to synergistic osteogenesis and osteoclastogenesis. Biomater. Sci., 2022, 10: 1697-1712.

[38] Wei H, Cui J, Lin K, Xie J, Wang X. Recent advances in smart stimuli-responsive biomaterials for bone therapeutics and regeneration. Bone Res, 2022, 10: 17.

[39] Holstein SA. A patent review of bisphosphonates in treating bone disease. Expert Opin. Ther. Pat., 2019, 29: 315-325.

[40] Miller PD, Derman RJ. What is the best balance of benefits and risks among anti-resorptive therapies for postmenopausal osteoporosis?. Osteoporos. Int., 2010, 21: 1793-1802.

[41] Place ES, Evans ND, Stevens MM. Complexity in biomaterials for tissue engineering. Nat. Mater., 2009, 8: 457-470.

[42] Chen Q, . Progranulin promotes regeneration of inflammatory periodontal bone defect in rats via anti-inflammation, osteoclastogenic inhibition, and osteogenic promotion. Inflammation, 2019, 42: 221-234.

[43] Tan J, . Sustained release of two bioactive factors from supramolecular hydrogel promotes periodontal bone regeneration. ACS Nano, 2019, 13: 5616-5622.

[44] Liang Q, Du L, Zhang R, Kang W, Ge S. Stromal cell-derived factor-1/Exendin-4 cotherapy facilitates the proliferation, migration and osteogenic differentiation of human periodontal ligament stem cells in vitro and promotes periodontal bone regeneration in vivo. Cell Prolif., 2021, 54: e12997.

[45] Zhang X, . Nanoscale materials-based platforms for the treatment of bone-related diseases. Matter, 2021, 4: 2727-2764.

[46] Zhang X, . Electroactive electrospun nanofibers for tissue engineering. Nano Today, 2021, 39: 101196.

[47] Detsch R, Boccaccini AR. The role of osteoclasts in bone tissue engineering. J. Tissue Eng. Regen. Med., 2015, 9: 1133-1149.

[48] Tuckermann J, Adams RH. The endothelium-bone axis in development, homeostasis and bone and joint disease. Nat. Rev. Rheumatol., 2021, 17: 608-620.

[49] Mao L, . The synergistic effects of Sr and Si bioactive ions on osteogenesis, osteoclastogenesis and angiogenesis for osteoporotic bone regeneration. Acta Biomater., 2017, 61: 217-232.

[50] Kim H, . Selenoprotein W ensures physiological bone remodeling by preventing hyperactivity of osteoclasts. Nat. Commun., 2021, 12

[51] Zhou Y, . Cytokine-scavenging nanodecoys reconstruct osteoclast/osteoblast balance toward the treatment of postmenopausal osteoporosis. Sci. Adv., 2021, 7: eabl6432.

[52] Bhattacharyya A, . Bioink homogeneity control during 3D bioprinting of multicomponent micro/nanocomposite hydrogel for even tissue regeneration using novel twin screw extrusion system. Chem. Eng. J., 2021, 415: 128971.

[53] Vallet-Regi M, Ruiz-Hernandez E. Bioceramics: from bone regeneration to cancer nanomedicine. Adv. Mater., 2011, 23: 5177-5218.

[54] Zheng K, Niu W, Lei B, Boccaccini AR. Immunomodulatory bioactive glasses for tissue regeneration. Acta Biomater., 2021, 133: 168-186.

[55] He F, . Novel extrusion-microdrilling approach to fabricate calcium phosphate-based bioceramic scaffolds enabling fast bone regeneration. ACS Appl. Mater. Interfaces, 2020, 12: 32340-32351.

[56] Qin C, . 3D bioprinting of multicellular scaffolds for osteochondral regeneration. Mater. Today, 2021, 49: 68-84.

[57] Awad K, . Ionic silicon protects oxidative damage and promotes skeletal muscle cell regeneration. Int. J. Mol. Sci., 2021, 22: 497.

[58] Gu H, . The stimulation of osteogenic differentiation of human adipose-derived stem cells by ionic products from akermanite dissolution via activation of the ERK pathway. Biomaterials, 2011, 32: 7023-7033.

[59] Xing M, Wang X, Wang E, Gao L, Chang J. Bone tissue engineering strategy based on the synergistic effects of silicon and strontium ions. Acta Biomater., 2018, 72: 381-395.

[60] Bai J, . Biomimetic osteogenic peptide with mussel adhesion and osteoimmunomodulatory functions to ameliorate interfacial osseointegration under chronic inflammation. Biomaterials, 2020, 255: 120197.

[61] Zhong Z, . Zn/Sr dual ions-collagen co-assembly hydroxyapatite enhances bone regeneration through procedural osteo-immunomodulation and osteogenesis. Bioact. Mater., 2021, 10: 195-206.

[62] Casarrubios L, . Silicon substituted hydroxyapatite/VEGF scaffolds stimulate bone regeneration in osteoporotic sheep. Acta Biomater., 2020, 101: 544-553.

[63] Park-Min K-H. Metabolic reprogramming in osteoclasts. Semin. Immunopathol., 2019, 41: 565-572.

[64] Zhang Y, . PGC1β organizes the osteoclast cytoskeleton by mitochondrial biogenesis and activation. J. Bone Miner. Res., 2018, 33: 1114-1125.

[65] Lemma S, . Energy metabolism in osteoclast formation and activity. Int. J. Biochem. Cell Biol., 2016, 79: 168-180.

[66] Izawa T, . ASXL2 regulates glucose, lipid, and skeletal homeostasis. Cell Rep., 2015, 11: 1625-1637.

[67] He Y, . Nanoporous titanium implant surface promotes osteogenesis by suppressing osteoclastogenesis via integrin beta1/FAKpY397/MAPK pathway. Bioact. Mater., 2021, 8: 109-123.

[68] Chen Z, . Osteoimmunomodulation for the development of advanced bone biomaterials. Mater. Today, 2016, 19: 304-321.

[69] Wang Z, . Autophagy mediated CoCrMo particle-induced peri-implant osteolysis by promoting osteoblast apoptosis. Autophagy, 2015, 11: 2358-2369.

[70] Chen FM, Zhang M, Wu ZF. Toward delivery of multiple growth factors in tissue engineering. Biomaterials, 2010, 31: 6279-6308.

[71] Donos N, Park JC, Vajgel A, de Carvalho Farias B, Dereka X. Description of the periodontal pocket in preclinical models: limitations and considerations. Periodontol 2000, 2018, 76: 16-34.

[72] He XT, . Suppression of histone deacetylases by SAHA relieves bone cancer pain in rats via inhibiting activation of glial cells in spinal dorsal horn and dorsal root ganglia. J. Neuroinflammation, 2020, 17

[73] Hu CH, . Sympathetic neurostress drives osteoblastic exosomal MiR-21 transfer to disrupt bone homeostasis and promote osteopenia. Small Methods, 2022, 6: e2100763.

[74] Li X, . M2 macrophages enhance the cementoblastic differentiation of periodontal ligament stem cells via the Akt and JNK pathways. Stem Cells, 2019, 37: 1567-1580.

[75] Li X, . LncRNA GACAT2 binds with protein PKM1/2 to regulate cell mitochondrial function and cementogenesis in an inflammatory environment. Bone Res, 2022, 10: 29.

Funding

National Natural Science Foundation of China (National Science Foundation of China)(82130026, 82170958, 82001102)

Natural Science Foundation of Shaanxi Province (Shaanxi Province Natural Science Foundation)(2020JQ-447)