Sustained zinc release in cooperation with CaP scaffold promoted bone regeneration via directing stem cell fate and triggering a pro-healing immune stimuli

Journal of Nanobiotechnology - Tập 19 Số 1 - 2021
Xin Huang1, Danfeng Huang1, Ting Zhu2, Xiaohua Yu1, Kaicheng Xu1, Hengyuan Li1, Hao Qu1, Zhiyuan Zhou3, Kui Cheng3, Wenjian Wen3, Zhaoming Ye1
1Department of Orthopedics, Musculoskeletal Tumor Center, The Second Affiliated Hospital of Zhejiang University School of Medicine, 88# Jiefang Road, Hangzhou, 310009, China
2Department of Thoracic Surgery, Shaoxing People’s Hospital, Shaoxing Hospital, Zhejiang University School of Medicine, No. 568 Zhongxing North Road, Yuecheng District, Shaoxing, 312000, China
3School of Materials Science and Engineering, Zhejiang University, Hangzhou 310027, China

Tóm tắt

AbstractMetal ions have been identified as important bone metabolism regulators and widely used in the field of bone tissue engineering, however their exact role during bone regeneration remains unclear. Herein, the aim of study was to comprehensively explore the interactions between osteoinductive and osteo-immunomodulatory properties of these metal ions. In particular, the osteoinductive role of zinc ions (Zn2+), as well as its interactions with local immune microenvironment during bone healing process, was investigated in this study using a sustained Zn2+ delivery system incorporating Zn2+ into β-tricalcium phosphate/poly(L-lactic acid) (TCP/PLLA) scaffolds. The presence of Zn2+ largely enhanced osteogenic differentiation of periosteum-derived progenitor cells (PDPCs), which was coincident with increased transition from M1 to M2 macrophages (M$$\varphi $$ φ s). We further confirmed that induction of M2 polarization by Zn2+ was realized via PI3K/Akt/mTOR pathway, whereas marker molecules on this pathway were strictly regulated by the addition of Zn2+. Synergically, this favorable immunomodulatory effect of Zn2+ further improved the osteogenic differentiation of PDPCs induced by Zn2+ in vitro. Consistently, the spontaneous osteogenesis and pro-healing osteoimmunomodulation of the scaffolds were thoroughly identified in vivo using a rat air pouch model and a calvarial critical-size defect model. Taken together, Zn2+-releasing bioactive ceramics could be ideal scaffolds in bone tissue engineering due to their reciprocal interactions between osteoinductive and immunomodulatory characteristics. Clarification of this synergic role of Zn2+ during osteogenesis could pave the way to develop more sophisticated metal-ion based orthopedic therapeutic strategies.

Từ khóa


Tài liệu tham khảo

Giannoudis PV, Harwood PJ, Tosounidis T, Kanakaris NK. Restoration of long bone defects treated with the induced membrane technique: protocol and outcomes. Injury. 2016;47(Suppl 6):S53-s61.

Yu Y, Wang Y, Zhang W, Wang H, Li J, Pan L, Han F, Li B. Biomimetic periosteum-bone substitute composed of preosteoblast-derived matrix and hydrogel for large segmental bone defect repair. Acta Biomater. 2020;113:317–27.

Zhou M, Yang X, Li S, Kapat K, Guo K, Perera FH, Qian L, Miranda P, Che Y. Bioinspired channeled, rhBMP-2-coated β-TCP scaffolds with embedded autologous vascular bundles for increased vascularization and osteogenesis of prefabricated tissue-engineered bone. Materials Biol Appl. 2021;118:111389.

Campana V, Milano G, Pagano E, Barba M, Cicione C, Salonna G, Lattanzi W, Logroscino G. Bone substitutes in orthopaedic surgery: from basic science to clinical practice. J Mater Sci Mater Med. 2014;25(10):2445–61.

Zhang M, Matinlinna JP, Tsoi JKH, Liu W, Cui X, Lu WW, Pan H. Recent developments in biomaterials for long-bone segmental defect reconstruction: A narrative overview. J Orthop Transl. 2020;22:26–33.

Koons GL, Diba M, Mikos AG. Materials design for bone-tissue engineering. Nat Rev Mater. 2020;5(8):584–603.

Martinez-Zelaya VR, Zarranz L, Herrera EZ, Alves AT, Uzeda MJ, Mavropoulos E, Rossi AL, Mello A, Granjeiro JM, Calasans-Maia MD, et al. In vitro and in vivo evaluations of nanocrystalline Zn-doped carbonated hydroxyapatite/alginate microspheres: zinc and calcium bioavailability and bone regeneration. Int J Nanomed. 2019;14:3471–90.

Xia Y, Fan X, Yang H, Li L, He C, Cheng C, Haag R. ZnO/Nanocarbons-modified fibrous scaffolds for stem cell-based osteogenic differentiation. Small (Weinheim an der Bergstrasse, Germany). 2020;16(38):e2003010.

Chandramohan Y, Jeganathan K, Sivanesan S, Koka P, Amritha TMS, Vimalraj S, Dhanasekaran A. Assessment of human ovarian follicular fluid derived mesenchymal stem cells in chitosan/PCL/Zn scaffold for bone tissue regeneration. Life Sci. 2021;264:118502.

Li P, Dai J, Schweizer E, Rupp F, Heiss A, Richter A, Klotz UE, Geis-Gerstorfer J, Scheideler L, Alexander D. Response of human periosteal cells to degradation products of zinc and its alloy. Mater Biol Appl. 2020;108:110208.

Bose S, Sarkar N, Vahabzadeh S. Sustained release of vitamin C from PCL coated TCP induces proliferation and differentiation of osteoblast cells and suppresses osteosarcoma cell growth. Mater Biol Appl. 2019;105:110096.

Dinarvand P, Seyedjafari E, Shafiee A, Jandaghi AB, Doostmohammadi A, Fathi MH, Farhadian S, Soleimani M. New approach to bone tissue engineering: simultaneous application of hydroxyapatite and bioactive glass coated on a poly(L-lactic acid) scaffold. ACS Appl Mater Inter. 2011;3(11):4518–24.

Best SM, Porter AE, Thian ES, Huang J. Bioceramics: Past, present and for the future. J Eur Ceram Soc. 2008;28(7):1319–27.

Feng P, Wu P, Gao C, Yang Y, Guo W, Yang W, Shuai C. A Multimaterial Scaffold With Tunable Properties: Toward Bone Tissue Repair. Adv Sci (Weinh). 2018;5(6):1700817.

Huang Q, Liu Y, Ouyang Z, Feng Q. Comparing the regeneration potential between PLLA/Aragonite and PLLA/Vaterite pearl composite scaffolds in rabbit radius segmental bone defects. Bioactive Mater. 2020;5(4):980–9.

Ge M, Xue L, Nie T, Ma H, Zhang J. The precision structural regulation of PLLA porous scaffold and its influence on the proliferation and differentiation of MC3T3-E1 cells. J Biomater Sci Polym Ed. 2016;27(17):1685–97.

Li B, Chen Y, He J, Zhang J, Wang S, Xiao W, Liu Z, Liao X. Biomimetic Membranes of Methacrylated Gelatin/Nanohydroxyapatite/Poly(l-Lactic Acid) for Enhanced Bone Regeneration. ACS Biomater Sci Eng. 2020;6(12):6737–47.

Abdelhamid HN, Dowaidar M, Hällbrink M, Langel Ü. Gene delivery using cell penetrating peptides-zeolitic imidazolate frameworks. Microporous Mesoporous Mater. 2020;300:110173.

Abdelhamid HN, Dowaidar M, Langel Ü. Carbonized chitosan encapsulated hierarchical porous zeolitic imidazolate frameworks nanoparticles for gene delivery. Microporous Mesoporous Mater. 2020;302:110200.

Lee J, Byun H, Madhurakkat Perikamana SK, Lee S, Shin H. Current Advances in Immunomodulatory Biomaterials for Bone Regeneration. Advanced healthcare materials. 2019;8(4):1e1801106.

He J, Chen G, Liu M, Xu Z, Chen H, Yang L, Lv Y. Scaffold strategies for modulating immune microenvironment during bone regeneration. Mater Biol Appl. 2020;108:110411.

Tiffany AS, Gray DL, Woods TJ, Subedi K, Harley BAC. The inclusion of zinc into mineralized collagen scaffolds for craniofacial bone repair applications. Acta Biomater. 2019;93:86–96.

Wang X, Chu W, Zhuang Y, Shi D, Tao H, Jin C, Dai K, Zhao J, Gan Y. Bone mesenchymal stem cell-enriched β-tricalcium phosphate scaffold processed by the screen-enrich-combine circulating system promotes regeneration of diaphyseal bone non-union. Cell Transplant. 2019;28(2):212–23.

Pereira RC, Benelli R, Canciani B, Scaranari M, Daculsi G, Cancedda R, Gentili C. Beta-tricalcium phosphate ceramic triggers fast and robust bone formation by human mesenchymal stem cells. J Tissue Eng Regen Med. 2019;13(6):1007–18.

Niu Y, Wang Z, Shi Y, Dong L, Wang C. Modulating macrophage activities to promote endogenous bone regeneration: Biological mechanisms and engineering approaches. Bioactive Mater. 2021;6(1):244–61.

Jamalpoor Z, Asgari A, Lashkari MH, Mirshafiey A, Mohsenzadegan M. Modulation of macrophage polarization for bone tissue engineering applications. Iran J Allergy Asthma Immunol. 2018;17(5):398–408.

Sadowska JM, Ginebra MP. Inflammation and biomaterials: role of the immune response in bone regeneration by inorganic scaffolds. J Mater Chem B. 2020;8(41):9404–27.

Zhu WQ, Shao SY, Xu LN, Chen WQ, Yu XY, Tang KM, Tang ZH, Zhang FM, Qiu J. Enhanced corrosion resistance of zinc-containing nanowires-modified titanium surface under exposure to oxidizing microenvironment. J Nanobiotechnol. 2019;17(1):55.

Chen B, You Y, Ma A, Song Y, Jiao J, Song L, Shi E, Zhong X, Li Y, Li C. Zn-Incorporated TiO(2) Nanotube Surface Improves Osteogenesis Ability Through Influencing Immunomodulatory Function of Macrophages. Int J Nanomed. 2020;15:2095–118.

Toledano M, Toledano-Osorio M, Osorio R, Carrasco-Carmona Á, Gutiérrez-Pérez JL, Gutiérrez-Corrales A, Serrera-Figallo MA, Lynch CD, Torres-Lagares D: Doxycycline and Zinc Loaded Silica-Nanofibrous Polymers as Biomaterials for Bone Regeneration. Polymers 2020, 12(5).

Kido T, Ishiwata K, Suka M, Yanagisawa H. Inflammatory response under zinc deficiency is exacerbated by dysfunction of the T helper type 2 lymphocyte-M2 macrophage pathway. Immunology. 2019;156(4):356–72.

Higashimura Y, Takagi T, Naito Y, Uchiyama K, Mizushima K, Tanaka M, Hamaguchi M, Itoh Y. Zinc deficiency activates the IL-23/Th17 axis to aggravate experimental colitis in mice. J Crohns Colitis. 2020;14(6):856–66.

Jia X, Xu H, Miron RJ, Yin C, Zhang X, Wu M, Zhang Y. EZH1 Is Associated with TCP-induced bone regeneration through macrophage polarization. Stem Cells Int. 2018;2018:6310560.

Chen Z, Wu C, Gu W, Klein T, Crawford R, Xiao Y. Osteogenic differentiation of bone marrow MSCs by β-tricalcium phosphate stimulating macrophages via BMP2 signalling pathway. Biomaterials. 2014;35(5):1507–18.

Li M, Guo X, Qi W, Wu Z, de Bruijn JD, Xiao Y, Bao C, Yuan H. Macrophage polarization plays roles in bone formation instructed by calcium phosphate ceramics. J Mater Chem B. 2020;8(9):1863–77.

Pajarinen J, Lin T, Gibon E, Kohno Y, Maruyama M, Nathan K, Lu L, Yao Z, Goodman SB. Mesenchymal stem cell-macrophage crosstalk and bone healing. Biomaterials. 2019;196:80–9.

Muñoz J, Akhavan NS, Mullins AP, Arjmandi BH. Macrophage Polarization and Osteoporosis: A Review. Nutrients 2020, 12(10).

Miao S, Cheng K, Weng W, Du P, Shen G, Han G, Yan W, Zhang S. Fabrication and evaluation of Zn containing fluoridated hydroxyapatite layer with Zn release ability. Acta Biomater. 2008;4(2):441–6.

Huang X, Yang D, Yan W, Shi Z, Feng J, Gao Y, Weng W, Yan S. Osteochondral repair using the combination of fibroblast growth factor and amorphous calcium phosphate/poly(L-lactic acid) hybrid materials. Biomaterials. 2007;28(20):3091–100.

Lai Y, Li Y, Cao H, Long J, Wang X, Li L, Li C, Jia Q, Teng B, Tang T, et al. Osteogenic magnesium incorporated into PLGA/TCP porous scaffold by 3D printing for repairing challenging bone defect. Biomaterials. 2019;197:207–19.

Yamamoto A, Honma R, Sumita M. Cytotoxicity evaluation of 43 metal salts using murine fibroblasts and osteoblastic cells. J Biomed Mater Res. 1998;39(2):331–40.

Yang HC, Park HC, Quan H, Kim Y. Immunomodulation of Biomaterials by Controlling Macrophage Polarization. Adv Exp Med Biol. 2018;1064:197–206.

Yin S, Zhang W, Zhang Z, Jiang X. Recent advances in scaffold design and material for vascularized tissue-engineered bone regeneration. Adv Healthcare Mater. 2019;8(10):1801433.

Wan B, Wang R, Sun Y, Cao J, Wang H, Guo J, Chen D. Building osteogenic microenvironments with strontium-substituted calcium phosphate ceramics. Front Bioeng Biotechnol. 2020;8:591467.

Vavken J, Mameghani A, Vavken P, Schaeren S. Complications and cancer rates in spine fusion with recombinant human bone morphogenetic protein-2 (rhBMP-2). European spine journal : official publication of the European Spine Society, the European Spinal Deformity Society, and the European Section of the Cervical Spine Research Society. 2016;25(12):3979–89.

Halloran D, Durbano HW, Nohe A. Bone Morphogenetic Protein-2 in Development and Bone Homeostasis. J Develop Biol 2020, 8(3).

Bose S, Fielding G, Tarafder S, Bandyopadhyay A. Understanding of dopant-induced osteogenesis and angiogenesis in calcium phosphate ceramics. Trends Biotechnol. 2013;31(10):594–605.

Amin N, Clark CCT, Taghizadeh M, Djafarnejad S. Zinc supplements and bone health: The role of the RANKL-RANK axis as a therapeutic target. J Trace Elem Med Biol . 2020;57:126417.

Huang T, Yan G, Guan M. Zinc Homeostasis in Bone: Zinc Transporters and Bone Diseases. Int J Mol Sci 2020, 21(4).

O'Connor JP, Kanjilal D, Teitelbaum M, Lin SS, Cottrell JA. Zinc as a Therapeutic Agent in Bone Regeneration. Materials (Basel, Switzerland) 2020, 13(10).

Fernandes MH, Alves MM, Cebotarenco M, Ribeiro IAC, Grenho L, Gomes PS, Carmezim MJ, Santos CF. Citrate zinc hydroxyapatite nanorods with enhanced cytocompatibility and osteogenesis for bone regeneration. Mater Biol Appl. 2020;115:111147.

Chopra V, Thomas J, Sharma A, Panwar V, Kaushik S, Sharma S, Porwal K, Kulkarni C, Rajput S, Singh H, et al. Synthesis and evaluation of a zinc eluting rGO/hydroxyapatite nanocomposite optimized for bone augmentation. ACS Biomater Sci Eng. 2020;6(12):6710–25.

Sun TW, Yu WL, Zhu YJ, Chen F, Zhang YG, Jiang YY, He YH. Porous nanocomposite comprising ultralong hydroxyapatite nanowires decorated with zinc-containing nanoparticles and chitosan: synthesis and application in bone defect repair. Chemistry. 2018;24(35):8809–21.

Yu W, Sun TW, Qi C, Ding Z, Zhao H, Zhao S, Shi Z, Zhu YJ, Chen D, He Y. Evaluation of zinc-doped mesoporous hydroxyapatite microspheres for the construction of a novel biomimetic scaffold optimized for bone augmentation. Int J Nanomed. 2017;12:2293–306.

Bostancioglu RB, Gurbuz M, Akyurekli AG, Dogan A, Koparal AS, Koparal AT. Adhesion profile and differentiation capacity of human adipose tissue derived mesenchymal stem cells grown on metal ion (Zn, Ag and Cu) doped hydroxyapatite nano-coated surfaces. Colloids Surf, B. 2017;155:415–28.

Chou J, Hao J, Hatoyama H, Ben-Nissan B, Milthorpe B, Otsuka M. Effect of biomimetic zinc-containing tricalcium phosphate (Zn-TCP) on the growth and osteogenic differentiation of mesenchymal stem cells. J Tissue Eng Regen Med. 2015;9(7):852–8.

Luo X, Barbieri D, Davison N, Yan Y, de Bruijn JD, Yuan H. Zinc in calcium phosphate mediates bone induction: in vitro and in vivo model. Acta Biomater. 2014;10(1):477–85.

Cama G, Nkhwa S, Gharibi B, Lagazzo A, Cabella R, Carbone C, Dubruel P, Haugen H, Di Silvio L, Deb S. The role of new zinc incorporated monetite cements on osteogenic differentiation of human mesenchymal stem cells. Mater Sci Eng, C Mater Biol Appl. 2017;78:485–94.

Xiong K, Zhang J, Zhu Y, Chen L, Ye J. Zinc doping induced differences in the surface composition, surface morphology and osteogenesis performance of the calcium phosphate cement hydration products. Materials Biol Appl. 2019;105:110065.

Grotenhuis N, De Witte SF, van Osch GJ, Bayon Y, Lange JF, Bastiaansen-Jenniskens YM. Biomaterials influence macrophage-mesenchymal stem cell interaction in vitro. Tissue Eng Part A. 2016;22(17–18):1098–107.

Huang R, Wang X, Zhou Y, Xiao Y. RANKL-induced M1 macrophages are involved in bone formation. Bone Res. 2017;5:17019.

Holloway WR, Collier FM, Herbst RE, Hodge JM, Nicholson GC. Osteoblast-mediated effects of zinc on isolated rat osteoclasts: inhibition of bone resorption and enhancement of osteoclast number. Bone. 1996;19(2):137–42.

Zheng M, Weng M, Zhang X, Li R, Tong Q, Chen Z: Beta-tricalcium phosphate promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells through macrophages. Biomedical materials (Bristol, England) 2021.

Gu Y, Xie X, Zhuang R, Weir MD, Oates TW, Bai Y, Zhao L, Xu HHK: A Biphasic Calcium Phosphate Cement Enhances Dentin Regeneration by Dental Pulp Stem Cells and Promotes Macrophages M2 Phenotype In Vitro. Tissue engineering Part A 2021.

Gu Y, Zhuang R, Xie X, Bai Y. Osteogenic stimulation of human dental pulp stem cells with self-setting biphasic calcium phosphate cement. J Biomed Mater Res B Appl Biomater. 2020;108(4):1669–78.

Tai S, Cheng JY, Ishii H, Shimono K, Zangiacomi V, Satoh T, Hosono T, Suzuki E, Yamaguchi K, Maruyama K. Effects of beta-tricalcium phosphate particles on primary cultured murine dendritic cells and macrophages. Int Immunopharmacol. 2016;40:419–27.

Chen Z, Yuen J, Crawford R, Chang J, Wu C, Xiao Y. The effect of osteoimmunomodulation on the osteogenic effects of cobalt incorporated β-tricalcium phosphate. Biomaterials. 2015;61:126–38.

Fernandes KR, Zhang Y, Magri AMP, Renno ACM, van den Beucken J. Biomaterial property effects on platelets and macrophages: An in vitro study. ACS Biomater Sci Eng. 2017;3(12):3318–27.

Chen X, Wang M, Chen F, Wang J, Li X, Liang J, Fan Y, Xiao Y, Zhang X. Correlations between macrophage polarization and osteoinduction of porous calcium phosphate ceramics. Acta Biomater. 2020;103:318–32.

Gammoh NZ, Rink L: Zinc in Infection and Inflammation. Nutrients 2017, 9(6).

Su Y, Cockerill I, Wang Y, Qin YX, Chang L, Zheng Y, Zhu D. Zinc-based biomaterials for regeneration and therapy. Trends Biotechnol. 2019;37(4):428–41.

Jarosz M, Olbert M, Wyszogrodzka G, Młyniec K, Librowski T. Antioxidant and anti-inflammatory effects of zinc. Zinc-dependent NF-κB signaling Inflammopharmacology. 2017;25(1):11–24.

Choi S, Liu X, Pan Z. Zinc deficiency and cellular oxidative stress: prognostic implications in cardiovascular diseases. Acta Pharmacol Sin. 2018;39(7):1120–32.

Velard F, Laurent-Maquin D, Braux J, Guillaume C, Bouthors S, Jallot E, Nedelec JM, Belaaouaj A, Laquerriere P. The effect of zinc on hydroxyapatite-mediated activation of human polymorphonuclear neutrophils and bone implant-associated acute inflammation. Biomaterials. 2010;31(8):2001–9.

Kim MH, Jeong HJ. Zinc oxide nanoparticles suppress LPS-induced NF-κB activation by inducing A20, a negative regulator of NF-κB, in RAW 2647 Macrophages. J Nanosci Nanotechnol. 2015;15(9):6509–15.

Makumire S, Chakravadhanula VS, Köllisch G, Redel E, Shonhai A. Immunomodulatory activity of zinc peroxide (ZnO2) and titanium dioxide (TiO2) nanoparticles and their effects on DNA and protein integrity. Toxicol Lett. 2014;227(1):56–64.

Lin CD, Kou YY, Liao CY, Li CH, Huang SP, Cheng YW, Liao WC, Chen HX, Wu PL, Kang JJ, et al. Zinc oxide nanoparticles impair bacterial clearance by macrophages. Nanomedicine (Lond). 2014;9(9):1327–39.

Ilves M, Palomäki J, Vippola M, Lehto M, Savolainen K, Savinko T, Alenius H. Topically applied ZnO nanoparticles suppress allergen induced skin inflammation but induce vigorous IgE production in the atopic dermatitis mouse model. Part Fibre Toxicol. 2014;11:38.

Ogle ME, Segar CE, Sridhar S, Botchwey EA. Monocytes and macrophages in tissue repair: Implications for immunoregenerative biomaterial design. Exp Biol Med (Maywood). 2016;241(10):1084–97.

Liu W, Li J, Cheng M, Wang Q, Yeung KWK, Chu PK, Zhang X. Zinc-modified sulfonated polyetheretherketone surface with immunomodulatory function for guiding cell fate and bone regeneration. Adv Sci (Weinh). 2018;5(10):1800749.

Bai X, Liu W, Xu L, Ye Q, Zhou H, Berg C, Yuan H, Li J, Xia W. Sequential macrophage transition facilitates endogenous bone regeneration induced by Zn-doped porous microcrystalline bioactive glass. J Mater Chem B. 2021;9(12):2885–98.

Cerqueira A, Romero-Gavilán F, García-Arnáez I, Martinez-Ramos C, Ozturan S, Iloro I, Azkargorta M, Elortza F, Izquierdo R, Gurruchaga M, et al. Bioactive zinc-doped sol-gel coating modulates protein adsorption patterns and in vitro cell responses. Mater Biol Appl. 2021;121:111839.

Dierichs L, Kloubert V, Rink L. Cellular zinc homeostasis modulates polarization of THP-1-derived macrophages. Eur J Nutr. 2018;57(6):2161–9.

Sun W, Yang J, Wang W, Hou J, Cheng Y, Fu Y, Xu Z, Cai L. The beneficial effects of Zn on Akt-mediated insulin and cell survival signaling pathways in diabetes. J Trace Elem . 2018;46:117–27.

Shao Y, Wolf PG, Guo S, Guo Y, Gaskins HR, Zhang B. Zinc enhances intestinal epithelial barrier function through the PI3K/AKT/mTOR signaling pathway in Caco-2 cells. J Nutr Biochem. 2017;43:18–26.

Ohashi K, Nagata Y, Wada E, Zammit PS, Shiozuka M, Matsuda R. Zinc promotes proliferation and activation of myogenic cells via the PI3K/Akt and ERK signaling cascade. Exp Cell Res. 2015;333(2):228–37.

McClung JP, Tarr TN, Barnes BR, Scrimgeour AG, Young AJ. Effect of supplemental dietary zinc on the mammalian target of rapamycin (mTOR) signaling pathway in skeletal muscle and liver from post-absorptive mice. Biol Trace Elem Res. 2007;118(1):65–76.

Barthel A, Ostrakhovitch EA, Walter PL, Kampkötter A, Klotz LO. Stimulation of phosphoinositide 3-kinase/Akt signaling by copper and zinc ions: mechanisms and consequences. Arch Biochem Biophys. 2007;463(2):175–82.

Kim S, Jung Y, Kim D, Koh H, Chung J. Extracellular zinc activates p70 S6 kinase through the phosphatidylinositol 3-kinase signaling pathway. J Biol Chem. 2000;275(34):25979–84.

Tang X, Shay NF. Zinc has an insulin-like effect on glucose transport mediated by phosphoinositol-3-kinase and Akt in 3T3-L1 fibroblasts and adipocytes. J Nutr. 2001;131(5):1414–20.

Linton MF, Moslehi JJ, Babaev VR: Akt Signaling in Macrophage Polarization, Survival, and Atherosclerosis. Int J Mol Sci 2019, 20(11).

Vergadi E, Ieronymaki E, Lyroni K, Vaporidi K, Tsatsanis C. Akt signaling pathway in macrophage activation and M1/M2 Polarization. J Immunol. 2017;198(3):1006–14.

Sun JL, Jiao K, Niu LN, Jiao Y, Song Q, Shen LJ, Tay FR, Chen JH. Intrafibrillar silicified collagen scaffold modulates monocyte to promote cell homing, angiogenesis and bone regeneration. Biomaterials. 2017;113:203–16.

Vasconcelos DM, Gonçalves RM, Almeida CR, Pereira IO, Oliveira MI, Neves N, Silva AM, Ribeiro AC, Cunha C, Almeida AR, et al. Fibrinogen scaffolds with immunomodulatory properties promote in vivo bone regeneration. Biomaterials. 2016;111:163–78.