nach oben

Erschienen in:

10.07.2023 | Research

Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo

verfasst von: Yaru Zhu, Wen Wang, Qiyu Chen, Tianshui Ren, Jianguang Yang, Gan Li, Yanbin Qi, Changyong Yuan, Penglai Wang

Erschienen in: Clinical Oral Investigations | Ausgabe 9/2023

Einloggen, um Zugang zu erhalten

Abstract

Objectives

We aimed to explore the osteogenic potential of periodontal ligament stem cells (PDLSCs) in bioprinted methacrylate gelatine (GelMA) hydrogels in vitro and in vivo.

Materials and methods

PDLSCs in GelMA hydrogels at various concentrations (3%, 5%, and 10%) were bioprinted. The mechanical properties (stiffness, nanostructure, swelling, and degradation properties) of bioprinted constructs and the biological properties (cell viability, proliferation, spreading, osteogenic differentiation, and cell survival in vivo) of PDLSCs in bioprinted constructs were evaluated. Then, the effect of bioprinted constructs on bone regeneration was investigated using a mouse cranial defect model.

Results

Ten percent GelMA printed constructs had a higher compression modulus, smaller porosity, lower swelling rate, and lower degradation rate than 3% GelMA. PDLSCs in bioprinted 10% GelMA bioprinted constructs showed lower cell viability, less cell spreading, upregulated osteogenic differentiation in vitro, and lower cell survival in vivo. Moreover, upregulated expression of ephrinB2 and EphB4 protein and their phosphorylated forms were found in PDLSCs in 10% GelMA bioprinted constructs, and inhibition of eprhinB2/EphB4 signalling reversed the enhanced osteogenic differentiation of PDLSCs in 10% GelMA. The in vivo experiment showed that 10% GelMA bioprinted constructs with PDLSCs contributed to more new bone formation than 10% GelMA constructs without PDLSCs and constructs with lower GelMA concentrations.

Conclusions

Bioprinted PDLSCs with high-concentrated GelMA hydrogels exhibited enhanced osteogenic differentiation partially through upregulated ephrinB2/EphB4 signalling in vitro and promoted bone regeneration in vivo, which might be more appropriate for future bone regeneration applications.

Clinical relevance

Bone defects are a common clinical oral problem. Our results provide a promising strategy for bone regeneration through bioprinting PDLSCs in GelMA hydrogels.

Genova T, Roato I, Carossa M, Motta C, Cavagnetto D, Mussano F (2020) Advances on bone substitutes through 3D bioprinting. Int J Mol Sci 21. https://doi.org/10.3390/ijms21197012

Tournier P, Guicheux J, Pare A, Maltezeanu A, Blondy T, Veziers J, Vignes C, Andre M, Lesoeur J, Barbeito A, Bardonnet R, Blanquart C, Corre P, Geoffroy V, Weiss P, Gaudin A (2021) A partially demineralized allogeneic bone graft: in vitro osteogenic potential and preclinical evaluation in two different intramembranous bone healing models. Sci Rep 11:4907. https://doi.org/10.1038/s41598-021-84039-6CrossRefPubMedPubMedCentral

Piard C, Baker H, Kamalitdinov T, Fisher J (2019) Bioprinted osteon-like scaffolds enhance in vivo neovascularization. Biofabrication 11:025013. https://doi.org/10.1088/1758-5090/ab078aCrossRefPubMedPubMedCentral

Du L, Qin C, Zhang H, Han F, Xue J, Wang Y, Wu J, Xiao Y, Huan Z and Wu C (2023) Multicellular bioprinting of biomimetic inks for tendon-to-bone regeneration. Adv Sci (Weinh) e2301309. https://doi.org/10.1002/advs.202301309

Madl CM, Heilshorn SC, Blau HM (2018) Bioengineering strategies to accelerate stem cell therapeutics. Nature 557:335–342. https://doi.org/10.1038/s41586-018-0089-zCrossRefPubMedPubMedCentral

Yang Y, Wang M, Yang S, Lin Y, Zhou Q, Li H, Tang T (2020) Bioprinting of an osteocyte network for biomimetic mineralization. Biofabrication 12:045013. https://doi.org/10.1088/1758-5090/aba1d0CrossRefPubMed

Heo DN, Ayan B, Dey M, Banerjee D, Wee H, Lewis GS, Ozbolat IT (2020) Aspiration-assisted bioprinting of co-cultured osteogenic spheroids for bone tissue engineering. Biofabrication 13. https://doi.org/10.1088/1758-5090/abc1bf

Ratheesh G, Vaquette C, Xiao Y (2020) Patient-specific bone particles bioprinting for bone tissue engineering. Adv Healthc Mater e2001323. https://doi.org/10.1002/adhm.202001323

Wang T, Li W, Zhang Y, Xu X, Qiang L, Miao W, Yue X, Jiao X, Zhou X, Ma Z, Li S, Ding M, Zhu J, Yang C, Wang H, Li T, Sun X, Wang J (2023) Bioprinted constructs that simulate nerve-bone crosstalk to improve microenvironment for bone repair. Bioact Mater 27:377–393. https://doi.org/10.1016/j.bioactmat.2023.02.013CrossRefPubMedPubMedCentral

10.

Hu X, Li H, Qiao L, Yang S, Wu H, Peng C, Zhang Y, Lan H, Yang H and Li K (2023) Fabrication of 3D gel-printed β-tricalcium phosphate/titanium dioxide porous scaffolds for cancellous bone tissue engineering. Int J Bioprinting 9. https://doi.org/10.18063/ijb.v9i2.673

11.

Yang X, Ma Y, Wang X, Yuan S, Huo F, Yi G, Zhang J, Yang B, Tian W (2023) A 3D-bioprinted functional module based on decellularized extracellular matrix bioink for periodontal regeneration. Adv Sci (Weinh) 10:e2205041. https://doi.org/10.1002/advs.202205041CrossRefPubMed

12.

Hao Y, Cao B, Deng L, Li J, Ran Z, Wu J, Pang B, Tan J, Luo D, Wu W (2023) The first 3D-bioprinted personalized active bone to repair bone defects: a case report. Int J Bioprint 9:654. https://doi.org/10.18063/ijb.v9i2.654CrossRefPubMed

13.

Yamada Y, Ito K, Nakamura S, Ueda M, Nagasaka T (2011) Promising cell-based therapy for bone regeneration using stem cells from deciduous teeth, dental pulp, and bone marrow. Cell Transplant 20:1003–1013. https://doi.org/10.3727/096368910X539128CrossRefPubMed

14.

Ma Y, Ji Y, Zhong T, Wan W, Yang Q, Li A, Zhang X, Lin M (2017) Bioprinting-based PDLSC-ECM screening for in vivo repair of alveolar bone defect using cell-laden, injectable and photocrosslinkable hydrogels. ACS Biomater Sci Eng 3:3534–3545. https://doi.org/10.1021/acsbiomaterials.7b00601CrossRefPubMed

15.

Yan C, Li N, Xiao T, Ye X, Fu L, Ye Y, Xu T, Yu J (2022) Extracellular vesicles from the inflammatory microenvironment regulate the osteogenic and odontogenic differentiation of periodontal ligament stem cells by miR-758-5p/LMBR1/BMP2/4 axis. J Transl Med 20:208. https://doi.org/10.1186/s12967-022-03412-9CrossRefPubMedPubMedCentral

16.

Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, Young M, Robey PG, Wang CY, Shi S (2004) Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 364:149–155. https://doi.org/10.1016/S0140-6736(04)16627-0CrossRefPubMed

17.

Yang H, Gao LN, An Y, Hu CH, Jin F, Zhou J, Jin Y, Chen FM (2013) Comparison of mesenchymal stem cells derived from gingival tissue and periodontal ligament in different incubation conditions. Biomaterials 34:7033–7047. https://doi.org/10.1016/j.biomaterials.2013.05.025CrossRefPubMed

18.

Thattaruparambil Raveendran N, Vaquette C, Meinert C, Samuel Ipe D, Ivanovski S (2019) Optimization of 3D bioprinting of periodontal ligament cells. Dent Mater 35:1683–1694. https://doi.org/10.1016/j.dental.2019.08.114CrossRefPubMed

19.

Liu W, Heinrich MA, Zhou Y, Akpek A, Hu N, Liu X, Guan X, Zhong Z, Jin X, Khademhosseini A, Zhang YS (2017) Extrusion bioprinting of shear-thinning gelatin methacryloyl bioinks. Adv Healthc Mater 6. https://doi.org/10.1002/adhm.201601451

20.

Billiet T, Gevaert E, De Schryver T, Cornelissen M, Dubruel P (2014) The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability. Biomaterials 35:49–62. https://doi.org/10.1016/j.biomaterials.2013.09.078CrossRefPubMed

21.

Pepelanova I, Kruppa K, Scheper T, Lavrentieva A (2018) Gelatin-Methacryloyl (GelMA) Hydrogels with defined degree of functionalization as a versatile toolkit for 3D cell culture and extrusion bioprinting. Bioengineering (Basel) 5. https://doi.org/10.3390/bioengineering5030055

22.

Nichol JW, Koshy ST, Bae H, Hwang CM, Yamanlar S, Khademhosseini A (2010) Cell-laden microengineered gelatin methacrylate hydrogels. Biomaterials 31:5536–5544. https://doi.org/10.1016/j.biomaterials.2010.03.064CrossRefPubMedPubMedCentral

23.

Xie M, Zheng Y, Gao Q, He Y (2020) Facile 3D cell culture protocol based on photocurable hydrogels. Bio-Design Manuf 4:149–153. https://doi.org/10.1007/s42242-020-00096-2CrossRef

24.

Zonderland J, Moroni L (2021) Steering cell behavior through mechanobiology in 3D: a regenerative medicine perspective. Biomaterials 268:120572. https://doi.org/10.1016/j.biomaterials.2020.120572CrossRefPubMed

25.

Kim C, Young JL, Holle AW, Jeong K, Major LG, Jeong JH, Aman ZM, Han DW, Hwang Y, Spatz JP, Choi YS (2020) Stem cell mechanosensation on gelatin methacryloyl (GelMA) stiffness gradient hydrogels. Ann Biomed Eng 48:893–902. https://doi.org/10.1007/s10439-019-02428-5CrossRefPubMed

26.

Lee HP, Gu L, Mooney DJ, Levenston ME, Chaudhuri O (2017) Mechanical confinement regulates cartilage matrix formation by chondrocytes. Nat Mater 16:1243–1251. https://doi.org/10.1038/nmat4993CrossRefPubMedPubMedCentral

27.

Liu N, Zhou M, Zhang Q, Yong L, Zhang T, Tian T, Ma Q, Lin S, Zhu B, Cai X (2018) Effect of substrate stiffness on proliferation and differentiation of periodontal ligament stem cells. Cell Prolif 51:e12478. https://doi.org/10.1111/cpr.12478CrossRefPubMedPubMedCentral

28.

Zhao C, Irie N, Takada Y, Shimoda K, Miyamoto T, Nishiwaki T, Suda T, Matsuo K (2006) Bidirectional ephrinB2-EphB4 signaling controls bone homeostasis. Cell Metab 4:111–121. https://doi.org/10.1016/j.cmet.2006.05.012CrossRefPubMed

29.

Toda H, Yamamoto M, Uyama H, Tabata Y (2017) Effect of hydrogel elasticity and ephrinB2-immobilized manner on Runx2 expression of human mesenchymal stem cells. Acta Biomater 58:312–322. https://doi.org/10.1016/j.actbio.2017.03.016CrossRefPubMed

30.

Wang P, Wang W, Geng T, Liu Y, Zhu S, Liu Z, Yuan C (2019) EphrinB2 regulates osteogenic differentiation of periodontal ligament stem cells and alveolar bone defect regeneration in beagles. J Tissue Eng 10:2041731419894361. https://doi.org/10.1177/2041731419894361CrossRefPubMedPubMedCentral

31.

Diercke K, Kohl A, Lux CJ, Erber R (2011) Strain-dependent up-regulation of ephrin-B2 protein in periodontal ligament fibroblasts contributes to osteogenesis during tooth movement. J Biol Chem 286:37651–37664. https://doi.org/10.1074/jbc.M110.166900CrossRefPubMedPubMedCentral

32.

Ouyang L, Yao R, Zhao Y, Sun W (2016) Effect of bioink properties on printability and cell viability for 3D bioplotting of embryonic stem cells. Biofabrication 8:035020. https://doi.org/10.1088/1758-5090/8/3/035020CrossRefPubMed

33.

Gonzalez-Fernandez T, Tenorio AJ, Campbell KT, Silva EA, Leach JK (2021) Alginate-based bioinks for 3D bioprinting and fabrication of anatomically accurate bone grafts. Tissue Eng Part A 27:1168–1181. https://doi.org/10.1089/ten.TEA.2020.0305CrossRefPubMedPubMedCentral

34.

Moghanian A, Portillo-Lara R, Shirzaei Sani E, Konisky H, Bassir SH, Annabi N (2020) Synthesis and characterization of osteoinductive visible light-activated adhesive composites with antimicrobial properties. J Tissue Eng Regen Med 14:66–81. https://doi.org/10.1002/term.2964CrossRefPubMed

35.

Roato I, Masante B, Putame G, Massai D, Mussano F (2022) Challenges of periodontal tissue engineering: increasing biomimicry through 3D printing and controlled dynamic environment. Nanomaterials (Basel) 12. https://doi.org/10.3390/nano12213878

36.

Celikkin N, Mastrogiacomo S, Jaroszewicz J, Walboomers XF, Swieszkowski W (2018) Gelatin methacrylate scaffold for bone tissue engineering: the influence of polymer concentration. J Biomed Mater Res A 106:201–209. https://doi.org/10.1002/jbm.a.36226CrossRefPubMed

37.

Hannink G, Arts JJ (2011) Bioresorbability, porosity and mechanical strength of bone substitutes: what is optimal for bone regeneration? Injury 42(Suppl 2):S22–S25. https://doi.org/10.1016/j.injury.2011.06.008CrossRefPubMed

38.

Lee HP, Stowers R, Chaudhuri O (2019) Volume expansion and TRPV4 activation regulate stem cell fate in three-dimensional microenvironments. Nat Commun 10:529. https://doi.org/10.1038/s41467-019-08465-xCrossRefPubMedPubMedCentral

39.

Kumar G, Waters MS, Farooque TM, Young MF, Simon CG Jr (2012) Freeform fabricated scaffolds with roughened struts that enhance both stem cell proliferation and differentiation by controlling cell shape. Biomaterials 33:4022–4030. https://doi.org/10.1016/j.biomaterials.2012.02.048CrossRefPubMedPubMedCentral

40.

Zheng L, Yang F, Shen H, Hu X, Mochizuki C, Sato M, Wang S, Zhang Y (2011) The effect of composition of calcium phosphate composite scaffolds on the formation of tooth tissue from human dental pulp stem cells. Biomaterials 32:7053–7059. https://doi.org/10.1016/j.biomaterials.2011.06.004CrossRefPubMed

41.

Wei W, Liu W, Kang H, Zhang X, Yu R, Liu J, Huang K, Zhang Y, Xie M, Hu Y, Dai H (2023) A one-stone-two-birds strategy for osteochondral regeneration based on a 3D printable biomimetic scaffold with kartogenin biochemical stimuli gradient. Adv Healthc Mater e2300108. https://doi.org/10.1002/adhm.202300108

42.

Shi Z, Zhong Q, Chen Y, Gao J, Pan X, Lian Q, Chen R, Wang P, Wang J, Shi Z, Cheng H (2021) Nanohydroxyapatite, nanosilicate-reinforced injectable, and biomimetic gelatin-methacryloyl hydrogel for bone tissue engineering. Int J Nanomedicine 16:5603–5619. https://doi.org/10.2147/IJN.S321387CrossRefPubMedPubMedCentral

43.

Pu X, Tong L, Wang X, Liu Q, Chen M, Li X, Lu G, Lan W, Li Q, Liang J, Sun Y, Fan Y, Zhang X (2022) Bioinspired hydrogel anchoring 3DP GelMA/HAp scaffolds accelerates bone reconstruction. ACS Appl Mater Interfaces 14:20591–20602. https://doi.org/10.1021/acsami.1c25015CrossRefPubMed

Titel: Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo
verfasst von: Yaru Zhu
Wen Wang
Qiyu Chen
Tianshui Ren
Jianguang Yang
Gan Li
Yanbin Qi
Changyong Yuan
Penglai Wang
Publikationsdatum: 10.07.2023
Verlag: Springer Berlin Heidelberg
Erschienen in: Clinical Oral Investigations / Ausgabe 9/2023
Print ISSN: 1432-6981
Elektronische ISSN: 1436-3771
DOI: https://doi.org/10.1007/s00784-023-05135-7

Bestellen Sie unseren kostenlosen Newsletter Update Zahnmedizin und bleiben Sie gut informiert – ganz bequem per eMail.

Newsletter bestellen

Klimawandel und Gesundheit: Was Sie in der Hausarztpraxis wissen müssen und tun können

Springer Medizin

Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo