Application of Inorganic Bioactive Materials in Organoid Research

doi:10.15541/jim20250007

Abstract

Abstract:

As an effective in vitro three-dimensional (3D) model, organoids can simulate the structure and function of corresponding tissues/organs, demonstrating broad application prospects in the biomedical field. The construction of organoids relies on the regulation of stem cell behaviors and multicellular interactions. Inorganic bioactive materials possess excellent biocompatibility and bioactivity, showing wide application in the field of biomedical research. Therefore, they can potentially regulate cell behaviors and cell-cell/cell-matrix interactions in the construction of organoids. In this review, the role of inorganic bioactive materials in organoid research was explored, emphasizing their contribution to organoid development and application, and summarizing the critical steps in organoid construction strategies. Subsequently, the biological functions of inorganic bioactive materials, particularly those compatible with key steps in organoid construction, were systematically elucidated, and the key mechanisms by which inorganic bioactive materials promote organoid development and application were emphasized, including their effects on key signaling pathways, regulations of matrix material properties and cellular energy metabolism. In addition, the application of organoids as auxiliary tools to promote the use of inorganic bioactive materials was reviewed. Finally, the strategies for further advancing organoid research by regulating various physical and biochemical clues provided by inorganic bioactive materials were prospected.

Key words: organoid, inorganic bioactive material, organoids development, bioactive ion, review

CLC Number:

TQ174

MA Wenping, HAN Yahui, WU Chengtie, LÜ Hongxu. Application of Inorganic Bioactive Materials in Organoid Research[J]. Journal of Inorganic Materials, 2025, 40(8): 888-900.

Figures/Tables 6

Fig. 1 Strategy for constructing cardiac organoids derived from iPSC[31] (a) Formation scheme of cardiac organoids; (b) Cardiac organoids forming three layers; (c) Fluorescent staining of myocardial marker MHC; (d) Staining images of cTnT and WT1; (e) Staining image of mesenchymal cell marker vimentin CHIR: laduviglusib (CHIR99021); MHC: myosin heavy chain; cTnT: cardiac troponin T; WT1: Wilms’ tumor 1

Fig. 2 LCS bioceramics regulating the behaviors of NSCs[51] (a) Schematic diagram of preparation and application of neural platform based on LCS; (b) SEM images of LCS; (c) Representative immunofluorescence staining images of Nestin and MAP2 expression in NSCs; (d) Representative immunofluorescence staining images of GFAP and Tuj1 protein expression in NSCs; (e) Representative dual immunofluorescence staining images of GFAP and Tuj1 in NSCs under different culture conditions; (f) Representative immunofluorescence staining images of matural neuron marker MAP. MAP2: microtubule-associated protein 2; GFAP: glial fibrillary acidic protein; Tuj1: neuronal class III β-tubulin clone

Fig. 3 MS nanospheres regulating the interaction between endothelial cells and dermal cells[55] (a) TEM image of MS nanospheres; (b) EDS mappings of elements including Mg, Si and O in MS nanospheres; (c) Fluorescence image displaying the biomimetic spatial distributions of human umbilical vascular endothelial cells (HUVECs, red fluorescence) and human hair dermal papilla cells (HHDPCs, green fluorescence); (d) Stained images of cell clusters during co-culture; (e, f) Expression of genes related to angiogenesis (e) and hair follicle formation (f) in co-culture and single culture systems; (g) Microscopic observation of hair growth in newborn skin area of nude mice; (h) Distribution of mature hair follicles in co-culture group. VEGF: vascular endothelial growth factor; HIF-1α: hypoxia-inducible factor 1-α; KDR: kinase insert domain receptor; VE-cad: Vascular endothelial cadherin; PDGF-α: platelet-derived growth factor receptor α; PDGF-β: platelet-derived growth factor receptor β; c-Myc: cellular myelocytomatosis oncogene; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

Fig. 4 Bioactive ions released by CS nanowires for promoting intestinal organoid development[47] (a) Proliferation of intestinal organoids treated with CS nanowires of different concentrations in culture medium; (b, c) Gene expression of four differentiated cell types treated with 0, 50 and 100 μg/mL CS nanowires for 3 (b) and 5 days (c); (d) Immunofluorescence staining of β-catenin in intestinal organoids treated with CS nanowires for 3 and 5 days; (e) Heat maps of gene expression related to Wnt/β-catenin signaling pathway in intestinal organoids after 3 and 5 days of CS nanowire treatment; (f) Relative glucose absorption of different groups; (g) Relative ATP production of different groups. Wnt3: Wingless-type MMTV integration site family, member 3; LRP6: Low-density lipoprotein receptor-related protein 6; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

Fig. 5 CNT for promoting intestinal organoid development[61] (a) Typical images of epithelial subpopulations in intestinal organoids treated with CNT (50 μg/mL) at different time periods; (b) Representative SEM images showing degradation of the matrix over time during development of intestinal organoids; (c) Statistical analysis of changes over time in phosphorylated p38 mitogen activated protein kinase (P-p38 MAPK), p38 mitogen activated protein kinase (p38 MAPK), yes-related protein (YAP), and phosphorylated yes-related protein (P-YAP) in intestinal organoids treated with CNT (50 μg/mL); (d) Quantification of YAP nuclear translocation over time in intestinal organoids treated with CNT (50 μg/mL); (e) Representative confocal images of mitochondrial membrane potential and relative mitochondrial activity in intestinal organoids over time; (f) Effects of different amino acids on the proliferation and differentiation of intestinal organoids; (g) Proposed mechanism of CNT induced intestinal organoid development. SWCNTs: single-walled carbon nanotubes; MWCNTs: multiwalled carbon nanotubes; L-FABP: liver fatty acid-binding protein; ISC: intestinal stem cell; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

Fig. 6 Application of organoids in exploring reduction of intestinal inflammatory damage by bioactive glass microspheres[71] (a) Concentration of TNF-α in macrophage culture medium after different treatments; (b) Expression of anti-inflammatory gene ARG in macrophage culture after different treatments; (c) CCK-8 testing of intestinal organoids cultured under different media conditions; (d) Immunofluorescence staining of Ki67 in intestinal organoids cultured under different conditions for 144 h; (e) Morphology and size of intestinal organoids cultured under different conditions for 72 and 144 h. BG: bioactive glass; LPS: lipopolysaccharide; CK: non-treated control; ARG: arginase; Conditioned media were collected from untreated RAW “CM-RAW-CK”, LPS-activated RAW “CM-LPS”, and LPS-activated RAW after pretreatment of BG extract liquids (1 : 100 dilution ratio) “CM-LPS/BG”; *, ** and *** indicate p<0.05, 0.01 and 0.001, respectively

References 77

[1]	HOFER M, LUTOLF M P. Engineering organoids. Nature Reviews Materials, 2021, 6(5): 402.
[2]	KRATOCHVIL M J, SEYMOUR A J, LI T L, et al. Engineered materials for organoid systems. Nature Reviews Materials, 2019, 4(9): 606. DOI
[3]	SATO T, VRIES R G, SNIPPERT H J, et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature, 2009, 459(7244): 262.
[4]	BARKER N, HUCH M, KUJALA P, et al. Lgr5^+ve stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. Cell Stem Cell, 2010, 6(1): 25.
[5]	CHO A N, JIN Y, AN Y, et al. Microfluidic device with brain extracellular matrix promotes structural and functional maturation of human brain organoids. Nature Communications, 2021, 12: 4730.
[6]	MORIZANE R, LAM A Q, FREEDMAN B S, et al. Nephron organoids derived from human pluripotent stem cells model kidney development and injury. Nature Biotechnology, 2015, 33(11): 1193. PMID
[7]	TAO T T, DENG P W, WANG Y Q, et al. Microengineered multi-organoid system from hiPSCs to recapitulate human liver-islet axis in normal and type 2 diabetes. Advanced Science, 2022, 9(5): 2103495.
[8]	LEWIS-ISRAELI Y R, WASSERMAN A H, GABALSKI M A, et al. Self-assembling human heart organoids for the modeling of cardiac development and congenital heart disease. Nature Communications, 2021, 12: 5142.
[9]	RICHARDS D J, LI Y, KERR C M, et al. Human cardiac organoids for the modelling of myocardial infarction and drug cardiotoxicity. Nature Biomedical Engineering, 2020, 4(4): 446. DOI PMID
[10]	DUTTA D, HEO I, CLEVERS H. Disease modeling in stem cell-derived 3D organoid systems. Trends in Molecular Medicine, 2017, 23(5): 393. DOI PMID
[11]	CRUZ-ACUÑA R, QUIRÓS M, FARKAS A E, et al. Synthetic hydrogels for human intestinal organoid generation and colonic wound repair. Nature Cell Biology, 2017, 19(11): 1326.
[12]	VLACHOGIANNIS G, HEDAYAT S, VATSIOU A, et al. Patient-derived organoids model treatment response of metastatic gastrointestinal cancers. Science, 2018, 359(6378): 920. DOI PMID
[13]	BROKESH A M, GAHARWAR A K.Inorganic biomaterials for regenerative medicine. ACS Applied Materials & Interfaces, 2020, 12(5): 5319.
[14]	ERMAKOV A, DAKS A, FEDOROVA O, et al. Ca²⁺-depended signaling pathways regulate self-renewal and pluripotency of stem cells. Cell Biology International, 2018, 42(9): 1086.
[15]	TAWFIK I, GOTTSCHALK B, JARC A, et al. T3-induced enhancement of mitochondrial Ca²⁺ uptake as a boost for mitochondrial metabolism. Free Radical Biology and Medicine, 2022, 181: 197.
[16]	HAN P P, WU C T, XIAO Y. The effect of silicate ions on proliferation, osteogenic differentiation and cell signalling pathways (WNT and SHH) of bone marrow stromal cells. Biomaterials Science, 2013, 1(4): 379. DOI PMID
[17]	AISENBREY E A, MURPHY W L. Synthetic alternatives to matrigel. Nature Reviews Materials, 2020, 5(7): 539. DOI PMID
[18]	ROSSI G, MANFRIN A, LUTOLF M P. Progress and potential in organoid research. Nature Reviews Genetics, 2018, 19(11): 671. DOI PMID
[19]	ROOKMAAKER M B, SCHUTGENS F, VERHAAR M C, et al. Development and application of human adult stem or progenitor cell organoids. Nature Reviews Nephrology, 2015, 11(9): 546. DOI PMID
[20]	LUKONIN I, SERRA D, MEYLAN L C, et al. Phenotypic landscape of intestinal organoid regeneration. Nature, 2020, 586(7828): 275.
[21]	BOUFFI C, WIKENHEISER-BROKAMP K A, CHATURVEDI P, et al. In vivo development of immune tissue in human intestinal organoids transplanted into humanized mice. Nature Biotechnology, 2023, 41(6): 824.
[22]	SORRENTINO G, REZAKHANI S, YILDIZ E, et al. Mechano- modulatory synthetic niches for liver organoid derivation. Nature Communications, 2020, 11: 3416.
[23]	CLEVERS H. Modeling development and disease with organoids. Cell, 2016, 165(7): 1586. DOI PMID
[24]	BARTFELD S, BAYRAM T, VAN DE WETERING M,et al. In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection. Gastroenterology, 2015, 148(1): 126.
[25]	XU J S, PATTON D, JACKSON S K, et al. In-vitro maintenance and functionality of primary renal tubules and their application in the study of relative renal toxicity of nephrotoxic drugs. Journal of Pharmacological and Toxicological Methods, 2013, 68(2): 269.
[26]	YU J Y, VODYANIK M A, SMUGA-OTTO K, et al. Induced pluripotent stem cell lines derived from human somatic cells. Science, 2007, 318(5858): 1917. DOI PMID
[27]	LEE J, SUTANI A, KANEKO R, et al. In vitro generation of functional murine heart organoids via FGF4 and extracellular matrix. Nature Communications, 2020, 11: 4283.
[28]	ROMITTI M, TOURNEUR A, DA FONSECA B D F,et al. Transplantable human thyroid organoids generated from embryonic stem cells to rescue hypothyroidism. Nature Communications, 2022, 13: 7057. DOI PMID
[29]	WANG S Y, WANG X, TAN Z L, et al. Human ESC-derived expandable hepatic organoids enable therapeutic liver repopulation and pathophysiological modeling of alcoholic liver injury. Cell Research, 2019, 29(12): 1009. DOI PMID
[30]	TAKAHASHI K, TANABE K, OHNUKI M, et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 2007, 131(5): 861. DOI PMID
[31]	DRAKHLIS L, BISWANATH S, FARR C M, et al. Human heart-forming organoids recapitulate early heart and foregut development. Nature Biotechnology, 2021, 39(6): 737. DOI PMID
[32]	FRENZ-WIESSNER S, FAIRLEY S D, BUSER M, et al. Generation of complex bone marrow organoids from human induced pluripotent stem cells. Nature Methods, 2024, 21(5): 868.
[33]	YOON S J, ELAHI L S, PAȘCA A M, et al. Reliability of human cortical organoid generation. Nature Methods, 2019, 16(1): 75.
[34]	GIGER S, HOFER M, MILJKOVIC-LICINA M, et al. Microarrayed human bone marrow organoids for modeling blood stem cell dynamics. APL Bioengineering, 2022, 6(3): 036101.
[35]	PRONDZYNSKI M, BERKSON P, TREMBLEY M A, et al. Efficient and reproducible generation of human iPSC-derived cardiomyocytes and cardiac organoids in stirred suspension systems. Nature Communications, 2024, 15: 5929. DOI PMID
[36]	BERGENHEIM F, FREGNI G, BUCHANAN C F, et al. A fully defined 3D matrix for ex vivo expansion of human colonic organoids from biopsy tissue. Biomaterials, 2020, 262: 120248.
[37]	HUCH M, KNOBLICH J A, LUTOLF M P, et al. The hope and the hype of organoid research. Development, 2017, 144(6): 938. DOI PMID
[38]	LIU H T, WANG Y Q, CUI K L, et al. Advances in hydrogels in organoids and organs-on-a-chip. Advanced Materials, 2019, 31(50): 1902042.
[39]	POLING H M, WU D, BROWN N, et al. Mechanically induced development and maturation of human intestinal organoids in vivo. Nature Biomedical Engineering, 2018, 2(6): 429.
[40]	HOMAN K A, GUPTA N, KROLL K T, et al. Flow-enhanced vascularization and maturation of kidney organoids in vitro. Nature Methods, 2019, 16(3): 255.
[41]	DE BOER J, SIDDAPPA R, GASPAR C, et al. Wnt signaling inhibits osteogenic differentiation of human mesenchymal stem cells. Bone, 2004, 34(5): 818. PMID
[42]	WEI P F, JING W, YUAN Z Y, et al. Vancomycin- and strontium-loaded microspheres with multifunctional activities against bacteria, in angiogenesis, and in osteogenesis for enhancing infected bone regeneration. ACS Applied Materials & Interfaces, 2019, 11(34): 30596.
[43]	CHEN L, DENG C J, LI J Y, et al. 3D printing of a lithium- calcium-silicate crystal bioscaffold with dual bioactivities for osteochondral interface reconstruction. Biomaterials, 2019, 196: 138.
[44]	WANG S, HASHEMI S, STRATTON S, et al. The effect of physical cues of biomaterial scaffolds on stem cell behavior. Advanced Healthcare Materials, 2021, 10(3): 2001244.
[45]	WANG P Y, CLEMENTS L R, THISSEN H, et al. Screening mesenchymal stem cell attachment and differentiation on porous silicon gradients. Advanced Functional Materials, 2012, 22(16): 3414.
[46]	GJOREVSKI N, SACHS N, MANFRIN A, et al. Designer matrices for intestinal stem cell and organoid culture. Nature, 2016, 539(7630): 560.
[47]	MA W P, ZHENG Y, YANG G Z, et al. A bioactive calcium silicate nanowire-containing hydrogel for organoid formation and functionalization. Materials Horizons, 2024, 11(12): 2957. DOI PMID
[48]	DAS S R, UZ M, DING S W, et al. Electrical differentiation of mesenchymal stem cells into schwann-cell-like phenotypes using inkjet-printed graphene circuits. Advanced Healthcare Materials, 2017, 6(7): 1601087.
[49]	SHENG R Y Y, MU J, CHERNOZEM R V, et al. Fabrication and characterization of piezoelectric polymer composites and cytocompatibility with mesenchymal stem cells. ACS Applied Materials & Interfaces, 2023, 15(3): 3731.
[50]	LIU Z R, WAN X Y, WANG Z L, et al. Electroactive biomaterials and systems for cell fate determination and tissue regeneration: design and applications. Advanced Materials, 2021, 33(32): 2007429.
[51]	ZHANG H J, QIN C, SHI Z, et al. Bioprinting of inorganic- biomaterial/neural-stem-cell constructs for multiple tissue regeneration and functional recovery. National Science Review, 2024, 11(4): nwae035.
[52]	DU L, QIN C, ZHANG H J, et al. Multicellular bioprinting of biomimetic inks for tendon-to-bone regeneration. Advanced Science, 2023, 10(21): 2301309.
[53]	HE D, LI H Y. Biomaterials affect cell-cell interactions in vitro in tissue engineering. Journal of Materials Science & Technology, 2021, 63: 62.
[54]	GAUTAM V, NAUREEN S, SHAHID N, et al. Engineering highly interconnected neuronal networks on nanowire scaffolds. Nano Letters, 2017, 17(6): 3369. DOI PMID
[55]	MA J G, QIN C, WU J F, et al. 3D multicellular micropatterning biomaterials for hair regeneration and vascularization. Materials Horizons, 2023, 10(9): 3773. DOI PMID
[56]	WANG X Y, GAO L, HAN Y, et al. Silicon-enhanced adipogenesis and angiogenesis for vascularized adipose tissue engineering. Advanced Science, 2018, 5(11): 1800776.
[57]	GAO C D, PENG S P, FENG P, et al. Bone biomaterials and interactions with stem cells. Bone Research, 2017, 5: 17059. DOI PMID
[58]	LUO J J, WALKER M, XIAO Y B, et al. The influence of nanotopography on cell behaviour through interactions with the extracellular matrix-a review. Bioactive Materials, 2022, 15: 145.
[59]	CHEN W C, TIAN B X, LIANG J Q, et al. Matrix stiffness regulates the interactions between endothelial cells and monocytes. Biomaterials, 2019, 221: 119362.
[60]	ZHANG Z, GAO S, HU Y N, et al. Ti₃C₂T_xMXene composite 3D hydrogel potentiates mTOR signaling to promote the generation of functional hair cells in cochlea organoids. Advanced Science, 2022, 9(32): 2203557.
[61]	BAO L, CUI X J, WANG X Y, et al. Carbon nanotubes promote the development of intestinal organoids through regulating extracellular matrix viscoelasticity and intracellular energy metabolism. ACS Nano, 2021, 15(10): 15858. DOI PMID
[62]	WANG J, WU Y, LI G F, et al. Engineering large-scale self-mineralizing bone organoids with bone matrix-inspired hydroxyapatite hybrid bioinks. Advanced Materials, 2024, 36(30): 2309875.
[63]	TAN Y, COYLE R C, BARRS R W, et al. Nanowired human cardiac organoid transplantation enables highly efficient and effective recovery of infarcted hearts. Science Advances, 2023, 9(31): eadf2898.
[64]	TIAN M, WEI J S, LV E G, et al. Drug evaluation platform based on non-destructive and real-time in situ organoid fate state monitoring by graphene field-effect transistor. Chemical Engineering Journal, 2024, 498: 155355.
[65]	ROTH J G, BRUNEL L G, HUANG M S, et al. Spatially controlled construction of assembloids using bioprinting. Nature Communications, 2023, 14: 4346. DOI PMID
[66]	POON W, ZHANG Y N, OUYANG B, et al. Elimination pathways of nanoparticles. ACS Nano, 2019, 13(5): 5785. DOI PMID
[67]	ZHANG R, LI D, ZHAO R B, et al. Spike structure of gold nanobranches induces hepatotoxicity in mouse hepatocyte organoid models. Journal of Nanobiotechnology, 2024, 22(1): 92. DOI PMID
[68]	BAEK A, KWON I H, LEE D H, et al. Novel organoid culture system for improved safety assessment of nanomaterials. Nano Letters, 2024, 24(3): 805. DOI PMID
[69]	WEISS P, TAYLOR A C. Reconstitution of complete organs from single-cell suspensions of chick embryos in advanced stages of differentiation. Proceedings of the National Academy of Scinences of the United States of America, 1960, 46(9): 1177.
[70]	MA P Q, CHEN Y, LAI X Y, et al. The translational application of hydrogel for organoid technology: challenges and future perspectives. Macromolecular Bioscience, 2021, 21(10): 2100191.
[71]	ZHU Y L, WANG Y W, XIA G G, et al. Oral delivery of bioactive glass-loaded core-shell hydrogel microspheres for effective treatment of inflammatory bowel disease. Advanced Science, 2023, 10(18): 2207418.
[72]	ZHANG H, LU Y, ZHANG R, et al. Synthesis of multifunctional plasmonic nanodarts through one-end deposition on gold nanobipyramids for tumor organoid ablation and antimicrobial applications. Advanced Functional Materials, 2024, 34(44): 2405588.
[73]	ISSA R, LOZANO N, KOSTARELOS K, et al. Functioning human lung organoids model pulmonary tissue response from carbon nanomaterial exposures. Nano Today, 2024, 56: 102254.
[74]	HEID S, BOCCACCINI A R. Advancing bioinks for 3D bioprinting using reactive fillers: a review. Acta Biomaterialia, 2020, 113: 1. DOI PMID
[75]	LI R T, LIU K, HUANG X, et al. Bioactive materials promote wound healing through modulation of cell behaviors. Advanced Science, 2022, 9(10): 2105152.
[76]	THUNDYIL J, PAVLOVSKI D, SOBEY C G, et al. Adiponectin receptor signalling in the brain. British Journal of Pharmacology, 2012, 165(2): 313. DOI PMID
[77]	PUSCHHOF J, PLEGUEZUELOS-MANZANO C, CLEVERS H. Organoids and organs-on-chips: insights into human gut-microbe interactions. Cell Host & Microbe, 2021, 29(6): 867.