To address the limitations associated with current bone graft materials, particularly the slow resorption of deproteinized bovine bone minerals and the adverse effects of high-dose BMP-2, we developed an innovative tripartite regeneration platform. This platform employs a decalcified dentin matrix (DDM) as a physiological carrier for the spatio-temporal co-delivery of BMP-2 and Nell-1, representing a novel combined strategy. The DDM particles, optimized to a size range of 500–1000 μm through EDTA hierarchical decalcification (achieving 70% decalcification), exhibited a marked increase in pulp tubule diameter, cross-sectional area, and porosity. Scanning electron microscopy (SEM) analysis confirmed that the absence of a smear layer and the presence of uniform tubules facilitated osteoblast infiltration. In a rat model with critical-sized skull defects, the synergistic DBN structure (comprising DDM, BMP-2, and NELL-1) resulted in nearly complete bone regeneration within 8 weeks, significantly enhancing bone volume and bone mineral density. This bionic platform addresses the “bone-induced inflammation paradox” by utilizing the hierarchical topological structure of DDM and the synergistic dynamics of two factors: the rapid release of BMP-2 and the sustained release of NELL-1. This approach surpasses the clinical gold standard and satisfies the FDA’s efficacy criteria for the repair of critical size defects.
WangYYamaguchiYHsuehP, et al.Influence of implant macrodesign and insertion load during implant placement on primary stability in artificial bone. Int. J. Implant Dent2025; 11: 56. https://doi.org/10.1186/s40729-025-00644-4
2.
JiangZKongDZhouC, et al.Stability of alveolar ridge following horizontal guided bone regeneration after implant loading for 1 -2 years: a retrospective comparative study. BMC Oral Health2025; 25: 673. https://doi.org/10.1186/s12903-025-06025-y
3.
RuesSSchmitterMKappelS, et al.Effect of bone quality and quantity on the primary stability of dental implants in a simulated bicortical placement. Clin Oral Invest2021; 25: 1265–1272. https://doi.org/10.1007/s00784-020-03432-z
4.
QianYWuJYangW, et al.FTO-associated osteoclastogenesis promotes alveolar bone resorption in apical periodontitis male rat via the HK1/USP14/RANK pathway. Nat Commun2025; 16: 1519. https://doi.org/10.1038/s41467-025-56615-1
5.
MingPLiKZhuW. Synergistic exacerbation of periprosthetic osteolysis by inflammatory bowel disease and titanium ions: impaired osteogenesis of BMSCs via PI3K/AKT signaling. Mater Today Bio2025; 34: 102236. https://doi.org/10.1016/j.mtbio.2025.102236
6.
YuDShenWDaiJ, et al.Treatment of large bone defects in load-bearing bone: traditional and novel bone grafts. J Zhejiang Univ - Sci B2025; 26: 421–447. https://doi.org/10.1631/jzus.B2300669
7.
XiaoPChenCShenX, et al.Bone volume and height changes for lateral window sinus floor elevation using two types of deproteinized bovine bone mineral: a retrospective cohort study of 1-4 years. Clin Oral Implants Res2024; 35: 1493–1505. https://doi.org/10.1111/clr.14337
8.
ChenRZhangWDingY, et al.Prominent alveolar bone graft substitute derived from silk fibroin/hyaluronic acid/demineralized dentin matrix hybrid hydrogel. Biomater Res2025; 29: 0243. https://doi.org/10.34133/bmr.0243
WangCNistalaRCaoM, et al.Repair of limb ischemia is dependent on hematopoietic stem cell specific-shp-1 regulation of tgf-β1. Arterioscler Thromb Vasc Biol2023; 43: 92–108. https://doi.org/10.1161/atvbaha.122.318205
11.
UmIWKuJKKimYK, et al.Histological review of demineralized dentin matrix as a carrier of rhBMP-2. Tissue Eng Part B2020; 26: 284–293. https://doi.org/10.1089/ten.TEB.2019.0291
12.
SohnDSKimJRKimHG, et al.Comparison of immunohistochemical analysis on sinus augmentation using demineralized tooth graft and bovine bone. J Korean Assoc Oral Maxillofac Surg2021; 47: 269–278. https://doi.org/10.5125/jkaoms.2021.47.4.269
13.
QiuPOuyangYLiuS, et al.Environmental response temporal release injectable hydrogel for controlled growth factor release to enhance inflammatory periodontal bone defect regeneration. Adv Mater2026; 38: e12531. https://doi.org/10.1002/adma.202512531
14.
ZaraJNSiuRKZhangX, et al.High doses of bone morphogenetic protein 2 induce structurally abnormal bone and inflammation in vivo. Tissue Eng2011; 17: 1389–1399. https://doi.org/10.1089/ten.TEA.2010.0555
ChenXWangHYuM, et al.Cumulative inactivation of nell-1 in wnt1 expressing cell lineages results in craniofacial skeletal hypoplasia and postnatal hydrocephalus. Cell Death Differ2020; 27: 1415–1430. https://doi.org/10.1038/s41418-019-0427-1
17.
ChenHZhangZZhangL, et al.miR-27a protects human mitral valve interstitial cell from TNF-α-induced inflammatory injury via up-regulation of NELL-1. Braz J Med Biol Res2018; 51: e6997. https://doi.org/10.1590/1414-431x20186997
18.
ZhangXZaraJSiuRK, et al.The role of NELL-1, a growth factor associated with craniosynostosis, in promoting bone regeneration. J Dent Res2010; 89: 865–878. https://doi.org/10.1177/0022034510376401
19.
SongHZhangYZhangZ, et al.Hydroxyapatite/NELL-1 nanoparticles electrospun fibers for osteoinduction in bone tissue engineering application. Int J Nanomed2021; 16: 4321–4332. https://doi.org/10.2147/IJN.S309567
20.
LiuLLamWMRNaiduM, et al.Synergistic effect of nell-1 and an ultra-low dose of bmp-2 on spinal fusion. Tissue Eng2019; 25: 1677–1689. https://doi.org/10.1089/ten.TEA.2019.0124
21.
ShenJJamesAWZhangX, et al.Novel wnt regulator nel-like molecule-1 antagonizes adipogenesis and augments osteogenesis induced by bone morphogenetic protein 2. Am J Pathol2016; 186: 419–434. https://doi.org/10.1016/j.ajpath.2015.10.011
22.
WuJWangQHanQ, et al.Effects of Nel-like molecule-1 and bone morphogenetic protein 2 combination on rat pulp repair. J Mol Histol2019; 50: 253–261. https://doi.org/10.1007/s10735-019-09822-2
23.
LiuSSunJYuanS, et al.Treated dentin matrix induces odontogenic differentiation of dental pulp stem cells via regulation of Wnt/β-catenin signaling. Bioact Mater2022; 7: 85–97. https://doi.org/10.1016/j.bioactmat.2021.05.026
24.
SultanNMowafeyBAtaF, et al.Enhanced bone regeneration using demineralized dentin matrix: a comparative study in alveolar bone repair. Int Dent J2025; 75: 100817. https://doi.org/10.1016/j.identj.2025.03.026
25.
KogaTMinamizatoTKawaiY, et al.Bone regeneration using dentin matrix depends on the degree of demineralization and particle size. PLoS One2016; 11: e0147235. https://doi.org/10.1371/journal.pone.0147235
26.
WangXZhangDPengH, et al.Optimize the pore size-pore distribution-pore geometry-porosity of 3D-printed porous tantalum to obtain optimal critical bone defect repair capability. Biomater Adv2023; 154: 213638. https://doi.org/10.1016/j.bioadv.2023.213638
27.
YangFChenCZhouQ, et al.Laser beam melting 3D printing of Ti6Al4V based porous structured dental implants: fabrication, biocompatibility analysis and photoelastic study. Sci Rep2017; 7: 45360. https://doi.org/10.1038/srep45360
28.
MacalesterWBoussahelAMoreno-TortoleroRO, et al.A 3D In-vitro model of the human dentine interface shows long-range osteoinduction from the dentine surface. Int J Oral Sci2024; 16: 37. https://doi.org/10.1038/s41368-024-00298-9
29.
FigueiredoMHenriquesJMartinsG, et al.Physicochemical characterization of biomaterials commonly used in dentistry as bone substitutes--comparison with human bone. J Biomed Mater Res B Appl Biomater2010; 92: 409–419. https://doi.org/10.1002/jbm.b.31529
30.
TanoueROhtaKMiyazonoY, et al.Three-dimensional ultrastructural analysis of the interface between an implanted demineralised dentin matrix and the surrounding newly formed bone. Sci Rep2018; 8: 2858. https://doi.org/10.1038/s41598-018-21291-3
31.
QinXZouFChenW, et al.Demineralized dentin as a semi-rigid barrier for guiding periodontal tissue regeneration. J Periodontol2015; 86: 1370–1379. https://doi.org/10.1902/jop.2015.150271
32.
ChenYKongMZhangT, et al.Advancing bioprinting technology utilizing portable bioprinters: from various device designs to dental applications. Ann Biomed Eng2025; 53: 2408–2425. https://doi.org/10.1007/s10439-025-03789-w
33.
EberliköseHYaylacıSKaçaroğluD, et al.Multifunctional peptide nanofiber coatings enhance bone regeneration on xenograft materials. Sci Rep2025; 15: 31541. https://doi.org/10.1038/s41598-025-15743-w
34.
DengAZhangHHuY, et al.Microsphere strategy to generate conformal bone organoid units with osteoimmunomodulation and sustainable oxygen supply for bone regeneration. Adv Sci2025; 12: e01437. https://doi.org/10.1002/advs.202501437
35.
MartinezDCDobkowskaAMarekR, et al.In vitro and in vivo degradation behavior of Mg-0.45Zn-0.45Ca (ZX00) screws for orthopedic applications. Bioact Mater2023; 28: 132–154. https://doi.org/10.1016/j.bioactmat.2023.05.004
36.
LianQLiuHLiJ, et al.Enhancing radiosensitivity of osteosarcoma by ITGB3 knockdown: a mechanism linked to enhanced osteogenic differentiation status through JNK/c-JUN/RUNX2 pathway activation. J Exp Clin Cancer Res2025; 44: 159. https://doi.org/10.1186/s13046-025-03417-4
ZouXShenJChenF, et al.NELL-1 binds to APR3 affecting human osteoblast proliferation and differentiation. FEBS Lett2011; 585: 2410–2418. https://doi.org/10.1016/j.febslet.2011.06.024
39.
ShenJJamesAWChungJ, et al.NELL-1 promotes cell adhesion and differentiation via Integrinβ1. J Cell Biochem2012; 113: 3620–3628. https://doi.org/10.1002/jcb.24253
40.
AsgarySShamszadehS. Immunomodulatory and angiogenic strategies in vital pulp therapy: a systematic review of scaffold-based, scaffold-free, and cell-laden interventions. Clin Oral Invest2026; 30: 73. https://doi.org/10.1007/s00784-025-06737-z
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
