Abstract An ideal tissue engineering scaffold should recapitulate the native extracellular matrix (ECM) microenvironment, providing structural support and biochemical cues that regulate cellular behavior and promote tissue regeneration. Decellularized extracellular matrix (dECM) preserves the compositional and biological features of native ECM, but its limited processability restricts direct fabrication into scaffolds with defined structures and functions. To overcome these limitations, a variety of strategies have been developed for building dECM-based scaffolds including hydrogels, scaffolds, membranes and other formats for broader application. In this review, we begin with a brief overview of the decellularization strategies and the compositional characteristics of dECM, and then we focus on summarizing the recent developments in processing strategies including enzymatic and non-enzymatic processing of dECM materials. Next, this review focuses on the design and application of dECM-based scaffolds with different forms by non-enzymatic processing treatment. Finally, we discuss the current challenges and future perspectives for non-enzymatic engineering of dECM-based scaffolds.
Abstract Craniomaxillofacial bone defects and implant-related infections require materials that support regeneration while providing localized therapeutic action. Quercetin offers osteogenic, antibacterial, and anticancer activity, but poor stability and bioavailability limit direct use. To address this, a Quercetin–zinc (Q–Zn2+) complex is loaded onto 3D-printed (3DP) tricalcium phosphate (TCP) scaffolds, where zinc coordination stabilizes Quercetin through metal-ligand interactions and enables controlled release. The release behavior of Q–Zn2+ from 3DP TCP and its antibacterial effects remain unexplored. Characterization of the complex shows that Zn2+ chelates with Quercetin’s hydroxyl and carbonyl groups. Release studies at pH 5.0 and pH 7.4 show sustained delivery of Quercetin over 24 days, reaching 16% release at pH 5.0 and 5% at pH 7.4 for complexed Quercetin. In vitro evaluation shows that Q–Zn2+-loaded scaffolds increase osteoblast viability by 42%, reduce osteosarcoma viability by ∼55%, and achieve 93% inhibition of Staphylococcus aureus compared to TCP control. Zn2+ chelation stabilizes Quercetin release while improving osteoblast viability and suppressing bacterial and osteosarcoma growth from 3DP TCP scaffolds. This approach offers a route for controlled therapeutic delivery of Quercetin in calcium phosphate bone graft materials.
Abstract Replicating the precise topographical guidance of native extracellular matrix is a key goal in functional tissue engineering. Textile-based scaffolds possess biomimetic fibrous architecture and clinical suitability, but have been largely confined to passive mechanical support, overlooking their bioactive surface topography. Here, we demonstrate that woven architecture itself can serve as a programmable bioactive cue. Using silk fibroin, representative plain (SFP), twill (SFT), and satin (SFS) weaves are fabricated. Systematic characterization reveals distinctly warp-float lengths and warp-to-weft float ratios: SFP with short floats (274.2 ± 25.9 μm, ratio ≈1), SFT with intermediate floats (492.7 ± 17.8 μm, ratio ≈2), and SFS with long continuous warp floats (830.8 ± 20.5 μm, ratio >19). In vitro multi-lineage cell assays demonstrate that SFS weave induces remarkable anisotropic guidance, yielding exceptional cell elongation (aspect ratio > 18), high alignment (> 84%), and accelerated directional migration. In vivo, the aligned topography of SFS scaffolds orchestrates a pro-regenerative immune microenvironment, as evidenced by enhanced collagen deposition, M2 macrophage polarization (upregulated CD206, downregulated CD86), and increased angiogenesis (CD31), synergistically accelerating tissue repair. These findings highlight woven architecture as a designable parameter that directly programs cellular organization and immune responses, advancing topology-driven strategies for regenerative biomaterials.
Abstract Osteomyelitis, a chronic bone infection, poses significant challenges in clinical management due to the risks of severe bone necrosis and the increasing prevalence of antibiotic-resistant bacteria. Conventional therapeutic strategies, including surgical debridement and systemic antibiotic therapy, often fall short of achieving optimal outcomes due to antibiotic resistance and failure to achieve high-quality bone regeneration post-surgery. To address these clinical challenges, this study developed an alkaline calcium phosphate cement enabling sequential osteomyelitis control and bone regeneration. This cement was formed through a stepwise reaction between calcium hydroxide (Ca(OH)2) and sodium polyphosphate ((NaPO3)n) in an aqueous environment, enabling provisional defect filling as well as immediate generation of a hyperalkaline environment upon implantation to effectively inhibit both Gram-positive and Gram-negative bacteria. The pH value of bone cement extract gradually decreased from a hyperalkaline state (>11.5) to a mildly alkaline microenvironment (7.8-8.5) over time with the progression of the reaction between Ca(OH)2 and (NaPO3)n in situ, subsequently fostering a favorable microenvironment for bone regeneration. Meanwhile, a stable and effective rat osteomyelitis model was established to verify the sequential antibacterial and osteogenic performance of the material in vivo. In vitro cellular experiments demonstrated that bone cement enhances the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs). Moreover, an in vivo osteomyelitis rat model also revealed the absence of infectious reactions or tissue necrosis, as well as bone formation around the cement post-implantation, confirming the feasibility of this material in treating osteomyelitis. Collectively, these results indicated that the as-developed cement can serve as an active bone repair material adapted to dynamic microenvironment regulation needs, offering a promising approach for the treatment of osteomyelitis and the promotion of bone regeneration in a sequential manner.
Abstract Osteosarcoma is the most common primary malignant bone tumor in adolescents, significantly impacting their quality of life. The difficulty in removing all residual tumor cells during surgery contributes to the recurrence of osteosarcoma. To provide an alternative strategy, we developed an iron-based polysaccharide hydrogel (gIONPs@CMOT) with immunomodulatory capabilities to activate antitumor immunity and remodel the postoperative tumor immune microenvironment. The hydrogel consisted of oxidized trichosanthes polysaccharide (oTPS), and carboxymethyl chitosan (CMCS), interspersed with glycosaminoglycan-modified Fe3O4 nanoparticles (gIONPs). In vitro and in vivo studies demonstrated that gIONPs in the hydrogel were capable of reprogramming macrophages towards an antitumor phenotype via the nuclear factor-kappa B (NF-κB) pathway. The gIONPs@CMOT hydrogel promoted the maturation of dendritic cells and enhanced their antigen presentation through the cyclic GMP–AMP synthase (cGAS) and stimulator of interferon genes (STING) pathway. The oTPS in the hydrogel not only played a role in crosslinking with CMCS, but also enhanced immune activation. These findings support gIONPs@CMOT as a local immunomodulatory platform for postoperative osteosarcoma by activating antitumor immunity to inhibit tumor growth and suppress tumor recurrence, without significant toxicity.
