Characteristic and Chondrogenic Differentiation Analysis of Hybrid Hydrogels Comprised of Hyaluronic Acid Methacryloyl (HAMA), Gelatin Methacryloyl (GelMA), and the Acrylate-Functionalized Nano-Silica Crosslinker

Abstract

Developing a biomaterial suitable for adipose-derived stem cell (ADSCs)-laden scaffolds that can directly bond to cartilage tissue surfaces in tissue engineering has still been a significant challenge. The bioinspired hybrid hydrogel approaches based on hyaluronic acid methacryloyl (HAMA) and gelatin methacryloyl (GelMA) appear to have more promise. Herein, we report the cartilage tissue engineering application of a novel photocured hybrid hydrogel system comprising HAMA, GelMA, and 0~1.0% (w/v) acrylate-functionalized nano-silica (AFnSi) crosslinker, in addition to describing the preparation of related HAMA, GelMA, and AFnSi materials and confirming their related chemical evidence. The study also examines the physicochemical characteristics of these hybrid hydrogels, including swelling behavior, morphological conformation, mechanical properties, and biodegradation. To further investigate cell viability and chondrogenic differentiation, the hADSCs were loaded with a two-to-one ratio of the HAMA-GelMA (HG) hybrid hydrogel with 0~1.0% (w/v) AFnSi crosslinker to examine the process of optimal chondrogenic development. Results showed that the morphological microstructure, mechanical properties, and longer degradation time of the HG+0.5% (w/v) AFnSi hydrogel demonstrated the acellular novel matrix was optimal to support hADSCs differentiation. In other words, the in vitro experimental results showed that hADSCs laden in the photocured hybrid hydrogel of HG+0.5% (w/v) AFnSi not only significantly increased chondrogenic marker gene expressions such as SOX-9, aggrecan, and type II collagen expression compared to the HA and HG groups, but also enhanced the expression of sulfated glycosaminoglycan (sGAG) and type II collagen formation. We have concluded that the photocured hybrid hydrogel of HG+0.5% (w/v) AFnSi will provide a suitable environment for articular cartilage tissue engineering applications.

Anatomy, molecular structures, and hyaluronic acid – Gelatin injectable hydrogels as a therapeutic alternative for hyaline cartilage recovery: A review

“…These findings allowed to investigate how the matrix viscoelasticity affected MSCs behavior and fate, suggesting that hydrogel microenvironment with increasing viscosity improve chondrogenesis 132 . Factors such as pore size, swelling ratio and compression modulus should be characterized to elucidate the suitable parameters to induce chondrogenesis; in fact, pore size (75 ± 0.2 μm), swelling ratio (35%), and compression modulus (35 ± 1.5 kPa) of hyaluronic acid methacryloyl and gelatin methacryloyl hydrogels promotes human adipose‐derived stem cells differentiation into a chondrocyte lineage, stimulating SOX9, aggrecan, Col2α1‐IIa and GAGs 133 . Similar cartilage specific genes have been up‐regulated in polyglutamic acid – sodium alginate hydrogels whit higher compression modulus (107 ± 8.6 kPa) and a toughness range of 481 ± 34 kJ/m 3 –2166 ± 63 kJ/m 3,133 .…”

Designing functional hyaluronic acid-based hydrogels for cartilage tissue engineering

“… Hydrogels Components Physicochemical properties Biofunctions Ref. Methacrylated HA (HA-MA) HA; methacrylic anhydride (MA) Viscoelasticity; controlled pore size and degradation rate Encapsulating cells; promoting MSCs proliferation, migration and chondrogenesis; promoting ECM deposition [ [30] , [31] , [32] , [33] , [34] , [35] ] HA-MA/MeLAHA HA, MA, MeLAHA Hydrolytic degradation and enzymatic degradation Promoting MSCs chondrogenesis and ECM deposition; ensuring stability of hydrogel scaffold [ 36 , 37 ] HA-MA/MMP7 HA, MA, matrix metalloproteinase 7 Tunable mechanical properties, swelling performance and degradation rates Promoting MSCs chondrogenesis [ 38 , 39 ] HA-MA/Gel-MA HA, MA, gelatin, Improving the mechanical properties Enhancing cell proliferation, aggregation and chondrogenesis; maintaining the chondrocyte phenotype; enhancing deposition and distribution of ECM; accelerating cartilage repair [ [40] , [41] , [42] , [43] , [44] ] HA-MA/Gel-MA/BC HA, MA, gelatin, bacterial cellulose Improving the mechanical properties and printing fidelity Facilitating chondrocyte proliferation and protein expression [ 45 ] HA-MA/Gel-MA/AFnSi HA, MA, gelatin, acrylate-functionalized nanosilica Controllable pore sizes, swelling ratios and mechanical properties Promoting the chondrogenic gene expression and ECM deposition [ 46 ] HA-MA/CNF HA, MA, methacrylated cellulose nanofibers Enhanced mechanical properties; decent restorability Enhancing BMSCs proliferation and chondrogenesis; accelerating full-thickness cartilage repair [ 48 ...…”

“…Furthermore, compared with porcine-derived Gel-MA, the HA-MA/Gel-MA hybrid hydrogel constructed using bovine-derived Gel-MA and HA-MA through irgacure 2959-induced photocrosslinking (365 ​nm) exhibited the best similarity to native articular cartilage after 28 days of chondrocyte cultivation [ 44 ]. The mechanical properties of HA-MA/Gel-MA hybrid hydrogels can be further optimized by incorporating organic polymers or inorganic nanoparticles [ 45 , 46 ]. For example, Sang et al.…”

“…used acrylate-functionalized nanosilica (AFnSi) to crosslink HA-MA/Gel-MA hybrid hydrogels, and the obtained hydrogels exhibited controllable pore sizes, swelling ratios, and mechanical properties. Moreover, the hydrogels could promote the gene expression of chondrogenic markers, including SOX9, proteoglycan, and collagen II, in human adipose-derived MSCs (AMSCs) and increase the formation of sulfated GAG (sGAG) and collagen II [ 46 ].…”

Photo-Crosslinkable Hydrogels for 3D Bioprinting in the Repair of Osteochondral Defects: A Review of Present Applications and Future Perspectives

“…It solidifies into gel within 10 s under visible light irradiation. Due to the portable forming method and good biocompatibility, materials based on HAMA have been applied to many biomedical fields, including in chondrocyte culture and cartilage regeneration, tumor model construction, drug-controlled release, microneedle preparation, wound dressing, biosensors, and postoperative anti-adhesion [ 45 , 46 , 47 , 48 ]. Galarraga et al used a norbornene-modified hyaluronic acid (NorHA) macromer as a representative bioink and varied the printing parameters (e.g., capillary length, flow rate, light intensity) to identify printing conditions that were optimal for the ink [ 49 ], and then marrow mesenchymal stem cells (MSCs) were encapsulated in the bioink and the compound bioink was fabricated using photo irradiation for cartilage repair.…”