Bone tissue engineering (BTE) is an ongoing field of research based on clinical needs to treat delayed and non-union long bone fractures. An ideal tissue engineering scaffold should have a biodegradability property matching the rate of new bone turnover, be non-toxic, have good mechanical properties, and mimic the natural extracellular matrix to induce bone regeneration. In this study, biodegradable chitosan (CS) scaffolds were prepared with combinations of bioactive ceramics, namely hydroxyapatite (HAp), tricalcium phosphate-α (TCP- α), and fluorapatite (FAp), with a fixed concentration of benzophenone photoinitiator (50 µL of 0.1% (w/v)) and crosslinked using a UV curing system. The efficacy of the one-step crosslinking reaction was assessed using swelling and compression testing, SEM and FTIR analysis, and biodegradation studies in simulated body fluid. Results indicate that the scaffolds had comparable mechanical properties, which were: 13.69 ± 1.06 (CS/HAp), 12.82 ± 4.10 (CS/TCP-α), 13.87 ± 2.9 (CS/HAp/TCP-α), and 15.55 ± 0.56 (CS/FAp). Consequently, various benzophenone concentrations were added to CS/HAp formulations to determine their effect on the degradation rate. Based on the mechanical properties and degradation profile of CS/HAp, it was found that 5 µL of 0.1% (w/v) benzophenone resulted in the highest degradation rate at eight weeks (54.48% degraded), while maintaining compressive strength between (4.04 ± 1.49 to 10.17 ± 4.78 MPa) during degradation testing. These results indicate that incorporating bioceramics with a suitable photoinitiator concentration can tailor the biodegradability and load-bearing capacity of the scaffolds.

A large bone defect is defined as a defect that exceeds the regenerative capacity of the bone. Nowadays, autologous bone grafting is still the gold standard treatment. In this study, a hybrid bone tissue engineering scaffold (BTE) was designed with biocompatibility, biodegradability and adequate mechanical strength as the primary objectives. Chitosan (CS) is a biocompatible and biodegradable polymer that can be used in a wide range of applications in bone tissue engineering. Hydroxyapatite (HAp) and fluorapatite (FAp) have the potential to improve the mechanical properties of CS. In the present work, different volumes of acetic acid (AA) and different ratios of HAp and FAp scaffolds were prepared and UV cross-linked to form a 3D structure. The properties of the scaffolds were characterised by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, swelling studies and compression testing. The cytotoxicity result was obtained by the MTT assay. The degradation rate was tested by weight loss after the scaffold was immersed in SBF. The results showed that a crosslinked structure was formed and that bonding occurred between different materials within the scaffold. Additionally, the scaffolds not only provided sufficient mechanical strength but were also cytocompatibility, depending on their composition. The scaffolds were degraded gradually within a 6-to-8-week testing period, which closely matches bone regeneration rates, indicating their potential in the BTE field.

Bone has the ability to heal itself; however, bone defects fail to heal once the damage exceeds a critical size. Bone regeneration remains a significant clinical challenge, with autograft considered the ideal bone graft material due to its sufficient porosity, osteogenic cells, and biological growth factors. However, limitations to bone grafting, such as limited bone stock and high resorption rates, have led to a great deal of research into developing bone graft substitutes. The P28 peptide is a small molecule bioactive biomimetic alternative to mimic the bone morphogenetic protein 2 (BMP-2). In this study, we investigated the potential of P28-loaded hybrid scaffolds to mimic the natural bone structure for enhancing the bone regeneration process. We hypothesized that the peptide-loaded scaffolds and nude scaffolds both have the potential to promote bone healing, and the bone healing process is accelerated by the release of the peptide. To verify our hypothesis, C2C12 cells were evaluated for the presence of calcium deposits by histological stain at 7 and 14 days in cultures with hybrid scaffolds. Total RNA was isolated from C2C12 cells cultured with hybrid scaffolds for 7 and 14 days to assess osteoblast differentiation. The project findings demonstrated that the hybrid scaffold could enhance osteoblast differentiation and significantly improve the therapeutic effects of the scaffold in bone regeneration.

Repairing large bone defects remains a significant clinical challenge due to the limitations of current treatments, including infection risk, donor site morbidity, and insufficient vascularization. The autograft is still the gold standard for large bone defects. In this study, we developed chitosan-based (CS-based) scaffolds, incorporating with hydroxyapatite (HAp) and fluorapatite (FAp) ceramics, fabricated by UV crosslinking and freeze-drying, and loaded with P28 peptide, alone or in combination with vascular endothelial growth factor (VEGF), to evaluate the effect of dual bioactive factor delivery. We hypothesized that CS-based scaffolds would optimize ceramic composition and co-delivery of P28 and VEGF, and can enhance early-stage osteogenic differentiation and support bone regeneration. The CS-based scaffolds were characterized by their physicochemical properties, including swelling behavior, mechanical strength, porosity, and in vitro degradation. Biological evaluations were performed including cell proliferation assays, ALP activity, ARS staining, and RT-qPCR, to assess osteogenic differentiation. The results showed that the scaffolds had high porosity, excellent swelling behavior, and degraded within 8 weeks. Dual delivery of P28 and VEGF significantly enhanced early osteogenic markers, indicating a complementary effect. These findings demonstrated that CS-based scaffolds with an optimized ceramic ratio and bioactive factor incorporation have the potential to facilitate bone regeneration.