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The development of bioactive injectable system as cell carrier with minimal impact on viability of encapsulated cells represents a great challenge. In the present work, we propose a new pH-responsive chitosan-hydroxyapatite-based hydrogel with sodium bicarbonate (NaHCO) as the gelling agent. The in situ synthesis of hydroxyapatite phase has resulted in stable composite suspension and final homogeneous hydrogel. The application of sodium bicarbonate has allowed non-cytotoxic fast gelation of chitosan-hydroxyapatite within 4min, and without excess of sodium ions concentration. Rheological properties of crosslinked hydrogel have demonstrated possible behaviour as 'strong physical hydrogel'. The live dead staining has confirmed good viability and dispersion, as well as proliferation of encapsulated cells by the culture time. Presented preliminary results show good potential of chitosan-hydroxyapatite/NaHCO as a cell carrier, whose impact on the cell differentiation need to be confirmed by encapsulation of other cell phenotypes.

Natural bone tissue is composed of calcium-deficient carbonated hydroxyapatite as the inorganic phase and collagen type I as the main organic phase. The biomimetic approach of scaffold development for bone tissue engineering application is focused on mimicking complex bone characteristics. Calcium phosphates are used in numerous studies as bioactive phases to mimic natural bone mineral. In order to mimic the organic phase, synthetic (e.g., poly(ε-caprolactone), polylactic acid, poly(lactide-co-glycolide acid) and natural (e.g., alginate, chitosan, collagen, gelatin, silk) biodegradable polymers are used. However, as materials obtained from natural sources are accepted better by the human organism, natural polymers have attracted increasing attention. Over the last three decades, chitosan was extensively studied as a natural polymer suitable for biomimetic scaffold development for bone tissue engineering applications. Different types of chitosan-based biomaterials (e.g., molded macroporous, fiber-based, hydrogel, microspheres and 3D-printed) with specific properties for different regenerative applications were developed due to chitosan’s unique properties. This review summarizes the state-of-the-art of biomaterials for bone regeneration and relevant studies on chitosan-based materials and composites.

Injectable hydrogels have emerged as promising biomaterials for tissue engineering applications. The goal of this study was to evaluate the potential of a pH-responsive chitosan-hydroxyapatite hydrogel to be used as a three-dimensional support for encapsulated mesenchymal stem cells (MSCs) osteogenic differentiation. In vitro enzymatic degradation of the hydrogel, during 28 days of incubation, in simulated physiological condiditons, was characterized by swelling measurements, molecular weight determination and SEM analysis of hydrogel microstructure. Osteogenic differentiation of encapsulated MSCs was confirmed by osteogenic Runx2, collagen type I and osteocalcin immunostaining and alkaline phosphatase quantification. The deposition of late osteogenic markers (calcium phosphates) detected by Alizarin red and von Kossa staining indicated an extracellular matrix mineralization.

The implementation of a powder bed selective laser processing (PBSLP) technique for bioactive ceramics, including selective laser sintering and melting (SLM/SLS), a laser powder bed fusion (L-PBF) approach is far more challenging when compared to its metallic and polymeric counterparts for the fabrication of biomedical materials. Direct PBSLP can offer binder-free fabrication of bioactive scaffolds without involving postprocessing techniques. This review explicitly focuses on the PBSLP technique for bioactive ceramics and encompasses a detailed overview of the PBSLP process and the general requirements and properties of the bioactive scaffolds for bone tissue growth. The bioactive ceramics enclosing calcium phosphate (CaP) and calcium silicates (CS) and their respective composite scaffolds processed through PBSLP are also extensively discussed. This review paper also categorizes the bone regeneration strategies of the bioactive scaffolds processed through PBSLP with the various modes of functionalization through the incorporation of drugs, stem cells, and growth factors to ameliorate critical-sized bone defects based on the fracture site length for personalized medicine.

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Nanocomposite structure of the bone can be mimicked by chitosan/hydroxyapatite (CS/HAp) composite scaffold. Biological hydroxyapatite (HAp) contains various ions, which have a crucial role in bone growth. The aim of the present work was to synthesize biomimetic hydroxyapatite and prepare composite scaffolds based on chitosan, where HAp was synthesised from hen eggshells, seashells and cuttlefish bone. The powders were composed of nano-structured calcium deficient HAp and amorphous calcium phosphate (ACP). In the as-prepared powders, Sr2+, Mg2+ and Na+ ions were detected as a result of using biogenic precursor of Ca2+ ions. Highly porous CS/HAp structures have been prepared by freeze-gelation technique. The CS/HAp scaffolds have shown highly porous structure with very well interconnected pores and homogeneously dispersed HAp particles. The MTT assay of CS/HAp scaffolds has shown no toxicity, and the live/dead assay has confirmed good viability and proliferation of seeded cells.