Double cross-linked interpenetrating polymer network (IPN) hydrogels of sodium alginate and gelatin (SA/G) reinforced with 50 wt % cellulose nanocrystals (CNC) have been prepared via the freeze-drying process. The IPNs were designed to incorporate CNC with carboxyl surface groups as a part of the network contribute to the structural integrity and mechanical stability of the hydrogel. Structural morphology studies of the hydrogels showed a three-dimensional (3D) network of interconnected pores with diameters in the range of 10-192 μm and hierarchical pores with a nanostructured pore wall roughness, which has potential benefits for cell adhesion. Significant improvements in the tensile strength and strain were achieved in 98% RH at 37 °C for CNC cross-linked IPNs. The high porosity of the scaffolds (>93%), high phosphate buffered saline (PBS) uptake, and cytocompatibility toward mesenchymal stem cells (MSCs) are confirmed and considered beneficial for use as a substitute for cartilage.
Fully bio-based 3-dimensional porous scaffolds based on freeze-dried cellulose nanofibers (70-90 wt%) stabilized using a genipin crosslinked matrix of gelatin and chitosan were prepared. Morphology studies using scanning electron microscopy showed that the scaffolds have interconnected pores with average pore diameters of 75-200 mm and nanoscaled pore wall roughness, both favorable for cell interactions with cartilage repair. X-ray tomography confirmed the 3-dimensional homogeneity and interconnectivity of the pores as well as the fibrillar structure of the scaffolds. The compression modulus of the scaffolds (1-3 MPa) at room conditions was higher than natural cartilage (z1 MPa). The lowered compression modulus of 10-60 kPa in phosphate buffered saline (PBS) at 37 C was considered favorable for chondrogenesis. The current study therefore successfully addressed the challenge of tailoring the pore structure and mechanical properties simultaneously for cartilage regeneration. Furthermore, the scaffolds' high porosity (z95%), high PBS uptake and good cytocompatibility towards chondrocytes are considered beneficial for cell attachment and extracellular matrix (ECM) production.
Cellulose nanofibrils (CNFs), unique and promising natural materials have gained significant attention recently for biomedical applications, due to their special biomechanical characteristics, surface chemistry, good biocompatibility and low toxicity. However, their long bio-persistence in the organism may provoke immune reactions and this aspect of CNFs has not been studied to date. Therefore, the aim of this work was to examine and compare the cytocompatibility and immunomodulatory properties of CNFs in vitro. CNFs (diameters of 10-70 nm; lengths of a few microns) were prepared from Norway spruce (Picea abies) by mechanical fibrillation and high pressure homogenisation. L929 cells, rat thymocytes or human peripheral blood mononuclear cells (PBMNCs) were cultivated with CNFs. None of the six concentrations of CNFs (31.25 µg/ml-1 mg/ml) induced cytotoxicity and oxidative stress in the L929 cells, nor induced necrosis and apoptosis of thymocytes and PBMNCs. Higher concentrations (250 µg/ml-1 mg/ml) slightly inhibited the metabolic activities of the L929 cells as a consequence of inhibited proliferation. The same concentrations of CNFs suppressed the proliferation of PBMNCs to phytohemaglutinine, a T-cell mitogen, and the process was followed by down-regulation of interleukin-2 (IL-2) and interferon-γ production. The highest concentration of CNFs inhibited IL-17A, but increased IL-10 and IL-6 production. The secretion of pro-inflammatory cytokines, IL-1β and tumor necrosis factor-α as well as Th2 cytokine (IL-4), remained unaltered. In conclusion, the results suggest that these CNFs are cytocompatible nanomaterial, according to current ISO criteria, with non-inflammatory and nonimmunogenic properties. Higher concentrations seem to be tolerogenic to the immune system, a characteristic very desirable for implantable biomaterials.
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