Nanocellulose‐Reinforced Hydroxyapatite Nanobelt Membrane as a Stem Cell Multi‐Lineage Differentiation Platform for Biomimetic Construction of Bioactive 3D Osteoid Tissue In Vitro

Abstract

Severe bone defects, especially accompanied by vascular and peripheral nerve injuries, remain a massive challenge. Most studies related to bone tissue engineering have focused on osteogenic differentiation of mesenchymal stem cells (MSCs), and ignored the formation of blood vessels and nerves in the newly generated bone owing to the lack of proper materials and methodology for tuning stem cells differentiated into osteogenic, neuronal, and endothelial cells (ECs) in the same scaffold system. Herein, a nanocellulose-reinforced hybrid membrane with good mechanical properties and control over biodegradation by assembling ultralong hydroxyapatite nanobelts in a bacterial nanocellulose hydrogel is designed and synthesized. Osteogenic, neuronal cells are successfully differentiated on this hybrid membrane. Based on the multi-lineage differentiation property of the membrane, a bioactive 3D osteoid tissue (osteogenic, neural, and ECs) is mimetically constructed in vitro using layer-by-layer culture and integration. The bone regeneration ability of the as-prepared bioactive osteoid tissue is assessed in vivo via heterotopic osteogenesis experiments for eight weeks. The rapid new bone growth and formation of blood capillaries and nerve fibers prove that the hybrid membrane can be universally applied as a stem cell multi-lineage differentiation platform, which has significant applications in bone tissue engineering.

Potassium Titanate Assembled Titanium Dioxide Nanotube Arrays Endow Titanium Implants Excellent Osseointegration Performance and Nerve Formation Potential

“…As shown in Figure 5C , the expression of MAP2 in the cells on the K 2 Ti 6 O 13 -TiO 2 hybrid nanorod arrays was 1.9 folds higher than that on the TCPs. Tuj1 and MAP2, which significantly affect neurogenesis, are the two most important types of representation forms during the neurogenic differentiation period ( Liu et al, 2021 ). The above experimental results show that NSCs cultured on K 2 Ti 6 O 13 -TiO 2 hybrid nanorod arrays present higher neuron-related gene expression levels than those on TCPs or TiO 2 arrays, indicating that the K 2 Ti 6 O 13 -TiO 2 hybrid nanorod arrays possess excellent acceleration functions for the neurogenic differentiation of NSCs.…”

“…As mentioned above, TiO 2 and nanostructures on the surface of titanium implants can regulate the osteogenic differentiation of MSCs. Notably, the incorporation of essential biological elements has been proven to be beneficial for activating the peripheral nervous system ( Liu et al, 2021 ) and regulating bone regeneration ( Li et al, 2020 ). Potassium is one of the most abundant cations in intracellular fluid and plays an essential role in cell function, particularly in excitable cells, such as muscles and nerves ( Fortin et al, 2008 ; Wang et al, 2020 ).…”

Nanohydroxyapatite as a Biomaterial for Peripheral Nerve Regeneration after Mechanical Damage—In Vitro Study

“…Ideal material for nerve regeneration should have physicochemical properties similar to the nerves [ 20 , 21 ]. The neuroregenerative activity of nHAp has been demonstrated in various scaffolding materials, e.g., in type I collagen gel, nanocellulose hydrogel, and carbon nanotubes [ 16 , 21 , 22 , 23 , 24 ]. Nanohydroxyapatite can also be used as a coating for other materials that are themselves neurotoxic, e.g., carbon nanotubes, which improve nerve regeneration by directing axon growth [ 23 ].…”

“…Moreover, nHAp did not neutralize the electrophysiological activity of neurons, as Liu et al demonstrated in primary cultures of rat neurons [ 23 ]. Currently, nanotubes made in 3D printing technology from combined materials containing hydroxyapatite are also being studied [ 24 , 25 ]. In vivo studies have shown that nHAp has a beneficial effect on nerve regeneration in rats after prior spinal distension.…”

Bacterial Nanocellulose as a Scaffold for In Vitro Cell Migration Assay

“…One of the best accepted clinical applications of BNC so far has been a topical wound dressing, while the potential of BNC application for medical implants, tissue engineering and drug delivery is great and still untapped [9][10][11][12]. Recently, BNC has emerged as a promising scaffold material for the adhesion and proliferation of cells and 3D bioprinting for tissue engineering purposes [13][14][15]. Biomedical applications of BNC rely largely on its network-like structure, which can be improved using chemical and genetic engineering tools [16][17][18][19].…”