Abstract
The ongoing research on biomaterials that support bone regeneration led to the quest for materials or material modifications that can actively influence the activity or balance of bone tissue cells. The bone biocompatibility of porous chitosan scaffolds was modified in the present study by the addition of calcium phosphates or hemocyanin. The first strategy comprised the incorporation of calcium phosphates into chitosan to create a biomimetic chitosan—mineral phase composite. The second strategy comprised dip-coating of chitosan scaffolds with hemocyanin extracted from crayfish hemolymph. The cytocompatibility was assessed in a mono-culture of human bone marrow stromal cells (hBMSCs) and their differentiation to osteoblasts; in a mono-culture of human monocytes (hMs) and their maturation to osteoclasts; and in a co-culture of hBMSC/osteoblasts—hM/osteoclasts. Mineral incorporation caused an increase in scaffold bioactivity, as shown by reduced calcium concentration in the cell culture medium, delayed differentiation of hBMSCs, and reduced osteoclastic maturation of hMs in mono-culture. Dip-coating with hemocyanin led to increased proliferation of hBMSCs and equivalent osteoclast maturation in mono-culture, while in co-culture, both an inhibitory effect of mineral incorporation on osteoblastogenesis and stimulatory effects of hemocyanin were observed. It was concluded that highly bioactive scaffolds (containing mineral phases) restrain osteoblast and osteoclast development, while hemocyanin coating significantly supports osteoblastogenesis. These influences on the osteoblasts/osteoclasts activity ratio may support scaffold-driven bone healing in the future.
Highlights
Human bone is a complex and dynamic system, and efforts to study any aspect of this system in vitro will require a framework that is both complex and dynamic
We investigated the use of calcium phosphates to alter the scaffolds bioactivity, in terms of influencing the calcium ion concentration in the surrounding liquid, and used hemocyanin to improve the cytocompatibility of chitosan scaffolds with respect to osteoblast–osteoclast interaction
The first material modification was performed by the incorporation of calcium phosphate into the chitosan scaffolds
Summary
Human bone is a complex and dynamic system, and efforts to study any aspect of this system in vitro will require a framework that is both complex and dynamic. Molecules 2020, 25, 4580 a variety of in vitro cell cultures have been tested, including osteoblast/osteoclast co-cultures. The use of this type of culture in studies on the influence of biomaterials on bone regeneration and their integration into the remodeling process of bone has been comprehensively reviewed by Zhu et al [1]. One of the most important functions of bone substitute materials for human bone is the stimulation of bone-forming cells, namely, osteoblasts, derived from human bone marrow stromal cells (hBMSC). No less important are the bone-resorbing cells, namely, osteoclasts derived from human monocytes (hM), which provide and enhance non-inflammatory biomaterial resorption [4,5,6]. In the context of biomaterial development, increased osteoclastic activity may result in indirect stimulation of osteoblasts via osteoblast/osteoclast cross-talk, which may ensure local bone regeneration as a consequence of biomaterial resorption
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