Cytotoxic effect of natural cuttlefish bone xenograft: an <em> in vitro </em> and <em> in vivo </em> study

Abstract

<jats:p>BACKGROUND Commercialized synthetic bone grafts are commonly used to replace the bone defect. Cuttlefish bone is naturally available and widely studied, but the specific cytotoxicity test has not been conducted. This study aimed to evaluate the cytotoxicity of the xenograft compared to commercial grafts. &#x0D; METHODS We performed an in vitro test evaluating the viability of human mesenchymal stem cells (hMSCs) when cultured for 48 hours with the tested materials (cuttlefish bone graft and fabricated PerOssal®). The trypsinized mitochondrial activity of the viable hMSC was assayed based on colorimetry of the formazan color change. The tested material was considered nontoxic if &gt;70% of the hMSCs were viable. The in vivo cytotoxic effect was evaluated by implanting the graft material in the femoral muscle of New Zealand (NZ) white rabbits. Nine rabbits were used in each test (cuttlefish bone, PerOssal®, and NaCl 0.9%). The systemic acute pyrogenic effect was evaluated based on 72 hours body weight changes and rectal temperature changes every 30 min in the first 3 hours and 72 hours post-implantation. &#x0D; RESULTS The mean percentage of hMSC viability when cultured with cuttlefish bone graft and PerOssal® was comparable (93.47% and 105.37%, respectively, p = 0.240). The in vivo cytotoxicity on NZ rabbit was similar between all tested materials, as shown by the minor changes in body weight (&lt;10% body weight, p = 0.391) and rectal temperature (&lt;0.5°C, p = 0.127). &#x0D; CONCLUSIONS Cuttlefish bone xenograft and fabricated PerOssal® have a similar non-cytotoxic effects on hMSCs and non-pyrogenic systemic effects on rabbits.</jats:p>

Characterization and Potential of Enhancing Bone Regeneration Using Raw Cuttlebone Aragonite Nanoparticles: In Vitro and In Vivo Studies.

“…Additionally, Irianto K A and colleagues confirmed the high viability of human mesenchymal stem cells when exposed to CB, thereby underscoring its potential safety for human applications. 24 Although some heavy metals in small amounts have been speculated to be found in CB, as mercury, copper, zinc, lead, and cadmium, they showed no cytotoxic effect on cell culture of human mesenchymal stem cells. Moreover, CB allowed the growth and attachment of these cells and maintained their regular fibroblast-like shape.…”

In Situ Construction of Morphologically Different Hydroxyapatite-Mineralized Structures on a Three-Dimensional Bionic Chitin Scaffold

“…10 Its special natural porous skeletal structure can promote the regeneration of newly formed bone tissue, and thus it has been widely used for bone tissue engineering. 11,12 β-Chitin from cuttlefish bone is the second most abundant natural polymeric polysaccharide after cellulose and consists of N-acetyl-D-glucosamine linked by β-1,4-glycosidic bonds, 13,14 which has a parallel arrangement of molecular chains resulting in weak intermolecular hydrogen bonding, high affinity for solvents, and high reactivity. 15−17 of mineralization (calcification).…”

“…Cuttlefish bone, mainly composed of aragonite inorganic mineral and β-chitin organic matter, has an exquisite three-dimensional interconnected pore channel structure, with a porosity of about 93% and pore diameters varying from 200 to 600 μm . Its special natural porous skeletal structure can promote the regeneration of newly formed bone tissue, and thus it has been widely used for bone tissue engineering. , β-Chitin from cuttlefish bone is the second most abundant natural polymeric polysaccharide after cellulose and consists of N -acetyl- d -glucosamine linked by β-1,4-glycosidic bonds, , which has a parallel arrangement of molecular chains resulting in weak intermolecular hydrogen bonding, high affinity for solvents, and high reactivity. − β-Chitin has received much attention in the field of tissue engineering due to its biodegradability, biocompatibility, nontoxicity, and promotion of mineralization (calcification). − However, the application of cuttlefish bone in tendon–bone healing is limited by the fact that the inorganic minerals in cuttlefish bone do not match the specific gradient structure of tendon–bone healing. Therefore, this work wanted to extract the organic matter while preserving the special natural porous skeletal structure of cuttlefish bone and use it as a template for the deposition of mineralized inorganic minerals to construct a gradient mineral volume scaffold with a special natural porous skeletal structure.…”