In vivo regeneration functionalities of experimental organo-biomaterials containing water-soluble nacre extract

BACKGROUND: Novel multifunctional biomaterials were recently designed to allow for an optimized tissue regeneration process. PURPOSE: To comprehensively assess (photographic, radiographic and histological) the in vivo functionality of demineralized bovine bone matrix (DBM) associated with an experimental marine organic extract (MOE) from nacre in a sheep ectopic grafting model. MATERIALS AND METHODS: Synthesis of MOE was based on mixing powdered nacre (0.05 g, particles average size <0.1 mm) with acetic acid (5 mL, pH 7) under constant stirring for 72 hours (25 °C). Polyethylene tubes (3/animal, n = 4, diameter: 5.0 mm × length: 10.0 mm) from the control (empty) or experimental groups (DBM or DBM + MOE) were then intramuscularly implanted into the lumbar regions of sheep (n = 8, 2-years old, ≈45 kg). Animals were euthanized at 3 and 6 months to allow for the collection of tissue samples. Tissue samples were fixed in formalin 10% (buffered, 7 days) in preparation for photographic, radiographic and histological assessments. Acquired images were then analyzed using digital image analysis software to quantify the amount of neoformed tissues, whereas radiographic and histological analyses were performed to determine radiopacity and classification of tissues deposited inside of the tubes. RESULTS: Photographic and radiographic analyses have shown that both pure (unaltered) and MOE-modified DBM were capable of depositing neoformed tissues (at 3 and 6 months), where higher levels of deposition and radiopacity were observed on groups treated with experimental materials. Histological results, however, demonstrated that tissues formed from both unaltered and MOE-modified DBM were only fibrous connective in origin. CONCLUSIONS: As an ectopic grafting in sheep, the experimental organo-biomaterial association applied did not reveal any osteoinductive property but led to a fibrous tissue repair only.