Radiographic and histological evaluation of ectopic application of deproteinized bovine bone matrix

Neto

et al. 2018

Heliyon

BackgroundNovel multifunctional biomaterials were recently designed to allow for an optimized tissue regeneration process.PurposeTo 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 methodsSynthesis 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.ResultsPhotographic 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.ConclusionsAs 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.

Section: Discussionmentioning

confidence: 99%

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

Neto

et al. 2018

Heliyon

“…The methods of preparing the HDB reference the related research [ 8 ]. The cancellous bones of the femora were collected from New Zealand rabbits; then, soft tissue, cartilage and bone marrow were removed.…”

Section: Methodsmentioning

confidence: 99%

Repair of bone defects in rat radii with a composite of allogeneic adipose-derived stem cells and heterogeneous deproteinized bone

et al. 2018

BackgroundIn the bone tissue engineering domain, seed cells, scaffold and cell-scaffold composites are three focuses. In this study, the feasibility of using allogeneic adipose-derived stem cells(ADSCs) combined with heterogeneous deproteinized bone (HDB) to repair segmental radial defects was investigated by observing the repair of the defect area.MethodsADSCs were cultured in vitro, purified, antigen-detected and osteogenic differentiation potency-measured; then, the ADSCs of the third generation were seeded into HDB to prepare an ADSCs-HDB composite partly with osteogenesis induced cells. Sixty Wistar rats were randomly divided into four groups with 15 in each group. A bone defect (4 mm in length) was created at the left radius in each rat. Two kinds of ADSCs-HDB composites were implanted in the ADSCs osteogenesis group or ADSCs group; HDB was implanted in the negative control group; nothing was filled in the blank control group. The bone defect repair was evaluated by gross observation, molybdenum target X-ray examination and histological analyses after surgery.ResultsGross observation: the bone defect area was completely filled and difficult to recognize in the ADSCs osteogenesis group. The connection of the ADSCs group was strong, but the implants were clearly identifiable. The joints of the negative control group were slightly thick but the connection was unstable. In the blank control group, kermesinus tissue was between the two ends and bones were not connected after 8 weeks. Molybdenum target X-ray examinations: In the ADSCs osteogenesis group, evident bridges in the graft were observed in the defects in the fourth week; the defects were filled with new bone completely and a marrow cavity appeared at 8 weeks. In the ADSCs group, there were some callus formations, but the radial defect was still obvious at 8 weeks. In the negative control group, fracture lines were clear. In the blank control group, no osseous bridges were observed, which resulted in bone nonunion eventually in 8 weeks. There were significant differences in the callus density between experimental groups and the blank control group at 4 and 8 weeks (P < 0.01). Histological measures showed that the rate and quality of the new bone formation and remodelling was significantly different between the experimental and control groups.ConclusionsA composite of ADSCs-HDB has a strong osteogenic ability. It can repair segmental bone defects well and is promising to serve as grafting material in bone tissue engineering.

“…The guidelines for the handling and usage of laboratory animals recommended by the European Community were followed in the present study. The present pilot study was conducted based on methodologies previously described by Le Nihouannen et al (2005), da Silva et al (2016) and Zielak et al (2018).…”

Section: Internal Review Board (Irb)mentioning

confidence: 99%

“…The experimental MOE was then separated from suspended particles by centrifugation and stored under refrigeration (4C) until further use. (da Silva et al, 2016;Zielak et al, 2018). Experimental groups (CC BY 4.0) | ISSN 2525-3409 | DOI: http://dx.doi.org/10.33448/rsd-v10i5.15017 G1 and G4 served as the negative control (empty tubes), while groups G2 and G5 served as the positive control (tubes containing unaltered IBBM [GenOx Inorg  , Baumer, Brazil]).…”

Section: Manufacturing Of Marine Organic Extract (Moe)mentioning

confidence: 99%

In vivo transdifferentiation, osteoconductive and osteoinductive properties of experimental water-soluble organo-biomaterials – A Pilot Study

Crivellaro

Spada

Judachesci

et al. 2021

RSD

Self Cite

Inorganic bovine bone matrix (IBBM) is a biomaterial with proven osteoconductive functionalities. The objective of this study was to assess the in vivo bone regeneration functionalities of IBBM modified or not by an experimental MOE in sheep. MOE synthesis was performed by suspending nacre particles (0.05 g, diameters < 0.01 mm) in anhydrous acetic acid (pH 7, 5 mL, 25°C, 72 hours) using magnetic stirring. Polyethylene carriers (d= 5.0 mm, l= 10.0 mm, open ends) of negative control (sham) or experimental groups (IBBM or MOE-modified IBBM) were placed (n=3 conditions /animal; intramuscularly) adjacent to the lower spine of adult sheep (8 animals, » 45 Kg, 2 years old). Tissues were harvested (at 3 or 6 months) after implantation in preparation for histological (H), morphometrical (MM) and immunohistochemical analyses (IH; Wnt-3a, CD34, Vimentin and PREF-1). MM data were tested for normality and variance homogeneity using the Shapiro-Wilk and Levene tests, and Mann Whitney and Kruskal-Wallis, respectively. IM data were analyzed using two-way ANOVA and Tukey tests. Differences (p < 0.05) were observed between experimental groups (IBBM and IBBM+MOE at both 3 and 6 months) and controls (sham) for total area; Differences were not found for presence of remnant particles among experimental groups. The highest formation of bone was observed with IBBM+MOE (6-months). No differences (p > 0.05) were found on IM analysis (CD34, Vimentin, PREF-1, Wnt3a). Results indicated that experimental materials (IBBM+MOE) display promising functionalities. Additional studies are necessary to define biomaterials’ longitudinal effects and long-term biocompatibility properties.