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E/bone scaffold alone, FB/ANG 0.5; FB scaffold PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/9282946 loaded with 0.5 g of ANG, FB/ANG 2.0; FB scaffold loaded with 2.0 g of ANG. Scale bar = 250 mKim et al. Biomaterials Research (2015) 19:Page 6 ofimplanted with FB and FB/ANG scaffolds. Extensive newly formed bone was observed in the FB/ANG groups, and the bone had coalesced with the host bone. Furthermore, extensive new bone was observed in the group implanted with FB/ANG 2.0 scaffolds (Fig. 6). Images of the central area of the calvarial defects collected after MT staining showed large amounts of fibroblastic connective tissue in the group with untreated defects, whereas mature new bone was clearly observed in the FB and FB/ANG groups. In addition, more newly formed bone was observed in the FB/ANG 2.0 group than in the FB/ANG 0.5 group (Fig. 7).Micro-computed tomography evaluationReconstructed three-dimensional micro-CT images of non-implanted (empty) and implanted FB, FB/ANG 0.5, and FB/ANG 2.0 scaffolds are shown in Fig. 8a. In all groups, bone regeneration occurred from the margins of the defect site. In particular, new bone was generated not only in the margin of the defect but also in theFig. 7 Goldner's Masson trichrome staining of Methyl 2-((4-nitro-1h-pyrazol-1-yl)methyl)benzoate regenerated bone in the central area of the calvarial defect after implantation for 4 weeks and 8 weeks. After implantation for 4 weeks, mature bone (arrow) was distinctly observed around the residual material bone particles in the angiogenin-containing groups. Chronic inflammatory cells were also observed to infiltrate all defect areas. After implantation for 8 weeks, the greatest extent of regenerated bone (arrow) and blood vessels (arrowhead) was observed in the FB/ANG 2.0 group. Asterisk: residual material bone particles. Scale bar = 250 mcentral area of the defect among the bone powder in the bone powder-containing scaffolds (FB, FB/ANG 0.5, and FB/ANG 2.0). Quantification of bone regeneration revealed s12998-016-0127-6 significant differences in the group with untreated defects and the groups implanted with FB and (S)-3-(tert-Butoxycarbonyl)-2,2-dimethyloxazolidine-4-carboxylic acid FB/ANG scaffolds. In particular, at 8 weeks, the newly formed bone volume was significantly higher in the FB/ANG 2.0 implanted group (34.67 ?1.25 ) than in the untreated defect (12.82 ?1.49 ), FB (20.12 ?1.08 ), and FB/ANG 0.5 (27.34 ?2.17 ) groups (Fig. 8b). The results of the micro-CT analysis were similar to those of the histological analysis.Fig. 6 Microscopic evaluation of calvarial sections stained with hematoxylin-eosin after 8 weeks. Remaining bone particle material (asterisk) was observed in all scaffold-implanted groups. Newly generated mineralized bone (black arrow) was observed near the margin of the defect site (arrowhead) in the calvarial bone. HB represents the host bone. Scale bar PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/12113769 = 500 mDiscussion Bone tissue engineering is promising for treating large bone defects in patients with severe bone loss. Therefore, many studies have attempted to improve bone tissue defect repair using biological scaffolds. One of the major factors in bone tissue engineering is angiogenesisKim et al. Biomaterials Research (2015) 19:Page 7 ofFig. 8 Three dimensional images of calvarial defects after implantation for 8 weeks (a). Newly regenerated bone covered the entire defect with scaffold treatment after implantation for 8 weeks. a; empty, b; FB, c; FB/ANG 0.5, d; FB/ANG 2.0. B. The bone volume generated in the bone defects. The amount of new bone significantly increased over the study duration in the FB/ANG-implanted groups (b). Furthermore, the FB.

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