Our previous researches have demonstrated the enhanced angiogenesis and ectopic bone formation by a single delivery system of stromal cell-derived fac

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2 Our previous researches have demonstrated the enhanced angiogenesis and ectopic bone formation by a single delivery system of stromal cell-derived factor-1 (SDF-1) and bone morphogenetic protein-2 (BMP-2) from gelatin hydrogels, respectively. In this study, when implanted with gelatin hydrogels incorporating mixed SDF-1 and BMP- 2, in vivo bone regeneration at an ulna critical-sized defect or a subcutaneous site was investigated to compare with that of hydrogels incorporating either SDF-1 or BMP-2. The release profiles of SDF-1 and BMP-2, the degradation profile of gelatin hydrogels, and the recruitment of bone marrow-derived mesenchymal stem and endothelial positive cells were assessed. The gelatin hydrogels incorporating mixed SDF-1 and BMP-2 enhanced bone regeneration to a significant extent, compared with those incorporating either SDF-1 or BMP-2. The release study showed the simultaneously controlled release of both SDF-1 and BMP-2 from gelatin hydrogels over 4 weeks. The time profiles of SDF-1 and BMP-2 release were correlated well with those of gelatin hydrogels degradation (R ). After implantation into green fluorescent protein (GFP)-positive Chimeric mice, the strongest osteocalcin (OCN)-positive cells were observed in the gelatin hydrogels incorporating mixed SDF-1 and BMP-2 4 weeks later. In addition, the same localization of OCN- and GFP-positive cells was observed, indicating the recruitment of bone marrow-derived mesenchymal stem cells to those hydrogels implanted and their osteogenic differentiation thereat. On the other hand, OCN-positive cells were not found for the hydrogels incorporating SDF-1 or phosphate-buffered saline (PBS). Furthermore, the highest expressions of CD31- and CD34-positive cells (endothelial markers) were clearly observed in the hydrogels incorporating mixed SDF-1 and BMP-2. An enhanced angiogenesis was observed around the tissue implanted with hydrogels incorporating mixed SDF-1 and BMP-2 or BMP-2. It is concluded that the dual release of SDF-1 and BMP-2 from gelatin hydrogels enhanced bone regeneration through an increased recruitment of osteoblast progenitor cells and an enhanced angiogenesis.

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6 Objectives: This study aimed to conduct a case-control investigation in maxilofacial morphogenesis, using a sample of Id2 homozygous knockout (KO) and wild-type (WT) mice. A special interest was to establish a relationship among Id2, BMP signals and chondrogenesis. Materials and Methods: Appropriate ethics approval has been received. Crania collected from mice at the age of 0 day, 2 weeks and 12 weeks were assessed with a Euclidean Distance Matrix Analysis method. Cultured synchondroses of the murine cranial base were inspected with a histological approach and examined with a reverse transcription polymerase chain reaction technique. Results: A shorter nasofrontal profile was seen in Id2 KO 12-week-olds but not 0-day-olds. The WT-KO gap in the skull length instead of the width increased with age. KO 2-week-olds showed a narrower hypertrophic zone and an inhibited proliferative zone in the presphenoid and the spheno-occipital synchondroses. Id2 expression in WT synchondroses was identified. Distribution of Type X collagen other than osteopontin was downregulated in Id2 KO samples. The WT murine cranial base showed a wider hypertrophic zone, a higher degree of ectopic hypertrophy and a larger number of proliferative chondrocytes in the presence of exogenous BMP2, BMP4 and BMP7, respectively. Such acquired growth was not detected in KO subjects. A 5-fold upregulation of Smad7 transcripts and a decrease in phosphorylation of Smad1-Smad5-and-Smad8-positive cells were identified in Id2 KO cartilage. Conclusions: Postnatal chondrogenesis was related to Id2 that acted downstream to enhance BMP signals by inhibiting Smad7 expression.

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