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Silk bilayer scaffolds can induce fast integration with subchondral bone and support cartilage repair

Abstract

Publicado em : J Tissue Eng Regen Med 2014; 8 (Suppl. 1)Introduction: Osteochondral defect (OCD) regeneration presents major challenges in orthopedics. Since healing of cartilage and bone should be simultaneously considered, ideal scaffolds should be those that can mimic both tissues properties. In this study, bilayered silk and silk-nano calcium phosphate (Silk/Silk-NanoCaP) scaffolds with tailored mechanical properties were developed for OCD tissue engineering application. Materials and methods: Aqueous silk solution (16%) was prepared.1 Nano calcium phosphate particles (16%) were synthesized in the silk solution (Silk-NanoCaP).2 The bony layer was prepared by addition of NaCl particles (500–1000 lm) into the Silk-NanoCaP suspension. After drying for 2 days and salt-leaching overnight, silk solution was added on top of the bony layer using the same procedure to produce the chondral layer. The !nal scaffolds were evaluated through in vitro culture of rabbit bone marrow stromal cells (RBMSCs) for 2 weeks, and in vivo implantation in a rabbit knee OCD for 4 weeks. Results: The RBMSCs cultured in the scaffolds presented increasing viability from day 1 to day 7 by MTS assay. Good adhesion and migration of the RBMSCs in the scaffolds were achieved, as observed under the scanning electron microscope. Cell proliferation was observed from day 7 to day 14 as determined by DNA quanti!cation. The bony layer induced higher alkaline phosphatase level as compared to the chondral layer, in osteogenic condition. Histological analysis (H&E) showed that the bilayered scaffolds integrated well with the host tissue, after 4 weeks of implantation in a critical size OC defect (Fig. 1). Abundant new bone formation was detected in the Silk-NanoCaP layer. Cartilage regeneration occurred in the silk layer. Discussion and conclusions: The bilayered scaffolds favored the attachment, proliferation, and differentiation of RBMSCs. The bony layer of the bilayered scaffolds possessed osteoconductive properties. The bilayered scaffolds were biocompatible in vitro and in vivo. These scaffolds also induced both subchondral bone regeneration and supported cartilage regeneration, thus showing great promise in OCD regeneration. Acknowledgments: The authors thank FCT projects Tissue2Tissue and OsteoCart, and the FP7 Programme POLARIS. Yan LP was awarded a FCT PhD scholarship. Investigador FCT program (IF/00423/2012) and (IF/00411/2013) are also greatly acknowledged. Disclosure: The authors declare that there is no con"ict of interest

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