Chinese Journal of Tissue Engineering Research ›› 2016, Vol. 20 ›› Issue (52): 7773-7780.doi: 10.3969/j.issn.2095-4344.2016.52.003

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Construction of a polycaprolactone/bone extracellular matrix scaffold with three-dimensional printing technology and its osteoinductivity in vitro

Yang Peng, Li Chun-de
  

  1. Department of Orthopaedics, Peking University First Hospital, Beijing 100034, China
  • Received:2016-09-23 Online:2016-12-16 Published:2016-12-16
  • Contact: Li Chun-de, Chief physician, Professor, Department of Orthopaedics, Peking University First Hospital, Beijing 100034, China
  • About author:Yang Peng, M.D., Department of Orthopaedics, Peking University First Hospital, Beijing 100034, China

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Abstract:

BACKGROUND: Scholars are still looking for ideal bone tissue-engineered scaffolds, and  three-dimensional (3D) printing technology is a novel construction method. In the meanwhile, bone extracellular matrix is becoming a hotspot in osteogenic induction.
OBJECTIVE: To construct the polycaprolactone/bone extracellular matrix scaffold using 3D printing technology and co-culture method, and to detect its osteogenic property.
METHODS: 216 3D-printed polycaprolactone scaffolds were divided into group A (96 pores, n=72) and group B(48 pores, n=144). Passage 5 bone marrow mesenchymal stem cells from Sprague-Dawley rats were seeded onto the two kinds of polycaprolactone scaffolds, and the group A was used for alizarin red staining and Masson staining, while the group B for collagen and glycosaminoglycan detection at 1, 2 and 3 weeks of incubation. Afterwards, the scaffolds at 1, 2 and 3 weeks of culture were decellularized and labeled as groups AE1, AE2, AE3, BE1, BE2 and BE3. Then passage 5 bone marrow mesenchymal stem cells from Sprague-Dawley rats were seeded onto each scaffold again, and the former three groups underwent alizarin red staining, and the latter three were used for calcium, alkaline phosphatase activity and DNA quantitative analysis at 1, 2 and 3 weeks of culture.
RESULTS AND CONCLUSION: Masson staining, glycosaminoglycan and hydroxyproline quantitative analysis showed that the extracellular matrix on the composite scaffold increased with time. Alkaline phosphatase activity revealed that the composite scaffold had a significantly stronger osteogenic differentiation than the normal polycaprolactone scaffold (P < 0.05). Alizarin red staining and calcium quantitative analysis showed that the mineralization of the composite scaffold was more obvious than that of the normal polycaprolactone scaffold (P < 0.05), but the total DNA analysis did not differ significantly between scaffolds. These results suggest that the composite scaffold with extracellular matrix is constructed successfully using the 3D technology and co-culture method and exhibits a better osteoinductivity.

Key words: Extracellular Matrix, Stem Cells, Tissue Engineering

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