Chinese Journal of Tissue Engineering Research ›› 2021, Vol. 25 ›› Issue (34): 5420-5426.doi: 10.12307/2021.233

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Osteogenesis using bone marrow mesenchymal stem cell sheets combined with three-dimensional printed Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol scaffold in vivo

Liu Xiaoyuan1, 2, Li Lei1, 2, Zhang Kai1, 2, Li Jun1, 2, Han Xiangzhen1, 2, He Huiyu1, 2   

  1. 1Department of Prosthodontics, the First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Received:2020-07-27 Revised:2020-07-29 Accepted:2020-09-15 Online:2021-12-08 Published:2021-07-26
  • Contact: He Huiyu, MD, Professor, Department of Prosthodontics, the First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Liu Xiaoyuan, Master candidate, physician, Department of Prosthodontics, the First Affiliated Hospital (Affiliated Stomatological Hospital) of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; Xinjiang Uygur Autonomous Region Institute of Stomatology, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    the Xinjiang Uygur Autonomous Region Science and Technology Support Project, No. 2018E02060 (to HHY)

Abstract: BACKGROUND: Cervus elaphus antler powder is a bone tissue engineering material with good biocompatibility and mechanical properties. The use of cell patch technology and three-dimensional (3D) printing technology to build tissue engineered bone can accomplish individualized treatment of limit bone defects.
OBJECTIVE: To explore the ability and osteogenesis effect of bone marrow mesenchymal stem cell sheet combined with 3D printed Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol bone scaffold to repair the limit defect of sheep mandible.
METHODS: The whole bone marrow method was used to cultivate sheep bone marrow mesenchymal stem cells, and the cell sheets technology and 3D printing technology were used to construct the bone marrow mesenchymal stem cell sheets combined with 3D printed Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol scaffold tissue engineered bone. Totally 12 Xinjiang Altay big-tail sheep were randomly divided into 1-, 2-, and 3-month groups (n=4 per group). An experimental bone defect area of 20 mm×3 mm×5 mm was prepared in the bilateral mandible edentulous area. Two sheep in each group were implanted with cell membrane combined with 3D printed Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol scaffold. Cell sheets combined with gelatin sponge was implanted on the other side. The other two sheep was composited with nano-hydroxyapatite/silk fibroin/polyvinyl alcohol scaffold, and the other side was implanted with cell sheets combined with gelatin sponge. The experimental animals were sacrificed at the end of 1, 2, and 3 months after implantation, and the mandible specimens were taken for gross observation, cone beam CT, histological observation, and RT-PCR to detect related osteogenic indicators. 
RESULTS AND CONCLUSION: (1) Cone beam CT: At the end of the 1st and 2nd months, the bone defect area in the cell sheets compounded with Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol group was thin and cloudy; the scaffold absorbed more. The absorption of the cell sheets compounded with nano-hydroxyapatite/silk fibroin/polyvinyl alcohol group was relatively low. At the end of the 3rd month, a large amount of new bone was formed in the cell sheets composite Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol group, and the density was close to the surrounding bone; however, the bone defect area in the composite nano-hydroxyapatite/silk fibroin/polyvinyl alcohol group was not overgrown and the bone density was lower. The cell sheets composite gelatin sponge group did not change significantly within 3 months, and there was no new bone formation. (2) Histological observation: At the end of the 3rd month, compared with the other two groups, the scaffold material of the cell sheets composite Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol group was more absorbed, showing the regular arrangement of bone trabeculae and mature plate-like bone. There was also a small amount of new bone formation in the composite nano-hydroxyapatite/silk fibroin/polyvinyl alcohol group, and the cell sheets composite gelatin sponge group did not change significantly within 3 months. (3) RT-PCR: The mRNA expression levels of osteopontin, osteocalcin and type I collagen were higher than those of the other two groups in the cell sheets composite Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol group, and the expression of osteogenic genes reached the highest level at the third month. (4) The results show that the bone marrow mesenchymal stem cell sheets combined with 3D printed Cervus elaphus antler powder/silk fibroin/polyvinyl alcohol scaffold tissue engineered bone can repair the limit bone defect of the sheep mandible, and can meet the repair and reconstruction of the experimental sheep jaw defect.


Key words: stem cells, bone marrow mesenchymal stem cells, cell sheets, 3D printing, Cervus elaphus antler powder, silk fibroin, polyvinyl alcohol, limit bone defect, sheep

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