Ectopic study of calcium phosphate cement seeded with antagomir-30d transfected BMSCs
CHEN Wei1, WANG Yan1, JIN Qiong2, HUANG Hui3
1.Department of Pediatric Dentistry, School & Hospital of Stomatology Wenzhou Medical University, Wenzhou 325000, China; 2.Department of Implant Prosthodontics, School & Hospital of Stomatology Wenzhou Medical University, Wenzhou 325000, China; 3.Department of Prosthodontics, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
CHEN Wei,WANG Yan,JIN Qiong, et al. Ectopic study of calcium phosphate cement seeded with antagomir-30d transfected BMSCs[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2023, 53(1): 1-6.
Abstract：Objective: To constructed a compound of antagomir-30d/bone marrow mesenchymal stem cells (BMSCs)/calcium phosphate cement (CPC) and implant s.c. in BALB/c-nu mice to demonstrate that antagomir-30d could promote ectopic osteogenesis. Methods: The BMSCs transfected with the optimum concentration of antagomir-30d/NC/blank was loaded on calcium phosphate cement scaffolds and s.c. was implanted into BALB/c-nu mice. Implants were removed after 2, 4, 8 week (wk) respectively. 2-wk implants were subjected to RNA extraction, and then gene expression of osteoblast marker genes alkaline phosphatase (ALP), osteocalcin (OC) and Runt-related transcription factor 2 (RUNX2) was analyzed. 4-wk and 8-wk implants were subjected to
histological staining and histomorphometric analysis was made to observe new bone formation. Results: Gene expression of osteoblast marker genes was analyzed after 2 wk implantation. RT-PCR analysis revealed up-regulation of ALP, OC and RUNX2 in the antagomir-30d-treated implants compared with implants transfected with miR negative control and the un-transfected group. The histological staining revealed that the antagomir-30d-treated implants had a small amount of new bone formation at 4 wk. Implants transfected with miR negative control and the un-transfected group showed new bone formation barely. Histomorphometric analysis showed that the percentage of new bone area of antagomir-30d-treated implants was 1.28%±0.19%. At 8 wk, new bone formation was clearly visible in all groups, while the new bone formation of antagomir-30d-treated group was more significant than NC and un-transfected groups (P<0.05). New bone area percentage was 17.79%±1.15%, 1.82%±0.53%, 2.33%±0.52% respectively. Conclusion: Antagomir-30d accelerates ectopic bone formation in vivo, and a compound of antagomir-30d/BMSCs/CPC is promising as tissue engineering complexes in repairing jaw bone defects.
