Background:

Autogenous bone graft is the gold standard bone grafting substrate available in spinal fusion because of its osteoconductive, osteogenic, and osteoinductive properties. However, several shortcomings including bleeding, infection, chronic pain, and nerve injury are known to be associated with the procedure. Bone tissue engineering has emerged as an alternative therapeutic strategy for bone grafts. New materials have been developed and tested that can substitute for the autogenous bone grafts used in the spinal fusion. The purpose of this study is to evaluate the role of a novel tissue-engineered bone graft with silicon-substituted calcium phosphate (Si-CaP), autogenous fine particulate bone powder (AFPBP), and bone marrow mesenchymal stem cells (BMSCs) using a rabbit posterolateral lumbar fusion model based on bone tissue engineering principles. The application of this graft can represent a novel choice for autogenous bone to reduce the amount of autogenous bone and promote spinal fusion.

Methods:

BMSCs from New Zealand white rabbits were isolated and cultured in vitro. Then, BMSCs were marked by the cell tracker chloromethyl-benzamidodialkylcarbocyanine (CM-Dil). A total of 96 New Zealand White rabbits were randomly divided into four groups: (a) AFPBP, (b) Si-CaP, (c) Si-CaP/AFPBP, (d) Si-CaP/AFPBP/BMSCs.The rabbits underwent bilateral posterolateral spine arthrodesis of the L5-L6 intertransverse processes using different grafts. Spinal fusion and bone formation were evaluated at 4, 8, and 12 weeks after surgery by manual palpation, radiology, micro-computed tomography (micro-CT), histology, and scanning electronic microscopy (SEM).

Results:

The rate of fusion by manual palpation was higher in the Si-CaP/AFPBP/BMSCs group than the other groups at 8 weeks. The fusion rates in the Si-CaP/AFPBP/BMSCs and the AFPBP groups both reached 100%, which was higher than the Si-CaP/AFPBP group (62.5%) (P ​> ​0.05) and Si-CaP group (37.5%) (P ​< ​0.05) at 12 weeks. New bone formation was observed in all groups after implantation by radiology and micro-CT. The radiographic and CT scores increased in all groups from 4 to 12 weeks, indicating a time-dependent osteogenetic process. The Si-CaP/AFPBP/BMSCs group showed a larger amount of newly formed bone than the Si-CaP/AFPBP and Si-CaP groups at 12 weeks. Bone formation in the Si-CaP/AFPBP/BMSCs group was similar to the AFPBP group. Histology showed that new bone formation continued and increased along with the degradation and absorption of Si-CaP and AFPBP from 4 to 12 weeks in the Si-CaP, Si-CaP/AFPBP, and Si-CaP/AFPBP/BMSCs groups. At 4 weeks, a higher proportion of bone was detected in the AFPBP group (23.49%) compared with the Si-CaP/AFPBP/BMSCs group (14.66%, P ​< ​0.05). In the Si-CaP/AFPBP/BMSCs group at 8 weeks, the area percentage of new bone formation was 28.56%, which was less than the AFPBP group (33.21%, P ​< ​0.05). No difference in bone volume was observed between the Si-CaP/AFPBP/BMSCs group (44.39%) and AFPBP group (45.06%) at 12 weeks (P ​> ​0.05). At 12 weeks, new trabecular were visible in the Si-CaP/AFPBP/BMSCs group by SEM. CM-Dil-positive cells were observed at all stages. Compared with histological images, BMSCs participate in various stages of osteogenesis by transforming into osteoblasts, chondrocytes, and osteocytes.

Conclusion:

This study demonstrated for the first time that Si-CaP/AFPBP/BMSCs is a novel tissue-engineered bone graft with excellent bioactivity, biocompatibility, and biodegradability. The graft could reduce the amount of autogenous bone and promote spinal fusion in a rabbit posterolateral lumbar fusion model, representing a novel alternative to autogenous bone.

The translational potential of this article:

The translational potential of this article lies in that this graft will be a novel spinal fusion graft with great potential for clinical applications.