Hydroxyapatite/beta-tricalcium phosphate scaffolds combined with adipose-derived stem cells for the treatment of spinal defects in rabbits

doi:10.3969/j.issn.2095-4344.0505

Abstract

Abstract:

BACKGROUND: Repair of bone defects is not only a clinical problem, but also a hot topic in the field of orthopedics. Although autologous bone grafting is considered as the “gold standard” for bone repair, its use is limited due to the limited source of autogenous bone, bone infections and pains that are easy to occur in the donor region. Allograft bones are always associated with immune rejection, slow healing, and infection. Therefore, it is imminent to develop new materials for bone repair.
OBJECTIVE: To explore the effect of rabbit adipose-derived stem cells (rADSCs) as seed cells and hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) composite as a carrier on the repair of rabbit vertebral defects.
METHODS: Thirty-eight 3-month-old New Zealand white rabbits were selected, and two of them were used to culture rADSCs in vitro. Passage 3 rADSCc were inoculated on HA/β-TCP scaffolds and then cultured in vitro for 2 weeks. A 5 mm×5 mm×3 mm bone defect was prepared at the anterior edge of L4/5 vertebral body in the remaining 36 rabbits. These model rabbits were then randomized into cell-scaffold composite group, scaffold group and control group with no intervention, with 12 rabbits in each group. rADSCs/HA/β-TCP composite and HA/β-TCP scaffold were implanted into the cell-scaffold and HA/β-TCP groups, respectively. Anteroposterior and lateral DR of the spine and Lane-Sandhu X-ray were performed at 4, 8, 12 postoperative weeks. All rabbits were sacrificed at 12 postoperative weeks and specimens were collected for gross and histopathological observations.
RESULTS AND CONCLUSION: Under the gross observation, bone defects in the cell-scaffold group were essentially replaced by new bone tissues, which was significantly better than that in the scaffold group and control group. At 12 postoperative weeks, the material implanted was basically absorbed in the cell-scaffold group, partially absorbed in the scaffold group and poorly absorbed in the control group in which there was a clear boundary with the surrounding tissues and patchy calcified shadows were visible. X-ray results showed that the repair effect in the cell-scaffold group was better than that in the scaffold group and control group (P < 0.05). Histopathological findings showed the marked absorption of the implant in the cell-scaffold group, partial residual in the scaffold group with some fibrous calluses and osteoid tissues, and a large amount of fiber tissues and a small amount of calluses in the control group. Overall, the rADSCs/HA/β-TCA has a good ability to repair bone defects.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Spinal Injuries, Adipose Tissue, Stem Cells, Hydroxyapatites, Calcium Phosphates, Tissue Engineering

CLC Number:

R394.2

Wang Teng-fei, Song Xing-hua, Maimaitiaili Abulikemu, Chen Jiang-tao, Tao Ying, Yang Yong. Hydroxyapatite/beta-tricalcium phosphate scaffolds combined with adipose-derived stem cells for the treatment of spinal defects in rabbits[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(13): 2081-2086.

Figures/Tables 3

2.1 脂肪干细胞形态学原代兔脂肪干细胞培养三四个小时开始贴壁，呈小圆形，部分细胞呈多边形等形态，24 h后基本完全贴壁，分布均匀，增殖能力强，传代后平均3 d即可达到90%以上融合(图2A)。将兔脂肪干细胞与HA/β-TCA复合培养7 d后，在扫描电镜下可见材料表面呈多孔状，兔脂肪干细胞黏附于材料表面和空隙中，并向四周延伸，邻居细胞紧密连接，细胞在复合体上生长良好，增殖明显(图2B)。 2.2 大体观察结果术后所有兔伤口愈合良好，三四天后活动、进食基本正常。术后12周时，A组椎体缺损区被骨组织取代，表面未见明显复合体材料，愈合较好(图3A)。B组椎体缺损区基本修复，边界不清，有新骨形成，少许纤维组织填充，触及硬度不高(图3B)。C组椎体缺损区见大量纤维组织，未见成骨形成(图3C)。 2.3 影像学检查结果术后4周：A组椎体缺损区有少量骨痂形成，复合体与周围骨组织紧密接触(图4A)。B组椎体缺损区材料与周围骨组织界线清晰，材料无明显变化(图4B)。C组椎体缺损区界线清晰，片状低密度影，未见骨痂形成(图4C)。术后8周：A组椎体缺损区较前缩小，椎体缺损区可见片状密度增高影，材料吸收，边界模糊(图5A)。B组椎体缺损区可见絮状高密度影，材料与周围骨组织尚清晰，有少量骨痂形成(图5B)。C组椎体缺损区明显，可见点状钙化影(图5C)。术后12周：A组椎体缺损区复合体材料基本吸收，材料与骨有骨性连接，两椎体间基本融合(图6A)。B组椎体缺损区大部分修复，部分骨痂形成，边界模糊(图6B)。C组椎体缺损区边界尚清晰，缺损周边可见片状钙化影(图6C)。根据X射线片结果，按照Lane-Sandhu X射线评分标准进行椎体缺损修复的放射学评分，见表1。 2.4 组织病理学观察结果术后12周时，A组材料吸收明显，可见少量成纤维细胞及成骨细胞(图7A)。B组部分材料残留，可见到纤维性骨痂及少许类骨样组织形成(图7B)。C组可见大量纤维结缔组织，可见少许骨痂形成(图7C)。"

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