纤维蛋白凝胶对上颌扩弓成骨矿化的影响

doi:10.19438/j.cjoms.2021.05.003

中国口腔颌面外科杂志 ›› 2021, Vol. 19 ›› Issue (5): 397-404.doi: 10.19438/j.cjoms.2021.05.003

纤维蛋白凝胶对上颌扩弓成骨矿化的影响

赵涵江¹, 王泽莹¹, 林丹^1,*, 沈国芳^1,2,*

1.上海交通大学医学院附属第九人民医院口腔颅颌面科,上海交通大学口腔医学院, 国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海 200011;
2.上海健康医学院,上海 201318

收稿日期:2021-03-18 修回日期:2021-05-11 出版日期:2021-09-20 发布日期:2021-10-20
通讯作者: 林丹,E-mail:d.lin@foxmail.com;沈国芳,E-mail:shengf@sumhs.edu.cn。^*共同通信作者
作者简介:赵涵江（1994-）,男,硕士研究生,E-mail:hanjiangzhao@sjtu.edu.cn
基金资助:
国家自然科学基金（81970973,81570947）; 上海市青年科技英才扬帆计划（19YF1426500）

Effect of fibrin glue on osteogenesis and mineralization during rapid maxillary expansion

ZHAO Han-jiang¹, WANG Ze-ying¹, LIN Dan¹, SHEN Guo-fang^1,2

1. Department of Oral and Craniomaxillofacial Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology. Shanghai 200011;
2. Shanghai University of Medicine and Health Sciences. Shanghai 201318, China

Received:2021-03-18 Revised:2021-05-11 Online:2021-09-20 Published:2021-10-20

摘要/Abstract

摘要： 目的研究原位注射纤维蛋白凝胶对SD大鼠上颌扩弓成骨矿化的影响。方法体外实验中,将大鼠骨髓间充质细胞（rBMSCs）在不同凝血酶浓度制备的纤维蛋白凝胶环境下进行共培养,通过CCK-8、 ALP染色和活性定量以及茜素红染色,检测梯度浓度凝血酶制备的纤维蛋白凝胶降解产物对rBMSCs细胞增殖和成骨分化的影响。将rBMSCs分别采用共培养、二维表面培养和三维培养方法与纤维蛋白凝胶进行结合,观察细胞形态和黏附情况。体内实验中,构建SD大鼠上颌扩弓模型（n=12）,以100 g作为扩弓力,扩弓7 d,随机分为2组,每组6只,将仅配戴14 d保持器作为对照组（R组）,将原位注射纤维蛋白凝胶联合配戴14 d保持器作为实验组（R+FG组）,通过显微CT（Micro-CT）三维重建和分析,比较新生骨量和骨矿化密度的差异。通过H-E染色、Masson三色染色、TRAP染色和顺序荧光染色,观察腭中缝的组织学差异。采用GraphPad Prism 8.0.1软件包对数据进行统计学分析。结果体外实验结果显示,纤维蛋白凝胶降解产物对rBMSCs细胞增殖和成骨分化无显著影响（P>0.05）,且以上述3种方式培养的rBMSCs均具有良好的细胞延展性。体内实验结果显示,实验组新生骨量（P<0.001）和骨小梁矿化密度（P<0.01）较对照组显著提升,腭中缝内可见大量矿化程度更高的新生骨小梁组织;破骨细胞数量减少,矿化沉积速率显著加快（P<0.001）。结论纤维蛋白凝胶降解产物对rBMSCs增殖、分化无影响。SD大鼠上颌扩弓后,腭中缝组织可通过原位注射纤维蛋白凝胶,促进新骨生成和矿化速率,抑制破骨细胞分化。

关键词: 上颌快速扩弓, 骨缝牵引成骨, 纤维蛋白凝胶, 骨再生

Abstract: PURPOSE: To investigate the effect of accelerating osteogenesis and mineralization during rapid maxillary expansion by in situ injection of fibrin glue. METHODS: In in vitro experiments, rat bone marrow stromal cells (rBMSCs) were co-cultured with fibrin glue fabricated with different concentrations of thrombin. The effects of degradation products of fibrin glue on the proliferation and osteogenic differentiation of rBMSCs were tested by CCK-8, alkaline phosphatase (ALP) staining and ALP activity quantification as well as alizarin red staining. rBMSCs were combined with fibrin glue by co-culture, two-dimensional surface culture and three-dimensional culture, respectively. Cell morphology and adhesion were observed by FITC-phalloidine fluorescence staining. In in vivo experiments, 12 6-week male SD rats were used to establish rapid maxillary expansion (RME) model with 100 g expanding force for 7 days. After 7-day expansion, they were divided into two groups: retention for 14 days as control group (R group) while in situ injection of fibrin glue combined with retention for 14 days as experimental group (R+FG group). Micro-CT scanning for three-dimensional reconstruction and analysis were conducted to compare the difference of new bone volume and bone mineral density. The osteogenesis and angiogenesis in mid-palatal suture were observed by H-E staining, Masson's trichrome staining, TRAP staining and sequential fluorescent labeling. GraphPad Prism 8.0.1 software package was used for statistical analysis. RESULTS: The degradation products of fibrin gel had little effect on cell proliferation and osteogenic differentiation of rBMSCs(P>0.05). In addition, rBMSCs cultured in three ways had great cell tractility. Compared with R group, the relative bone volume(P<0.001) and bone mineral density of trabeculae (P<0.001) were significantly increased in R+FG group. A large number of new trabeculae with higher mineralization degree were observed and the number of osteoclasts was decreased in mid-palatal suture. Mineral apposition rate was significantly accelerated in R+FG group(P<0.001). CONCLUSIONS: The degradation products of fibrin glue had no effect on proliferation and differentiation of rBMSCs. Fibrin glue could promote new bone formation, mineralized rate and inhibit osteo-clastogenesis in mid-palatal suture after RME in SD rats, which might be related to regulation of osteogenic differentiation by the surface structure of fibrin glue.

Key words: Rapid maxillary expansion, Sutural distraction osteogenesis, Fibrin glue, Bone regeneration

中图分类号:

R782.2

赵涵江, 王泽莹, 林丹, 沈国芳. 纤维蛋白凝胶对上颌扩弓成骨矿化的影响[J]. 中国口腔颌面外科杂志, 2021, 19(5): 397-404.

ZHAO Han-jiang, WANG Ze-ying, LIN Dan, SHEN Guo-fang. Effect of fibrin glue on osteogenesis and mineralization during rapid maxillary expansion[J]. China J Oral Maxillofac Surg, 2021, 19(5): 397-404.

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纤维蛋白凝胶对上颌扩弓成骨矿化的影响

Effect of fibrin glue on osteogenesis and mineralization during rapid maxillary expansion

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