Guo Ke, Wang Wenchao, Liu Zonglin, Xu Weifeng, Zhang Shanyong, Yang Chi
Department of Oral and Maxillofacial Surgery, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology Shanghai 200011, China.
Int J Clin Exp Pathol. 2020 May 1;13(5):837-845. eCollection 2020.
The present study aimed to investigate the reliability of acellular decalcified teeth in the development of bone scaffolds and in bone regeneration in rats.
(1) Forty-eight human teeth were divided into two groups in vitro: twenty-four were decalcified, while the remaining twenty-four were decalcified and decellularized, following which a conventional scanning-electron microscope analysis was performed. (2) In another experiment, six male SD rats aged 10-12 weeks were selected, then decalcified and acellular decalcified teeth were embedded subcutaneously in the abdomen of the rats. After 4 weeks, the rats were sacrificed for H-E staining and immunohistochemical staining to observe the inflammatory reaction around the two materials. (3) In the ectopic osteogenesis experiment, bone defects were simulated in bilateral craniotectal areas of 12 male SD rats (age 10-12 weeks), following which acellular decalcified teeth were implanted in the right bone defect. The non-implanted left side was used as blank control. At week 4 and week 8, 6 rats were randomly selected for execution, complete specimens were obtained, and micro-CT scan was performed to compare the bone mass from gross morphology. H-E staining was performed at 4 and 8 weeks to observe the surrounding inflammatory response and immunohistochemistry was performed at 8 weeks to observe the degree of new bone formation. SPSS 23.0 software package was used for statistical processing.
(1) Under scanning electron microscope, cells in the teeth subjected to acellular decalcification completely disappeared, leaving only inorganic scaffolds. (2) After 4 weeks, the amount of inflammatory reaction in the tissues surrounding acellular decalcified teeth was significantly lower than that in the tissues surrounding decalcified teeth. (3) After four and eight weeks, the amount of bone formation in the bone defects was significantly higher in rats implanted with acellular decalcified teeth than in those in the blank control group (P<0.05). After four and eight weeks, hematoxylin-eosin staining revealed that the degree of inflammatory response was similar around acellular decalcified teeth and blank controls. Immunohistochemistry indicated that the osteocalcin levels were significantly higher around acellular decalcified teeth than that around blank controls.
Acellular decalcified teeth show significantly decreased inflammatory reaction, better biocompatibility, better osteogenic potential, and better plasticity than decalcified teeth alone.
本研究旨在探讨脱细胞脱钙牙在大鼠骨支架构建及骨再生中的可靠性。
（1）48颗人牙在体外分为两组：24颗进行脱钙处理，其余24颗进行脱钙及脱细胞处理，随后进行常规扫描电子显微镜分析。（2）在另一实验中，选取6只10 - 12周龄的雄性SD大鼠，将脱钙牙和脱细胞脱钙牙皮下植入大鼠腹部。4周后，处死大鼠进行苏木精 - 伊红（H-E）染色和免疫组织化学染色，观察两种材料周围的炎症反应。（3）在异位成骨实验中，在12只10 - 12周龄雄性SD大鼠的双侧颅骨区域模拟骨缺损，将脱细胞脱钙牙植入右侧骨缺损处。未植入的左侧作为空白对照。在第4周和第8周，随机选取6只大鼠处死，获取完整标本，进行显微CT扫描以从大体形态上比较骨量。在第4周和第8周进行H-E染色观察周围炎症反应，在第8周进行免疫组织化学染色观察新骨形成程度。使用SPSS 23.0软件包进行统计处理。
（1）扫描电子显微镜下，经脱细胞脱钙处理的牙齿中细胞完全消失，仅留下无机支架。（2）4周后，脱细胞脱钙牙周围组织的炎症反应程度明显低于脱钙牙周围组织。（3）4周和8周后，植入脱细胞脱钙牙的大鼠骨缺损处的骨形成量明显高于空白对照组（P<0.05）。4周和8周后，苏木精 - 伊红染色显示脱细胞脱钙牙周围和空白对照周围的炎症反应程度相似。免疫组织化学表明，脱细胞脱钙牙周围的骨钙素水平明显高于空白对照周围。
与单纯脱钙牙相比，脱细胞脱钙牙的炎症反应明显降低，生物相容性更好，成骨潜力更好，可塑性更强。
