Experimental study of acellular and demineralized human tooth in repair of calvarial bone defect in rats

doi:10.19438/j.cjoms.2018.05.003

Abstract

Abstract: PURPOSE: To investigate the feasibility of acellular decalcified human teeth as bone scaffold materials and their ability to induce bone defect and osteogenesis in rats. METHODS: Twenty-four isolated human teeth were treated with decalcification and decellularization respectively. In experiment 1, 6 male SD rats aged 10-12 weeks were used to embed decalcified teeth and acellular and demineralized teeth under the skin of the abdomen of the rats. After 4 weeks, the rats were sacrificed and H-E staining was performed to observe the inflammatory reaction around the two materials. In experiment 2, 12 male SD rats of 10-12 weeks old were selected to prepare bilateral calvarial bone defects,the left side of the bone defect was implanted with decalcified and acellular teeth, while the right side of the bone defect without any material was used as a blank control. Six rats were randomly selected for 4 weeks and 8 weeks respectively to be sacrificed. Complete specimens were obtained and Micro- CT scan was performed to compare bone mass from the general morphology. Rats at 4 weeks were taken for H-E staining and immunohistochemistry to observe the peripheral inflammatory response and the extent of newly formed bone. SPSS 23.0 software package was used for ANOVA of repeated measurement. RESULTS: After 4 weeks, H-E staining showed that the inflammatory reaction between acellular and demineralized teeth and surrounding tissues was significantly less than that of decalcified teeth. Micro-CT scanning and quantitative analysis of 4 and 8 weeks of rat calvarial bone showed that the bone mass in the bone defect implanted with acellular and demineralized teeth was significantly higher than that in the blank control (P<0.05). The results of H-E staining of rats at 4 weeks showed that the inflammatory response around the acellular and demineralized teeth was similar to the blank control. Immunohistochemical results showed that the number of new osteoblasts around the acellular and demineralized teeth was significantly higher than that of the blank control. CONCLUSIONS: Decalcified and decellularized teeth are a kind of biological scaffold material with good biocompatibility,osteogenic induction ability and plasticity.

Key words: Calvarial bone defect, Demineralized bone matrix, Acellular bone extracellular matrix, Extracted human teeth, Rat

CLC Number:

R318.08

LIU Zong-lin, GUO Ke, WANG Wen-chao, ZHANG Yao-sheng, XU Wei-feng, ZHANG Shan-yong, LIU Xiu-ming, JIN Jia-min. Experimental study of acellular and demineralized human tooth in repair of calvarial bone defect in rats[J]. China Journal of Oral and Maxillofacial Surgery, 2018, 16(5): 397-401.

References

[1] 杨天府, 汪颖. 同种异体骨移植的免疫反应[J]. 中国修复重建外科杂志, 1994, 8(3): 181-184.
[2] Gzitrom AA, Axelrod T, Ternandes B, et al.Antigen presenting cells and bone allotransplantation[J]. Clin Orthop,1985,197(1):27-41.
[3] Lin X, Zhao C, Zhu P, et al.Periosteum extracellular-matrix-mediated acellular mineralization during bone formation[J]. Adv Healthc Mater, 2018, 7(4): 1700660.
[4] Xue J, He A, Zhu Y, et al.Repair of articular cartilage defects with acellular cartilage sheets in a swine model[J]. Biomed Mater, 2018, 13(2): 025016.
[5] Tang K, Wu J, Xiong Z, et al.Human acellular amniotic membrane: A potential osteoinductive biomaterial for bone regeneration[J]. J Biomater Appl, 2018, 32(6): 754-764.
[6] 杨胜银, 陈平, 鲍济波, 等. 脱矿牙本质基质骨诱导性及对相关细胞鉴定的实验研究[J]. 华西口腔医学杂志, 2018, 36(1): 33-38.
[7] Ren B, Hu X, Cheng J, et al.Synthesis and characterization of polyphosphazene microspheres incorporating demineralized bone matrix scaffolds controlled release of growth factor for chondrogenesis applications[J]. Oncotarget, 2017, 8(69): 114314-114327.
[8] Zhu W, Qiu Y, Sheng F, et al.An effective delivery vehicle of demineralized bone matrix incorporated with engineered collagen-binding human bone morphogenetic protein-2 to accelerate spinal fusion at low dose[J]. J Mater Sci Mater Med, 2017, 29(1): 2.
[9] Mahyudin F, Utomo DN, Suroto H, et al.Comparative effectiveness of bone grafting using xenograft freeze-dried cortical bovine,allograft freeze-dried cortical new zealand white rabbit,xenograft hydroxyapatite bovine, and xenograft demineralized bone matrix bovine in bone defect of femoral diaphysis of white rabbit:experimental study in vivo [J]. Int J Biomater, 2017, 2017: 7571523.
[10] Urist MR.Bone: transplants, implants, derivatives and substitutes-a survey of research of the past decade[J]. Instr Course Lect, 1960, 17: 184-195.
[11] Bath-balogh M, Fehrenbach MJ. Illustrated dental embryology, histology, and anatomy[M]. St. Louis, Mo: Elsevier Saunders, 2006: 839-841.
[12] Yeomans JD, Urist MR.Bone induction by demineralized dentin implanted into oral, osseous and muscle tissues[J]. Arch Oral Biol, 1967, 12(8): 999-1008.
[13] Fugazzotto PA, De Paoli S, Benfenati SP.The use of allogenic freeze-dried dentin in the repair of periodontal osseous defects in humans[J]. Quintessence Int, 1986, 17(8): 461-477.
[14] Su-Gwan K, Hak-Kyun K, Sung-Chul L.Combined implantation of particulate dentine, plaster of Paris, and a bone xenograft (Bio-Oss) for bone regeneration in rats[J]. J Craniomaxillofac Surg, 2001, 29(5): 282-288.
[15] Finkelman RD, Mohan S, Jennings JC, et al.Quantitation of growth factors IGF-I,SGF/IGF-II, and TGF-beta in human dentin[J]. J Bone Miner Res, 1990, 5(7): 717-723.
[16] Guo W, He Y, Zhang X, et al.The use of dentin matrix scaffold and dental follicle cells for dentin regeneration[J]. Biomaterials, 2009, 30(35): 6708-6723.
[17] Li R, Guo W, Yang B.Human treated dentin matrix as a natural scaffold for complete human dentin tissue regeneration[J]. Biomaterials, 2011, 32(20): 4525-4538.
[18] Kim YK, Kim SG, Byeon JH, et al.Development of a novel bone grafting material using autogenous teeth[J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2010, 109(4): 496-503.
[19] Lee JY, Kim YK, Yi YJ, et al.Clinical evaluation of ridge augmentation using autogenous tooth bone graft material: case series study[J]. J Korean Assoc Oral Maxillofac Surg, 2013, 39(4): 156-160.
[20] Nakanishi T, Kokubun K, Oda H, et al.Bioluminescence imaging of bone formation using hairless osteocalcin-luciferase transgenic mice[J]. Bone, 2012, 51(3): 369-375.
[21] 张春丽, 赵彦涛, 白玉龙, 等. 一种高效制备工艺对脱钙骨基质性能的影响[J]. 中国骨与关节损伤杂志, 2015, 30(1): 79-81.
[22] 高君昭, 张雪健, 侯佳丽, 等. 脱细胞猪心包膜在引导骨组织再生术中的屏障作用[J].上海口腔医学, 2017, 26(1): 167-170.
[23] Tang Q, Chen L, Wei F, et al.Effect of 15-deoxy-Δ-prostaglandin JNanocapsules on inflammation and bone regeneration in a rat bone defect model[J]. Chin Med J (Engl), 2017, 130(3): 347-356.
[24] Sidney LE, Heathman TR, Britchford ER, et al.Investigation of localized delivery of diclofenac sodium from poly(D,L-lactic acid-co-glycolic acid)/poly(ethylene glycol) scaffolds using an in vitro osteoblast inflammation model[J]. Tissue Eng Part A, 2015, 21(1-2): 362-373.