不同接种条件对牙周膜干细胞体内外矿化的影响
Differentiation Ability of Periodontal Ligament Stem Cells in Different Scaffolds

作者: 常秀梅 :广东医学院,广东 东莞; 金 岩 :第四军医大学口腔医学院口腔组织病理教研室,陕西 西安;

关键词: 牙周膜干细胞冻干胶原膜陶瓷化骨分化Periodontal Ligament Stem Cells Lyophilization Collagen Membrane Ceramic Bovine Bone Differentiation

摘要:
目的:分离培养人牙周膜干细胞(periodontal ligament stem cells,PDLSCs),观察不同接种条件下成牙能力。方法:体外培养人牙周膜干细胞,细胞与胶原凝胶混合,实验组,接种至陶瓷化骨(ceramic bovine bone, CBB),对照组接种至冻干胶原膜(lyophilization collagen membrane, LCM),矿化液连续培养2 w后,移植入免疫缺陷小鼠皮下上,培养6周取材。扫描电镜观察与材料的贴附情况,细胞计数观察细胞在材料上的粘附和增殖情况,组织化学方法检测不同支架上矿化培养后PDLSCs的碱性磷酸酶(ALP)。扫描电镜观察体外培养条件下细胞附着,基质分泌,组织学观察牙周组织形成能力。结果:扫描电镜下可见PDLSCs与CBB相容性好,细胞生长密集,长入向BBC空隙内,体内条件下,CBB组形成牙周膜–牙骨质复合体结构,LCM组未形成牙周组织。结论:体内外培养条件下,CBB均具有较好诱导作用,是牙齿组织再生研究的优势支架材料。

Abstract: Objective: To investigate the performance of combine collagen gen and ceramic bovine bone for human periodontal ligament stem cells. Methods: Human periodontal ligament stem cells were cultured in vitro. The subsequent cell passaging was conducted in conditioned medium containing dexamethasone, beta-sodium glycerophosphate and ascorbid acid. Then the human periodontal ligament stem cells were divided into two groups: one was cultured in BBC and the other was cul-tured in LCM. Attachment was observed by SEM and proliferation was evaluated by cell counting, ALP was measured in vitro histochemical method, and differentiation was observed by HE stain after transplant into nude mice. Results: BBC improved the proliferation and differentiation of cells cultured in vitro, showing no difference in attachment between two groups. Conclusion: BBC can improve the proliferation and differentiation of human periodontal stem cells.

文章引用: 常秀梅 , 金 岩 (2015) 不同接种条件对牙周膜干细胞体内外矿化的影响。 生物医学, 5, 23-31. doi: 10.12677/HJBM.2015.53004

参考文献

[1] Saygin, N.E., Giannobile, W.V., Somerman, M.J., et al. (2000) Molecular and cell biology of cementum. Periodontology, 24, 73-98.
http://dx.doi.org/10.1034/j.1600-0757.2000.2240105.x

[2] 常秀梅, 刘宏伟, 金岩, 等 (2009) 犬牙周膜干细胞体外分离培养和鉴定的实验研究. 华西口腔医学杂志, 27, 79-83.

[3] 许永华, 施新猷, 胡蕴玉, 等 (2000) 牛陶瓷化骨与体外培养兔骨膜成骨细胞的相容性. 第四军医大学学报, 21, 512-514.

[4] 程晓兵, 薛振恂, 周树夏, 等 (2002) 多孔块状B一礴酸三钙陶瓷兔颅骨骨膜下埋下植可吸收性的定量研究. 中国临床康复, 6, 189.

[5] Minamide, A., Tamaki, T., Kawakami, M., et al. (1999) Experimental spinal fusion using sintered bovine bone coated with type I collagen and recombinant human bone morphogenetic protein-2. Spine, 24, 1863-1871.
http://dx.doi.org/10.1097/00007632-199909150-00002

[6] Matsumoto, T., Kawakami, M., Kuribayashi, K., et a1. (1999) Effects of sintered bovine bone on cell proliferation, collagen synthesis, and osteoblastic expression in MC3T3-E1 ostoblast-like cells. Journal of Orthopaedic Research, 17, 586-592.

[7] Handa, K., Saito, M., Yamauchi, M., et al. (2002) Cementum matrix formation in vivo by cultured dental follicle cells. Bone, 31, 606-611.
http://dx.doi.org/10.1016/S8756-3282(02)00868-2

[8] Pitaru, S., Savion, N., Hekmati, H., et al. (1992) Binding of a cementum attachment protein to extracellular matrix components and to dental surfaces. Journal of Periodontal Research, 27, 640-646.
http://dx.doi.org/10.1111/j.1600-0765.1992.tb01748.x

[9] Liu, H.W., Yacobi, R., Savion, N.A., et al. (1997) Collagenous cementum-derived attachment protein is a marker for progenitors of the mineralized tissue-forming cell lineage of the periodontal ligament. Journal of Bone and Mineral Research, 12, 1691-1699.
http://dx.doi.org/10.1359/jbmr.1997.12.10.1691

[10] Hunter, G.K. and Goldberg, H.A. (1993) Nucleation of hy-droxyapatite by bone sialoprotein. Proceedings of the National Academy of Sciences of the United States of America, 90, 8562-8565.
http://dx.doi.org/10.1073/pnas.90.18.8562

[11] Falla, N., Van Vlasselaer, Bierkens, J., et a1. (1993) Characterization of a enriched ostioprogenitor population of the routine bone iTiRrrow. Blood, 82, 3580.

[12] BarKana, I., Narayanan, A.S., Grosskop, A., Savion, N. and Pitaru, S. (2000) Cementum attachment protein enriches putative cementoblastic populations on root surfaces in vitro. Journal of Dental Research, 79, 1482-1488.
http://dx.doi.org/10.1177/00220345000790070901

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