人牙髓干细胞与脱细胞猪小肠黏膜下层的生物相容性

doi:10.16098/j.issn.0529-1356.2018.06.021

摘要/Abstract

摘要：

目的探讨体外培养的人牙髓干细胞（DPSCs）与脱细胞猪小肠黏膜下层(SIS)的生物相容性，为组织工程角膜上皮构建寻找理想的种子细胞。方法改良酶组织块法，培养人DPSCs并鉴定。细胞计数试剂盒（CCK-8）实验分别检测脱细胞SIS浸提液及脱细胞SIS对DPSCs增殖活性的影响。实验分组：（1）脱细胞SIS浸提液培养组为实验组，含10%胎牛血清的DMEM培养基培养组为对照组；（2）复合脱细胞SIS培养组作为实验组，非复合脱细胞SIS培养组为对照组。HE染色检测DPSCs接种于脱细胞SIS后的生长情况。结果成功获取DPSCs。脱细胞SIS浸提液及脱细胞SIS对DPSCs增殖无明显影响，与对照组相比差异无统计学意义（P>0.05）。DPSCs接种第3天和第7天后，能够在脱细胞SIS表面生长，支架纤维有断裂，细胞间有一定的连接。结论 DPSCs与脱细胞SIS具有一定的生物相容性，有望用于组织工程角膜上皮的构建。

关键词: 小肠黏膜下层, 牙髓干细胞, 生物相容性, 组织工程, 流式细胞术, 人

Abstract:

Objective To investigate the biocompatibility of dental pulp stem cells (DPSCs) cultured in vitro with acellular porcine small intestinal submucosa (SIS), and to find an ideal seed cell for tissue engineering corneal epithelium construction. Methods To culture human DPSCs by modified enzyme tissue block method and identificate DPSCs. control groups: DPSCs were cultured in DMEM medium containing 10% fetal bovine serum. The compound acellular SIS culture group served as the experimental group, and the non-compound acellular SIS culture group was the control group. HE staining was used to detect the growth of DPSCs inoculated with acellular SIS. Results DPSCs were successfully obtained. The acellular SIS extract and acellular SIS had no significant effect on the proliferation of DPSCs, but there was no significant difference compared with the control group (P>0.05). DPSCs grew monolayer 3 days，7 days after inoculation with acellular SIS. The scaffold fibers were broken and the cells were connected to each other. Conclusion DPSCs has certain biocompatibility with acellular SIS, which is expected to be used in the construction of tissue engineered corneal epithelium.

Key words: Small intestinal submucosa, Dental pulp stem cell, Biocompatibility, Tissue engineering, Flow cytometry, Human

葛芳杜立群. 人牙髓干细胞与脱细胞猪小肠黏膜下层的生物相容性[J]. 解剖学报, 2018, 49(6): 819-824.

GE Fang DU Li-qun . Biocompatibility of human dental pulp stem cells with acellular porcine intestinal submucosa[J]. Acta Anatomica Sinica, 2018, 49(6): 819-824.

参考文献

［1］Nakamura T, Inatomi T, Sotozono C, et al. Ocular surface reconstruction using stem cell and tissue engineering［J］. Prog Retin Eye Res, 2016,51:187-207.

［2］Wirostko B, Rafii M, Sullivan DA, et al. Novel therapy to treat corneal epithelial defects: a hypothesis with growth hormone［J］. Ocul Surf, 2015,13(3):204-212.

［3］Menzel-Severing J, Kruse FE, Schl?tzer-Schrehardt U. Stem cell-based therapy for corneal epithelial reconstruction: present and future［J］. Can J Ophthalmol, 2013,48(1):13-21.

［4］Gronthos S, Mankani M, Brahim J, et al. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo［J］. Proc Natl Acad Sci USA, 2000,97(25):13625-13630.

［5］Gronthos S, Brahim J, Li W, et al. Stem cell properties of human dental pulp stem cells［J］. J Dent Res, 2002,81(8):531-535.

［6］Chai Y, Jiang X, Ito Y, et al. Fate of the mammalian cranial neural crest during tooth and mandibular morphogenesis［J］. Development, 2000,127(8):1671-1679.

［7］Nuti N, Corallo C, Chan BM, et al. Multipotent differentiation of human dental pulp stem cells: a literature review［J］. Stem Cell Rev, 2016,12(5):511-523.

［8］Umemura N, Ohkoshi E, Tajima M, et al. Hyaluronan induces odontoblastic differentiation of dental pulp stem cells via CD44［J］. Stem Cell Res Ther, 2016,7(1):135.

［9］Guzalinuer A, Muhetaer H, Wu H, et al. Experimental study on the transforming growth factor β3 combined with dental pulp stem cells in early bone integration of implant［J］. Chinese Journal of Stomatology, 2018,53(4):259-263. (in Chinese)

古扎丽努尔·阿巴拜克力, 木合塔尔〖DK〗·霍加, 仵韩, 等. 转化生长因子β3联合牙髓干细胞在种植体周围早期骨结合中的作用［J］. 中华口腔医学杂志, 2018,53(4):259-263.

［10］Ghasemi Hamidabadi H, Rezvani Z, Nazm Bojnordi M, et al. Chitosan-intercalated montmorillonite/poly(vinyl alcohol) nanofibers as a platform to guide neuronlike differentiation of human dental pulp stem cells［J］. ACS Appl Mater Interfaces, 2017,9(13):11392-11404.

［11］Wang FY, Du LQ. Preparation and histological evaluation of the decellularized scaffold for porcine small intestinal submucosa［J］. Acta Anatomica Sinica, 2016,47(6):824-828. (in Chinese)

王付燕, 杜立群. 猪小肠黏膜下层脱细胞支架的制备及组织学评价［J］. 解剖学报, 2016,47(6):824-828.

［12］Ouyang H, Xue Y, Lin Y, et al. WNT7A and PAX6 define corneal epithelium homeostasis and pathogenesis［J］. Nature, 2014,511(7509):358-361.

［13］Yam GH, Peh GS, Singhal S, et al. Dental stem cells: a future asset of ocular cell therapy［J］. Expert Rev Mol Med, 2015,17:e20.

［14］Potdar PD. Human dental pulp stem cells: Applications in future regenerative medicine［J］. World J Stem Cells, 2015,7(5):839-851.

［15］Botelho J, Cavacas MA, Machado Ⅴ, et al. Dental stem cells: recent progresses in tissue engineering and regenerative medicine［J］. Ann Med, 2017,49(8):644-651.

［16］Tsai CL, Chuang PC, Kuo HK, et al. Differentiation of Stem Cells From Human Exfoliated Deciduous Teeth Toward a Phenotype of Corneal Epithelium In Vitro［J］. Cornea, 2015,34(11):1471-1477.

［17］Monteiro BG, Serafim RC, Melo GB, et al. Human immature dental pulp stem cells share key characteristic features with limbal stem cells［J］. Cell Prolif, 2009,42(5):587-594.

［18］Kushnerev E, Shawcross SG, Sothirachagan S, et al. Regeneration of corneal epithelium with dental pulp stem cells using a contact lens delivery system［J］. Invest Ophthalmol Vis Sci, 2016,57(13):5192-5199.

［19］Syed-Picard FN, Du Y, Lathrop KL, et al. Dental pulp stem cells: a new cellular resource for corneal stromal regeneration［J］. Stem Cells Transl Med, 2015,4(3):276-285.

［20］Roozafzoon R, Lashay A, Vasei M, et al. Dental pulp stem cells differentiation into retinal ganglion-like cells in a three dimensional network［J］. Biochem Biophys Res Commun, 2015,457(2):154-160.

［21］Gonmanee T, Thonabulsombat C, Vongsavan K, et al. Differentiation of stem cells from human deciduous and permanent teeth into spiral ganglion neuron-like cells［J］. Arch Oral Biol, 2018,88:34-41.

［22］D’Aquino R, De Rosa A, Lanza Ⅴ, et al. Human mandible bone defect repair by the grafting of dental pulp stem/progenitor cells and collagen sponge biocomplexes［J］. Eur Cell Mater, 2009,18:75-83.

［23］Gomes JáP, Geraldes Monteiro B, Melo GB, et al. Corneal reconstruction with tissue-engineered cell sheets composed of human immature dental pulp stem cells［J］. Invest Opthalmol Vis Sci, 2010,51(3):1408-1414.

［24］Davis NF, Cunnane EM, O’Brien FJ, et al. Tissue engineered extracellular matrices (ECMs) in urology: Evolution and future directions［J］. Surgeon, 2018,16(1):55-65.

［25］Goulle F. Use of porcine small intestinal submucosa for corneal reconstruction in dogs and cats: 106 cases［J］. J Small Anim Pract, 2012,53(1):34-43.

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