重组人釉原蛋白促进人脐静脉内皮细胞成血管分化作用的研究

doi:10.13591/j.cnki.kqyx.2022.09.006

Abstract

Abstract: Objective To investigate the effects of recombinant human amelogenin(rhAm) on angiogenesis of human umbilical vein endothelial cell(HUVEC) in vitro. Methods HUVEC were cultured in vitro. The effect of different concentrations of rhAm(0.1, 10.0, 50.0, 100.0 μg/mL rhAm) were compared with control group. The proliferation of HUVEC was investigated via MTT assay. The migration of HUVEC was investigated via scratch assay. Expression of angiogenesis-related genes and proteins were measured via real-time PCR and western blotting. The angiogenic structure formation of HUVEC was investigated via tube formation assay. Results The proliferation of HUVEC was stimulated by rhAm at concentration of 0.1 μg/mL and 10.0 μg/mL(P＜0.05), while inhibited at concentration of 100.0 μg/mL (P＜0.01). At concentration of 10.0 μg/mL, rhAm stimulated HUVEC migration in scratch assay(P＜0.01). According to the result of real-time PCR, the mRNA expression of intercellular cell adhesion molecule-1 (ICAM-1)(P＜ 0.01) and E-selectin (P＜0.05) were increased by rhAm at concentration of 10.0 μg/mL; the mRNA expression of vascular endothelial growth factor receptor(VEGFR)-1 and VEGFR-2 had a trend of up-regulation but no statistical difference. According to the result of western blotting, the protein expression of ICAM-1(P＜0.01)was increased by rhAm at concentration of 10.0 μg/mL, while the protein expression of VEGFR-1, VEGFR-2 and E-selectin had a trend of up-regulation but no statistical difference. RhAm induced the formation of angiogenic structure in vitro. Conclusion Results of the present in vitro study show the potential influence of rhAm on the angiogenic activity of HUVEC at a lower concentration, which may play an important role in periodontal tissue regeneration.

Key words: recombinant human amelogenin, angiogenesis, endo-thelial cell, periodontal tissue regeneration

CLC Number:

R781.4

ZHUANG Qixiang, XIA Yiru, DONG Jiachen, XIE Yufeng, SHU Rong. Effect of recombinant human amelogenin on angiogenesis of human umbilical vein endothelial cell in vitro[J]. Stomatology, 2022, 42(9): 796-801.

References

[1] Vaquette C, Pilipchuk SP, Bartold PM, et al. Tissue engineered constructs for periodontal regeneration:Current status and future perspectives[J]. Adv Healthc Mater, 2018, 7(21):e1800457.
[2] Félétou M. The endothelium:Part 1:Multiple functions of the endothelial cells—focus on endothelium-derived vasoactive mediators[M]. San Rafael: Morgan & Claypool life sciences, 2011.
[3] Nowak-Sliwinska P, Alitalo K, Allen E, et al. Consensus guidelines for the use and interpretation of angiogenesis assays[J]. Angiogenesis, 2018, 21(3):425-532.
[4] Sturtzel C. Endothelialcells[J]. Adv Exp Med Biol, 2017, 1003(9):71-91.
[5] Miron RJ, Sculean A, Cochran DL, et al. Twenty years of enamel matrix derivative:The past, the present and the future[J]. J Clin Periodontol, 2016, 43(8):668-683.
[6] 林智恺, 束蓉, 宋忠臣. 重组釉原蛋白的研究进展[J]. 牙体牙髓牙周病学杂志, 2011, 21(1):48-51.
[7] 陈发明, 高丽娜, 陈芳. 牙周再生治疗现状和进展[J]. 口腔疾病防治, 2019, 27(1):9-16.
[8] Tavelli L, McGuire MK, Zucchelli G, et al. Biologics-based regenerative technologies for periodontal soft tissue engineering[J]. J Periodontol, 2020, 91(2):147-154.
[9] 林智恺, 束蓉, 程岚, 等. 重组全长人釉原蛋白对人成纤维细胞黏附、增殖、迁移的影响[J]. 上海口腔医学, 2013, 22(1):7-12.
[10] Hakki SS, Bozkurt SB, Türkay E, et al. Recombinant amelogenin regulates the bioactivity of mouse cementoblasts in vitro[J]. Int J Oral Sci, 2018, 10(2):15.
[11] 廖蔚文, 宋忠臣, 束蓉, 等. 炎症微环境下重组人釉原蛋白对人牙周膜成纤维细胞炎症因子表达的影响[J]. 上海交通大学学报(医学版), 2016, 36(1):33-37.
[12] Lamalice L, le Boeuf F, Huot J. Endothelial cell migration during angiogenesis[J]. Circ Res, 2007, 100(6):782-794.
[13] Amin HD, Olsen I, Knowles J, et al. A tyrosine-rich amelogenin peptide promotes neovasculogenesis in vitro and ex vivo[J]. Acta Biomater, 2014, 10(5):1930-1939.
[14] Andrukhov O, Gemperli AC, Tang Y, et al. Effect of different enamel matrix derivative proteins on behavior and differentiation of endothelial cells[J]. Dent Mater, 2015, 31(7):822-832.
[15] Jonke E, Gemperli AC, Zhang TW, et al. Effect of tyrosine-rich amelogenin peptide on behavior and differentiation of endothelial cells[J]. Clin Oral Investig, 2016, 20(8):2275-2284.
[16] 廖蔚文, 宋忠臣, 束蓉. 炎症微环境对牙周组织再生的影响[J]. 口腔医学, 2015, 35(9):786-788.
[17] Bui TM, Wiesolek HL, Sumagin R. ICAM-1:A master regulator of cellular responses in inflammation, injury resolution, and tumorigenesis[J]. J Leukoc Biol, 2020, 108(3):787-799.
[18] Zhong L, Simard MJ, Huot J. Endothelial microRNAs regulating the NF-κB pathway and cell adhesion molecules during inflammation[J]. FASEB J, 2018, 32(8):4070-4084.
[19] Melincovici CS, Boşca AB, Şuşman S, et al. Vascular endothelial growth factor (VEGF)-key factor in normal and pathological angiogenesis[J]. Rom J MorpholEmbryol, 2018, 59(2):455-467.
[20] Afacan B, Öztürk VÖ, PaşalıÇ, et al. Gingival crevicular fluid and salivary HIF-1α, VEGF, and TNF-α levels in periodontal health and disease[J]. J Periodontol, 2019, 90(7):788-797.

Effect of recombinant human amelogenin on angiogenesis of human umbilical vein endothelial cell in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 11

Recommended Articles

Metrics

Comments

[1]	WANG Yiyu, HUANG Jiaping, DING Peihui, DONG Yan. Research progress of periodontal tissue regeneration by combined biomaterials of dental derived mesenchymal stem cells [J]. Stomatology, 2023, 43(3): 261-266.
[2]	YAO Minhui, WU Jintao, ZHOU Yu, CHU Fengqing, JIANG Jiajia, CHEN Yue, ZHOU Lili, LI Zehan. Advances in the study on cytokines related to dental pulp regeneration [J]. Stomatology, 2023, 43(3): 282-288.
[3]	SUN Dian, XIAO Yijing, SHEN Ming. Study on the effect of circBIRC6 on promoting angiogenesis [J]. Stomatology, 2022, 42(6): 508-514.
[4]	. Research progress of exosome functions in salivary adenoid cystic carcinoma [J]. , 2021, 41(7): 664-668.
[5]	. Research progress of miRNA-21 in promoting angiogenesis differentiation of stem cells [J]. , 2019, 39(5): 446-449.
[6]	. The role of TNF-α/ NF κB signaling pathway and its antagonism in periodontitis pathogenesis and periodontal tissue regeneration [J]. , 2019, 39(1): 1-5.
[7]	. Advance on the role of GDF15 in the angiogenesis and bone metabolism [J]. , 2016, 36(5): 453-457.
[8]	. Application of adipose derived stem cells in wound healing [J]. , 2016, 36(1): 81-83.
[9]	Fang WANG. Research progress on the osteogenic mechanisms of parathyroid hormone and its effect on craniomaxillofacial and periodontal tissue regeneration [J]. , 2015, 35(12): 1076-1079.
[10]	. Expression of osteoclastic factors in regenerated periodontal tissue under orthodontic force [J]. , 2014, 34(9): 641-645.
[11]	. Wnt Signaling and Periodontal Ligament [J]. , 2014, 34(1): 65-67.