Chinese Journal of Tissue Engineering Research ›› 2022, Vol. 26 ›› Issue (20): 3164-3172.doi: 10.12307/2022.615
Previous Articles Next Articles
Li Tengyan1, 2, Nie Minhai1, 2, Liu Xuqian1, 2
Received:
2021-04-06
Accepted:
2021-06-09
Online:
2022-07-18
Published:
2022-01-19
Contact:
Liu Xuqian, MD, Associate professor, Department of Periodontal Mucosal Diseases, Affiliated Hospital of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China; Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, Southwest Medical University, Luzhou 646000, Sichuan Province, China
About author:
Li Tengyan, Master, Physician, Department of Periodontal Mucosal Diseases, Affiliated Hospital of Stomatology, Southwest Medical University, Luzhou 646000, Sichuan Province, China; Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, Southwest Medical University, Luzhou 646000, Sichuan Province, China
Supported by:
CLC Number:
Li Tengyan, Nie Minhai, Liu Xuqian. Effects of concentrated growth factor combined with epidermal growth factor on the proliferation and aging of oral mucosa equivalents[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(20): 3164-3172.
Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks
2.3 浓缩生长因子、表皮生长因子最佳作用浓度的筛选 2.3.1 浓缩生长因子最佳作用浓度的筛选 浓缩生长因子对人牙龈成纤维细胞、上皮细胞、正常人皮肤黑色素细胞增殖的影响与其浓度有关,其中50%浓缩生长因子对人牙龈成纤维细胞增殖的促进作用最强,30%浓缩生长因子对上皮细胞和正常人皮肤黑色素细胞增殖的促进作用最强(P < 0.01)。在3种细胞与浓缩生长因子共培养中均观察到,当浓缩生长因子浓度低于最佳浓度时对细胞增殖起促进作用,且随浓缩生长因子浓度升高而增强,当浓缩生长因子浓度高于最佳浓度时,其对细胞增殖的促进作用逐渐减弱,甚至抑制细胞增殖(P < 0.01或P < 0.05)。遂分别采用50%浓缩生长因子用于人牙龈成纤维细胞后续实验,采用30% 浓缩生长因子用于上皮细胞、正常人皮肤黑色素细胞后续实验,见图4。"
2.3.2 表皮生长因子最佳作用浓度的筛选 表皮生长因子对人牙龈成纤维细胞、上皮细胞、正常人皮肤黑色素细胞增殖的影响与其浓度有关,其中,10 μg/L 表皮生长因子对人牙龈成纤维细胞和上皮细胞增殖的促进作用最强,20 μg/L 表皮生长因子对正常人皮肤黑色素细胞增殖的促进作用最强(P < 0.01)。在3种细胞与表皮生长因子条件培养基共培养中均观察到以最佳表皮生长因子作用浓度为峰值点,当表皮生长因子浓度低于最佳浓度时,其对细胞增殖起促进作用,且强度随表皮生长因子浓度升高而增强,而当表皮生长因子浓度高于最佳浓度之后,其对细胞增殖的促进作用逐渐减弱,甚至高浓度表皮生长因子抑制细胞增殖(P < 0.01或P < 0.05)。所以,采用10 μg/L表皮生长因子用于人牙龈成纤维细胞和上皮细胞后续实验,20 μg/L 表皮生长因子用于正常人皮肤黑色素细胞后续实验,见图5。"
2.3.3 浓缩生长因子 + 表皮生长因子最佳作用浓度的筛选 在既定最佳浓缩生长因子作用浓度基础上协同不同浓度表皮生长因子与细胞共培养观察其对细胞增殖的影响,结果显示其对细胞增殖的影响与表皮生长因子浓度有关,其中50%浓缩生长因子 + 10 μg/L 表皮生长因子对人牙龈成纤维细胞增殖的促进作用最强,30%浓缩生长因子 + 10 μg/L表皮生长因子对上皮细胞和正常人皮肤黑色素细胞增殖的促进作用最强(P < 0.01)。当表皮生长因子浓度低于10 μg/L时,与浓缩生长因子联合对细胞增殖发挥了促进作用,且随表皮生长因子浓度上升而增强,当表皮生长因子浓度高于10 μg/L时,其对细胞增殖的促进作用逐渐弱于单纯浓缩生长因子作用和对照组,表皮生长因子浓度过高甚至抑制细胞增殖(P < 0.01或P < 0.05)。综上,采用10 μg/L 表皮生长因子联合最佳作用浓度浓缩生长因子用于后续实验,即人牙龈成纤维细胞50%浓缩生长因子+ 10 μg/L表皮生长因子,上皮细胞、正常人皮肤黑色素细胞30% 浓缩生长因子 + 10 μg/L 表皮生长因子,见图6。"
2.4 浓缩生长因子联合表皮生长因子3种细胞增殖的影响 CCK-8实验结果显示浓缩生长因子、表皮生长因子、浓缩生长因子 + 表皮生长因子均促进细胞增殖,在3种细胞中效应分别呈现为:人牙龈成纤维细胞10 μg/L表皮生长因子 > 50%浓缩生长因子+10 μg/L 表皮生长因子 > 50%浓缩生长因子(P < 0.01或P < 0.05);上皮细胞10 μg/L表皮生长因子> 30%浓缩生长因子+ 10 μg/L 表皮生长因子> 30% 浓缩生长因子(P < 0.01或P < 0.05);正常人皮肤黑色素细胞20 μg/L表皮生长因子> 30%浓缩生长因子+ 10 μg/L表皮生长因子> 30%浓缩生长因子+ 20 μg/L表皮生长因子> 30%浓缩生长因子 (P < 0.01或P < 0.05),见图7-9。"
2.5 浓缩生长因子联合表皮生长因子对3种细胞迁移的影响 与CCK-8实验结果相对应,浓缩生长因子、表皮生长因子、浓缩生长因子 + 表皮生长因子均促进3种细胞的迁移,其效应分别呈现为:人牙龈成纤维细胞10 μg/L表皮生长因子 > 50%浓缩生长因子+ 10 μg/L表皮生长因子> 50%浓缩生长因子(P < 0.01);上皮细胞10 μg/L表皮生长因子> 30% 浓缩生长因子 + 10 μg/L表皮生长因子> 30%浓缩生长因子(P < 0.01);正常人皮肤黑色素细胞20 μg/L表皮生长因子> 30%浓缩生长因子+ 10 μg/L表皮生长因子> 30% 浓缩生长因子+20 μg/L 表皮生长因子> 30% 浓缩生长因子(P < 0.01),见图10-12。"
2.6 浓缩生长因子联合表皮生长因子对3种细胞凋亡的影响 浓缩生长因子、表皮生长因子、浓缩生长因子 + 表皮生长因子均抑制可抑制3种细胞凋亡,其效应分别呈现为:人牙龈成纤维细胞10 μg/L表皮生长因子> 50%浓缩生长因子+ 10 μg/L表皮生长因子> 50% 浓缩生长因子(P < 0.01或P < 0.05);上皮细胞10 μg/L 表皮生长因子> 30%浓缩生长因子+ 10 μg/L表皮生长因子> 30% 浓缩生长因子(P < 0.01);正常人皮肤黑色素细胞20 μg/L 表皮生长因子> 30% 浓缩生长因子+ 10 μg/L表皮生长因子> 30% 浓缩生长因子+ 20 μg/L 表皮生长因子> 30% 浓缩生长因子(P < 0.01),见图13-15。"
[1] 陈谦明.口腔黏膜病学 [M]. 4版.北京:人民卫生出版社,2012: 1-175. [2] LIU N, GUAN S, WANG H, et al. The Antimicrobial Peptide Nal-P-113 Exerts a Reparative Effect by Promoting Cell Proliferation, Migration, and Cell Cycle Progression. Biomed Res Int. 2018;2018:7349351. [3] CHEN J, BEKALE LA, KHOMTCHOUK KM, et al. Locally administered heparin-binding epidermal growth factor-like growth factor reduces radiation-induced oral mucositis in mice. Sci Rep. 2020;10(1):17327. [4] ZHAO QM, GAO J, HUANG XX, et al. Concentrated Growth Factors Extracted from Blood Plasma Used to Repair Nasal Septal Mucosal Defect After Rhinoplasty. Aesthetic Plast Surg. 2020;44(2):511-516. [5] QI L, LIU L, HU Y, et al. Concentrated growth factor promotes gingival regeneration through the AKT/Wnt/β-catenin and YAP signaling pathways. Artif Cells Nanomed Biotechnol. 2020;48(1):920-932. [6] 张慧,王蕊,王芹,等.重组人表皮生长因子联合糖皮质激素治疗糜烂型口腔扁平苔藓 [J]. 中国临床医生杂志,2019,47(12):1492-1494. [7] RODELLA LF, FAVERO G, BONINSEGNA R, et al. Growth factors, CD34 positive cells, and fibrin network analysis in concentrated growth factors fraction. Microsc Res Tech. 2011;74(8):772-777. [8] 魏中武, 刘双喜, 陈灼庚, 等. 比较浓缩生长因子提取液和富血小板纤维蛋白提取液对成骨细胞MC3T3-E1增殖的影响[J]. 中南大学学报(医学版),2020,45(8):901-908. [9] QIN J, WANG L, SUN Y, et al. Concentrated growth factor increases Schwann cell proliferation and neurotrophic factor secretion and promotes functional nerve recovery in vivo. Pubmed. 2016;37(2):493-500. [10] ZHANG L, AI H. Concentrated growth factor promotes proliferation, osteogenic differentiation, and angiogenic potential of rabbit periosteum-derived cells in vitro. BioMed Central. 2019;14(1):146. [11] CHEN J, JIAO D, ZHANG M, et al. Concentrated Growth Factors Can Inhibit Photoaging Damage Induced by ultraviolet A (UVA) on the Human Dermal Fibroblasts In Vitro. Med Sci Monit. 2019;25:3739-3749. [12] AGHAMOHAMADI Z, KADKHODAZADEH M, TORSHABI M, et al. A compound of concentrated growth factor and periodontal ligament stem cell-derived conditioned medium. Tissue Cell. 2020;65:101373. [13] ÖZVERI KB, IŞıK G, ÖZDEN YM, et al. Effect of concentrated growth factor (CGF) on short-term clinical outcomes after partially impacted mandibular third molar surgery: A split-mouth randomized clinical study. J Stomatol Oral Maxillofac Sur. 2020;121(2):118-123. [14] KAO CH.Use of concentrate growth factors gel or membrane in chronic wound healing: Description of 18 cases. Int Wound J. 2020;17(1):158-166. [15] KAMAL A, SALMAN B, ABDUL RNH, et al. The Efficacy of Concentrated Growth Factor in the Healing of Alveolar Osteitis: A Clinical Study. Int J Dent. 2020;2020:9038629. [16] 李媛姣子, 罗赛, 徐渴鑫, 等. 注射浓缩生长因子改善面部炎性衰老的临床观察 [J]. 中国美容整形外科杂志,2019,30(4):236-239. [17] ZHAO LL, XI QC, FANG MS, et al. Observation on the Clinical Effect of Breast Augmentation by Transplanting Concentrated Growth Factor and Autologous Fat. CJPRS. 2019;1(3):18-23. [18] HARDWICKE J, SCHMALJOHANN D, BOYCE D, et al. Epidermal growth factor therapy and wound healing--past, present and future perspectives. Surgeon. 2008;6(3):172-177. [19] AMIN DN, HIDA K, BIELENBERG DR, et al. Tumor endothelial cells express epidermal growth factor receptor (EGFR) but not ErbB3 and are responsive to EGF and to EGFR kinase inhibitors. Cancer Res. 2006;66(4):2173-2180. [20] GALVEZ-CONTRERAS AY, GONZALEZ-CASTANEDA RE, CAMPOS-ORDONEZ T,et al. Phenytoin enhances the phosphorylation of epidermal growth factor receptor and fibroblast growth factor receptor in the subventricular zone and promotes the proliferation of neural precursor cells and oligodendrocyte differentiation. Eur J Neurosci. 2016;43(2):139-147. [21] XU H, LIU L, CONG M, et al. EGF neutralization antibodies attenuate liver fibrosis by inhibiting myofibroblast proliferation in bile duct ligation mice. Histochem Cell Biol. 2020;154(1):107-116. [22] 陈洁, 金绍林. 应用纳米银敷料联合表皮生长因子凝胶治疗小面积骨外露创面 [J]. 生物医学工程与临床,2021,25(1):62-65. [23] KOZER N, CLAYTON AHA. In-cell structural dynamics of an EGF receptor during ligand-induced dimer-oligomer transition. Eur Biophys J. 2020;49(1):21-37. [24] 刘志荣. 重组人表皮生长因子治疗口腔黏膜损伤的疗效观察 [J]. 山西医药杂志(下半月刊),2012,41(6):586-587. [25] 王汉明, 朱晓密, 赵雅君. 口疮1号方治疗大鼠口腔溃疡的实验研究 [J].口腔医学研究,2014,30(7):627-629+634. [26] DHARMANI P, DE SIMONE C, CHADEE K. The probiotic mixture vsl#3 accelerates gastriculcer healing by stimulating vascular endothelial growth factor. Plos One. 2013;8(3):e58671. [27] 孙浩博, 吕国忠. 重组人表皮生长因子对深Ⅱ度烧伤患者创面愈合效果及相关炎症因子水平的影响 [J]. 中国美容医学,2020,29(10):75-78. [28] 刘鹏, 徐全臣, 许晓燕,等.人牙龈成纤维细胞原代培养方法比较 [J]. 齐鲁医学杂志,2016,31(1):18-20+23. [29] 张建伟, 徐铖, 杨慧雅, 等. 口腔黏膜色素痣4例临床及组织病理分析 [J]. 中国皮肤性病学杂志,2012,26(11):993-994+996. [30] YING J, WANG Q, JIANG M, et al. Hydrogen Sulfide Promotes Cell Proliferation and Melanin Synthesis in Primary Human Epidermal Melanocytes. Skin Pharmacol Physiol. 2020;33(2):61-68. [31] PARK KY, KIM J. Synthesis and Biological Evaluation of the Anti-Melanogenesis Effect of Coumaric and Caffeic Acid-Conjugated Peptides in Human Melanocytes. Front Pharmacol. 2020;11:922. [32] ANDO H, YOSHIMOTO S, YOSHIDA M, et al. Dermal Fibroblasts Internalize Phosphatidylserine-Exposed Secretory Melanosome Clusters and Apoptotic Melanocytes. Int J Mol Sci. 2020;21(16):5789. [33] 项锦敏, 赵琼瑜, 远航, 等.乌鳖黑色素理化性质及其抗氧化活性研究 [J/OL].天然产物研究与开发:1-15[2021-01-16]. [34] BRISSETT AE, HOM DB. The effects of tissue sealants, platelet gels, and growth factors on wound healing. Curr Opin Otolaryngol Head Neck Surg.2003;11(4):245-250. [35] CHEN J, JIANG H. Clinical Application of Concentrated Growth Factor Fibrin Combined With Bone Repair Materials in Jaw Defects. CJOMS. 2020;78(7):1041. [36] JIN R, SONG G, CHAI J, et al. Effects of concentrated growth factor on proliferation, migration, and differentiation of human dental pulp stem cells in vitro. J Tissue Eng. 2018;9:2041731418817505. [37] MORIKAWA M, DERYNCK R, MIYAZONO K. TGF-β and the TGF-β Family: Context-Dependent Roles in Cell and Tissue Physiology. CSH Perspect Biol. 2016;8(5):a021873. [38] HRUBI E, IMRE L, ROBASZKIEWICZ A, et al. Diverse effect of BMP-2 homodimer on mesenchymal progenitors of different origin. Hum Cell. 2018;31(2):139-148. [39] MELINCOVICI CS, BOŞCA AB, ŞUŞMAN S, et al. Vascular endothelial growth factor (VEGF) - key factor in normal and pathological angiogenesis. Rom J Morphol Embryol. 2018;59(2):455-467. [40] MIHAYLOVA Z, TSIKANDELOVA R, SANIMIROV P, et al. Role of PDGF-BB in proliferation, differentiation and maintaining stem cell properties of PDL cells in vitro. Arch Oral Biol. 2018;85:1-9. [41] 江飞玉. FGF21在高糖状态下对前列腺癌细胞增殖、自噬的影响及其机制研究 [D]. 重庆:重庆医科大学,2020. [42] ZHAO X, DAI W, ZHU H, et al. Epidermal growth factor (EGF) induces apoptosis in a transfected cell line expressing EGF receptor on its membrane. Cell Biol Int. 2006;30(8): 653-658. [43] ARE A, PINAEV G, BUROVA E, et al. Attachment of A-431 cells on immobilized antibodies to the EGF receptor promotes cell spreading and reorganization of the microfilament system. Cell Motil Cytoskel. 2001;48(1):24-36. [44] 罗雅馨, 毕浩然, 陈晓旭, 等. 细胞外基质与组织的再生与修复 [J]. 中国组织工程研究,2021,25(11):1785-1790. [45] 陈盈哲, 全知怎, 姜葳, 等. 口腔黏膜创伤修复中生长因子对牙龈成纤维细胞的影响及调节机制 [J]. 广西医科大学学报,2018,35(10): 1335-1340. [46] 任科伟, 范卫民, 姜雪峰,等. 周期性机械应力通过激活EGFR-ERK1/2信号通路促进大鼠软骨细胞增殖和细胞外基质合成 [J]. 江苏医药,2013,39(21):2526-2528. [47] ARıCAN G, ÖZMERIÇ A, FıRAT A, et al. Micro-ct findings of concentrated growth factors (cgf) on bone healing in masquelet’s technique-an experimental study in rabbits.Arch Orthop Trauma Surg.2020;10.1007/s00402-020-03596-z. |
[1] | LIU Danni, SUN Guanghua, ZHOU Guijuan, LIU Hongya, ZHOU Jun, TAN Jinqu, HUANG Xiarong, PENG Ting, FENG Wei-bin, LUO Fu. Effect of electroacupuncture on apoptosis of neurons in cerebral cortex of rats with cerebral ischemia-reperfusion injury at "Shuigou" and "Baihui" points [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(在线): 1-6. |
[2] | Pan Baoshun, Fang Zhen, Gao Mingjie, Fang Guiming, Chen Jinshui. Design for posterior atlantoaxial internal fixation system with fusion cage based on imaging data [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(9): 1372-1376. |
[3] | Yao Xiaoling, Peng Jiancheng, Xu Yuerong, Yang Zhidong, Zhang Shuncong. Variable-angle zero-notch anterior interbody fusion system in the treatment of cervical spondylotic myelopathy: 30-month follow-up [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(9): 1377-1382. |
[4] | Wu Cong, Jia Quanzhong, Liu Lun. Relationship between transforming growth factor beta1 expression and chondrocyte migration in adult articular cartilage after fragmentation [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(8): 1167-1172. |
[5] | Wang Jifang, Bao Zhen, Qiao Yahong. miR-206 regulates EVI1 gene expression and cell biological behavior in stem cells of small cell lung cancer [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1027-1031. |
[6] | Wen Dandan, Li Qiang, Shen Caiqi, Ji Zhe, Jin Peisheng. Nocardia rubra cell wall skeleton for extemal use improves the viability of adipogenic mesenchymal stem cells and promotes diabetes wound repair [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1038-1044. |
[7] | Zhu Bingbing, Deng Jianghua, Chen Jingjing, Mu Xiaoling. Interleukin-8 receptor enhances the migration and adhesion of umbilical cord mesenchymal stem cells to injured endothelium [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1045-1050. |
[8] | Zhang Yujie, Yang Jiandong, Cai Jun, Zhu Shoulei, Tian Yuan. Mechanism by which allicin inhibits proliferation and promotes apoptosis of rat vascular endothelial cells [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1080-1084. |
[9] | Zhang Jinglin, Leng Min, Zhu Boheng, Wang Hong. Mechanism and application of stem cell-derived exosomes in promoting diabetic wound healing [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1113-1118. |
[10] | Huang Chenwei, Fei Yankang, Zhu Mengmei, Li Penghao, Yu Bing. Important role of glutathione in stemness and regulation of stem cells [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 1119-1124. |
[11] | Tian Chuan, Zhu Xiangqing, Yang Zailing, Yan Donghai, Li Ye, Wang Yanying, Yang Yukun, He Jie, Lü Guanke, Cai Xuemin, Shu Liping, He Zhixu, Pan Xinghua. Bone marrow mesenchymal stem cells regulate ovarian aging in macaques [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(7): 985-991. |
[12] | Xu Kuishuai, Zhang Liang, Chen Jinli, Ren Zhongkai, Zhao Xia, Li Tianyu, Yu Tengbo. Effect of force line changes on lower limb joints after medial open wedge high tibial osteotomy [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(6): 821-826. |
[13] | Deng Shuang, Pu Rui, Chen Ziyang, Zhang Jianchao, Yuan Lingyan . Effects of exercise preconditioning on myocardial protection and apoptosis in a mouse model of myocardial remodeling due to early stress overload [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(5): 717-723. |
[14] | Yang Sidi, Wang Qian, Xu Nuo, Wang Ronghan, Jin Chuanqi, Lu Ying, Dong Ming. Biodentine enhances the proliferation and differentiation of osteoblasts through upregulating bone morphogenetic protein-2 [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(4): 516-520. |
[15] | Shen Jiahua, Fu Yong. Application of graphene-based nanomaterials in stem cells [J]. Chinese Journal of Tissue Engineering Research, 2022, 26(4): 604-609. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||