镁黄长石浸提液对人成纤维细胞增殖和迁移的影响
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摘要
目的观察不同浓度的镁黄长石浸提液对成纤维细胞增殖和迁移的影响。方法将成纤维细胞培养于不同浓度的镁黄长石浸提液中,用CCK-8法和流式细胞学检测细胞的增殖情况;通过细胞划痕实验观察细胞迁移能力变化。RT-q-PCR法及Western blot检测I、Ⅲ型胶原蛋白及部分整合素亚基mRNA和蛋白的表达。结果镁黄长石浸提液促进诱导成纤维细胞增殖的作用具有时间及浓度依赖性。1/64浓度浸提液提高细胞活力和增殖能力最明显。RT-q-PCR及Western blot定量检测Ⅰ、Ⅲ型胶原蛋白,mRNA及蛋白表达均有提高。划痕实验结果显示1/64浓度浸提液相对于阴性对照组迁移能力明显增强,RT-q-PCR及Western blot定量检测结果提示迁移能力的增强与细胞内的整合素α5β1和α3β1上调有关。结论镁黄长石浸提液在1/64浓度能显著提高成纤维细胞的增殖和迁移能力。
Objective To observe akermanite extracts at different concentrations on the proliferation and migration of human fibroblast. Methods CCK-8 assay and flow cytometry assay were used to detect the proliferation of human fibroblast treated with different concentrations of akermanite extracts(1/4, 1/16, 1/64,1/256). A scratch assay was performed to evaluate the effects of akermanite on fibroblast migration. Furthermore, Collagen I, Ⅲ and integrin subunits were investigated using RT-q-PCR and Western blot assay. Results Akermanite extract promoted the proliferation of fibroblast in a time-and concentration-dependent manner. The cell viability and proliferation in 1/64 concerntration group increased most significantly. RT-q-PCR and Western blot assays showed that the mRNA and protein expressions of collagen Ⅰ and Ⅲ were up-regulated. A scratch assay showed that the cell migration ability of 1/64 concerntration group was significantly enhanced. Quantitative results of RT-q-PCR and Western blot indicated that the enhancement of migration ability was related to the up-regulation of the expressions of integrin α5β1 and α3β1. Conclusion The akermanite extract with concentration of 1/64 can significantly improve the proliferation and migration ability of fibroblast.
Objective To observe akermanite extracts at different concentrations on the proliferation and migration of human fibroblast. Methods CCK-8 assay and flow cytometry assay were used to detect the proliferation of human fibroblast treated with different concentrations of akermanite extracts(1/4, 1/16, 1/64,1/256). A scratch assay was performed to evaluate the effects of akermanite on fibroblast migration. Furthermore, Collagen I, Ⅲ and integrin subunits were investigated using RT-q-PCR and Western blot assay. Results Akermanite extract promoted the proliferation of fibroblast in a time-and concentration-dependent manner. The cell viability and proliferation in 1/64 concerntration group increased most significantly. RT-q-PCR and Western blot assays showed that the mRNA and protein expressions of collagen Ⅰ and Ⅲ were up-regulated. A scratch assay showed that the cell migration ability of 1/64 concerntration group was significantly enhanced. Quantitative results of RT-q-PCR and Western blot indicated that the enhancement of migration ability was related to the up-regulation of the expressions of integrin α5β1 and α3β1. Conclusion The akermanite extract with concentration of 1/64 can significantly improve the proliferation and migration ability of fibroblast.
引文
1 Govindaraju P,Todd L,Shetye S,et al.CD44-dependent inflammation,fibrogenesis,and collagenolysis regulates extracellular matrix remodeling and tensile strength during cutaneous wound healing[J].Matrix Biol,2019,75-76:314-330.
2 Takeo M,Lee W,Ito M.Wound healing and skin regeneration[J].Cold Spring Harb Perspect Med,2015,5(1):a023267.
3 Sharafabadi AK,Abdellahi M,Kazemi A,et al.A novel and economical route for synthesizing akermanite(Ca2MgSi2O7)nanobioceramic[J].Mater Sci Eng C Mater Biol Appl,2017,71:1072-1078.
4 Gao C,Peng S,Feng P,et al.Bone biomaterials and interactions with stem cells[J/OL].http://europepmc.org/abstract/MED/29285402.
5 Fernandes JS,Gentile P,Pires RA,et al.Multifunctional bioactive glass and glass-ceramic biomaterials with antibacterial properties for repair and regeneration of bone tissue[J].Acta Biomater,2017,59:2-11.
6 Huang Y,Wu C,Zhang X,et al.Regulation of immune response by bioactive ions released from silicate bioceramics for bone regeneration[J].Acta Biomater,2018,66:81-92.
7 Xia L,Yin Z,Mao L,et al.Akermanite bioceramics promote osteogenesis,angiogenesis and suppress osteoclastogenesis for osteoporotic bone regeneration[J].Sci Rep,2016,6:22005.
8 Lv F,Wang J,Xu P,et al.A conducive bioceramic/polymer composite biomaterial for diabetic wound healing[J].Acta Biomater,2017,60:128-143.
9杜庆华,曹均凯,董溪溪,等.镁黄长石浸提液诱导多潜能干细胞的成骨分化[J].中国组织工程研究,2015,19(14):2236-2242.
10 Rognoni E,Watt FM.Skin Cell Heterogeneity in Development,Wound Healing,and Cancer[J].Trends Cell Biol,2018,28(9):709-722.
11 Woodley DT.Distinct Fibroblasts in the Papillary and Reticular Dermis:Implications for Wound Healing[J].Dermatol Clin,2017,35(1):95-100.
12 Martin PE,O'Shaughnessy EM,Wright CS,et al.The potential of human induced pluripotent stem cells for modelling diabetic wound healing in vitro[J].Clin Sci(Lond),2018,132(15):1629-1643.
13 Kallis PJ,Friedman AJ.Collagen Powder in Wound Healing[J].JDrugs Dermatol,2018,17(4):403-408.
14 Torres P,Castro M,Reyes M,et al.Histatins,wound healing,and cell migration[J].Oral Dis,2018,24(7):1150-1160.
15 Stunova A,Vistejnova L.Dermal fibroblasts-A heterogeneous population with regulatory function in wound healing[J].Cytokine Growth Factor Rev,2018,39:137-150.
16 Cole MA,Quan T,Voorhees JJ,et al.Extracellular matrix regulation of fibroblast function:redefining our perspective on skin aging[J].J Cell Commun Signal,2018,12(1):35-43.
17 Rippa AL,Vorotelyak EA,Vasiliev AV,et al.The role of integrins in the development and homeostasis of the epidermis and skin appendages[J].Acta Naturae,2013,5(4):22-33.
18 Barroca V,Lewandowski D,Jaracz-Ros A,et al.Paternal Insulinlike Growth Factor 2(Igf2)Regulates Stem Cell Activity During Adulthood[J].EBioMedicine,2017,15:150-162.
19 Varanasi VG,Odatsu T,Bishop T,et al.Enhanced osteoprogenitor elongated collagen fiber matrix formation by bioactive glass ionic silicon dependent on Sp7(osterix)transcription[J].J Biomed Mater Res A,2016,104(10):2604-2615.
20 Shuai C,Han Z,Feng P,et al.Akermanite scaffolds reinforced with boron nitride nanosheets in bone tissue engineering[J].JMater Sci Mater Med,2015,26(5):188.
2 Takeo M,Lee W,Ito M.Wound healing and skin regeneration[J].Cold Spring Harb Perspect Med,2015,5(1):a023267.
3 Sharafabadi AK,Abdellahi M,Kazemi A,et al.A novel and economical route for synthesizing akermanite(Ca2MgSi2O7)nanobioceramic[J].Mater Sci Eng C Mater Biol Appl,2017,71:1072-1078.
4 Gao C,Peng S,Feng P,et al.Bone biomaterials and interactions with stem cells[J/OL].http://europepmc.org/abstract/MED/29285402.
5 Fernandes JS,Gentile P,Pires RA,et al.Multifunctional bioactive glass and glass-ceramic biomaterials with antibacterial properties for repair and regeneration of bone tissue[J].Acta Biomater,2017,59:2-11.
6 Huang Y,Wu C,Zhang X,et al.Regulation of immune response by bioactive ions released from silicate bioceramics for bone regeneration[J].Acta Biomater,2018,66:81-92.
7 Xia L,Yin Z,Mao L,et al.Akermanite bioceramics promote osteogenesis,angiogenesis and suppress osteoclastogenesis for osteoporotic bone regeneration[J].Sci Rep,2016,6:22005.
8 Lv F,Wang J,Xu P,et al.A conducive bioceramic/polymer composite biomaterial for diabetic wound healing[J].Acta Biomater,2017,60:128-143.
9杜庆华,曹均凯,董溪溪,等.镁黄长石浸提液诱导多潜能干细胞的成骨分化[J].中国组织工程研究,2015,19(14):2236-2242.
10 Rognoni E,Watt FM.Skin Cell Heterogeneity in Development,Wound Healing,and Cancer[J].Trends Cell Biol,2018,28(9):709-722.
11 Woodley DT.Distinct Fibroblasts in the Papillary and Reticular Dermis:Implications for Wound Healing[J].Dermatol Clin,2017,35(1):95-100.
12 Martin PE,O'Shaughnessy EM,Wright CS,et al.The potential of human induced pluripotent stem cells for modelling diabetic wound healing in vitro[J].Clin Sci(Lond),2018,132(15):1629-1643.
13 Kallis PJ,Friedman AJ.Collagen Powder in Wound Healing[J].JDrugs Dermatol,2018,17(4):403-408.
14 Torres P,Castro M,Reyes M,et al.Histatins,wound healing,and cell migration[J].Oral Dis,2018,24(7):1150-1160.
15 Stunova A,Vistejnova L.Dermal fibroblasts-A heterogeneous population with regulatory function in wound healing[J].Cytokine Growth Factor Rev,2018,39:137-150.
16 Cole MA,Quan T,Voorhees JJ,et al.Extracellular matrix regulation of fibroblast function:redefining our perspective on skin aging[J].J Cell Commun Signal,2018,12(1):35-43.
17 Rippa AL,Vorotelyak EA,Vasiliev AV,et al.The role of integrins in the development and homeostasis of the epidermis and skin appendages[J].Acta Naturae,2013,5(4):22-33.
18 Barroca V,Lewandowski D,Jaracz-Ros A,et al.Paternal Insulinlike Growth Factor 2(Igf2)Regulates Stem Cell Activity During Adulthood[J].EBioMedicine,2017,15:150-162.
19 Varanasi VG,Odatsu T,Bishop T,et al.Enhanced osteoprogenitor elongated collagen fiber matrix formation by bioactive glass ionic silicon dependent on Sp7(osterix)transcription[J].J Biomed Mater Res A,2016,104(10):2604-2615.
20 Shuai C,Han Z,Feng P,et al.Akermanite scaffolds reinforced with boron nitride nanosheets in bone tissue engineering[J].JMater Sci Mater Med,2015,26(5):188.