Int J Stomatol

Int J Stomatol ›› 2021, Vol. 48 ›› Issue (5): 520-527.doi: 10.7518/gjkq.2021075

• Original Articles • Previous Articles     Next Articles

Effects of platelet-rich plasma and concentrated growth factor on the proliferation and osteogenic differentiation of human periodontal cells

Liu Juan(),Chen Bin,Yan Fuhua()   

  1. Dept. of Periodontology, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing 210008, China
  • Received:2021-02-03 Revised:2021-06-02 Online:2021-09-01 Published:2021-09-10
  • Contact: Fuhua Yan E-mail:1044042989@qq.com;yanfh@nju.edu.cn
  • Supported by:
    National Natural Science Foundation of China(81771078);Project of Nanjing Clinical Research Center for Oral Diseases(2019060009);Key Project of Science and Technology Bureau of Jiangsu Province(BL2013002)

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Abstract:

Objective The study aimed to analyze the effects of platelet-rich plasma (PRP) and concentrated growth factor (CGF) on the proliferation and osteogenic differentiation of human periodontal membrane cells (hPDLCs). Metho-ds The hPDLCs were cultured and isolated from periodontal ligament tissue. PRP and CGF were prepared in accordance with the standard protocols, and platelet count was performed separately. The hPDLCs were cultured to three groups: whole blood (control group), PRP (PRP group), and CGF (CGF group) at various time points. Cell proliferation was detected by the CCK-8 method, and cell migration was detected by the cell scratch assay. The hPDLCs were treated with 1%, 5%, and 10% of PRP and CGF for 24, 48, and 72 h, respectively, for osteogenic induction. Expression levels of osteogenic transcription factors Runx2, Osx, Dlx5, and Msx2 were detected by Western blot. Results Both PRP and CGF could promote the proliferation and migration of the hPDLCs (P<0.05). The expression levels of osteogenic factors Runx2, Osx, and Dlx5 increased significantly; the expression of Msx2, which inhibited osteogenesis, decreased significantly (P<0.05). Conclusion PRP and CGF could promote the proliferation and migration of hPDLCs and induce the expression of osteogenic differentiation-related transcription factors.

Key words: platelet-rich plasma, concentrated growth factor, human periodontal ligament cells, cell proliferation, osteogenic differentiation

CLC Number: 

  • Q25

TrendMD: 

Fig 1

The preparation of CGF from the whole blood"

Fig 2

The isolation culture and identification of hPDLCs"

Tab 1

The platelet count of whole blood, PRP and CGF"

样本编号 志愿者基本资料 血小板计数/（109·L-1）
性别 年龄/岁 全血 PRP CGF
1 男 22 278 2 459 –
2 女 22 190 1 305 –
3 女 23 193 2 558 –
4 男 26 263 2 553 –
5 女 25 210 3 127 –
6 男 24 269 1 953 –
7 女 23 237 2 044 –

Fig 3

The effect of PRP and CGF on the proliferation ability of hPDLCs"

Fig 4

The effect of PRP and CGF on the migration ability of hPDLCs crystal violet staining"

Fig 5

The effect of PRP and CGF on the expression of Msx2, Osx, Dlx5 and Runx2 detected with Western blot"

Fig 6

The effect of PRP and CGF on the expression levels of Runx2, Dlx5, Osx and Msx2"

[1] 闫福华, 李丽丽. 牙周再生治疗研究进展[J]. 口腔医学研究, 2018, 34(3):217-222.
Yan FH, Li LL. Development in periodontal regene-rative treatment[J]. J Oral Sci Res, 2018, 34(3):217-222.
[2] Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human pe-riodontal ligament[J]. Lancet, 2004, 364(9429):149-155.
doi: 10.1016/S0140-6736(04)16627-0
[3] Marx RE, Carlson ER, Eichstaedt RM, et al. Platelet-rich plasma: growth factor enhancement for bone grafts[J]. Oral Surg Oral Med Oral Pathol Oral Ra-diol Endod, 1998, 85(6):638-646.
[4] Weibrich G, Hansen T, Kleis W, et al. Effect of platelet concentration in platelet-rich plasma on peri-implant bone regeneration[J]. Bone, 2004, 34(4):665-671.
pmid: 15050897
[5] Anitua E, Sánchez M, Orive G, et al. The potential impact of the preparation rich in growth factors (PR-GF) in different medical fields[J]. Biomaterials, 2007, 28(31):4551-4560.
doi: 10.1016/j.biomaterials.2007.06.037
[6] 张宇, 林野, 邱立新, 等. 富血小板血浆促进口腔种植骨再生的临床应用研究[J]. 中华口腔医学杂志, 2004, 39(4):269-272.
Zhang Y, Lin Y, Qiu LX, et al. Using platelet-rich plasma (PRP) to improve bone regeneration in im-plant bone defect[J]. Chin J Stomatol, 2004, 39(4):269-272.
[7] Choi BH, Zhu SJ, Kim BY, et al. Effect of platelet-rich plasma (PRP) concentration on the viability and proliferation of alveolar bone cells: an in vitro study[J]. Int J Oral Maxillofac Surg, 2005, 34(4):420-424.
doi: 10.1016/j.ijom.2004.10.018
[8] Bozkurt Doğan Ş, Öngöz Dede F, Ballı U, et al. Concentrated growth factor in the treatment of adjacent multiple gingival recessions: a split-mouth randomi-zed clinical trial[J]. J Clin Periodontol, 2015, 42(9):868-875.
doi: 10.1111/jcpe.12444 pmid: 26269089
[9] Pirpir C, Yilmaz O, Candirli C, et al. Evaluation of effectiveness of concentrated growth factor on osseointegration[J]. Int J Implant Dent, 2017, 3(1):7.
doi: 10.1186/s40729-017-0069-3
[10] Honda H, Tamai N, Naka N, et al. Bone tissue engineering with bone marrow-derived stromal cells integrated with concentrated growth factor in Rattus norvegicus calvaria defect model[J]. J Artif Organs, 2013, 16(3):305-315.
doi: 10.1007/s10047-013-0711-7
[11] 魏中武, 黄谢山, 陈灼庚. 浓缩生长因子在口腔临床中的应用及研究进展[J]. 国际口腔医学杂志, 2020, 47(2):235-243.
Wei ZW, Huang XS, Chen ZG. Application and re-search progress on concentrated growth factor in oral clinic[J]. Int J Stomatol, 2020, 47(2):235-243.
[12] Loi F, Córdova LA, Pajarinen J, et al. Inflammation, fracture and bone repair[J]. Bone, 2016, 86:119-130.
doi: 10.1016/j.bone.2016.02.020
[13] Zhang XL, Shi KQ, Jia PT, et al. Effects of platelet-rich plasma on angiogenesis and osteogenesis-associated factors in rabbits with avascular necrosis of the femoral head[J]. Eur Rev Med Pharmacol Sci, 2018, 22(7):2143-2152.
[14] Yu B, Wang Z. Effect of concentrated growth factors on beagle periodontal ligament stem cells in vitro[J]. Mol Med Rep, 2014, 9(1):235-242.
doi: 10.3892/mmr.2013.1756
[15] Fontana S, Olmedo DG, Linares JA, et al. Effect of platelet-rich plasma on the peri-implant bone respon-se: an experimental study[J]. Implant Dent, 2004, 13(1):73-78.
doi: 10.1097/01.ID.0000116455.68968.29
[16] Bartold PM, Gronthos S. Standardization of criteria defining periodontal ligament stem cells[J]. J Dent Res, 2017, 96(5):487-490.
doi: 10.1177/0022034517697653 pmid: 28425840
[17] 刘娟, 赵红宇, 轩东英, 等. 人牙周膜细胞群多向分化潜能的实验研究[J]. 华西口腔医学杂志, 2010, 28(2):185-189.
Liu J, Zhao HY, Xuan DY, et al. Differentiation characteristics of human periodontal ligament cell population in vitro[J]. West China J Stomatol, 2010, 28(2):185-189.
[18] 廖海清, 曹正国. 经典Wnt信号通路在牙周膜细胞成骨分化过程中的调控[J]. 口腔医学研究, 2016, 32(3):224-227.
Liao HQ, Cao ZG. Effect of canonical Wnt signalling pathway on the osteogenic differentiation and proli-feration of PDLCs[J]. J Oral Sci Res, 2016, 32(3):224-227.
[19] Tamiya H, Ikeda T, Jeong JH, et al. Analysis of the Runx2 promoter in osseous and non-osseous cells and identification of HIF2A as a potent transcription activator[J]. Gene, 2008, 416(1/2):53-60.
doi: 10.1016/j.gene.2008.03.003
[20] Nakashima K, Zhou X, Kunkel G, et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation[J]. Cell, 2002, 108(1):17-29.
pmid: 11792318
[21] Lee MH, Kim YJ, Kim HJ, et al. BMP-2-induced Runx2 expression is mediated by Dlx5, and TGF-beta 1 opposes the BMP-2-induced osteoblast differentiation by suppression of Dlx5 expression[J]. J Biol Chem, 2003, 278(36):34387-34394.
doi: 10.1074/jbc.M211386200
[22] 乔静, 欧阳翔英, 曹采方. 富血小板血浆对人牙周膜细胞增殖及分化的影响[J]. 上海口腔医学, 2008, 17(1):60-63.
Qiao J, Ouyang XY, Cao CF. The effect of different concentrations of platelet-rich plasma on human pe-riodontal ligament cells in vitro[J]. Shanghai J Sto-matol, 2008, 17(1):60-63.
[23] 钟声, 闫福华, 卢友光, 等. 富血小板血浆对牙周膜成纤维细胞损伤模型影响的研究[J]. 口腔医学研究, 2005, 21(5):504-506.
Zhong S, Yan FH, Lu YG, et al. Effect of platelet-rich plasma (PRP) on periodontal ligament fibroblasts in an in vitro wound healing model[J]. J Oral Sci Res, 2005, 21(5):504-506.
[24] Durmuşlar MC, Balli U, Dede FÖ, et al. Histological evaluation of the effect of concentrated growth factor on bone healing[J]. J Craniofac Surg, 2016, 27(6):1494-1497.
doi: 10.1097/SCS.0000000000002873 pmid: 27428921
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[9] . [J]. Foreign Med Sci: Stomatol, 2004, 31(02): 126 -128 .
[10] . [J]. Inter J Stomatol, 2008, 35(S1): .