Protective effect of low-level laser irradiation on lipopolysaccharide-mediated inflammatory injury of human periodontal ligament fibroblasts

doi:10.7518/hxkq.2021.03.003

Abstract

Abstract: Objective

To study the effect and mechanism of low-level laser irradiation (LLLI) on lipopolysaccharide (LPS)-induced inflammatory injury of human periodontal ligament fibroblasts (hPDLFs).

Methods

hPDLFs were inoculated into well plates and randomly divided into the normal group, LPS group, and LPS+LLLI group. The cells in the normal group were cultured in conventional medium. The hPDLFs in the LPS and LPS+LLLI groups were cultured in RPMI1640 medium containing 1 mg·L^-1 LPS. The three subgroups of the LPS+LLLI group were exposed to different LLLI. After 4 days, the cell apoptosis, viability, and intracellular free Ca²⁺ concentration of each group were measured. The contents of tumor necrosis factor-α (TNF-α), interleukin (IL)-8, IL-1β, and IL-6 were measured by enzyme linked immunosorbent assay (ELISA). Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of matrix metalloproteinase (MMP)-2, MMP-3, and MMP-9 genes and proteins of hPDLFs in each group.

Results

Compared with the normal group, the LPS group showed increased apoptosis rate of hPDLFs and intracellular free Ca²⁺concentration and decreased cell viability (P<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels were higher in the cell supernatant (P<0.05), and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs was significantly increased (P<0.05). Compared with the LPS group, the LPS+LLLI group showed significantly decreased apoptosis rate and intracellular free Ca²⁺ concentration and significantly increased cell viability (P<0.05). The TNF-α, IL-8, IL-1β, and IL-6 levels in the supernatant of cells and the expression of MMP-2, MMP-3, and MMP-9 genes and proteins of hPDLFs were significantly decreased (P<0.05).

Conclusion

LLLI has a protective effect on the inflammatory injury of hPDLFs induced by LPS, and the effect is most obvious when the irradiation intensity is 4 J·cm^-2.

Key words: low-level laser irradiation, human periodontal ligament fibroblasts, lipopolysaccharide, inflammatory injury, effect

CLC Number:

Q 786

Jin Xiaolan, Zhang Yanan, Sun Chengrui, Zou Zhaohui. Protective effect of low-level laser irradiation on lipopolysaccharide-mediated inflammatory injury of human periodontal ligament fibroblasts[J]. West China Journal of Stomatology, 2021, 39(3): 260-266.

Figures/Tables 6

Fig 1

Tab 1

Fig 2

Tab 2

Fig 3

Fig 4

References 14

1	Wang F, Guan M, Wei L, et al. IL-18 promotes the secretion of matrix metalloproteinases in human periodontal ligament fibroblasts by activating NF-κB signaling[J]. Mol Med Rep, 2019, 19(1): 703-710.
2	Zhang L, Chen W, Li Y, et al. Effect of 650-nm low-level laser irradiation on c-Jun, c-Fos, ICAM-1, and CCL2 expression in experimental periodontitis[J]. Lasers Med Sci, 2020, 35(1): 31-40.
3	隋华欣, 吕培军, 王宇光, 等. 低能量激光照射对人脂肪基质细胞增殖分化的影响[J]. 北京大学学报(医学版), 2017, 49(2): 337-343.
	Sui HX, Lü PJ, Wang YG, et al. Effect of low-level laser irradiation on proliferation and osteogenic differentiation of human adipose-derived stromal cells[J]. J Peking Univ (Health Sci), 2017, 49(2): 337-343.
4	朱建国, 张菊香, 闫海燕, 等. 低能量氦-氖激光血管内照射与西比灵联合治疗糖尿病性周围神经病[J]. 中华物理医学与康复杂志, 2005, 27(3): 180-181.
	Zhu JG, Zhang JX, Yan HY, et al. Low-energy helium-neon laser intravascular irradiation combined with Sibelin for the treatment of diabetic peripheral neuropathy[J]. Chin J Phys Med Rehabil, 2005, 27(3): 180-181.
5	赵燕娟, 李荣华, 任刚, 等. 低能量激光联合牙周基础治疗对种植体周围炎龈沟液中IL-8、b-FGF及IL-1β水平的影响[J]. 国际生物医学工程杂志, 2019, 42(2): 130-133.
	Zhao YJ, Li RH, Ren G, et al. Effect of low level laser treatment combined with periodontal initial therapy on IL-8, b-FGF and IL-1β content in gingival crevicular fluid on peri-implantitis[J]. Int Biomed Eng J, 2019, 42(2): 130-133.
6	李凤, 徐华顺. 低能量激光配合牙周基础治疗对糖尿病合并慢性牙周炎患者龈沟液细胞因子及LPS、leptin的影响[J]. 上海口腔医学, 2018, 27(6): 637-640.
	Li F, Xu HS. Effects of low level laser combined with basic periodontal therapy on cytokines and LPS, leptin in gingival crevicular fluid of diabetes mellitus complicated with chronic periodontitis patients[J]. Shanghai J Stomatol, 2018, 27(6): 637-640.
7	Lee JH, Chiang MH, Chen PH, et al. Anti-inflammatory effects of low-level laser therapy on human periodontal ligament cells: in vitro study[J]. Lasers Med Sci, 2018, 33(3): 469-477.
8	Wu X, Zhang G, Feng X, et al. Transcriptome analysis of human periodontal ligament fibroblasts exposed to Porphyromonas gingivalis LPS[J]. Arch Oral Biol, 2020, 110: 104632.
9	Lu WL, Song DZ, Yue JL, et al. NLRP3 inflammasome may regulate inflammatory response of human periodontal ligament fibroblasts in an apoptosis-associated speck-like protein containing a CARD (ASC)-dependent manner[J]. Int Endod J, 2017, 50(10): 967-975.
10	Tanaka K, Miyake Y, Hanioka T, et al. The IL18 promoter polymorphism, rs1946518, is associated with the risk of periodontitis in Japanese women: the Kyushu Okinawa maternal and child health study[J]. Tohoku J Exp Med, 2017, 243(3): 159-164.
11	Almeida-Junior LA, Marques NCT, Prado MTO, et al. Effect of single and multiple doses of low-level laser therapy on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED)[J]. Lasers Med Sci, 2019, 34(9): 1917-1924.
12	任刚, 刘毅, 李荣华, 等. 低能量激光治疗对临床患者种植体周围炎的疗效研究[J]. 国际生物医学工程杂志, 2018, 41(5): 439-442.
	Ren G, Liu Y, Li RH, et al. Effect of low energy laser therapy on peri-implantitis in clinical patients[J]. Int J Biomed Eng, 2018, 41(5): 439-442.
13	Gürsoy M, Könönen E, Tervahartiala T, et al. Longitudinal study of salivary proteinases during pregnancy and postpartum[J]. J Periodontal Res, 2010, 45(4): 496-503.
14	Tiranathanagul S, Pattamapun K, Yongchaitrakul T, et al. MMP-2 activation by Actinobacillus actinomycetemcomitans supernatant in human PDL cells was corresponded with reduction of TIMP-2[J]. Oral Dis, 2004, 10(6): 383-388.

组别	细胞存活率/%	细胞内Ca²⁺浓度/（nmol·L^-1）
正常组	100*	226.69±19.45*
LPS组	34.52±7.86^#	312.38±28.08^#
2 J·cm^-2组	52.36±11.38^#*^@	257.25±26.32^#*^@
4 J·cm^-2组	72.5±12.25^#*	281.42±24.37^#*
6 J·cm^-2组	55.54±9.46^#*^@	251.24±21.32^#*^@

组别	TNF-α	IL-8	IL-1β	IL-6
正常组	0.79±0.14*	0.92±0.02*	1.03±0.04*	0.84±0.03*
LPS组	5.27±0.08^#	6.38±0.06^#	7.14±0.06^#	3.36±0.05^#
2 J·cm^-2组	3.92±0.05^#*^@	5.06±0.07^#*^@	5.86±0.05^#*^@	2.47±0.04^#*^@
4 J·cm^-2组	2.82±0.02^#*	3.56±0.04^#*	3.75±0.06^#*	1.38±0.02^#*
6 J·cm^-2组	3.48±0.03^#*^@	5.01±0.06^#*^@	5.64±0.03^#*^@	2.35±0.03^#*^@

[1]	Tang Xiaoxue, Zhou Zheng, Li Qiqi, Jiang Dandan. Effects of sitagliptin activation of the stromal cell-derived factor-1/CXC chemokine receptor 4 signaling pathway on the proliferation, apoptosis, inflammation, and osteogenic differentiation of human periodontal ligament stem cells induced by lipopolysaccharide [J]. West China Journal of Stomatology, 2024, 42(1): 37-45.
[2]	Lu Ting, Zhu Jiahao, Yang Shihe, Shen Zhe, Zhong Liangjun. Effects of Foxp3 gene silencing on the expression of inflammatory cytokines and the proliferation and migration of human periodontal ligament fibroblasts in an inflammatory environment [J]. West China Journal of Stomatology, 2023, 41(3): 269-275.
[3]	Liu Xiaolin, Wang Yu, Feng Xiaowei, Liu Wenjing, Li Xiangjun.. Comparison of surgical effects between the modified West China method and Millard method in repairing unila-teral cleft lip [J]. West China Journal of Stomatology, 2022, 40(3): 309-313.
[4]	Liao Shengnan, Weitong Lü, Tang Quan, Ma Yuwen, Liu Lijia, Wang Liang, Peng Xian. Study on the inhibitory effect of selective estrogen receptor modulators on Streptococcus mutans [J]. West China Journal of Stomatology, 2022, 40(2): 218-224.
[5]	Zhang Xiujuan, Wu Wei, Chen Xin, Fang Zhichun, Ye Jinxiang, Ou Xiaoyan.. Effects of grape seed proanthocyanidins on the expression of inflammatory mediators in gingival epithelial cells [J]. West China Journal of Stomatology, 2022, 40(1): 39-44.
[6]	Tang Lina, Liu Gang, Yang Chao, Sha Xiaowei, Wang Siyu. Short-term efficacy of superficial X-ray treatment for infantile maxillofacial hemangioma [J]. West China Journal of Stomatology, 2021, 39(4): 464-468.
[7]	Zhu Jianhui, Zheng Xin, Peng Xian, Xu Xin, Margolskee Robert, Zhou Xuedong. Regulation effect of lipopolysaccharide on the alternative splicing and function of sweet taste receptor T1R2 [J]. West China Journal of Stomatology, 2021, 39(4): 469-474.
[8]	Liu Xiang, Kang Wenyan, Shang Lingling, Ge Shaohua. Sitagliptin inhibits lipopolysaccharide-induced inflammatory response in human gingival fibroblasts by blocking nuclear factor-κB signaling pathway [J]. West China Journal of Stomatology, 2021, 39(2): 153-163.
[9]	Si Yuting, Song Jinhua, Fang Zhen, Han Xiaozhe, Jiang Shaoyun. MicroRNA-146a regulates the production of cytokines in lymphocytes stimulated by Porphyromonas gingivalis lipopolysaccharide [J]. West China Journal of Stomatology, 2021, 39(1): 26-31.
[10]	Yun Yang,Shanshan Chen,Chunmei Xu,Yafei Wu,Lei Zhao. Expression of triggering receptors expressed by myeloid cells-1 in macrophages stimulated by Porphyromonas gingivalis-lipopolysaccharide [J]. West China Journal of Stomatology, 2018, 36(5): 475-481.
[11]	Min Zhou, Zuolin. Wang. Protective effects of hydrogen-rich medium on lipopolysaccharides-induced injury in human periodontal ligament cells [J]. West China Journal of Stomatology, 2018, 36(2): 123-127.
[12]	Jiayi Wu, Ruijie Huang. Research progress on QMix properties in root canal irrigation [J]. West China Journal of Stomatology, 2017, 35(5): 543-548.
[13]	Jinmeng Li, Guomin. Ou. Accuracy of computer-guided implant placement and influencing factors [J]. West China Journal of Stomatology, 2017, 35(1): 93-98.
[14]	Shen Daonan, Cheng Wei, Jia Yue, Zhao Lei, Wu Yafei. Effect of glutaredoxin on oxidative stress of umbilical vein endothelial cell exposed to Porphyromonas gingivalis lipopolysaccharide [J]. West China Journal of Stomatology, 2015, 33(6): 613-616.
[15]	Zhan Xueling, Gao Jie, Liu Ying, Hu Jiao, Xue Yanxiang, Wu Buling. Lactoferrin downregulates the expression of Toll like receptor 4 stimulated by lipopolysaccharide in human periodontal ligament cells [J]. West China Journal of Stomatology, 2014, 32(2): 166-170.