Journal of Otolaryngology and Ophthalmology of Shandong University ›› 2025, Vol. 39 ›› Issue (3): 141-147.doi: 10.6040/j.issn.1673-3770.0.2023.433

• Original Article • Previous Articles    

Therapeutic effects of human umbilical cord mesenchymal stem cells induced by TNF-α on limbal stem cell deficiency in mice

YU Haonan1, ZHONG Yingying2, WANG Xinmeng1, ZHANG Min3, JIANG Qingmin2, LI Nan2, LI Yan2   

  1. 1. Affiliated Hospital of Weifang Medical University , School of Clinical Medicine , Weifang Medical University, Weifang 261031, Shandong, China2. The Eye Center, Affiliated Hospital of Weifang Medical University, Weifang 261031, Shandong, China3. The First Clinical Medical College, Jinan University, Guangzhou Panyu District Campus, Guangzhou 511436, Guangdong, China
  • Published:2025-06-04

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Abstract: Objective To investigate the repairing effect of local application of tumor necrosis factor alpha(TNF-α)-induced human umbilical cord mesenchymal stem cell conditioned medium(hUCMSC-CM)on limbal stem cell deficiency(LSCD)in mice. Methods Primary human umbilical cord mesenchymal stem cells were isolated and cultured in vitro by the adherent tissue block culture method, the fourth generation hUCMSCs were induced with TNF-α, and the TNF-α-induced human umbilical cord mesenchymal stem cell conditioned medium(hUCMSC-CMT)was collected. Forty-eight healthy 6-week-old C57 male mice were randomly divided into the control group(group N)and LSCD experimental group, and the LSCD experimental group model was established by limbal alkali burn and divided into negative PBS injection control group(group A), hUCMSC-CM injection treatment group(group B), and the injection treatment group hUCMSC-CMT(group C)according to the different components of subconjunctival injection after modeling, and the subconjunctival injection was once a day for 7 consecutive days. On the 3rd and 7th days after LSCD modeling, the degree of corneal epithelial defect was observed and scored by slit-lamp microscopy, and corneal neovascularization was observed and scored by slit-lamp microscopy on the 7th and 21st days after LSCD modeling. Immunohistochemical staining was used to detect the expression of α-smooth muscle actin(α-SMA)and ocular surface mucin 5AC(Muc-5AC). Results The mice in group A showed large areas of corneal epithelial defect on day 3 after modeling; obvious congested limbal blood vessels were observed on day 7 after modeling. The corneal epithelial defect developed in more than 2/3 of the cornea. On the 21st day after modeling, more neovascularization in the corneal limbus was observed, and some could reach 2/3 of the corneal diameter. According to the statistical results, both group B and group C had significant improvements in reducing corneal neovascularization and repairing corneal epithelial defect, and the curative effect was stronger in group C than in group B. Immunohistochemical staining showed that there was no expression of α-SMA in the corneal stroma and no expression of Muc-5AC in the corneal epithelial layer of the normal group. At 7 and 21 days after modeling, the cornea of each experimental group showed positive expression of α-SMA. The expression of α-SMA was significantly lower in groups B and C than in group A. The average optical density values of Muc-5AC in three experimental groups on the 7th day after modeling were not significantly different. Conclusion Subconjunctival injection of hUCMSC-CMT or hUCMSC-CM could enhance the function of limbal stem cells, promote corneal epithelial repair, and protect ocular surface tissues. In addition, hUCMSC-CMT had stronger repairing and therapeutic effects on limbal stem cell deficiency in mice than hUCMSC-CM.

Key words: Tumor necrosis factor alpha, Human umbilical cord mesenchymal stem cells, Limbal stem cell deficiency, Conditioned medium

CLC Number: 

  • R772.2
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