1. Academic Validation
  2. Nicotinamide improves in vitro lens regeneration in a mouse capsular bag model

Nicotinamide improves in vitro lens regeneration in a mouse capsular bag model

  • Stem Cell Res Ther. 2022 May 12;13(1):198. doi: 10.1186/s13287-022-02862-8.
Xiaomin Liu 1 2 Qingjun Zhou 1 Yusen Huang 1 2 Zheng Fan 1 3 Haoyun Duan 1 Menghan Wang 1 3 Zongyi Li 4 5 Lixin Xie 6 7
Affiliations

Affiliations

  • 1 State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Eye Institute of Shandong First Medical University, Qingdao, 26600, China.
  • 2 Qingdao Eye Hospital of Shandong First Medical University, Qingdao, 26600, China.
  • 3 Qingdao University Medical College, Qingdao, 26600, China.
  • 4 State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Eye Institute of Shandong First Medical University, Qingdao, 26600, China. lizongyi119@163.com.
  • 5 Qingdao Eye Hospital of Shandong First Medical University, Qingdao, 26600, China. lizongyi119@163.com.
  • 6 State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Eye Institute of Shandong First Medical University, Qingdao, 26600, China. xielixin@hotmail.com.
  • 7 Qingdao Eye Hospital of Shandong First Medical University, Qingdao, 26600, China. xielixin@hotmail.com.
Abstract

Background: Mammalian lens regeneration holds great potential as a cataract therapy. However, the mechanism of mammalian lens regeneration is unclear, and the methods for optimization remain in question.

Methods: We developed an in vitro lens regeneration model using mouse capsular bag culture and improved the transparency of the regenerated lens using nicotinamide (NAM). We used D4476 and SSTC3 as a Casein Kinase 1A inhibitor and agonist, respectively. The expression of lens-specific markers was examined by Real-Time PCR, immunostaining, and western blotting. The structure of the in vitro regenerated lens was investigated using 3,3'-dihexyloxacarbocyanine iodide (DiOC6) and methylene blue staining, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL), and transmission electron microscopy.

Results: The in vitro lens regeneration model was developed to mimic the process of in vivo mammalian lens regeneration in a mouse capsular bag culture. In the early stage, the remanent lens epithelial cells proliferated across the posterior capsule and differentiated into lens fiber cells (LFCs). The regenerated lenses appeared opaque after 28 days; however, NAM treatment effectively maintained the transparency of the regenerated lens. We demonstrated that NAM maintained lens epithelial cell survival, promoted the differentiation and regular cellular arrangement of LFCs, and reduced lens-related cell Apoptosis. Mechanistically, NAM enhanced the differentiation and transparency of regenerative lenses partly by inhibiting Casein Kinase 1A activity.

Conclusion: This study provides a new in vitro model for regeneration study and demonstrates the potential of NAM in in vitro mammalian lens regeneration.

Keywords

Capsular bag culture; Differentiation; Lens regeneration; Nicotinamide; Transparency.

Figures
Products