Effect of recombinant human amelogenin-loaded PCLA-PEG-PCLA hydrogels on biological properties of human periodontal ligament fibroblasts
10.3969/j.issn.1674-8115.2019.03.005
- Author:
Hang NING
1
Author Information
1. Department of Periodontology, College of Stomatology, Shanghai Jiao Tong University School of Medicine
- Publication Type:Journal Article
- Keywords:
Amelogenin;
Attachment;
Hydrogel;
Migration;
Osteogenic differentiation;
Periodontal tissue regeneration;
Proliferation
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2019;39(3):244-252
- CountryChina
- Language:Chinese
-
Abstract:
Objective • To determine the effect of recombinant human amelogenin (rhAm)-loaded PCLA-PEG-PCLA hydrogels on cell proliferation, immigration, attachment and osteogenic differentiation of human periodontal ligament fibroblasts (HPDLFs). Methods • HPDLFs were obtained by tissue block method in vitro from extracted premolars and the 3rd-5th passages of HPDLFs were treated with DMEM medium (control group), 20 μg/mL rhAm (rhAm group) or rhAm-loaded PCLA-PEG-PCLA hydrogels (rhAm-loaded hydrogel group). Proliferation activity was measured by CCK-8, while cell migration was assayed both by wound-healing experiment in vitro and Transwell experiment. Cell attachment was measured by hemocytometer and observed by scanning electron microscope. Osteogenic differentiation was measured by real-time PCR, with ALP, Runx2, CEMP1 and CAP as the target genes. Results • RhAm-loaded PCLA-PEG-PCLA hydrogels had no significant effect on cell growth curve of HPDLFs, but promoted cell proliferation after 3 days (P<0.05). RhAm accelerated cell migration mostly both in wound-healing experiment and Transwell experiment, with rhAm-loaded hydrogels in the second place. RhAm-loaded hydrogels promoted cell attachment, and in the 4th hour the promotion was of statistic significance (P<0.05). Meanwhile cells of rhAm group and rhAm-loaded hydrogel group had a better stretch condition than control group under the scanning electron microscope. After culture with rhAm-loaded hydrogels, ALP, Runx2, CEMP1, and CAP mRNA were upregulated in different time points. Conclusion • Recombinant human amelogenin-loaded PCLA-PEG-PCLA hydrogels can significantly improve proliferation, attachment and osteogenic differentiation of HPDLFs, but has no effect on cell migration on a statistical scale.
