Reversal of senescence-associated beta-galactosidase expression during in vitro three-dimensional tissue-engineering of human chondrocytes in a polymer scaffold.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Biopolymers* ; Gene Expression Regulation* ; Tissue Engineering*/methods ; Tissue Scaffolds*; Cellular Senescence/*genetics ; Chondrocytes/*metabolism ; beta-Galactosidase/*genetics; Biomarkers ; Cell Culture Techniques ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Flow Cytometry ; Fluorescent Antibody Technique ; Humans ; beta-Galactosidase/metabolism

Regenerative medicine applications require cells that are not inflicted with senescence after in vitro culture for an optimal in vivo outcome. Methods to overcome replicative senescence include genomic modifications which have their own disadvantages. We have evaluated a three-dimensional (3D) thermo-reversible gelation polymer (TGP) matrix environment for its capabilities to reverse cellular senescence. The expression of senescence-associated beta-galactosidase (SA-βgal) by human chondrocytes from osteoarthritis-affected cartilage tissue, grown in a conventional two-dimensional (2D) monolayer culture versus in 3D-TGP were compared. In 2D, the cells de-differentiated into fibroblasts, expressed higher SA-βgal and started degenerating at 25 days. SA-βgal levels decreased when the chondrocytes were transferred from the 2D to the 3D-TGP culture, with cells exhibiting a tissue-like growth until 42-45 days. Other senescence associated markers such as p16 ^INK4a and p21 were also expressed only in 2D cultured cells but not in 3D-TGP tissue engineered cartilage. This is a first-of-its-kind report of a chemically synthesized and reproducible in vitro environment yielding an advantageous reversal of aging of human chondrocytes without any genomic modifications. The method is worth consideration as an optimal method for growing cells for regenerative medicine applications.

Regen Ther. 2020 May 15;14:234-237. (PMID: 32435676)
Int J Rheum Dis. 2016 Mar;19(3):226-32. (PMID: 26112901)
J Orthop. 2017 Feb 03;14(2):223-225. (PMID: 28203047)
Life Sci. 2021 Aug 1;278:119553. (PMID: 33932445)
J Endocrinol. 2016 Aug;230(2):R59-76. (PMID: 27325241)
FEBS J. 2021 Jan;288(1):56-80. (PMID: 32961620)
Aging (Albany NY). 2016 Jan;8(1):3-11. (PMID: 26805432)
Biomed Res Int. 2014;2014:925121. (PMID: 24719895)
J Am Geriatr Soc. 2017 Oct;65(10):2297-2301. (PMID: 28869295)
BMC Geriatr. 2013 Sep 24;13:99. (PMID: 24063536)
J Exp Med. 2021 Mar 1;218(3):. (PMID: 33231616)
Genome Med. 2016 Jun 20;8(1):69. (PMID: 27323951)
Int J Mol Sci. 2020 Mar 29;21(7):. (PMID: 32235300)
J Vis Exp. 2017 May 1;(123):. (PMID: 28518126)
Aging Cell. 2002 Oct;1(1):57-65. (PMID: 12882354)
Nat Rev Rheumatol. 2021 Jan;17(1):47-57. (PMID: 33208917)
Nat Rev Mol Cell Biol. 2007 Sep;8(9):729-40. (PMID: 17667954)
Exp Cell Res. 1965 Mar;37:614-36. (PMID: 14315085)
J Orthop. 2021 Jan 16;23:138-141. (PMID: 33510554)
Aging Cell. 2006 Apr;5(2):187-95. (PMID: 16626397)
Biochem Biophys Res Commun. 2004 May 14;317(4):1103-7. (PMID: 15094382)
Tissue Eng. 2006 May;12(5):1237-45. (PMID: 16771637)
Tissue Eng Part A. 2009 Feb;15(2):407-15. (PMID: 18724830)
Indian J Ophthalmol. 2014 Feb;62(2):130-5. (PMID: 24008800)
J Gerontol A Biol Sci Med Sci. 2017 Jun 1;72(6):780-785. (PMID: 27516624)
Front Physiol. 2018 Jun 25;9:706. (PMID: 29988615)
Int J Urol. 2019 Feb;26(2):253-257. (PMID: 30468021)

0 (Biomarkers)
0 (Biopolymers)
EC 3.2.1.23 (beta-Galactosidase)

Date Created: 20210708 Date Completed: 20211123 Latest Revision: 20240402

20240402

PMC8263703

10.1038/s41598-021-93607-9

34234261