Layering Poly (lactic-co-glycolic acid)-based electrospun membranes and co-culture cell sheets for engineering temporomandibular joint disc

Abstract

The temporomandibular joint disk (TMJD) lacks blood vessels and is characterized by slow self-repair. Qualitative lesions in TMJD are difficult to repair. In this study, electrospun poly (lactic-co-glycolic acid) (PLGA) scaffolds were used to reconstruct temporomandibular joint discs by tissue engineering. Rabbit temporomandibular joint disc cells (TMJDCs) and rabbit synovium-derived mesenchymal stem cells (SMSCs) were co-cultured in 1:1 ratios. Cell sheets were induced by ascorbic acid incubated with electrospun PLGA scaffolds for 14 days in the presence (10 ng/ml in culture medium) or absence of TGF-β3. Dimethylmethylene Blue Assay (DMMB) was used to determine the content of glycosaminoglycans in the extracellular matrix. The expression of Col1a1, Col2a1, Sox-9 and Runx-2 was quantified by RT-PCR, and the expression of type II collagen was observed by immunofluorescent staining. After 14 days of cultivation, the electrospun PLGA scaffold-loaded cell sheets could form an articular disc tissue with certain morphological characteristics. The expression of chondrogenic-related genes (Col2a1, Sox-9) and the secretion of extracellular matrix (GAG, type II collagen) in the co-culture group were close to those in the TMJDC group alone. The results suggest that PLGA electrospun scaffold-loaded co-cultured cell membrane could be used in the tissue engineering reconstruction of the temporomandibular joint disc