Study On The Tissue-engineering Temporomandibular Joint Disc By Layering Electrospun Membranes And Co-culture Cell Sheet

Objective:The project aims to establishe a model for the regeneration of temporomandibular joint disc by co-culture cell sheet of temporomandibular joint disc cells and synovium-derived mesenchymals on PLGA electrospun scaffolds.The aim is to investigate the interaction between growth factors and co-culture in a 3D culture enviroment and to elighten future works for sustainable development of temporomandibular joint disc tissue engineering.Methods:Temporomandibular joint disc cells and synovium-derived mesenchymals were cultured alone and in a 1:1 ratio.The ascorbic acid was used to induce the cell sheet.The cell sheets were cultured on the PLGA electrospun scaffolds like‘Sandwich’.Then the samples were cultured in the chongenic medium with TGF-β(10ng/ml)or wiehout TGF-β(0ng/ml) for 14 days.Cell proliferation,extracellular matrix production, and chondrogenic gene expression were evaluated after two weeks.Results:The group with TGF-β presented more overall GAG than that without TGF-β(P<0.01).The GAG/DNA ratio of co-culture group had achieved an equivalent level with TMJDCs group(P>0.05). The group with TGF-β had induced more type II collagen than the group without TGF-β.And the relative mRNA level of Col2a1 and Sox-9 showed an increase in TMJDCs or SMSCs monoculrure with TGF-β( P <0.01).In the co-culture group with TGF-βthe relative mRNA level of Col2a1 incresed(P<0.01),but the level of SOX9 decreased(P<0.01).Conclusion:The ‘Sandwich’model witch co-culture cell sheet of temporomandibular joint disc cells and synovium-derived mesenchymals on PLGA electrospun scaffolds can form a tissue that similar to temporomandibular joint disc.And TGF-β can promote proliferation and increase the extracelluar matrix synthesis.As mechanism isn`t clear,we need further study to solve it.