| Background:Due to its unique anatomical and organizational structure, Tendon tissue is lack of self-repair capacity. Tendon injury frequently results in scar tissue formation. In order to solve this problem, tissue engineering has become a hotspot for tendon injury. However,the artificial biological material carrier has shortcomings such as aging, exclusion. Bone marrow mesenchymal stem cells (BMSCs) have been widely used in the field of tissue regeneration as a veterinary seed cell, but it has been reported that ectopic calcification and stem cell formation occurred after application. Tendon-derived stem cells (TDSCs)have more advantages in theory for tendon repair due to their origins. Connective tissue growth factor (CTGF) can induce the differentiation of both cells into tendon cells. So,we use CTGF and these two cells to construct engineered tendon tissue without other materials, and compare the two kinds of seed cells to choose the more suitable one for tendon repair.Objective:1. To confirm the ability of connective tissue growth factor (CTGF) to induce bone marrow mesenchymal stem cells (BMSCs) and tendon-derived stem cells (TDSCs)to differentiate into tendon cells.2. To construct engineered tendon tissue using CTGF together with BMSCs or TDSCs.3. To explore and compare the engineered tendon tissue constructed by BMSCs and TDSCs.Methods:1. BMSCs and TDSCs from GFP transgenic rat were isolated and characterized.2. BMSCs and TDSCs at passage 3 were added into 25ng/ml CTGF and cultured for 2 weeks. Two tenogenic related genes (scleraxis and tenomodulin) were selected and RT-qPCR was used to test the mRNA expression of the two genes.3. BMSCs and TDSCs at passage 3 were cultured with 25ng/ml CTGF for 2 weeks.The engineered tendon tissue was constructed in vitro.4. The engineered tendon tissues constructed by BMSCs and TDSCs were implanted subcutaneously into nude mice. the engineered tendon tissue samples were collected at week 8 and 12. Histomorphological examination and immunohistochemical staining were performed.Results:1. The RT-qPCR results showed that after induced by CTGF, the mRNA expression of tenomodulin and scleraxis were significantly increased in BMSCs and TDSCs(p<0.05 ) . Compared to BMSCs, mRNA expression of tenomodulin and scleraxis in TDSCs was significantly higher (p<0.05 ).2. Histology results showed that engineered tendon tissue constructed by both stem cells showed an immature tissue structure. Compared to BMSCs, TDSCs engineered tendon tissue had more suitable structure.3. Histology and immunohistochemistry results showed that engineered tendon tissue constructed by both stem cells formed tendon like tissue in the nude mouse. TDSCs engineered tendon tissue had more mature structure and more similar extracellular matrix than which constructed by BMSCs.Conclusions:1. The tenogenic differentiational potential of BMSCs and TDSCs can be promoted by CTGF in vitro, furthermore, TDSCs have better potential on the tenogenic differentiation.2. Using only CTGF together with BMSCs or TDSCs, engineered tendon tissue can be constructed in vitro.3. Both the engineered tendon tissue can form tendon like tissue in nude mouse model.TDSCS group had better results. This indicated that TDSCs more suitable for tendon regeneration in tissue engineering field. |
PDF Full Text Request