In Vitro Generation of a Scaffold-Free Tissue-Engineered Construct (TEC) Derived from Human Synovial Mesenchymal Stem Cells: Biological and Mechanical Properties and Further Chondrogenic Potential

Abstract

The purpose of this study was to characterize a tissue-engineered construct (TEC) generated with human synovial mesenchymal stem cells (MSCs). MSCs were cultured in medium with ascorbic acid 2-phosphate (Asc-2P) and were subsequently detached from the substratum. The detached cell/matrix complex spontaneously contracted to develop a basic TEC. The volume of the TEC assessed by varying initial cell density showed that it was proportional to initial cell densities up to 4 x 10(5) cells/cm(2). Assessment of the mechanical properties of TEC using a custom device showed that the load at failure and stiffness of the constructs significantly increased with time of culture in the presence of Asc-2P, while in the absence of Asc-2P, the constructs were mechanically weak. Thus, the basic TEC possesses sufficiently self-supporting mechanical properties in spite of not containing artificial scaffolding. TEC further cultured in chondrogenic media exhibited positive alcian blue staining with elevated expression of chondrogenic marker genes. Based on these findings, such human TEC may be a promising method to promote cartilage repair for future clinical application.

Synovia-Derived Mesenchymal Stem Cell Application in Musculoskeletal Injuries: A Review

“…This group demonstrated cells' ability to adhere to human chondral fragments [56], as it was previously demonstrated in the pig model. They also applied a xeno-free technique, by using human serum [5,56].…”

“…This group demonstrated cells' ability to adhere to human chondral fragments [56], as it was previously demonstrated in the pig model. They also applied a xeno-free technique, by using human serum [5,56]. Autologous human serum is reported to be more effective in promoting SM-MSCs' proliferation, in comparison to other MSC sources [5,10,57].…”

“…Scaffoldfree tissue-engineered constructs (TECs) were developed in order to overcome the previous drawbacks. MSCs are cultured in monolayer with addition of ascorbic acid and are then submitted to shear stress, resulting in their detachment and spontaneous contraction to form the 3D structure, similar to what is observed with collagen gels [56].…”

“…The TEC showed to have stable adhesion to a porcine cartilage matrix, in an explant culture, possibly due to the adhesion properties of fibronectin. When cultured in chondrogenic medium, enhanced expression of glycosaminoglycans and chondrogenic matrix genes, as collagen II and aggrecan, was observed [33,56], suggesting that SM-MSCs in the TEC retain their chondrogenic potential [56]. They also proposed a xenofree system for the development of this technique, as the TEC is produced without an exogenous scaffold, with autologous serum and MSCs.…”

Cell-based Meniscal Tissue Engineering: A Case for Synoviocytes

“…Scaffolds may further be defined as ''smart scaffolds'' if they carry with them some type of signal that can direct appropriate tissue formation either from the implanted cell type or from the surrounding resident cellular population. With respect to the delivery of synoviocytes for cartilage (hyaline or fibrocartilage) engineering, a number of different systems have been investigated including hydrogels (alginate [53,66] collagen [51,63,94], and gellan [28]), pellet cultures [63,80], micromasses [3,66], small intestinal submucosa (SIS) [84], hyaluronan-based scaffolds (Hyaff-111; FAB, Abano Terme, Padova, Italy) [56], polyglycolic acid (PGA) [67,74], PGA/poly (L) lactic acid (PLLA) combinations [34], and scaffold-free cell infusions [44,60]. With relatively few studies directly comparing delivery methods, elucidating an optimal carrier or scaffold from the literature alone is difficult.…”