Tenocyte-imprinted substrate: a topography-based inducer for tenogenic differentiation in adipose tissue-derived mesenchymal stem cells

Abstract

Tendon tissue engineering based on stem cell differentiation has attracted a great deal of attention in recent years. Previous studies have examined the effect of cell-imprinted polydimethylsiloxane (PDMS) substrate on induction differentiation in stem cells. In this study, we used tenocyte morphology as a positive mold to create a tenocyte-imprinted substrate on PDMS. The morphology and topography of this tenocyte replica on PDMS was evaluated with scanning electron microscopy and atomic force microscopy. Then the tenogenic differentiation induction capacity of tenocytes replica in ADSCs was investigated and compared with other groups including tissue replica (Which was produced similar to the tenocyte replica and was evaluated by SEM), decellularized tendon, and BMP-12 as other potentially inducers. This comparison gives us an estimate of the ability of tenocyte-imprinted PDMS (which called cell replica in the present study) to induce differentiation compared to other inducers. For this reason ADSCs were divided into 5 groups including control, cell replica, tissue replica, decellularized tendon and BMP-12.ADSCs were seeded on each group seperately and investigated by real-time RT-PCR after 7 and 14 days. Our results showed that in spite of the higher effect of growth factor on tenogenic differentiation, cell replica can also induce tenocyte marker expression (Scleraxis and Tenomodulin) in ADSCs. Moreover, tenogenic differentiation induction capacity of cell replica was greater than tissue replica. Immunocytochemistry analysis revealed that ADSCs seeding on cell replica for 14 days led to Scleraxis and Tenomodulin expression at the protein level. Also, immunohistochemistry indicated that contrary to the promising results in vitro, there was little difference between ADSCs cultured on tenocyte-imprinted PDMS and untreated ADSCs. The results of such studies could lead to the production of inexpensive cell culture plates or biomaterials that can induce differentiation in stem cells without growth factors or other supplements.

In Vitro Innovation of Tendon Tissue Engineering Strategies

“…This cell type's tenogenic ability has been shown including when exposed to tendon extracellular matrix and TGFβ3 [207]. Moreover, ADSCs seeded on a tropoelastin-coated biomimetic scaffold, after 21 days, showed an increased protein expression of tendon-related markers such as Scx and Tnmd and were able to secrete extracellular matrix components such as Col I, Col III, Tnc, and Dcn [203].…”

“…MSCs are an important source in regenerative medicine and have received great attention in the tenogenic differentiation and regeneration of functional tendons [203]. When cultured in normoxic conditions, MSCs display enhanced proliferation rates, retention of stem cell properties, inhibition of senescence, and increased differentiation ability [249,250].…”

“…A fibrin hydrogel merged with an elastic braided hyaluronated band scaffold was also described as committing human BMSC into a tenogenic phenotype under cyclic strain [ 198 , 202 ]. Dai et al [ 203 ] showed that BMSCs are more responsive to bone morphogenetic protein-12 (BMP-12) stimulation compared to ADSCs [ 204 ].…”

Molecular Imprinting Strategies for Tissue Engineering Applications: A Review

“…On the other hand, the use of stem cells suffers from the problems of their correct differentiation, as well as keeping their phenotype over time. In this context, the use of MI substrates, although with less differentiation potential when compared to GFs, represents an inexpensive and simple way to expand and differentiate stem cells for TE purposes [ 113 ]. In a recent example, Kashani et al proposed the combination of MIPs and microfluidics devices by using a two-step approach: create a chondrocytes-imprinted substrate using a first microfluidic device, and then culture stem cells over it using a second device.…”

P75 and S100 gene expression induced by cell‐imprinted substrate and beta‐carotene to nerve tissue engineering

“…19 Furthermore, studies have shown that fabricating artificial microenvironments with PDMS can mechanically, physically, and chemically mimic the ECM of target cells. 20,21 Implanting the suitable cells on the patterned polymeric surfaces is of high importance. Human mesenchymal stem cells (hMSCs) are known as multipotent stem cells in which, with controlled supervision, they may undergo differentiation into several mesenchymal tissue lineages, including fat, muscle, bone, cartilage, and neural cells.…”