Bionic Silk Fibroin Film Induces Morphological Changes and Differentiation of Tendon Stem/Progenitor Cells

Abstract

Purpose. Tendon injuries are common musculoskeletal system disorders, but the ability for tendon regeneration is limited. Silk fibroin (SF) film may be suitable for tendon regeneration due to its excellent biocompatibility and physical properties. This study is aimed at evaluating the application value of bionic SF film in tendon regeneration. Methods. Tendon stem/progenitor cells (TSPCs) were isolated from rat Achilles tendon and characterized based on their surface marker expression and multilineage differentiation potential. SF films with smooth or bionic microstructure surfaces (5, 10, 15, 20 μm) were prepared. The morphology and mechanical properties of natural tendons and SF films were characterized. TSPCs were used as the seed cells, and the cell viability and cell adhesion morphology were analyzed. The tendongenesis-related gene expression of TSPCs was also evaluated using quantitative polymerase chain reaction. Results. Compared to the native tendon, only the 10, 15, and 20 μm SF film groups had comparable maximum loading and ultimate stress, with the exception of the breaking elongation rate. The 10 μm SF film group had the highest percentage of oriented cells and the most significant changes in cell morphology. The most significant upregulations in the expression of COL1A1, TNC, TNMD, and SCX were also observed in the 10 μm SF film group. Conclusion. SF film with a bionic microstructure can serve as a tissue engineering scaffold and provide biophysical cues for the use of TSPCs to achieve proper cellular adherence arrangement and morphology as well as promote the tenogenic differentiation of TSPCs, making it a valuable customizable biomaterial for future applications in tendon repair.

N-Acetyl-L-cysteine facilitates tendon repair and promotes the tenogenic differentiation of tendon stem/progenitor cells by enhancing the integrin α5/β1/PI3K/AKT signaling

“…TSPCs were isolated from tendon tissues of three 8-week old male Sprague-Dawley (SD) rats as three independent biological samples and identified for their cell surface marker expression and multi-directional differentiation potential, as our previous study [ 33 , 53 ]. All procedures and experiments were approved by the Ethics Committee of Army Medical University (Chongqing, China) and performed, according to the guidelines of the National Institutes of Health for the Care and Use of Laboratory Animals [ 53 , 54 ]. The TSPCs were cultured in F12 complete media, which were changed every three days and the cells were passaged every xxx days.…”

Micropattern Silk Fibroin Film Facilitates Tendon Repair In Vivo and Promotes Tenogenic Differentiation of Tendon Stem/Progenitor Cells through the α2β1/FAK/PI3K/AKT Signaling Pathway In Vitro

“…Preparation and Characterization of SF Films. SF films were prepared as previously described [21,22]. Briefly, the SF solution was dried to form smooth or micropattern films on polydimethylsiloxane (PDMS) plate, and their stability was improved by water-annealing treatment.…”

“…Thus, bionics has emerged as a new direction in the design and preparation of biomaterials for tissue repair [14,[18][19][20]. In our previous studies, we demonstrated that SF films mimicking the physical surface micropattern of fiber morphology of native tendon collagen can promote the tenogenic differentiation of tendon stem/progenitor cells (TSPCs) [21,22]. However, the repairing effect of micropattern SF films in tendon injury and their effect on regulating the intrinsic TSPC signaling pathways remain unclear.…”

Extraction of Silk Fibroin with Several Sericin Removal Processes and its Importance in Tissue Engineering: A Review

“…Tissue engineering is a promising therapeutic modality for the reconstruction of ligaments and tendons. Silk fibroin scaffold has found as an appropriate biomaterial being utilized for this purpose [134,135]. Tellado et al, was constructed a dual characteristic silk fibroin scaffold that contains two diverse pore arrangements.…”

“…Lu et al investigated the mechanical attributes of silk fibroin in tendon reconstruction. They concluded that the artificial micro-fabrication on the surface of silk fibroin film contributes remarkable biological supervision in the growth of tendon stem/progenitor cells (TSPCs) that quality makes silk fibroin film an effective ingredient for the reconstruction of the tendon tissue [135].…”