Combination of biochemical and mechanical cues for tendon tissue engineering

Abstract

Tendinopathies negatively affect the life quality of millions of people in occupational and athletic settings, as well as the general population. Tendon healing is a slow process, often with insufficient results to restore complete endurance and functionality of the tissue. Tissue engineering, using tendon progenitors, artificial matrices and bioreactors for mechanical stimulation, could be an important approach for treating rips, fraying and tissue rupture. In our work, C3H10T1/2 murine fibroblast cell line was exposed to a combination of stimuli: a biochemical stimulus provided by Transforming Growth Factor Beta (TGF‐β) and Ascorbic Acid (AA); a three‐dimensional environment represented by PEGylated‐Fibrinogen (PEG‐Fibrinogen) biomimetic matrix; and a mechanical induction exploiting a custom bioreactor applying uniaxial stretching. In vitro analyses by immunofluorescence and mechanical testing revealed that the proposed combined approach favours the organization of a three‐dimensional tissue‐like structure promoting a remarkable arrangement of the cells and the neo‐extracellular matrix, reflecting into enhanced mechanical strength. The proposed method represents a novel approach for tendon tissue engineering, demonstrating how the combined effect of biochemical and mechanical stimuli ameliorates biological and mechanical properties of the artificial tissue compared to those obtained with single inducement.

Tendon Tissue Engineering: Effects of Mechanical and Biochemical Stimulation on Stem Cell Alignment on Cell‐Laden Hydrogel Yarns

“…Indeed, previous studies demonstrated that BMP‐12 plays an important role in chemical cellular stimulation as well as tissue homeostasis, healing and repair, affects cell proliferation and migration, collagen production, angiogenesis, and ECM proteins deposition . Moreover, since tendon undergoes mechanical loading during its physiological operation, several studies suggested the effective role of mechanical stimulation on cellular differentiation and alignment . For this reason, the hydrogel bundles were cultured in four different conditions—growth medium (GA yarns used as control), culture medium modified with BMP‐12 (GA yarns BS), growth medium + mechanical static stimulation (GA yarns MS), and culture medium modified with BMP‐12 + mechanical static stimulation (GA yarns MBS)—so as to assess a beneficial effect induced by mechanical stimulation or mechanical stimuli combined with biochemical stimulation (Figure e,f).…”

“…Each cDNA sample was amplified in triplicate using KAPA SYBR FAST qPCR kit Master Mix (Kapa Biosystems). Relative mRNA levels were calculated by the delta delta CT method . Primer specificity was confirmed by melting curve analysis.…”

Supporting Cell-Based Tendon Therapy: Effect of PDGF-BB and Ascorbic Acid on Rabbit Achilles Tenocytes In Vitro

“…One way to improve cell-based tendon therapy is to perform a pre-treatment on the cell culture in order to increase cell proliferation and matrix synthesis before an application. Strategies for pre-treatment of tenocyte or MSCs cell culture include mechanical stimulation [8,22], three-dimensional spheroid cultures [23], macromolecular crowding [24] or biochemical supplementation of the culture medium [12,25]. From biochemical supplementation, stimulation by different growth factors, including transforming growth factor-beta (TGF-β) [8], bone morphogenetic protein-12 (BMP-12) [26], connective tissue growth factor (CTGF) [27], and platelet-derived growth factor-BB (PDGF-BB) [28] has been explored.…”

Skeletal muscle-derived Human Mesenchymal Stem Cells: influence of different culture conditions on proliferative and myogenic capabilities

“…Cells were fixed with 4% PFA in PBS for 10 minutes at 4°C and processed for immunofluorescence analysis as previously described (Testa et al, 2017…”