TGFβ2 differentially modulates smooth muscle cell proliferation and migration in electrospun gelatin-fibrinogen constructs

Abstract

A main goal of tissue engineering is the development of scaffolds that replace, restore and improve injured tissue. These scaffolds have to mimic natural tissue, constituted by an extracellular matrix (ECM) support, cells attached to the ECM, and signaling molecules such as growth factors that regulate cell function. In this study we created electrospun flat sheet scaffolds using different compositions of gelatin and fibrinogen. Smooth muscle cells (SMCs) were seeded on the scaffolds, and proliferation and infiltration were evaluated. Additionally, different concentrations of Transforming Growth Factor-beta2 (TGFβ2) were added to the medium with the aim of elucidating its effect on cell proliferation, migration and collagen production. Our results demostrated that a scafold with a composition of 80% gelatin-20% fibrinogen is suitable for tissue engineering applications since it promotes cell growth and migration. The addition of TGFβ2 at low concentrations (≤1ng/ml) to the culture medium resulted in an increase in SMC proliferation and scaffold infiltration, and in the reduction of collagen production. In contrast, TGFβ2 at concentrations >1ng/ml inhibited cell proliferation and migration while stimulating collagen production. According to our results TGFβ2 concentration has a differential effect on SMC function and thus can be used as a biochemical modulator that can be beneficial for tissue engineering applications.

Biomechanical Comparison of Glutaraldehyde-Crosslinked Gelatin Fibrinogen Electrospun Scaffolds to Porcine Coronary Arteries

“…However, we are currently conducting experiments to determine the effects of SMC-mediated gelatin/ fibrinogen remodeling and ECM deposition on construct mechanical properties. Finally, our previous paper [44] demonstrated that our gelatin/fibrinogen materials do promote cell division, migration, and collagen deposition ex vivo. Even though ex vivo experiments do not always translate into in vivo experiments, we do believe that if our acellular constructs were implanted in vivo, cell infiltration and native construct remodeling would occur.…”

“…Gelatin extracted from porcine skin (Sigma-Aldrich, St. Louis, MO) and fraction I bovine fibrinogen (Sigma-Aldrich) was dissolved in 1,1,1,3,3,3-hexafluoro-2-propanol (HFP) (Sigma-Aldrich) at a 80:20 ratio to create a 10% (w/v) solution. Constructs with this composition have been shown by our group to promote cell adherence and proliferation compared to alternative ratios [44]. In a ventilation hood, the solution was loaded into a 5 ml BD syringe with a 23 gauge stainless steel dispensing needle and attached to a NE-100 single syringe pump (New Era Pump Systems, Farmingdale, NY).…”

“…Researchers have electrospun gelatin/fibrinogen sheets and successfully cultured human cardiomyocytes with brief mechanical characterization [43]. Along those lines, our group investigated the feasibility of growing porcine vascular smooth muscle cells (SMCs) on electrospun gelatin/fibrinogen scaffolds [44]. Our results showed that SMCs proliferated and migrated within the scaffold.…”

Endothelial angiogenesis is directed by RUNX1T1-regulated VEGFA, BMP4 and TGF-β2 expression

“…The expression of TGF-β2 in indispensable for proliferation of smooth muscle cells (SMCs) [58] and recruitment of SMCs to lymphatic vessels stimulates the secretion of endothelial-derived Reelin for lymphatic vessel morphogenesis [59]. Here, we identified VEGFA, BMP4 and TGF-β2 as RUNX1T1-regulated secretory proteins in human and their expression positively associated with RUNX1T1 both in vitro and in vivo (Fig 6B–6G).…”

Simple Approach to Generate Fluorescent Quantum Dots/Gelatin Composite with Thermo-responsive and Reversible Sol-gel Transition

“…Cadmium sulfide (CdS) QDs, a kind of extensively utilized QDs, exhibit a diverse range of applications for biological imaging, light-emitting diode, photoelectrodes and solar cell (7)(8)(9)(10). It is noteworthy that some studies have been recently dedicated to the conjunction of CdS QDs and biopolymers, in order to reduce the toxicity or Gelatin, a widely used biopolymer derived from hydrolytic degradation of collagen, has a number of advantages such as biocompatibility, biodegradability, no toxicity and low cost (13)(14)(15)(16), which allow it to be an excellent candidate for numerous applications in pharmaceutical, food industry and biomedical material (13,(17)(18)(19)(20)(21). More interestingly, gelatin can undergo a reversible sol-gel transition which exhibits free-flowing liquid and free-standing gel states triggered by temperature (22).…”