Analysis of the Physico-Chemical, Mechanical and Biological Properties of Crosslinked Type-I Collagen from Horse Tendon: Towards the Development of Ideal Scaffolding Material for Urethral Regeneration

Abstract

Urethral stenosis is a pathological condition that consists in the narrowing of the urethral lumen because of the formation of scar tissue. Unfortunately, none of the current surgical approaches represent an optimal solution because of the high stricture recurrence rate. In this context, we preliminarily explored the potential of an insoluble type-I collagen from horse tendon as scaffolding material for the development of innovative devices for the regeneration of injured urethral tracts. Non-porous collagen-based substrates were produced and optimized, in terms of crosslinking density of the macromolecular structure, to either provide mechanical properties compliant with the urinary tract physiological stress and better sustain tissue regeneration. The effect of the adopted crosslinking strategy on the protein integrity and on the substrate physical–chemical, mechanical and biological properties was investigated in comparison with a decellularized matrix from porcine small intestinal submucosa (SIS patch), an extensively used xenograft licensed for clinical use in urology. The optimized production protocols allowed the preservation of the type I collagen native structure and the realization of a substrate with appealing end-use properties. The biological response, preliminarily investigated by immunofluorescence experiments on human adult renal stem/progenitor cells until 28 days, showed the formation of a stem-cell monolayer within 14 days and the onset of spheroids within 28 days. These results suggested the great potential of the collagen-based material for the development of scaffolds for urethral plate regeneration and for in vitro cellular studies.

Development of L-Lysine-Loaded PLGA Microparticles as a Controlled Release System for Angiogenesis Enhancement

“…The aqueous extract was collected as the supernatant after centrifugation at 4000 rpm for 5 min at room temperature. The LYS quantity present in the extract was spectrophotometrically determined by means of a slightly modified TNBS test [ 42 , 43 ]. About 0.2 mL of extract was added to 0.3 mL of 4% ( w / v ) NaHCO 3 solution and 0.5 mL of 0.05% ( w / v ) TNBS.…”

“…About 0.2 mL of extract was added to 0.3 mL of 4% ( w / v ) NaHCO 3 solution and 0.5 mL of 0.05% ( w / v ) TNBS. Then samples were hermetically sealed and heated at 40 °C for 2 h [ 43 ]. After that, 1.5 mL of 6 M HCl was added before heating the samples at 60 °C for 90 min [ 42 , 43 ].…”

“…Then samples were hermetically sealed and heated at 40 °C for 2 h [ 43 ]. After that, 1.5 mL of 6 M HCl was added before heating the samples at 60 °C for 90 min [ 42 , 43 ]. The reaction mixture was thus was brought to a final volume of 5 mL with distilled water and cooled down to room temperature.…”

An Update on the Clinical Efficacy and Safety of Collagen Injectables for Aesthetic and Regenerative Medicine Applications

“…The possibility to define specific scaffolds properties (i.e., by tuning protein concentration, solvent type and concentration, protein molecular weight, superficial morphology, 3D organization, and by pre- and post-production processing) offers a great opportunity to modulate the structure-related biological activity of the scaffolds in order to optimize their capability to induce and sustain tissue regeneration [ 20 , 35 , 43 , 49 , 50 , 51 , 52 , 53 , 54 , 55 ]. Moreover, the use of collagen is advantageous for regenerative medicine and tissue engineering applications because, being recognized as a self-molecule, it is metabolized by the natural body enzymatic apparatus that gradually breaks down collagen molecules and substitutes it with newly synthetized one.…”

Investigations on the Influence of Collagen Type on Physicochemical Properties of PVP/PVA Composites Enriched with Hydroxyapatite Developed for Biomedical Applications

“…Furthermore, the presence of collagen of fish or bovine origin increases the innovativeness of the developed composite materials. Collagen is a main component of extracellular matrix which makes it an interesting biopolymer for applications in tissue engineering and regeneration medicine [ 31 , 32 ].…”