Generation of a novel human dermal substitute functionalized with antibiotic-loaded nanostructured lipid carriers (NLCs) with antimicrobial properties for tissue engineering

Abstract

Background Treatment of patients affected by severe burns is challenging, especially due to the high risk of Pseudomonas infection. In the present work, we have generated a novel model of bioartificial human dermis substitute by tissue engineering to treat infected wounds using fibrin-agarose biomaterials functionalized with nanostructured lipid carriers (NLCs) loaded with two anti-Pseudomonas antibiotics: sodium colistimethate (SCM) and amikacin (AMK). Results Results show that the novel tissue-like substitutes have strong antibacterial effect on Pseudomonas cultures, directly proportional to the NLC concentration. Free DNA quantification, WST-1 and Caspase 7 immunohistochemical assays in the functionalized dermis substitute demonstrated that neither cell viability nor cell proliferation were affected by functionalization in most study groups. Furthermore, immunohistochemistry for PCNA and KI67 and histochemistry for collagen and proteoglycans revealed that cells proliferated and were metabolically active in the functionalized tissue with no differences with controls. When functionalized tissues were biomechanically characterized, we found that NLCs were able to improve some of the major biomechanical properties of these artificial tissues, although this strongly depended on the type and concentration of NLCs. Conclusions These results suggest that functionalization of fibrin-agarose human dermal substitutes with antibiotic-loaded NLCs is able to improve the antibacterial and biomechanical properties of these substitutes with no detectable side effects. This opens the door to future clinical use of functionalized tissues.

Antimicrobial Peptide Dendrimers and Quorum-Sensing Inhibitors in Formulating Next-Generation Anti-Infection Cell Therapy Dressings for Burns

“…The development of antimicrobial scaffolds for burn wound regeneration is an active research domain, including dermal substitutes incorporating nano-carriers loaded with antibiotics for skin engineering [25] as an example. Replacement of classical antibiotics by non-antibiotic antimicrobial agents in such formulations is currently considered as a promising strategy to prevent infections and, at the same time minimizing the probability of inducing antibiotic resistance in burn patients.…”

Usefulness of a Nanostructured Fibrin-Agarose Bone Substitute in a Model of Severely Critical Mandible Bone Defect

“…On the one hand, novel biofabrication methods should be applied to the HTEB in order to generate a product with higher analogy to the native mandible bone. In this regard, previous reports from our research group demonstrate that fibrin-agarose biomaterials can be improved and functionalized by chemical cross-linking [42], or by combination with magnetic nanoparticles [43] and bioactive nanostructured lipid carriers [44]. Future works should determine whether these methods can enhance the biological and biomechanical properties of the HTEB used in the present work.…”

Development of secretome-based strategies to improve cell culture protocols in tissue engineering

“…In brief, fixed tissues were embedded in paraffin and histological sections were obtained. Sections were deparaffinized and stained with hematoxylin–eosin and Masson trichrome staining (for global morphology), picrosirius red (for collagen fibers) and alcian blue (for proteoglycans) using routine histological and histochemical methods previously described 8 , 9 , 12 , 20 .…”

“…Despite important advances in the microsurgical field in recent decades, treatment of these diseases may be challenging, and the results remain suboptimal 6 , 7 . In this context, tissue engineering offers a promising strategy that combines cells, extracellular matrix, and signaling molecules to improve regeneration and clinical outcomes 8 , 9 .…”