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- New prospects in skin regeneration and repair using nanophased hydroxyapatite embedded in collagen nanofibersPublication . Ribeiro, Nilza; Sousa, Aureliana; Cunha-Reis, Cassilda; Oliveira, Ana Leite; Granja, Pedro L.; Monteiro, Fernando J.; Sousa, SusanaThis study reflects an exploitation of a composite matrix produced by electrospinning of collagen and electrospraying of nanophased hydroxyapatite (nanoHA), for skin regeneration applications. The main goal was to evaluate the effect of nanoHA, as source of localized calcium delivery, on human dermal fibroblasts, keratinocytes, and human mesenchymal stem cells (hMSCs) growth, proliferation, differentiation, and extracellular matrix production. This study revealed that calcium ions provided by nanoHA significantly enhanced cellular growth and proliferation rates and prevented adhesion of pathogenic bacteria strains typically found in human skin flora. Moreover, hMSCs were able to differentiate in both osteogenic and adipogenic lineages. Rat subcutaneous implantation of the membranes also revealed that no adverse reaction occurred. Therefore, the mechanically fit composite membrane presents a great potential to be used either as cell transplantation scaffold for skin wound regeneration or as wound dressing material in plastic surgery, burns treatment or skin diseases.
- Antimicrobial properties of gallium (III)-and iron (III)-loaded polysaccharides affecting the growth of escherichia coli, staphylococcus aureus, and pseudomonas aeruginosa, in vitroPublication . Best, Mark G.; Cunha-Reis, Cassilda; Ganin, Alexey Y.; Sousa, Aureliana; Johnston, Jenna; Oliveira, Ana L.; Smith, David G. E.; Yiu, Humphrey H. P.; Cooper, Ian R.Antimicrobial resistance (AMR) has become a global concern as many bacterial species have developed resistance to commonly prescribed antibiotics, making them ineffective to treatments. One type of antibiotics, gallium(III) compounds, stands out as possible candidates due to their unique “Trojan horse” mechanism to tackle bacterial growth, by substituting iron(III) in the metabolic cycles of bacteria. In this study, we tested three polysaccharides (carboxymethyl cellulose (CMC), alginate, and pectin) as the binding and delivery agent for gallium on three bacteria (Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus) with a potential bioresponsive delivery mode. Two types of analysis on bacterial growth (minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC)) were carried out while iron(III)-loaded polysaccharide samples were also tested for comparison. The results suggested that gallium showed an improved inhibitory activity on bacterial growth, in particular gallium(III)-loaded carboxymethyl cellulose (Ga-CMC) sample showing an inhibiting effect on growth for all three tested bacteria. At the MIC for all three bacteria, Ga-CMC showed no cytotoxicity effect on human dermal neonatal fibroblasts (HDNF). Therefore, these bioresponsive gallium(III) polysaccharide compounds show significant potential to be developed as the next-generation antibacterial agents with controlled release capability.
- In situ crosslinked electrospun gelatin nanofibers for skin regenerationPublication . Dias, J. R.; Baptista-Silva, S.; Oliveira, C. M. T. de; Sousa, A.; Oliveira, Ana L.; Bártolo, P. J.; Granja, P. L.Due to its intrinsic similarity to the extracellular matrix, gelatin electrospun nanofibrous meshes are promising scaffold structures for wound dressings and tissue engineering applications. However, gelatin is water soluble and presents poor mechanical properties, which generally constitute relevant limitations to its applicability. In this work, gelatin was in situ crosslinked with 1,4-butanediol diglycidyl ether (BDDGE) at different concentrations (2, 4 and 6 wt%) and incubation time-points (24, 48 and 72 h) at 37 °C. The physico-chemical and biological properties of BDDGE-crosslinked electrospun gelatin meshes were investigated. Results show that by changing the BDDGE concentration it is possible to produce nanofibers crosslinked in situ with well-defined morphology and modulate fiber size and mechanical properties. Crosslinked gelatin meshes show no toxicity towards fibroblasts, stimulating their adhesion, proliferation and synthesis of new extracellular matrix, thereby indicating the potential of this strategy for skin tissue engineering.
- Biomechanical performance of hybrid electrospun structures for skin regenerationPublication . Dias, J. R.; Baptista-Silva, S.; Sousa, A.; Oliveira, A. L.; Bártolo, P. J.; Granja, P. L.Wound dressings made by electrospun nanofibers have been demonstrating great potential to regenerate skin tissue as compared to the conventional membrane products available in the market. Until today most of the developed dressings have only demonstrated the capability to regenerate the dermis or epidermis. In this study we propose new hybrid electrospun meshes combining polycaprolactone and gelatin. Several approaches, multilayer, coating and blend were stablished to investigate the most appropriate hybrid structure with potential to promote skin regeneration in its full thickness. The structures were evaluated in terms of physico-chemical properties (porosity, water vapor permeability, contact angle and swelling degree) and according to its mechanical and biological performance. Multilayer and blend structures demonstrated to fit most of native skin requirements. However, looking to all the performed characterization we considered multilayer as the most promising hybrid structures, due its high porosity which contributed to an ideal water vapor permeability rate and good mechanical and biological properties. Based on this multilayer structure is a promisor wound dressing.