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

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.

[Incremental innovation in healthcare in Spain during the COVID-19 pandemic]. / La innovación incremental en salud en España durante la pandemia de la COVID-19.

OBJECTIVE: It is important to understand the impact of the COVID-19 pandemic on incremental innovation and its protection through industrial property rights, in order to acquiring valuable insights to develop effective public policies and corporate strategies. The objective was to analyze incremental innovations in response to the pandemic that have been protected by industrial property rights, and to examine whether the COVID-19 pandemic had a positive or negative effect on incremental innovation, promoting or inhibiting it. METHODS: Utility models in the health patent class have been used as indicators (01.01.20 to 31.12.21), since the information they provide and their characteristics (requirements of applications and publication) allowed us to obtain preliminary conclusions in the short term. Their frequency of application during the pandemic months was analyzed and compared with an equivalent period immediately before (01.01.18 to 31.12.19). RESULTS: The analysis showed that there had been greater activity in healthcare innovation by all agents (individuals, companies and the public sector). In the pandemic period of 2020-2021, 754 utility models were requested, representing a nearly 40% increase compared to the equivalent period of 2018-2019, of which 284 were identified as pandemic-related innovations, with 59.7% of rights holders being individuals, 36.4% being companies, and only 3.9% being public entities. CONCLUSIONS: In general, incremental innovations require less investment and shorter technology maturation times, which had made it possible to respond, in some cases successfully, to situations of initial shortages of many medical devices, such as ventilators and protective equipment.

OBJETIVO: Es importante comprender cómo la pandemia de COVID-19 ha afectado a la innovación incremental y su protección mediante derechos de propiedad industrial, con el fin de obtener información valiosa para desarrollar políticas públicas y estrategias empresariales. El objetivo de este estudio fue analizar las innovaciones incrementales como respuesta a la pandemia que han sido protegidas por derechos de propiedad industrial, y examinar si la pandemia de la COVID-19 había tenido un efecto positivo o negativo en la innovación incremental, fomentándola o inhibiéndola. METODOS: Se emplearon como indicadores los modelos de utilidad de la clase de patentes del ámbito salud en el periodo entre el 1 de enero de 2020 y el 31 de diciembre de 2021, puesto que la información que proporcionan y sus características (requisitos de solicitud y publicación) permitían extraer algunas conclusiones incipientes en el corto plazo. Se analizó su frecuencia de solicitud durante los meses de pandemia y se comparó con un periodo temporal equivalente inmediatamente anterior (1 de enero de 2018 a 31 de diciembre de 2019). RESULTADOS: El análisis mostró que hubo una mayor actividad de innovación sanitaria por parte de todos los agentes (particulares, empresas y sector público). En el período de la pandemia de 2020-2021, se solicitaron 754 modelos de utilidad, lo que representó un aumento de casi el 40% en comparación con el período equivalente de 2018-2019, de los cuales se identificaron 284 como innovaciones relacionadas con la pandemia, con el 59,7% de los titulares de los derechos siendo personas individuales, el 36,4% siendo empresas y solo el 3,9% siendo entidades públicas. CONCLUSIONES: En general, las innovaciones incrementales requieren menor inversión y plazos de maduración de la tecnología más cortos, lo que permitía responder, en algunos casos exitosamente, a las situaciones de desabastecimiento inicial de muchos productos sanitarios, como respiradores y equipos de protección.

A novel 3D biofabrication strategy to improve cell proliferation and differentiation of human Wharton's jelly mesenchymal stromal cells for cell therapy and tissue engineering.

Purpose: Obtaining sufficient numbers of cells in a short time is a major goal of cell culturing in cell therapy and tissue engineering. However, current bidimensional (2D) culture methods are associated to several limitations, including low efficiency and the loss of key cell differentiation markers on cultured cells. Methods: In the present work, we have designed a novel biofabrication method based on a three-dimensional (3D) culture system (FIBRIAGAR-3D). Human Wharton's jelly mesenchymal stromal cells (HWJSC) were cultured in 3D using 100%, 75%, 50%, and 25% concentrations of fibrin-agarose biomaterials (FA100, FA75, FA50 and FA25 group) and compared with control cells cultured using classical 2D systems (CTR-2D). Results: Our results showed a significant increase in the number of cells generated after 7 days of culture, with cells displaying numerous expansions towards the biomaterial, and a significant overexpression of the cell proliferation marker KI67 was found for the FA75 and FA100 groups. TUNEL and qRT-PCR analyses demonstrated that the use of FIBRIAGAR-3D was not associated with an induction of apoptosis by cultured cells. Instead, the 3D system retained the expression of typical phenotypic markers of HWJSC, including CD73, CD90, CD105, NANOG and OCT4, and biosynthesis markers such as types-I and IV collagens, with significant increase of some of these markers, especially in the FA100 group. Finally, our analysis of 8 cell signaling molecules revealed a significant decrease of GM-CSF, IFN-g, IL2, IL4, IL6, IL8, and TNFα, suggesting that the 3D culture system did not induce the expression of pro-inflammatory molecules. Conclusion: These results confirm the usefulness of FIBRIAGAR-3D culture systems to increase cell proliferation without altering cell phenotype of immunogenicity and opens the door to the possibility of using this novel biofabrication method in cell therapy and tissue engineering of the human cornea, oral mucosa, skin, urethra, among other structures.

Generation and Evaluation of Novel Biomaterials Based on Decellularized Sturgeon Cartilage for Use in Tissue Engineering.

Because cartilage has limited regenerative capability, a fully efficient advanced therapy medicinal product is needed to treat severe cartilage damage. We evaluated a novel biomaterial obtained by decellularizing sturgeon chondral endoskeleton tissue for use in cartilage tissue engineering. In silico analysis suggested high homology between human and sturgeon collagen proteins, and ultra-performance liquid chromatography confirmed that both types of cartilage consisted mainly of the same amino acids. Decellularized sturgeon cartilage was recellularized with human chondrocytes and four types of human mesenchymal stem cells (MSC) and their suitability for generating a cartilage substitute was assessed ex vivo and in vivo. The results supported the biocompatibility of the novel scaffold, as well as its ability to sustain cell adhesion, proliferation and differentiation. In vivo assays showed that the MSC cells in grafted cartilage disks were biosynthetically active and able to remodel the extracellular matrix of cartilage substitutes, with the production of type II collagen and other relevant components, especially when adipose tissue MSC were used. In addition, these cartilage substitutes triggered a pro-regenerative reaction mediated by CD206-positive M2 macrophages. These preliminary results warrant further research to characterize in greater detail the potential clinical translation of these novel cartilage substitutes.

Successful development and clinical translation of a novel anterior lamellar artificial cornea.

Blindness due to corneal diseases is a common pathology affecting up to 23 million individuals worldwide. The tissue-engineered anterior human cornea, which is currently being tested in a Phase I/II clinical trial to treat severe corneal trophic ulcers with preliminary good feasibility and safety results. This bioartificial cornea is based on a nanostructured fibrin-agarose biomaterial containing human allogeneic stromal keratocytes and cornea epithelial cells, mimicking the human native anterior cornea in terms of optical, mechanical, and biological behavior. This product is manufactured as a clinical-grade tissue engineering product, fulfilling European requirements and regulations. The clinical translation process included several phases: an initial in vitro and in vivo preclinical research plan, including preclinical advice from the Spanish Medicines Agency followed by additional preclinical development, the adaptation of the biofabrication protocols to a good manufacturing practice manufacturing process, including all quality controls required, and the design of an advanced therapy clinical trial. The experimental development and successful translation of advanced therapy medicinal products for clinical application has to overcome many obstacles, especially when undertaken by academia or SMEs. We expect that our experience and research strategy may help future researchers to efficiently transfer their preclinical results into the clinical settings.