Mimicking bone microenvironment: 2D and 3D in vitro models of human osteoblasts.

Understanding the in vitro biology and behavior of human osteoblasts is crucial for developing research models that reproduce closely the bone structure, its functions, and the cell-cell and cell-matrix interactions that occurs in vivo. Mimicking bone microenvironment is challenging, but necessary, to ensure the clinical translation of novel medicines to treat more reliable different bone pathologies. Currently, bone tissue engineering is moving from 2D cell culture models such as traditional culture, sandwich culture, micro-patterning, and altered substrate stiffness, towards more complex 3D models including spheroids, scaffolds, cell sheets, hydrogels, bioreactors, and microfluidics chips. There are many different factors, such cell line type, cell culture media, substrate roughness and stiffness that need consideration when developing in vitro models as they affect significantly the microenvironment and hence, the final outcome of the in vitro assay. Advanced technologies, such as 3D bioprinting and microfluidics, have allowed the development of more complex structures, bridging the gap between in vitro and in vivo models. In this review, past and current 2D and 3D in vitro models for human osteoblasts will be described in detail, highlighting the culture conditions and outcomes achieved, as well as the challenges and limitations of each model, offering a widen perspective on how these models can closely mimic the bone microenvironment and for which applications have shown more successful results.

Targeting lung macrophages for fungal and parasitic pulmonary infections with innovative amphotericin B dry powder inhalers.

The incidence of fungal pulmonary infections is known to be on the increase, and yet there is an alarming gap in terms of marketed antifungal therapies that are available for pulmonary administration. Amphotericin B (AmB) is a highly efficient broad-spectrum antifungal only marketed as an intravenous formulation. Based on the lack of effective antifungal and antiparasitic pulmonary treatments, the aim of this study was to develop a carbohydrate-based AmB dry powder inhaler (DPI) formulation, prepared by spray drying. Amorphous AmB microparticles were developed by combining 39.7 % AmB with 39.7 % γ-cyclodextrin, 8.1 % mannose and 12.5 % leucine. An increase in the mannose concentration from 8.1 to 29.8 %, led to partial drug crystallisation. Both formulations showed good in vitro lung deposition characteristics (80 % FPF < 5 µm and MMAD < 3 µm) at different air flow rates (60 and 30 L/min) when used with a DPI, but also during nebulisation upon reconstitution in water.

A novel formulation of solubilised amphotericin B designed for ophthalmic use.

Amphotericin B (AmB) is a wide spectrum antifungal with low incidence of clinical resistance. However, there are no licensed topical formulations with AmB in most developed countries. Extemporaneous preparations of AmB are frequently prepared from available marketed parenteral formulations. Herein, a solution of AmB with γ-cyclodextrin is described as suitable for topical administration as eye drops. This novel formulation is characterised by its ability to solubilise AmB and to maintain its antifungal activity, physicochemical stability and sterility over 30 days. Antifungal activity against Candida albicans was significantly higher (35%) for the new formulation than that obtained with Fungizone(®) based extemporaneous prepared suspension. Optimal 0.1% AmB-10% cyclodextrin formulation remained sterile and with an acceptable osmolarity, pH and particle size for ophthalmic use over 4 weeks. Complexation with γ-cyclodextrins improved AmB chemical stability compared to the reference eye drops suspension based on Fungizone(®). These results illustrate the feasibility of an ophthalmic AmB formulation easy enough to be licensed or prepared in community and hospital pharmacies.

Avaliação da atividade tóxica em Artemia salina e Biomphalaria glabrata de extratos de quatro espécies do gênero Eleocharis (Cyperaceae) / Toxic evaluation of four species of the genus Eleocharis (Cyperaceae) in Artemia salina and Biomphalaria glabrata

O gênero Eleocharis R. Br. compreende cerca de 200 espécies, ocorrendo em ambientes úmidos tais como brejos e margens de rios e lagos. Procurando novos agentes moluscicidas, os extratos de Eleocharis acutangula (Roxb.) Schult., Eleocharis interstincta (Vahl) Roem. & Schult., Eleocharis maculosa (Vahl) Roem. & Schult. e Eleocharis sellowiana Kunth foram testados para atividade moluscicida, contra caramujos adultos e desovas, e toxicidade (ensaio de letalidade com Artemia salina). O extrato hexânico de Eleocharis acutangula (parte subterrânea fresca) foi ativo contra Artemia salina (CL50 = 476,00 mg/mL), enquanto os demais extratos apresentaram CL50 >> 10³ mg/mL, sugerindo baixa toxicidade. O extrato hidro-etanólico de Eleocharis sellowiana (parte subterrânea fresca) foi ativo contra desovas de Biomphalaria glabrata (CL50 = 24,27 mg/mL) mas inativo contra indivíduos adultos. Os demais extratos testados não apresentaram atividade moluscicida.

The genus Eleocharis R. Br. comprises about 200 species, occurring in wet environments like swamps, lakes and river margins. In the search for new molluscicides, extracts from Eleocharis acutangula (Roxb.) Schult., Eleocharis interstincta (Vahl) Roem. & Schult., Eleocharis maculosa (Vahl) Roem. & Schult. and Eleocharis sellowiana Kunth were tested for molluscicidal activity (spawns and adult snails) and toxicity (Brine Shrimp Lethality - BSL - bioassay). The hexane extract of Eleocharis acutangula (fresh subterraneous parts) was active in the BSL bioassay (LC50 = 476 mg/mL), while the other extracts showed LC50 >> 10³ mg/mL, suggesting they have low toxicity. The aqueous ethanol extract of Eleocharis sellowiana (fresh subterraneous parts) was active against Biomphalaria glabrata spawns (LC50 = 24.27 mg/mL) but it was not lethal to adult snails. No other plant extract tested in this study showed molluscicidal activity.