Segurança clínica da aplicação intra-ovariana bovina de células-tronco mesenquimais alogênicas

A basic premise of reproductive physiology is that females have a finite and non-renewable stock of germinative cells, which results in a decrease in reproductive capacity over time. For female bovines, a further factor associated with this decrease is follicular puncture (ovum pickup - OPU), a technique widely used for production of embryos in vitro. As such, it is necessary to seek therapeutic or preventive options for fertility problems, and one potential option is treatment with mesenchymal stem cells (MSC), which exercise a paracrine effect in combating inflammatory and degenerative processes. However, as important as evaluating the efficacy of such treatments is an evaluation of safety. In this context, the current study was carried out with the application of 2.5 x 106 allogenic MSC derived from adipose tissue, to the ovarian cortex of healthy nelore (n = 5) and girolando (n = 5) cows. The animals were subsequently evaluated by ultrasonography, clinical examination, number of viable oocytes collected, and rate of embryo production. None of the animals presented any clinical alteration or any alteration on ultrasonography after receiving the MSC. Furthermore, comparison between the number of viable oocytes, embryos produced, and rate of embryo production before and after MSC application did not show a difference. Based on these data, it can be concluded that intraovarian application of 2.5 x 106 adipose-derived MSC is safe, and this technique represents a potential for study as a therapy in cases of ovarian degeneration or lesions.Una premisa básica de la fisiología reproductiva es que las hembras tienen un stock finito y no renovable de células germinativas, lo que resulta en una disminución de la capacidad reproductiva con el tiempo. Para las hembras bovinas, otro factor asociado a esta disminución es la punción folicular (ovum pickup - OPU), técnica ampliamente utilizada para la producción de embriones in vitro. Por tanto, es necesario buscar opciones terapéuticas o preventivas para los problemas de fertilidad, y una opción potencial es el tratamiento con células madre mesenquimales (MSC), que ejercen un efecto paracrino en la lucha contra los procesos inflamatorios y degenerativos. Sin embargo, tan importante como evaluar la eficacia de tales tratamientos es una evaluación de la seguridad. En este contexto, el presente estudio se llevó a cabo con la aplicación de 2,5 x 106 MSC alogénicas derivadas de tejido adiposo, a la corteza ovárica de vacas nelore (n = 5) y girolando (n = 5) sanas. Posteriormente, los animales se evaluaron mediante ultrasonografía, examen clínico, número de ovocitos viables recogidos y tasa de producción de embriones. Ninguno de los animales presentó alteración clínica ni alteración ecográfica tras recibir la MSC. Además, la comparación entre el número de ovocitos viables, los embriones producidos y la tasa de producción de embriones antes y después de la aplicación de MSC no mostró ninguna diferencia. En base a estos datos, se puede concluir que la aplicación intraovárica de 2,5 x 106 MSC derivadas de tejido adiposo es segura, y esta técnica representa un potencial de estudio como terapia en casos de degeneración o lesiones ováricas.Uma premissa básica da fisiologia reprodutiva é que as fêmeas possuem um estoque finito e não renovável de células germinativas, o que resulta em uma diminuição da capacidade reprodutiva ao longo do tempo. Para as fêmeas bovinas, outro fator associado a essa diminuição é a punção folicular (ovo pickup - OPU), técnica amplamente utilizada para produção de embriões in vitro. Assim, é necessário buscar opções terapêuticas ou preventivas para problemas de fertilidade, sendo uma opção potencial o tratamento com células-tronco mesenquimais (CTM), que exercem efeito parácrino no combate a processos inflamatórios e degenerativos. No entanto, tão importante quanto avaliar a eficácia de tais tratamentos é a avaliação da segurança. Nesse contexto, o presente estudo foi realizado com a aplicação de 2,5 x 106 MSC alogênicas derivadas do tecido adiposo, no córtex ovariano de vacas nelore (n = 5) e girolando (n = 5) hígidas. Os animais foram posteriormente avaliados por ultrassonografia, exame clínico, número de oócitos viáveis coletados e taxa de produção de embriões. Nenhum dos animais apresentou qualquer alteração clínica ou ultrassonográfica após receber o MSC. Além disso, a comparação entre o número de oócitos viáveis, embriões produzidos e taxa de produção de embriões antes e depois da aplicação de MSC não mostrou diferença. Com base nesses dados, pode-se concluir que a aplicação intraovariana de MSC de origem adiposa 2,5 x 106 é segura, e essa técnica representa um potencial para estudo como terapia em casos de degeneração ou lesões ovarianas

Pouteria macrophylla fruit extract microemulsion for cutaneous depigmentation : evaluation using a 3D pigmented skin model

Here, we verify the depigmenting action of Pouteria macrophylla fruit extract (EXT), incorporate it into a safe topical microemulsion and assess its effectiveness in a 3D pigmented skin model. Melanocytes-B16F10- were used to assess the EXT effects on cell viability, melanin synthesis, and melanin synthesis-related gene transcription factor expression, which demonstrated a 32% and 50% reduction of intra and extracellular melanin content, respectively. The developed microemulsion was composed of Cremophor EL®/Span 80 4:1 (w/w), ethyl oleate, and pH 4.5 HEPES buffer and had an average droplet size of 40 nm (PdI 0.40 ± 0.07). Skin irritation test with reconstituted epidermis (Skin Ethic RHETM) showed that the formulation is non-irritating. Tyrosinase inhibition was maintained after skin permeation in vitro, in which microemulsion showed twice the inhibition of the conventional emulsion (20.7 ± 2.2% and 10.7 ± 2.4%, respectively). The depigmenting effect of the microemulsion was finally confirmed in a 3D culture model of pigmented skin, in which histological analysis showed a more pronounced effect than a commercial depigmenting formulation. In conclusion, the developed microemulsion is a promising safe formulation for the administration of cutite fruit extract, which showed remarkable depigmenting potential compared to a commercial formulation

Skin regenerative potential of Cupuaçu seed extract (Theobroma grandiflorum), a native fruit from the Amazon : development of a topical formulation based on chitosan-coated nanocapsules

Scarless skin regeneration is a challenge in regenerative medicine. Herein, we explore the regenerative potential of a Cupuaçu seed extract (Theobroma grandiflorum) to develop an innovative skin regeneration formulation based on chitosan-coated nanocapsules. Cupuaçu seed extract significantly stimulated cell proliferation and migration. A reparative gene expression profile could be verified following extract treatment, which included high levels of MKI67, a cellular proliferation marker, and extracellular matrix genes, such as ELN and HAS2, which code for elastin and hyaluronic acid synthase 2. Formulations with Cupuaçu seed extract successfully entrapped into nanocapsules (EE% > 94%) were developed. Uncoated or coated nanocapsules with low-molecular-weight chitosan presented unimodal size distribution with hydrodynamic diameters of 278.3 ± 5.0 nm (PDI = 0.18 ± 0.02) and 337.2 ± 2.1 nm (PDI = 0.27 ± 0.01), respectively. Both nanosystems were physically stable for at least 120 days and showed to be non-irritating to reconstructed human epidermis. Chitosan coating promoted active penetration into undamaged skin areas, which were still covered by the stratum corneum. In conclusion, the present study demonstrated for the first time the biotechnological potential of the frequently discarded Cupuaçu seed as a valuable pharmaceutical ingredient to be used in regenerative skin products