Tag Archives: fibroblasts

Perspectives on the Combined Effects of Ocimum basilicum and Trifolium pratense Extracts in Terms of Phytochemical Profile and Pharmacological Effects

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By A. I. Antonescu, Simona Cavalu et al

Common and distinct therapeutic effects of Ocimum and Trifolium species.
Copyright I.A. Antonescu, Simona Cavalu et al.
Different species of Trifolium according to the amount of cyanogenic glycosides and isoflavones and identifying the species with rich chemical composition.
Copyright I.A. Antonescu, Simona Cavalu et al.
Pharmacological activities of extracts from Trifolium and Ocimum species after internal and external administration. Copyright I. A. Antonescu, Simona Cavalu et al.
Stages of the wound healing process (adapted from [121]).
Copyright I.A. Antonescu, Simona Cavalu et al.

The ability to promote wound healing by synergic effect of Trifolium Pratense and Ocimum basilicum mixt extract has not been studied yet, being the central point for future studies. So far, the anti-inflammatory, antimicrobial, antifungal and anticancer properties have been demonstrated for each extract individually, obtaining promising results, and for these reasons, in the future, the mixture of both extracts are of great interest to be studied, expecting for a potential synergistic effect.

To the best of our knowledge, there are no studies in the literature dealing with any in vitro tests of the mixture Trifolium pretense and Ocimum basilicum extracts. Our research group performed a preliminary study using the “scratch test” assay on human fibroblasts, by applying the extract mixture in different concentrations on fibroblasts culture, in order to evaluate the optimum concentration to promote the stimulation and proliferation of the cells. Within this test, which is an in vitro model of wound healing, human fibroblasts were primarly grown to a confluent monolayer, and then was scraped in a straight line with a pipette tip, in order to simulate a wound. The fibroblasts migration into the wound area was monitored during 48 h incubation in the presence of different concentrations of mixed plant extracts along with the control (no treatment). Copyright I.A. Antonescu, Simona Cavalu et al.

The spontaneous migration of dermal fibroblasts is evidenced under light microscopy, along with the control samples, showing the progressive covering of the pseudo-wound monitored at different times intervals. Copyright I.A. Antonescu, Simona Cavalu et al.
Wound healing percent express as fibroblast migration to cover the scratched area. Copyright I.A. Antonescu, Simona Cavalu et al.

The antioxidant, antimicrobial, antiviral, antifungal and anti-inflammatory activity of Ocimum and Trifolium species are summarized in this review in order to explore the therapeutic potential of Ocimum basilicum and Trifolium pretense in relation with their phytochemical profile and to highlight the pharmacological activity of aqueous or ethanol extracts. Special attention was devoted to the dermal pathology and wound healing effects, in the context of multiple skin conditions such as acne, eczema boils, psoriasis and rashes. Both extracts (Trifolium sp. and Ocimum sp.) are characterized by high content of antioxidant compounds, which are also responsible for the radiance and resistance of the skin and the slowing down of the aging process by maintaining estrogen levels. Moreover, the potential combined effect of the mixed extract is pointed out in terms of future applications for wound healing, based on some preliminary results obtained from a “scratch tests” assay performed with respect to human dermal fibroblasts. Copyright I.A. Antonescu, Simona Cavalu et al.

Full text at https://www.mdpi.com/2223-7747/10/7/1390/htm

The influence of propolis nanoparticles on dermal fibroblasts migration: premises for development of propolis-based collagen dermal patches

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By P. M. Pasca and Simona Cavalu

https://chalcogen.ro/929_PascaPM.pdf

Knowing the biological and pharmacological properties of propolis, the first goal of our study was to prepare and characterize a propolis nano-formulation (NPs) in order to be used for wound healing applications. The ability of propolis NPs to stimulate the migration of dermal fibroblasts in vitro was assessed by scratch test assay. The concentration of 200 μg/mL propolis NPs was found to have similar effect as the positive control. The second goal was to prepare a propolis-collagen membrane and to investigate the morphological and nanoindentation properties by AFM. The ultrastructure network of collagen fibrils was not affected by incorporation of propolis NPs, showing a nano-porous structure, favorable for soft tissue regeneration applications. Enzymatic degradation assay indicated a reduced degradation rate upon incorporation of propolis NPs in collagen matrix.

Ionotropic gelation method was applied for the preparation of propolis NPS. The nanoparticles were formed spontaneously due to ionic interaction between the protonated amine groups in chitosan and the negatively charged counter-ion TPP, being stabilized by Tween 80.

TEM image of propolis NPs; b) size distribution and c) EDX corresponding spectrum.
Copyright Simona Cavalu
Spontaneous evolution of human fibroblasts in cell culture medium, monitored at different time intervals (6, 12, 24, 48, 96 and 110 hours) until the confluence was achieved (Phase contrast image, scale bar 50 μm). Copyright Simona Cavalu
Fibroblasts migration monitored after different time intervals and wound closure under the treatment with propolis NPs at two different concentrations, compared to the positive and negative control. The initial area of the scratch (t=o) is represented by the red rectangle (Phase contrast image, scale bar 100 μm). Copyright Simona Cavalu.
The percent of restored fibroblasts monolayer upon migration of the cells into the free area, monitored during 48 h (Statistical relevance p<0.05). Copyright Simona Cavalu.
AFM images of neat collagen membrane (a,b) and collagen membrane with propolis NPs incorporated (c, d), in 3D and 2D configurations.

The tridimensional network of collagen fibrils is visible in both specimens (with or without propolis NPs incorporated) emphasizing the details of repetitive structure of the D-bands pattern of a single collagen fibril, with periodic gaps and grooves, in concordance with some previous published work [32, 33]. The periodicity of D-bands is less visible after propolis NPs incorporation. Moreover, after propolis NPs incorporation and freeze drying procedure, an obvious porous ultrastructure formation was noticed, as a result of fibers self-assembly.

Collagenase degradation test of neat collagen membrane and collagen-propolis NPs membrane (statistical relevance p<0.05). Copyright Simona Cavalu.

A collagen-based membrane was prepared and investigated by AFM in terms of morphological features and nanoindentation. The network of collagen fibrils was not affected by propolis NPs, showing a nano-porous structure, favorable for soft tissue regeneration applications. Enzymatic degradation assay indicated a reduced degradation rate upon incorporation of propolis NPs in collagen matrix. Corroborating the above mentioned results, we consider that modified-collagen membrane by adding propolis NPs in a controlled concentration, might represent a promising natural alternative to synthetic bandages for wound healing applications. Of course, further in vitro and in vivo tests are necessary to evaluate the biological performances of collagen-modified membranes, in terms of tissue adaptation and integration. (Simona Cavalu, PM Pasca, Digest Journal of Nanomaterials and Biostructures, Volume 16, Issue 3, Pages 929 – 938July-September 2021).