Abstract
Objective: In this study, we aimed to determine the action of the tinctures of Hamamelis virginiana, Maytenus ilicifolia, and Casearia sylvestris on tissues. For this, we investigated the histological sections of fixed skin tissue of Wistar rats for the changes in collagen and elastic fibers, epithelial cells, conjunctive cells, epidermal attachments, pigments, and granules using the optical microscopy technique. Since in the literature and published articles, the use of in vivo models, such as Wistar rats, is predominant to evaluate the healing action of herbal medicines. Methods: The tinctures of Hamamelis virginiana, Maytenus ilicifolia, and Casearia sylvestris, and ethanol 70% (blank) were used. The tinctures were obtained at 10% (w/v) through percolation using 70% ethanol (v/v) as the extraction liquid. This study was conducted in duplicate for each tincture and different incubation times (4, 24, and 48 h) at 37 °C in an oven. The slides used in this study (Wistar rat skin) were previously processed at the histology laboratory since the waste material was donated to conduct this experiment. The Research Ethics Committee approved the use of animals of the Life Sciences Center of the Pontifical Catholic University of Campinas (PUC-CAMPINAS) under the protocol approval number 367/08. Before initiating the staining process, the slides were removed from the oven, and the tincture deposited on the slides was removed. Weigert staining was performed subsequently. For semi-quantitative analysis, the histological sections were carefully observed, and the number of collagen and elastic fibers was evaluated based on the following scale: (+) normal fiber presence, (−) slight decrease, (− −) moderate decrease, and (− − −) intense decrease. The images were digitally captured to obtain the results using a photomicroscope. Results: The degradation of collagen fibers was best evidenced upon using Hamamelis virginiana tincture, which is concordant with the existing reports in the literature on its healing action via the precipitation of dermal proteins. Maytenus ilicifolia and Casearia sylvestris tinctures exhibited low proteolytic capacity as they only caused degradation of elastic fibers that are more delicate and very different from collagen in their constitution. Therefore, the application of the latter two as healing agents (which acts through precipitating proteins) is limited. Conclusions: This experimental histological model using the optical microscopy technique exhibits the advantage of agility and high efficiency as a simple and powerful approach.
Highlights
It is not known exactly who invented the microscope; there are reports suggesting that this invention was contributed by Hans Lippershey and Zacharias Janssen in 15901
In epithelial and conjunctive cells, epidermal attachments, pigments, and granules, the changes were non-existent or imperceptible, and semi-quantitative analyses were performed focusing on these two fibers
It was observed that only Hamamelis virginiana tincture exhibited proteolytic activity on collagen fibers after 24 and 48 h of incubation, as shown in Table 2 and Figures 1, 2, and 3
Summary
It is not known exactly who invented the microscope; there are reports suggesting that this invention was contributed by Hans Lippershey and Zacharias Janssen in 15901. Other facts attest that Galileo invented the microscope in 1609, which was named by Giovanni Faber in 16251 It was van Leeuwenhoek who used it for the first time to observe in vivo samples, and Hobert Hooke improved the equipment and compiled all his analyses in 1665, which is described in the book Micrographia[1]. Light optical microscopy has become an essential aspect of the biological, pharmaceutical, medical, and histological studies, allowing numerous scientific discoveries and advances. In this context, the microscope is essential, contributing to scientific knowledge to better understand the survival mechanism of living organisms through developing the ability to replace the damaged or dead cells, in addition to tissue repair[2]. Regeneration results in the restoration of damaged tissues, and cicatrization leads to the restoration of the original tissue but involves collagen deposition and scar formation[2]
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