Bone grafts that mimic the properties of the extracellular matrix (ECM) are well-
known for their ability to facilitate bone regeneration. Biomaterials can be designed

with a porous structure that mimics the natural architecture of bone tissue,
promoting cellular migration and differentiation. The development of biomaterials
often involves combining polymers and incorporating nanoparticles to create
composites with superior properties compared to individual components. In the
pursuit of enhanced composites, this study focused on developing collagen
scaffolds (2 mg) with varying amounts of alginate (1 mg, 2 mg, and 4 mg),
combined with three concentrations of TiO2 nanoparticles (2 μg, 4 μg, and 6 μg).
The scaffolds were prepared using type I collagen and alginate cross-linked with
CaCl2 through the casting and lyophilization technique, resulting in cylindrical
structures approximately 10 mm in length. FTIR analyses revealed the integrity of
collagen molecules, with slight shifts in the amide I and II groups due to
interactions with the C=O groups of alginate. AFM-QNM tests highlighted an
increase in the Young's Modulus (YM) in proportions equivalent to alginate and
collagen, with the 2:2 - 2 μg group achieving the highest YM at 3.52 ± 0.095. In this
proportion, the average pore size was 24.6 ± 2.10 μg, and the degradation rate

was 96% (2:2 - 4 μg) and 97% (2:2 – 6 μg). In vitro assays utilized human adipose-
derived mesenchymal stem cells (hASCs), demonstrating viability across all

scaffolds. The 4:2 proportion, with higher concentrations of alginate, stood out with
the best results by the end of the 3rd day. However, mineralization assessments
indicated lower mineralization in the 4:2 groups, while once again, the 2:2 groups
showed greater mineralized areas. The outcomes of this study contribute to
advancing the development of biomimetic scaffolds for bone regeneration. These
findings establish a solid foundation for future research and clinical applications in
the fields of tissue engineering and regenerative medicine.

The skin is an organ that has great reparative capacity, but some acute and chronic injuries
can trigger significant loss of its architecture. In these cases, tissue engineering can be applied
through the use of scaffolds. Biomaterials composed of decellularized extracellular matrix
(dECM) present low immunogenic response and provide a favorable environment for cell
adhesion and proliferation. Natural polymers, such as nano-chitosan, have been gaining
prominence in biomedical applications because they present healing and antimicrobial
properties. This work proposes the development of a dECM biomaterial functionalized with
nanochitosan for use as a biodressing. To obtain the dECM, fragments of bovine skin were
decellularized by agitation in 1% SDS medium, EDTA, Aprotinin and ATB. The samples
were submitted to histological analysis with H&E, Gomori's Trichrome and Alcian Blue
stains, besides DNA extraction and immunofluorescence. The nano-chitosan was obtained
from the ionic gelation method, and characterized by MET, DLS, SEM and antibacterial
activity. The decellularized fragments were functionalized with the nanoparticles by
adsorption. Characterization of the scaffold was done by histological analysis, AFM, FTIR,
SEM and EDS. The size of the nanochitosan structures ranged between 75.48 ± 13.73nm, and
the images showed a shapeless morphology. The polydispersity index was between 0.393 ±
0.015, being within the recommended parameters for natural polymers. The antimicrobial
activity of nano-chitosan was observed at all concentrations tested (0.26 μg/ml-1
to 19.7

μg/ml-1
) for the bacteria Escherichia coli, Enterobacter cloacae, Salmonella sp. and
Staphylococus aureus. The effectiveness of the matrix decellularization process was proven
by DNA extraction, with no characteristic bands in the decellularized sample compared to the
control. The immunofluorescence showed the absence of cell nuclei in the decellularized
tissue, leaving only some debris. The absence of nuclei was also visualized by histological
analysis, in which it was possible to observe the maintenance of much of the matrix structure
through qualitative evaluation of collagen fibers and preservation of glycosaminoglycans,
with preserved topographical architecture. The EDS and FTIR tests demonstrated good
functionalization of the matrix with the nano-chitosan. The SEM analysis demonstrated that
the dECM scaffold presents adequate structure and porosity to favor cell adhesion and
growth. Thus, this work generates perspectives of nanotechnological biodressing from
xenogenic sources for tissue repair.

Ovarian cancer (OC) is the second most common neoplasm related to the female
reproductive system. Currently, precision medicine in oncology has sought earlier and
more precise diagnostic methodologies, since a better understanding of aspects of tumor
growth dynamics considerably increases the chances of obtaining more satisfactory
results in the therapeutic approach. Personalized treatment, however, requires
increasingly specific diagnostic techniques, early detection of the disease and real-time
monitoring of tumor development, all of this, combined with less invasive procedures,
without exposing the patient to clinical risks. of a surgical biopsy. In this context,
extracellular vesicles (EVs) have demonstrated an interesting potential for a liquid
biopsy approach, since the characterization of microvesicles (MVs) circulating in
peripheral blood has been presented as a promising alternative for understanding the
dynamics of the development of tumors in various types of cancer. This study carried
out the phenotypic characterization of subpopulations of circulating microvesicles
(MVs) originating from cells of the immune system and tumor cells (CTCs) by flow
cytometry, evaluating their therapeutic potential as biomarkers for the diagnosis of
epithelial ovarian cancer (EOC). Peripheral blood was collected from 9 volunteers
without detectable ovarian disease (Control group) and from 21 patients with DSC. The
data demonstrated that patients with OC have a greater number of circulating MVs than
women without evidence of ovarian disease. Furthermore, the results obtained
demonstrate that MVs derived from eosinophils, platelets and CTCs-EpCAM are
biomarkers capable of identifying patients with EOC through peripheral blood with the
potential to be used in screening and diagnostic campaigns, in association with dosages.
of serum CA125 values. The data indicate that the combined use of measurements of
serum CA125 values associated with measurements of CTCs-EpCAM MVs can classify
patients as early, advanced, or free stage of epithelial ovarian cancer.

The skin is constantly exposed to stimuli from the external environment that can cause some injury and interfere with its barrier function. Therefore, mechanisms guarantee the repair of wounds, but some cases can cause pathological scars that involve, among other factors, impaired function and aesthetics. As a result, numerous therapies are being developed to improve healing. Collagen is a biomaterial with potential to be used because it is abundant, biocompatible, biodegradable and capable of aiding cell migration and proliferation, in addition to improving matrix deposition. Collagen may also be able to influence the immune response through inhibitory receptors and the indirect effects of tolerance that influence the inflammatory phase of skin lesion repair. Therefore, the objective of this work was to produce a heterologous collagen biogel (PEPTAN®) and verify its topical application on the wound on the day the injury was performed and its influence on inflammation and extracellular matrix deposition during the wound closure process. in the skin of Swiss mice. First, the morphological and physicochemical characterization of collagen biogels was performed. Biogels were produced: PEPTAN 1[1], PEPTAN 1[2], PEPTAN 2[1] and PEPTAN 2[2], which differ according to the heterologous source, porcine or bovine, and two different molecular weights. Morphological characterization by SEM showed that porcine collagen granules and gelatins, porcine and bovine, have different shapes, but most have a rounded shape. Bovine collagen granules have a filamentous shape. When comparing the size of the granules, the porcine collagen 5000, porcine 2000 and bovine 5000 presented statistically similar sizes. Bovine collagen 2000 presented smaller granules when compared to porcine collagen 5000, however, it presented granules with a similar size to porcine collagen 2000 and bovine collagen 5000. The size of gelatin granules did not show statistically significant differences. The biogels produced from these biomaterials were structured and porous, with the pores formed from overlapping plates. When comparing the pore size, it was observed that the PEPTAN 1[2] biogel has larger pores when compared to the PEPTAN 2 biogel[2]. The other biogels have pores of similar sizes. XRD analysis showed that collagen peptides and gelatins maintain the helical structure of collagen. These data were confirmed with the analysis of AFM, where it was possible to observe the formation of collagen fibers. Finally, the elemental analysis by EDS showed that in all collagens and gelatines used for the production of biogels, C, O and N were detected, with C being the greatest amount, followed by O and N, typical components of proteins. To evaluate the effects of topical application of PEPTAN® heterologous collagen biogels on skin wound repair, in vivo analyzes were performed. The experimental model consisted of performing two excisional lesions and an incisional lesion on the skin of the back, under anesthesia. Then, the treatment was performed according to the experimental group. The micropore that maintained the topical treatment on the lesions remained for 5 days. The animals were euthanized 7 and 60 days after the injury and the healing was analyzed macro and microscopically after staining with H&E, Toluidine Blue and Gomori Trichrome. In the incisional lesions, macroscopically at 7d, the PEPTAN 1[1] group presented an aesthetically better lesion, reduction of fibroblasts in the PEPTAN 2[1] and PEPTAN 2[2] groups, reduction of mast cells in the PEPTAN 1[2] group when compared with the PEPTAN 2[1] group and At 60d, the incisional lesions of the PEPTAN 1[1] and PEPTAN 1[2] groups present clearer and esthetically better scars when compared to saline. The histomorphometric analysis shows that the PEPTAN 2[2] group showed a higher deposition when compared to the Saline group. In excisional lesions, at 7d, the PEPTAN 1[2] group presented a smaller area, while the PEPTAN 1[1] group presented a larger area. Microscopically, quantitative analysis showed no statistical difference in leukocyte and mast cell counts. It was observed that the PEPTAN 2[1] group had a higher number of fibroblasts when compared to the PEPTAN 1[2] and PEPTAN 2[2] groups. At 60d, the PEPTAN 1[1] and PEPTAN 1[2] groups showed better collagen deposition when compared to the Saline group. The saline group presented a more marked and perceptible transition area between the lesion and the adjacent dermis. While the PEPTAN 1[1] and PEPTAN 1[2] groups showed this smoother transition. We conclude that the application of a dressing containing the PEPTAN® heterologous collagen scaffold improves the healing of skin lesions, with better remodeling of the extracellular matrix. 

Introduction: Parkinson's Disease (PD) affects the cardiorespiratory system causing greater left ventricular mass in the heart and respiratory muscle weakness compared to healthy individuals. Objective: Analyze the histomorphometric alterations of cardiac tissue and respiratory muscles, in addition to the functional performance of rats with PD submitted to TIAI. Materials and Methods: We used 70 male Wistar rats, 40 days old, divided into groups with Parkinson's (PA) and Sham (SH); subdivided into groups that performed High Intensity Interval Training Before Surgery (TIAIa); TIAI After Surgery (TIAId) and TIAI Before and After Surgery (TIAIad). Physical exercise was performed before and/or after PD induction. The rats performed the exercise, 5 times a week, for 25 minutes/day, for 4 or 8 weeks. The functional performance of the animals was evaluated by of false step and parallel bars tests. For morphometric analysis of the heart, the relative weight of the heart, diameter and thickness of the left ventricle were calculated. The histochemistry of the substantia nigra of the midbrain was performed using the Nissl method. Myocardial, diaphragm, intercostal and rectus abdominis muscles were stained with Hematoxylin and Eosin (HE). The histomorphometric study was performed with the Image J program to analyze the cross-sectional area of the muscles and the number of muscle fibers. Data analysis was performed using the GraphPad Prism 9.3 statistical program, one-way ANOVA test, followed by Tukey's post-hoc test (p<0.05). Results: The analysis of the hind legs with the parallel bars test showed that the trained animals showed improvement in functional performance compared to the animals in the sedentary group. The relationship between the body weight, heart weight and number of cardiomyocytes showed no significant difference in PA and SH trained animals. There was an increase in the areas of the diaphragm, intercostal and rectus abdominis muscles in the PA and SH trained groups compared to the sedentary ones. Conclusion: The TIAI provided hypertrophy of the respiratory muscles and left ventricle of the heart, in addition to improving the functional performance of the hind legs of animals with PD.

Parkinson's disease (PD) affects dopaminergic neurons located in the pars compacta of the substantia nigra. The practice of resistance physical activity is a non-pharmacological intervention that has been tested in the treatment of PD. Thus, the present project aimed to investigate the effects of high-intensity exercise on neuronal density and astrocyte expression in the brain of rats with PD that exercised before and after disease induction. Seventy Wistar rats were used, distributed as follows: 09 animals trained before PD induction (DP-Exa), 09 trained after PD induction (DP-Exd); 10 trained before and after PD induction (DP-Exad) and 09 sedentary animals (DP-Sed). The same groups were distributed to the group without PD (Sham). High-intensity physical training was performed on the vertical ladder before and/or after PD induction. It was performed for 5 days/week, 30 to 45 minutes, for 4 weeks. PD induction was performed using the electrolytic lesion model in the coordinates: AP equal to -4.9, ML equal to 1.7 and DV equal to 8.1. The animals' motor performance was evaluated at the beginning and at the end of training with false step the test, parallel bars test and open field tests. At the end of the experiment, the brain was removed for histochemistry, staining by the Nissl method, and immunohistochemistry by expression of Glial Fibrillary Acid Protein (GFAP) from the substantia nigra and striatum. The histomorphometric study was performed with the Image J program for counting neurons and astrocytes (GFAP). The GraphPad Prism 9.3 program was used, one-way ANOVA test, followed by Tukey's post-hoc (p<0.05). The analysis showed better performance of the animals that trained compared to the sedentary group, in the parallel bars and open field tests. Data on neuron counts in the striatum, substantia nigra and GFAP in the animals of the DP-Exa, Sham-Exa, DP-Exad and Sham-Exad groups showed a greater number of neurons and greater expression of GFAP, when compared to the groups DP-Exd, Sham-Exd, DP-Sed and Sham-Sed. in conclusion, the groups of animals that performed high-intensity training before, before and after PD induction had higher densities of neurons and astrocytes.

Plant species are a rich source for the discovery of active principles and the development of
new therapies. The species Solanum lycocarpum A.St.-Hil has antioxidant, anti-inflammatory
and antimicrobial effects confirmed in some experimental models. In addition, several
phytochemicals present in the extracts used were found in other species of the genus, such as
some phenolic compounds, which were related to the effects of improving the healing
process, with individual or synergistic effects on one or more stages of wound healing. The
research project aimed to analyze the effects of the use of ethanolic extract and the
hydroethanolic fraction of the ripe fruit of Solanum lycocarpum on the healing process of skin
wounds in mice. Seventy-two animals were randomly divided into three groups, a Control
Group that received topical application of physiological saline solution, a group treated with
ethanol extract and a group treated with the hydroethanolic fraction of the ripe fruit of
Solanum lycocarpum. Each group was subdivided into four other subgroups according to the
treatment period: 1, 5, 7 and 21 days, with n=6 in each subgroup. To make the excisional
wounds, a metallic dermatological punch, 7 mm in diameter, was used. Macroscopic
evaluations of each lesion were performed daily to verify the presence of inflammatory signs
(edema, hyperemia), the presence of crusts and secretions and to calculate the rate of
contraction of the wounds. After 1, 5, 7 or 21 days after the excisional lesion, the animals
were euthanized by a lethal dose of anesthetic. A fragment containing the wound was
removed, cleaved and fixed in a 10% neutral buffered formalin solution. Subsequently, the
material was histologically processed and stained by histological and histochemical methods,
for histopathological and morphometric evaluation of the inflammatory infiltrate, fibroblasts,
re-epithelialization and collagen deposition, after daily topical treatment with ethanolic extract
of the ripe fruit of Solanum lycocarpum and its hydroethanolic fraction. The results show that
macroscopically the treatments did not interfere with the closure of excisional wounds over
the 21 days. Topical application of ethanolic extract from the ripe fruit of Solanum
lycocarpum reduced the inflammatory infiltrate and areas of edema after 1, 5 or 7 days of treatment. No differences were observed in fibroblast, mast cell and blood vessel counts. In the treated groups, the remodeling process was in progress, however, a better evolution was observed in the group that received ethanol extract, where the epithelium was more similar to that of the areas without lesion.

There are several proposals for interferences in the immune response, either preventive or therapeutic. One of these very promising forms of interference that has been studied is the injection of immunologically tolerated proteins. Oral tolerance is a phenomenon that inhibits the immune response, such as the formation of specific antibodies, after immunization with antigen plus adjuvant in animals previously exposed to the protein via the oral route. In this work, we studied whether subcutaneous application (s.c.) in mice, of gold nanoparticles (AuNPs) associated with Ovalbumin (OVA) would contribute to the establishment of oral tolerance, as an adjuvant material. In this sense, we synthesized spherical AuNPs, with approximately 7 nm, functionalized with OVA and made the physical-chemical characterization of the material, for that, we used the Ultraviolet-Visible Spectroscopy (UV-Vis), Transmission Electron Microscopy (MET), Scattering Dynamic Light (DLS) and the Zeta Pontential. Subsequently, an oral treatment with OVA was performed for 5 consecutive days in Swiss, male, 8-week-old mice. Seven days after oral treatment, all animals received their primary injections (s.c) at the base of the tail and twenty-one days after the primary injections, the animals received secondary injections (s.c). On the day of the primary injections, lesions were made in one of the mice's ears. We evaluated the immunization potential, as well as the establishment of the OVA tolerance of the animals, verifying the total anti-OVA antibody titer with the Enzyme-linked Immunosorbent Assay (ELISA). The base of the tail was also histologically evaluated, at the place where the animals received injections and ear injuries, to assess the indirect effects of oral tolerance. The results demonstrated that AuNP + OVA have an adjuvant role and are able to maintain the indirect effects of oral tolerance. Subsequently, to understand
16
which intracellular signaling pathways AuNPs and AuNPs + OVA can interfere with immune responses, we did a modeling in silico interactions between AuNPs and AuNPs + OVA with OVA, LC3 proteins; ASC PYCARD; Glutathione; TLR4; PI3K and NF-Kβ. That way, we verified that AuNP has greater affinity with the OVA molecule, followed by Glutathione and ASC PYCARD domain, whereas AuNP + OVA has greater affinity with the Glutathione molecule, followed by NF-Kβ. These results suggest that our AuNPs and AUNPs + OVA may be interfering in cell autophagy pathways, AuNPs in inflammatory and apoptotic cell pathways and AUNPs + OVA in inflammatory pathways. However investigation is needed to confirm and understand these trials.

Medicinal plants are widely used in folk medicine, among them we have the species Ageratum conyzoides, popularly known as "mentrasto". This is a tropical plant, which has a wide variety of secondary metabolites that include terpenes, steroids and alkaloides, plus essential oils. A. conyzoides is used in folk medicine for the treatment of fevers, ulcers and inflammatory diseases. Some studies have shown the beneficial effect of aqueous fraction of A. conyzoides on benign prostatic hyperplasia, antitumor effect and relaxation of the jejunum, duodenum and uterus. There is no work in the literature demonstrating the effect of the essential oil of A. conyzoides, so the objective this work was to evaluate the effect of the essential oil of A. conyzoides on the contraction of smooth muscle organs, as well as to try to describe possible mechanism of action. To perform the experiments, vas deferens and mouse jejunum were used. These organs were stimulated respectively with KCl, adrenaline and KCl and carbacol. Increasing doses of essential oil of A. conyzoides were preincubated in the presence and absence of nifedipine. To compare the effect of the oil with the effect of nifedipine, IC50 was used. Phytochemical analysis showed that precocen I is in greater quantity in our oil. The essential oil of A. conyzoides was able to inhibit both the phasic component and the tonic component of KCl or adrenaline-induced vas deferens contraction. Nifedipine also inhibited contraction. The effect of A. conyzoides was enhanced in the presence of nifedipine, being the phasic component KCl contraction and the phasic and tonic components of the most sensitive adrenaline-induced contraction than the effect only of A. conyzoides on its components. In the jejunum, the phasic composition of KCl-induced contraction is more sensitive to the action of A. conyzoides than the tonic. In the presence of nifedipine the effect of the oil was enhanced, but only on the phasic component. In this way, our results suggest that the essential oil of A. conyzoides can inhibit voltage-gated calcium channels and may be acting in another mechanism responsible for inhibiting muscle contraction of smooth muscle organs.

Zika virus (ZIKV) is an emergent pathogen of the Flaviviridae family, transmitted by

Aedes mosquitoes. In the last years, ZIKV has caused neurological abnormalities and

Guillain Barré Syndrome (GBS) in adult individuals, and fetal developmental

complications in pregnant women, including Congenital Zika Syndrome (CZS).

Vasoactive intestinal peptide (VIP) is an antiinflammatory and imunomodulator peptide.

Through its membrane receptors VPAC 1 and VPAC 2, VIP influences the activity of a

diverse range of cells of the immune system. The distribution of VPAC receptors is

crucial to understand how VIP can act in different cells and tissues. The spleen is the

main secundary lymphoid organ, and houses T and B cells, and macrophages that

respond in an antiinflammatory manner to cholinergic vagal innervation. In this study,

newborn mice from ZIKV-infected and uninfected mothers were studied through

immunohistochemistry to quantify VPAC1 and VPAC2, CD11b, which is a

macrophage marker, and the nicotinic cholinergic receptor α7nAChR in the spleen of

the offspring. The results demonstrate that maternal infection by ZIKV significantly

increases the expression of VPAC 2 receptor in the spleen of newborn mice.

Surprisingly, it was found a dramatic increase in the population of megakaryocytes in

the spleen of ZIKV+ animals, when compared to the healthy group, suggesting that

ZIKV maternal infection influences the behaviour of hematopoetic cells in the offspring.

Nonetheless, mast cell precursors were found in the spleen of both groups PBS and

ZIKV+. Maternal ZIKV infection caused a significant decrease in the expression of

VPAC 1 in these mast cell precusors. Taken together, these data reveal the importance

of the differential distribution of VPAC receptors in immune cells during the

immunological response to ZIKV. Studies are needed to unreveal the functional relation

of VPAC receptors during ZIKV infection, how VIP can modulate the behaviour and

phenotype of cells in the spleen, and what is the functional consequences to

development of the offspring.

The healing of skin wounds is a process that involves a serie of events, cells and factors, aimed at repairing the tissue that initially suffered a trauma or injury. Many researches are currently focused on the serach for possible therapies and drugs that can contribute to the repair process. Medicinal plants are the subject of much research, given their range of pharmacological properties, including studies of their use in wounds in experimental models. Basil (Ocimum basilicum) is a plant that has been described with many properties, including anti-inflammatory and antioxidante. The aim of the presente work was to analyze the effects of intraperitoneal (i.p.) and the topical administration of the ethanolic extract of Ocimum basilicum (EEOB) in the healing process of incisional and excision cutaneous wounds in mice. Swiss mice were used, divided into groups according to treatment and type of wound. In the animals that made the incisional wound, the treatment was performed with intraperitoneal EEOB (i.p.) at 150 mg/Kg daily or saline i.p., for 5 or 7 days post-injury, depending on the group, and later the animals were euthanized. In the animals that made the excision wounds, the treatment was carried out in the groups treated with topical EEOB at 38 mg/Kg in the left lesion and topical EEOB at 100 mg/Kg in the right lesion, three times a day, while the control group received topical saline; the animals were subsequently euthanized 1, 3, 5 or 21 days after injury and treatment, depending on the experimental group. Thereafter, the lesions underwent histological processing and microscopic analysis, and the excisional lesions were also assessed macroscopically before euthanasia. Histopathological analyzes in animals with incisional lesion and treated with EEOB injection i.p. in relation to the group that received saline i.p. daily, point to a decrease in the number of leukocytes at 5 days and mast cells at 7 days post-injury. Regarding excision wounds, the analysis indicates that within 1 day there was a decrease in the number of leukocytes in the treated groups in relation to the saline group and a decrease in the area of edema and adipose tissue. At 3 days, the grouptreated with EEOB at 100 mg/Kg showed a larger area of macroscopic wound compared to the other groups. At 5 days, there was a decrease in new blood vessels in the EEOB treated group at 38 mg/Kg compared to saline; and the EEOB group at 100 mg/Kg had less adipose tissue compared to the other groups and a greater area of granulation tissue. Finally, at 21 days, reepithelization was complete in most animals of all groups, and collagen deposition in the scar region was better in the treated groups, with more similarity to the intact skin adjacent to the lesion area. The results point to an antiinflammatory action of EEOB in the groups that received the extract, mainly in the initial phase of the process. Furthermore/besides, the extract may have influenced the better collagen deposition in the tissue remodeling phase. It is suggested that EEOB has potentially effective therapeutic properties in the treatment of skin wounds healing and further in-depth studies on how the EEOB administration pathways work are needed to validate a possible plant-based treatment for the pathology in question.

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