Stem Cells and Tissue Engineering-Based Therapeutic Interventions: Promising Strategies to Improve Peripheral Nerve Regeneration.

Unlike the central nervous system, the peripheral one has the ability to regenerate itself after injury; however, this natural regeneration process is not always successful. In fact, even with some treatments, the prognosis is poor, and patients consequently suffer with the functional loss caused by injured nerves, generating several impacts on their quality of life. In the present review we aimed to address two strategies that may considerably potentiate peripheral nerve regeneration: stem cells and tissue engineering. In vitro studies have shown that pluripotent cells associated with neural scaffolds elaborated by tissue engineering can increase functional recovery, revascularization, remyelination, neurotrophin expression and reduce muscle atrophy. Although these results are very promising, it is important to note that there are some barriers to be circumvented: the host's immune response, the oncogenic properties attributed to stem cells and the duration of the pro-regenerative effects. After all, more studies are still needed to overcome the limitations of these treatments; those that address techniques for manipulating the lesion microenvironment combining different therapies seem to be the most promising and proactive ones.

Central giant cell granulomas of the jaws stromal cells harbour mutations and have osteogenic differentiation capacity, in vivo and in vitro.

BACKGROUND: Central giant cell granulomas (CGCG) of the jaws are osteolytic lesions that may behave aggressively and respond poorly to surgery. Microscopically, in addition to giant cells, there is a mononuclear cell population composed of macrophage/monocytic cells and spindle-shaped cells of mesenchymal origin. Seventy two percent of these tumours harbour mutually exclusive TRPV4, KRAS and FGFR1 mutations. We aimed to assess the mutational status of mononuclear and giant cells and the osteogenic potential of stromal cells in vitro and in vivo. METHODS AND RESULTS: We screened CGCG for signature mutations and used laser-capture microdissection to demonstrate that the mutations are restricted to the mononuclear cells. Additionally, we established CGCG primary cell culture and observed that the cells retained the mutations throughout passages. By flow cytometry, we observed predominance of CD14- CD51- CD61- cells, consistent with the expected profile for stromal cells. Considering the mesenchymal origin of stromal cells, we assessed the osteogenic differentiation potential of CGCG cells in culture by cytochemistry (von Kossa and alizarin red staining), alkaline phosphatase (ALP) activity assay and gene expression of osteogenic markers. CGCG cells presented self-capacity to increase ALP levels in a time-dependent manner and under osteogenic induction presented increasing number of calcium deposits, and overall higher expression of osteocalcin, RUNX2, ALPL and osteopontin than cells without osteogenic induction. A patient-derived xenograft model for CGCG was established, and osteoid material deposition was observed. CONCLUSION: Collectively, the results confirm that the signature mutations are restricted to stromal cells in CGCG, and the in vitro and in vivo results support that these cells have the capacity to differentiate into osteoblasts, in line with the bone formation often observed in the stroma of these lesions.

Vaginal stretching therapy and class IIIB vaginal laser treatment for pelvic floor myofascial pain: a randomized clinical trial.

PURPOSE: Several treatment options are proposed for the management of pelvic floor myofascial pain (PFMP). Manual therapy, such as vaginal stretching (VS), is one of these options. Photobiomodulation therapy (PBMT) with a laser device is a treatment option for PFMP that has been tested on other muscles. The aim of this study was to evaluate the effect of VS combined or not with PBMT for PFMP treatment. METHODS: One hundred three women with PFMP were enrolled in a double-blind randomized trial and assigned to VS+PBMT (10 treatments over 2 weeks with 100 mw delivering 12 joules to surface intravaginally, using near-infrared light 808 nm) and VS+shamPBMT treatment groups. Pain severity was assessed by Visual Analog Scale (VAS). Pelvic floor muscle function was assessed by Oxford Scale and surface electromyography. Urinary symptoms were evaluated by ICIQ-OAB and ICIQ-SF questionnaires, and intestinal constipation was assessed by ROMA criteria. RESULTS: There was a significant improvement in pain intensity (VAS) after treatment in both groups, with no difference between groups (p = 0.46). More than 50% of the women complained of severe pain before treatment, and after treatments, it was reported by less than 20% of women (p < 0.001), with no difference between groups (p = 0.08). Urinary symptoms improved in both groups (p < 0.001) with no difference between groups (p = 0.37). Intestinal constipation improved in the VS+PBMT group only (p = 0.01). CONCLUSION: VS and VS with near-infrared vaginal laser therapy were equally effective at decreasing myofascial pelvic pain and reducing urinary symptoms TRIAL REGISTRATION: REBEC (Registro Brasileiro de Ensaios Clínicos; Brazilian Registry of Clinical Trials) under no.RBR-2TDCQ4 (November 11, 2018).

Depression, anxiety, and stress in women with urinary incontinence with or without myofascial dysfunction in the pelvic floor muscles: A cross-sectional study.

AIMS: To compare the prevalence of psychological symptoms (depression, anxiety, and stress) in women with urinary incontinence (UI), according to the presence or absence of myofascial dysfunction (MD) in the pelvic floor muscles (PFMs). METHODS: Cross-sectional study, with women with UI who are 18 years old and over. The diagnosis of MD was defined by the pain of any intensity during the palpation of PFM. All participants answered the International Consultation on Incontinence Questionnaire-Short Form and the International Consultation on Incontinence Questionnaire Overactive Bladder for urinary symptoms and the Depression Anxiety and Stress Scale-Short Form-21 to check for the presence and degrees of depression, anxiety, and stress. RESULTS: Two hundred-thirty-four women with a mean age of 52.5 (±9.2) years were included. Almost half (51.7%) of women had MD. Women with MD showed higher mild and moderate anxiety scores (p = .005) and higher mild, moderate, and severe stress scores (p = .027) than women without MD. Depression scores were not associated with MD; however, women with and without MD reported severe or extremely severe depression, anxiety, and stress. CONCLUSIONS: The risk for depression, anxiety, and stress is high among women with UI regardless of the presence of MD. However, women with MD had higher scores for anxiety and stress than women with UI without MD.

Validation and cultural translation for the Brazilian Portuguese version of the Questionnaire for Urinary Incontinence Diagnosis.

INTRODUCTION AND HYPOTHESIS: Short questionnaires are important for validating the clinical diagnosis of urinary incontinence (UI). We sought to validate and culturally translate the Questionnaire for Urinary Incontinence Diagnosis (QUID) for the Brazilian Portuguese language. METHODS: A cross-sectional study with 457 women (330 with urinary incontinence and 127 controls) was performed in a Southeastern Brazilian outpatient clinic. Patients answered a pilot-tested, notarized, six-item questionnaire (QUID) for internal consistency as well as a control questionnaire (ICIQ-SF and ICIQ-OAB) for construct validity. In both groups, floor and ceiling effects were calculated. Within UI women, test-retest (n = 41) and responsiveness to conservative treatment (n = 74) were also analyzed. RESULTS: Internal consistency (Cronbach's alpha) from the QUID was adequate between the UI (0.845-0.850) and control (0.724-0.775) groups. Mean QUID scores were statistically different between UI and control groups (p < 0.05). No ceiling or floor effects were observed in incontinent patients. Test-retest reliability after 4 weeks (intraclass correlation coefficient [ICC]: 0.780-0.814) and responsiveness (0.867-0.889) were also adequate within UI women. Construct validity was adequate at all correlations between QUID and ICIQ-SF and ICIQ-OAB (r: 0.19-0.58; p <0.05). Responsiveness was demonstrated by a statistically significant difference in questions/subscale sores after physical therapy. CONCLUSION: The QUID presented adequate cultural translation, reliability, and good responsiveness to treatment in the Brazilian Portuguese language.

The Solanum glaucophyllum Desf. extract reduces mineralized matrix synthesis in osteogenically differentiated rat mesenchymal stem cells in vitro.

The Solanum glaucophyllum Desf. has been used to treat and prevent diseases in human and veterinary medicine. On the other hand, plant poisoning causes several bone diseases, among them osteoporosis, which is characterized by osteoblastic hypoplasia. Because the osteoblast is a cell derived from the differentiation of mesenchymal stem cells (MSCs) from bone marrow, the hypothesis is that the plant reduces the osteogenic differentiation of MSCs. The objective of this study was to evaluate the effects of S. glaucophyllum Desf. extract on MSCs cultured in osteogenic differentiation medium. We determined by liquid chromatography that 1 ml of plant extract contained 3.8 µl of 1,25(OH)2 D3 (calcitriol). Four groups of MSCs cultivated in osteogenic medium were evaluated as follows: (a) treated with 100 µl of extract/L containing 0.4 µg/L of calcitriol; (b) treated with 1 ml of extract/L containing 4 µg/L of calcitriol; (c) treated with 5 ml of extract/L containing 20 µg/L of calcitriol; and (d) a control group without extract. We performed alkaline phosphatase activity assay, analysis of MTT conversion to formazan, and evaluated the percentage of cells, and number and diameter of mineralization nodules. The expression of gene transcripts for osteopontin, bone sialoprotein and BMP-2 was analysed by RT-qPCR. After 21 days, there was a significant reduction in MTT conversion to formazan in treated groups, of the cellularity in the group with 5 ml of extract/L, and in the number and size of mineralization nodules in the groups treated with 1 and 5 ml of extract/L. The 5 ml extract/L concentration also reduced transcript expression of osteopontin. It is concluded that S. glaucophyllum Desf. at concentrations of 1 and 5 ml extract/L reduced mineralized matrix synthesis in MSCs cultivated in osteogenic differentiation medium, which suggests that this is one of the mechanisms by which osteoporosis occurs in intoxicated animals.

Didelphis albiventris: an overview of unprecedented transcriptome sequencing of the white-eared opossum.

BACKGROUND: The white-eared opossum (Didelphis albiventris) is widely distributed throughout Brazil and South America. It has been used as an animal model for studying different scientific questions ranging from the restoration of degraded green areas to medical aspects of Chagas disease, leishmaniasis and resistance against snake venom. As a marsupial, D. albiventris can also contribute to the understanding of the molecular mechanisms that govern the different stages of organogenesis. Opossum joeys are born after only 13 days, and the final stages of organogenesis occur when the neonates are inside the pouch, depending on lactation. As neither the genome of this opossum species nor its transcriptome has been completely sequenced, the use of D. albiventris as an animal model is limited. In this work, we sequenced the D. albiventris transcriptome by RNA-seq to obtain the first catalogue of differentially expressed (DE) genes and gene ontology (GO) annotations during the neonatal stages of marsupial development. RESULTS: The D. albiventris transcriptome was obtained from whole neonates harvested at birth (P0), at 5 days of age (P5) and at 10 days of age (P10). The de novo assembly of these transcripts generated 85,338 transcripts. Approximately 30% of these transcripts could be mapped against the amino acid sequences of M. domestica, the evolutionarily closest relative of D. albiventris to be sequenced thus far. Among the expressed transcripts, 2077 were found to be DE between P0 and P5, 13,780 between P0 and P10, and 1453 between P5 and P10. The enriched GO terms were mainly related to the immune system, blood tissue development and differentiation, vision, hearing, digestion, the CNS and limb development. CONCLUSIONS: The elucidation of opossum transcriptomes provides an out-group for better understanding the distinct characteristics associated with the evolution of mammalian species. This study provides the first transcriptome sequences and catalogue of genes for a marsupial species at different neonatal stages, allowing the study of the mechanisms involved in organogenesis.

Inhibition of the osteogenic differentiation of mesenchymal stem cells derived from the offspring of rats treated with caffeine during pregnancy and lactation.

Caffeine is an alkaloid that is widely consumed due to its presence in drugs, coffee, tea, and chocolate. This compound passes to offspring through the placenta and milk; can cause teratogenic mutations; and reduces the formation, growth, and mass of bone. Because mesenchymal stem cells (MSCs) are responsible for generating the entire skeleton, we hypothesized that these cells are targets of caffeine. This study evaluated the osteogenic differentiation of MSCs derived from the offspring of rats treated with caffeine during pregnancy and lactation. Twenty-four adult Wistar rats were randomly divided into four groups, including one control group and three experimental groups treated with 25, 50, or 100 mg/kg of caffeine. At weaning, three 21-day-old pups from each dam in each group were euthanized for extraction of bone marrow cells for in vitro tests. Caffeine doses of 50 and 100 mg/kg significantly reduced the activity of alkaline phosphatase at 7, 14, and 21 days and the expression of collagen I at 21 days. However, the expression of gene transcripts for alkaline phosphatase, Runx-2, and bone sialoprotein, as well as the synthesis of mineralization nodules, decreased significantly in all groups treated with caffeine. The expression of osteocalcin was significantly reduced only in the group treated with 50 mg/kg caffeine. The caffeine that passes from the mother to the offspring during pregnancy and lactation reduces the osteogenic differentiation of MSCs. We propose that this reduction in the osteogenic potential of MSCs may be involved in the pathogenesis of osteopenia resulting from caffeine consumption.

Experience of health professionals in care of the homeless population with tuberculosis.

OBJECTIVE: Analyzing statements of health professionals from a Street Clinic regarding care of a homeless population with tuberculosis. METHOD: This is a qualitative research, conducted in the central region of São Paulo at three basic health units in the period of November to December 2014. A semi-structured interview guideline was implemented for data collection and all interviews were recorded using a digital recorder. RESULTS: Six health professionals were interviewed. According to the Discourse Analysis perspective, three discursive segments emerged: experiences on care in the streets; weaknesses inherent to the treatment process; and incentives as a means of maintaining sick people in treatment. CONCLUSION: Caring for a the homeless population with tuberculosis constitutes a new and challenging experience. It involves difficulties in dealing with the reality of a miserable social context, a lack and inadequacy of services, as well as care limitations for treatment and treatment dropout, which reinforces multiresistance. However, the investigated Street Clinic teams seek to expand access to health and social care services to this population. OBJETIVO: Analisar os discursos dos profissionais de saúde do Consultório na Rua em relação ao cuidado à pessoa em situação de rua com tuberculose. MÉTODO: Trata-se de uma pesquisa qualitativa, realizada na região central do município de São Paulo, em três Unidades Básicas de Saúde, no período de novembro a dezembro de 2014. Utilizou-se de um roteiro de entrevista semiestruturada para a coleta de dados e todas as entrevistas foram gravadas com recurso a um gravador digital. RESULTADOS: Foram entrevistados seis profissionais de saúde. Segundo a perspectiva da Análise de Discurso, emergiram três blocos discursivos: experiência sobre o cuidar na rua; fragilidades inerentes ao processo de tratamento e incentivos como meio para a permanência do sujeito doente no tratamento. CONCLUSÃO: Cuidar da pessoa com tuberculose e em situação de rua constitui uma experiência nova e desafiadora, implica dificuldades em lidar com a realidade de um contexto social miserável, falta e inadequação de serviços, bem como limitações do cuidado para a cura e abandono do tratamento, podendo reforçar a multirresistência. Contudo, as equipes de Consultório na Rua investigadas buscam ampliar o acesso aos serviços de saúde e assistência social a essa população.

Osteogenic potential of osteoblasts from neonatal rats born to mothers treated with caffeine throughout pregnancy.

BACKGROUND: Caffeine is an active alkaloid that can cause damage to bones in formation during prenatal life into adulthood. This compound can pass across the placenta and into the mother's milk, causing a reduction in bone formation, growth and mass. The objective of this study was to examine the osteogenic potential of osteoblasts extracted from neonatal rats born to mothers treated with caffeine throughout pregnancy. METHODS: Twenty-four adult Wistar rats were randomly divided into four groups, consisting of one control group and three groups that were treated with 25, 50, or 100 mg/kg of caffeine by an oral-gastric probe throughout the duration of the experimental period (pregnancy). At birth, three puppies from each dam in each group were euthanized, and osteoblasts were extracted from the calvaria of these pups for in vitro testing. RESULTS: The osteoblasts extracted from the pups of rats that received 50 mg/kg caffeine during pregnancy exhibited increased expression of osteocalcin, osteopontin, sialoprotein, runx-2, alkaline phosphatase and type I collagen transcripts, resulting in increased synthesis of mineralization nodules. CONCLUSIONS: Neonates from rats treated with 50 mg/kg caffeine during pregnancy contained osteoblasts with a higher osteogenic potential characterized by increased expression of osteocalcin, osteopontin, sialoprotein, runx-2, alkaline phosphatase and type I collagen and increased synthesis of mineralization nodules.

Bone development in fetuses with intrauterine growth restriction caused by maternal endocrine-metabolic dysfunctions.

Intrauterine growth restriction (IUGR) affects a large proportion of infants, particularly in underdeveloped countries. Among the main causes of IUGR, maternal endocrine-metabolic dysfunction is highlighted, either due to its high incidence or due to the severity of the immediate and mediated changes that these dysfunctions cause in the fetus and the mother. Although the effects of endocrine and metabolic disorders have been widely researched, there are still no reviews that bring together and summarize the effects of these conditions on bone development in cases of IUGR. Therefore, the present literature review was conducted with the aim of discussing bone changes observed in fetuses with IUGR caused by maternal endocrine-metabolic dysfunction. The main endocrine dysfunctions that occur with IUGR include maternal hyperthyroidism, hypothyroidism, and hypoparathyroidism. Diabetes mellitus, hypertensive disorders, and obesity are the most important maternal metabolic dysfunctions that compromise fetal growth. The bone changes reported in the fetus are, for the most part, due to damage to cell proliferation and differentiation, as well as failures in the synthesis and mineralization of the extracellular matrix, which results in shortening and fragility of the bones. Some maternal dysfunctions, such as hyperthyroidism, have been widely studied, whereas conditions such as hypoparathyroidism and gestational hypertensive disorders require further study regarding the mechanisms underlying the development of bone changes. Similarly, there is a gap in the literature regarding changes related to intramembranous ossification, as most published articles only describe changes in endochondral bone formation associated with IUGR. Furthermore, there is a need for more research aimed at elucidating the late postnatal changes that occur in the skeletons of individuals affected by IUGR and their possible relationships with adult diseases, such as osteoarthritis and osteoporosis.

Periodontitis and diabetes in pregnant rats: Maternal-fetal outcomes.

Aim: To evaluate the repercussions of periodontitis and diabetes association on rat pregnancy and newborns. Methods: Diabetes was induced in female Wistar rats 24 h after birth through the administration of Streptozotocin. The diabetic condition of the rats was further confirmed in adulthood. After mating, the pregnant rats were distributed into four experimental groups (n = 12 rats/group): nondiabetic and diabetic with and without periodontitis. Periodontitis was induced by a ligature inserted into the first molar on day 0 of pregnancy. Body weight, water and feed consumption were evaluated weekly, and an oral glucose tolerance test was performed on day 17 of pregnancy. On day 21 of pregnancy, the animals were anesthetized and killed for organ removal. The hemimandibles were collected to analyze alveolar bone loss. Immunological and biochemical parameters were evaluated in the maternal blood samples, and reproductive performance was analyzed. The newborns were weighed, and anomalies evaluated. Results: The group with diabetes and periodontitis had a greater degree of alveolar bone loss, along with higher relative pancreatic weight, blood glucose levels, triglyceride and inflammatory cytokine levels, hepatic transaminase activity, and embryonic losses. In addition, these newborns had increased body weight, placental weight, a greater number of ossification centers, and a higher rate of visceral and skeletal anomalies. Conclusion: The combination of maternal diabetes and periodontitis negatively impacts maternal parameters and fetal development. The findings reinforce the importance of maintaining maternal oral health to ensure the general health of the offspring, especially in cases where diabetes is present.

Evaluation of Glomerular Filtration Rate Trends in People Living With HIV Corrected by the Baseline Glomerular Filtration Rate.

BACKGROUND: Chronic kidney disease, for which estimated glomerular filtration rate (eGFR) trajectories are early markers, is frequent in people living with HIV. SETTING: Identify eGFR trajectory patterns according to kidney function and assess associated factors over a 13-year follow-up period. METHODS: We evaluated longitudinal changes and its associated factors in eGFR of 3366 participants according to kidney function with a 2-level, linear, mixed model. RESULTS: Participants with initial kidney dysfunction experienced a slight eGFR increase, whereas others showed a slight decrease. A weak relationship was observed between baseline eGFR and its variation over time. Baseline eGFR was affected by age, CD4 + count, viral load, hypertension, hyperlipidemia, AIDS-defining illness and tenofovir (TDF) with integrase inhibitor (INSTI) or efavirenz. Significant factors for eGFR change included the following: in kidney dysfunction, CD4 + cell count of >350 cells per cubic millimeter and undetectable viral load increased eGFR, whereas TDF + protease inhibitor decreased eGFR; in mildly decreased kidney function, CD4 + cell count of >350 cells per cubic millimeter, AIDS-defining illness, and TDF + efavirenz increased eGFR, whereas age, hypertension, hyperlipidemia, and TDF + INSTI decreased eGFR; in normal kidney function, age, CD4 + cell count of > 350 cells per cubic millimeter, undetectable viral load, hypertension, hyperlipidemia, and TDF + INSTI decreased eGFR, whereas TDF + efavirenz increased eGFR (all P value for interaction < 0.05). CONCLUSION: Our findings suggest that eGFR trajectories varied widely between individuals in people living with HIV. In the lower eGFR group, virus-related factors were more relevant, whereas traditional risk factors for renal dysfunction were more prominent in the highest eGFR group.

Effect of the addition of different concentrations of Solanum glaucophyllum desf. extract on chondrocyte cultures from the growth cartilage of newborn rats.

Solanum glaucophyllum Desf. is a calcinogenic plant responsible for enzootic calcinosis that affects ruminants and causes alterations in bone and cartilaginous tissues, among others. It is believed that changes in cartilage tissue, with reduced bone growth, are due to hypercalcitoninism, caused by excess vitamin D. However, we hypothesized that S. glaucophyllum Desf. can act directly on chondrocytes and therefore, chondrocyte cultures from the epiphysis of the long bones of newborn rats were used as a model to elucidate the direct effects of S. glaucophyllum Desf. on bone growth. Plant samples were collected from Cañuelas, Argentina. An aliquot of the plant extract was used to quantify vitamin D (1,25(OH)2D3). The effects of the three concentrations of the plant extract were tested in cultures of chondrocytes extracted from the epiphyses of the long bones of 32 three-day-old Wistar rats. A control group (without extract), and three groups treated with different concentrations of plant extract were formed: group 1 (100 µL/L); group 2 (1 mL/L), and group 3 (5 mL/L), containing respectively 1 × 10-9 M, 1 × 10-8 M, and 5 × 10-8 M of 1,25(OH)2D3. After 7, 14, and 21 days of culture, MTT assay for cell viability, alkaline phosphatase activity, and quantification of the percentage of areas with glycosaminoglycans (GAG) stained with periodic acid-Schiff (PAS) were performed. On day 7, all chondrocytes in group 3, that is, those with the highest concentration of plant extract, died. On days 14 and 21, groups 1 and 2 showed a significant reduction in chondrocyte viability compared to the control. At 7, 14, and 21 days, groups 1 and 2 showed significantly lower alkaline phosphatase activity than the control. On day 21, group 2 showed a significant reduction in areas with PAS + GAGs. There were no significant differences between the groups in the expression of gene transcripts for Sox9, Col2, ColX, and aggrecan. The S. glaucophyllum Desf. extract directly affected growing rat chondrocytes by reducing viability, alkaline phosphatase activity, and GAG synthesis without altering the expression of gene transcripts for Sox9, Col2, ColX, and aggrecan, which may be one of the mechanisms by which there is a reduction in bone growth in animals intoxicated by the plant.

Chemometric Analysis for the Prediction of Biochemical Compounds in Leaves Using UV-VIS-NIR-SWIR Hyperspectroscopy.

Reflectance hyperspectroscopy is recognised for its potential to elucidate biochemical changes, thereby enhancing the understanding of plant biochemistry. This study used the UV-VIS-NIR-SWIR spectral range to identify the different biochemical constituents in Hibiscus and Geranium plants. Hyperspectral vegetation indices (HVIs), principal component analysis (PCA), and correlation matrices provided in-depth insights into spectral differences. Through the application of advanced algorithms-such as PLS, VIP, iPLS-VIP, GA, RF, and CARS-the most responsive wavelengths were discerned. PLSR models consistently achieved R2 values above 0.75, presenting noteworthy predictions of 0.86 for DPPH and 0.89 for lignin. The red-edge and SWIR bands displayed strong associations with pivotal plant pigments and structural molecules, thus expanding the perspectives on leaf spectral dynamics. These findings highlight the efficacy of spectroscopy coupled with multivariate analysis in evaluating the management of biochemical compounds. A technique was introduced to measure the photosynthetic pigments and structural compounds via hyperspectroscopy across UV-VIS-NIR-SWIR, underpinned by rapid multivariate PLSR. Collectively, our results underscore the burgeoning potential of hyperspectroscopy in precision agriculture. This indicates a promising paradigm shift in plant phenotyping and biochemical evaluation.

Kisspeptin (Kp-10) inhibits in vitro osteogenic differentiation of multipotent mesenchymal stromal cells extracted from the bone marrow of adult rats.

Kisspeptin (Kp-10) is a neuropeptide that binds to GPR54 receptors, exerting several functions mainly in the nervous and reproductive systems of the body. However, its effects and mechanisms of action on the skeletal system remain poorly understood. This study evaluated the effects of different concentrations of Kp-10 on in vitro osteogenic differentiation of multipotent mesenchymal stromal cells (MSCs) extracted from the bone marrow (BM) of adult Wistar rats. Two-month-old female rats were euthanized to extract BM from long bones to obtain MSCs. Four experimental groups were established in vitro: a control and Kp-10 at concentrations of 0.01, 0.05 and, 0.1 µg/mL. After induction of osteogenic differentiation, cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide (MTT) assay, alkaline phosphatase activity, collagen synthesis, percentage of area covered by MSCs/field and mineralized nodules/field, and immunocytochemistry of the GPR54 receptor tests. Furthermore, evaluation of gene transcripts for type I collagen, Runx-2, Bmp-2, bone sialoprotein, osteocalcin and osteopontin was performed using real-time RT-qPCR. It was observed that MSCs expressed GPR54 receptor to which Kp-10 binds during osteogenic differentiation, promoting a negative effect on osteogenic differentiation. This effect was observed at all the Kp-10 concentrations in a concentration-dependent manner, characterized by a decrease in the activity of alkaline phosphatase, collagen synthesis, mineralized nodules, and decreased expression of gene transcripts for type I collagen, osteocalcin, osteopontin, and Runx-2. Thus, Kp-10 inhibits in vitro osteogenic differentiation of MSCs extracted from the BM of adult Wistar rats.

Treatment of critical defects produced in calvaria of mice with mesenchymal stem cells.

Mesenchymal stem cells (MSC) are present in specialized niches in perivascular regions of adult tissues and are able to differentiate into various cell types, such as those committed to repairing. Bone marrow derived MSC from eight young mice C57BL/ 6 gfp(+) were expanded in culture for repairing critical defects in calvarial bone produced in twenty-four young isogenic adult C57BL/6 mice. The animals were subjected to a cranial defect of 6.0mm diameter and divided into two equal experimental groups. Control group did not receive any treatment and the treated group received a MSC pellet containing 1.0 x 10(7) cells/mL into the defects. The group treated with MSC showed increased angiogenesis and amount of new bone deposited on the defect limits than that observed in the control group. The results demonstrated that transplantation of bone marrow-derived MSC of C57BL/6 gfp(+) mice to bone critical defects produced in mice calvarial contributes positively to the bone repair process. MSC presets ability to influence the correct functioning of osteoblasts, increases the amount of mobilized cells for the repairing process, speeds up growth, and increases deposition of bone matrix.

Reactive treponemal and non-treponemal tests in pregnant women and associated factors.

OBJECTIVE: to identify the rate of reactive treponemal and non-treponemal tests in pregnant women during childbirth and to analyze the factors associated with this seroreactivity. METHOD: this is a cross-sectional, quantitative study with secondary sources of sociodemographic and clinical data on 2,626 pregnant women treated at a public maternity hospital in the interior of São Paulo, in 2020. For statistical analysis, Fisher's exact test, Mann-Whitney test and the logistic regression model were used. A difference of p < 0.05 was considered statistically significant. RESULTS: the rate of seropositivity for syphilis among pregnant women in this series was 2.74%. Among the groups with positive and non-reactive tests, marital status, occupation, place of residence and use of licit drugs indicated significant differences, but, in the final model, only unmarried marital status was associated with reactive tests (Odds Ratio: 0.169; Confidence Interval: 0.04-0.72; and p: 0.016). CONCLUSION: in this study, unmarried marital status was the only independent factor associated with seroreactivity for syphilis. Therefore, it is necessary to create strategies aimed at women in this condition, potentially reducing the rate of congenital syphilis.

Maternal Hyperthyroidism in Rats Alters the Composition and Gene Expression of the Matrix Produced In Vitro by Chondrocytes from Offspring with Intrauterine Growth Restriction.

Herein, we aimed to evaluate cultures of femoral chondrocytes from offspring of rats with intrauterine growth restriction (IUGR) induced by maternal hyperthyroidism. Fourteen adult female Wistar rats were divided into two groups, a control group and a group treated with daily L-thyroxine administration using an orogastric tube (50 µg/animal/day) during pregnancy. Three days after birth, the offspring were euthanized for chondrocyte extraction. At 7, 14, and 21 days, viability and alkaline-phosphatase (ALP) activity were assessed using the MTT assay and BCIP/NBT method, respectively, in a 2D culture. Pellets (3D cultures) were stained with periodic acid Schiff (PAS) to assess the morphology and percentage of PAS+ areas. The gene transcripts for Col2, Col10, Acan, Sox9, and Runx2 were evaluated by qRT-PCR. The MTT and ALP-assay results showed no significant differences between the groups. Maternal hyperthyroidism did not alter the chondrocyte morphology, but significantly reduced the percentage of PAS+ areas, decreased the expression of the gene transcripts of Col2 and Acan, and increased Sox9 expression. Maternal hyperthyroidism in rats alters the composition and gene expression of the matrix produced by chondrocytes from offspring with IUGR. This may be one of the mechanisms through which excess maternal thyroid hormones reduce offspring bone growth.

Factors associated with inadequate treatment of syphilis during pregnancy: an integrative review.

OBJECTIVES: to analyze the evidence available in literature on factors associated with inadequate treatment of syphilis in pregnant women. METHODS: an integrative review, carried out in the LILACS, CINAHL, Web of Science, Scopus, PubMed and EMBASE databases, with controlled descriptors therapeutic and prenatal syphilis. RESULTS: nine publications composed the interpretative analysis, in which low education, income and maternal age, temporary lack of medication and HIV infection were associated with inadequate treatment of syphilis during pregnancy, in addition to delay or absence of prenatal care and receiving the 1st dose of penicillin, lack of tests or treatment less than 30 days before childbirth, and partners' low compliance with treatment. FINAL CONSIDERATIONS: among the main factors associated with inadequate treatment, clinical and sociodemographic aspects stand out, as well as failures in drug dispensing, prescription and monitoring of treatment of pregnant women and their partners by the health system.