Immunohistochemistry Screening of Different Tyrosine Kinase Receptors in Canine Solid Tumors-Part I: Proposal of a Receptor Panel to Predict Therapies.

The use of tyrosine kinase inhibitors (TKI) has been growing in veterinary oncology and in the past few years several TKI have been tested in dogs. However, different from human medicine, we lack strategies to select patients to be treated with each TKI. Therefore, this study aimed to screen different tumor subtypes regarding TKI target immunoexpression as a predictor strategy to personalize the canine cancer treatment. It included 18 prostatic carcinomas, 36 soft tissue sarcomas, 20 mammary gland tumors, 6 urothelial bladder carcinomas, and 7 tumors from the endocrine system. A total of 87 patients with paraffin blocks were used to perform immunohistochemistry (IHC) of human epidermal growth factor receptor 2 (HER-2), epidermal growth factor receptors 1 (EGFR1), vascular endothelial growth factor receptor 2 (VEGFR-2), platelet derived growth factor receptor beta (PDGFR-ß), c-KIT, and extracellular signal-regulated kinase 1/2 (ERK1/ERK2). The immunohistochemical screening revealed a heterogeneous protein expression among histological types with mesenchymal tumors showing the lowest expression level and carcinomas the highest expression. We have demonstrated by IHC screening that HER2, EGFR1, VEGFR-2, PDGFR-ß and ERK1/ERK2 are commonly overexpressed in dogs with different carcinomas, and KIT expression is considered relatively low in the analyzed samples.

Clinicopathological Characteristics and the First Mutational Analysis of Gastrointestinal Stromal Tumors From Mexico: A Single Institution Experience.

Background Gastrointestinal stromal tumors (GISTs) arise from Cajal's interstitial cell precursors and display a variety of genetic mutations, primarily in the KIT and PDGFRA genes. These mutations are linked to tumor location, prognosis, and response to treatment. This study delves into the mutational patterns of GISTs in a Mexican population and their impact on overall survival (OS) and disease-free survival (DFS). Methodology This retrospective study examined 42 GIST cases diagnosed at the Oncology Hospital of the National Medical Center XXI Century between January 2018 and December 2020. Clinical, histological, and immunohistochemical data were gathered, and mutational analysis of KIT and PDGFRA genes was conducted using second-generation sequencing. Results The study group consisted of 52.4% females and 47.6% males, with an average age of 62.6 years. The most common tumor site was the stomach (59.5%), followed by the small intestine (26.2%). KIT mutations were detected in 71.4% of cases, predominantly involving exon 11. PDGFRA mutations were observed in 7.1% of cases. Recurrence was noted in 9.5% of patients, all with high-risk tumors. No significant link was identified between specific mutations and OS or DFS. Conclusions This investigation sheds light on the genetic landscape of GISTs in the Mexican population. While no significant association was established between particular mutations and survival outcomes, the study emphasizes the importance of molecular profiling in treatment decision-making. Further studies with larger sample sizes and longer follow-up periods are necessary to validate these results and explore their clinical relevance.

Effects of plasma rich in growth factors on wound healing in patients with venous ulcers.

Introduction: Significant evidence suggests that plasma-rich in growth factors (PRGF) favor the repair of chronic wounds, enabling a rapid return to functionality. However, components of PRGF and their effects on persistent ulcers and epithelial tissues are not well characterized. The goals of this research were to analyze the biological properties of platelet-derived factors, to examine their effectiveness on healing of venous ulcers, and to establish a correlation with clinical and sociodemographic data. Methods: For the preparation of PRGF, the centrifugation technique was used, obtaining a 100 % autologous and biocompatible blood sample that was treated with sodium citrate and calcium chloride. The patients were attended weekly at the outpatient clinic for nursing consultation and wound dressing changes, with PRGF application every 15 days. The treatment protocols are described, and follow-up results are reported. Results: Initially, the patients' ulcers ranged in sizes from 4 to 84 cm2. After 12 weeks of treatment, there was a significant mean reduction of 46.2 % in ulcer area. At baseline, epithelial tissue was absent in all venous ulcers, but its presence grew significantly by the treatment period. However, the reduction of the area of the ulcers did not show significant correlation with the concentrations of the patient's growth factors. Conclusions: Using the established protocol for PRGF isolating, it was possible to obtain a product with the presence of the six growth factors related to tissue regeneration and observed a positive response on wound healing following treatment of venous ulcers, with capacity to accelerate re-epithelialization and restore the skin functional integrity.

Alterations of Plasmatic Biomarkers of Neurodegeneration in Mucopolysaccharidosis Type II Patients Under Enzyme Replacement Therapy.

Mucopolysaccharidosis type II (MPS II) is a disorder caused by a deficient activity of iduronate-2-sulfatase, a lysosomal enzyme responsible for degrading glycosaminoglycans (GAGs). The abnormal storage of GAGs within lysosomes disrupts cellular homeostasis and leads to a severe symptomatology. Patients present neuropsychiatric impairment characterized by mental retardation and impaired cognition. The aim of this study was to quantify four neurodegeneration biomarkers in plasma: brain-derived neurotrophic factor (BDNF), platelet-derived growth factor (PDGF-AA), neural cell adhesion molecule (NCAM) and cathepsin-D, as well as to identify possible correlations with urinary GAGs in seven patients undergoing treatment with ERT (Elaprase® 0.5 mg/kg of body weight). Patients with both severe and attenuated forms of MPS II showed signs of neurodegeneration in neuroimaging exams. Patients have a decrease in BDNF and PDGF-AA concentrations, and an increase in NCAM level compared to controls. No alterations in cathepsin-D concentration were seen. GAGs levels were higher in patients than in controls, but no significant correlations between GAGs and biomarkers were observed. These results evidence that patients have neurodegeneration and that monitoring these biomarkers might be useful for assessing this process. To this date, this is the first work to analyze these plasmatic markers of neurodegeneration in patients.

The effect of cilostazol on the platelet-derived growth factor-beta/beta isoform reduction on venous hyperplasia in an experimental balloon-induced injury model.

BACKGROUND: Intimal hyperplasia is the response to endothelial injury. Platelet-derived growth factor is released early and favors the formation of intimal hyperplasia. Although multiple treatments, from open surgery to endovascular techniques, have been used they remain controversial. There is currently interest in developing pharmacological strategies to address this pathology. Local vascular inflammation induced by vessel barotrauma generates intimal hyperplasia due to mechanical stress over the venous endothelium. Cilostazol is a selective phosphodiesterase type 3 (PDE3) selective inhibitor with a regulatory effect over intimal hyperplasia. The objective was to investigate cilostazol's role in inhibiting smooth muscle cell proliferation due to changes in the expression and release of PDGF-BB isoform and the effect on developing IH using an experimental model of vascular barotrauma (balloon-induced injury model). METHODS: We included 12 New Zealand rabbits. The balloon-induced injury model (BIIM) and experimental group cilostazol (20 mg/kg/day) included 6 rabbits each. Contralateral veins from 6 rabbits used in BIIM model has been taken as control group. We measured and compared the expression of PDGF-BB and the development of IH. A pathologist board chooses a PDGFRα antibody to localized its expression by immunohistochemistry analysis. Subsequently, using an automated immunohistochemical staining machine, the PDGFR expression was evaluated using a Zeiss Primo Star 4 light microscope. RESULTS: The measurement obtained in the intimal layer was: 126.12 µm2 in the CG, 232 µm2 in the BIIM group, and 178 µm2 in the EG. A statistically significant difference was observed. Baseline serum concentrations of PDGF-BB in the BIIM group were 0.22 pg/mL. At 12 h 0.42 pg/mL, and 0.17 pg/mL at seven days. In the experimental group, the basal levels were 0.33 pg/mL. With the use of cilostazol, a lower peak was obtained at 12 h (0.08 pg/mL). This difference was statistically significant. CONCLUSIONS: Cilostazol induced a significant reduction of IH caused by barotrauma in the venous endothelium, which correlates with decrease in the PDGF-BB in serum. This could be attributed to the pharmacologic effect on PDGFR expression.

Effectiveness of Platelet-Rich Fibrin with Decalcified Freeze-Dried Bone Allograft Compared to Decalcified Freeze-Dried Bone Allograft Alone in Mandibular Grade-II Furcation Defects: A Quasi-Experimental Study

ABSTRACT Objective: To assess the effectiveness of platelet-rich fibrin (PRF) with decalcified freeze-dried bone allograft (DFDBA) compared to DFDBA alone in mandibular grade-II furcation defects. Material and Methods: A quasi-experimental study was conducted on nine patients with chronic periodontitis, each having two almost identical mandibular grade II furcation defects. Test sites (left mandibular first molars) were treated with open flap debridement (OFD), DFDBA, and PRF, whereas control sites (right mandibular first molars) received OFD and DFDBA alone. Clinical parameters (plaque index (PI), gingival index (GI), vertical clinical attachment level (VCAL) and horizontal clinical attachment level (HCAL) into the furcation defect) and radiographic measurements (mean alveolar bone defect) were done at baseline and after six months postoperatively. Results: The gain in relative horizontal clinical attachment level (RHCAL) in the test sites was 2.94±0.52 mm compared to 1.33±0.35 mm in control sites (p=0.01). Improvement in mean alveolar bone defect (MABD) (was 1.21±0.5 mm2 at test sites compared to 1.15±0.7 mm2 at control sites) probing pocket depth (PPD), recession, relative vertical attachment level (RVCAL), and percentage of bone fill was found in the test sites compared to control, which statistically insignificant. Conclusion: The test sites had better outcomes than control sites, which was significant for the parameter RHCAL. Therefore, combining the biological benefits of autologous PRF with DFDBA is an efficient and economical treatment modality for the management of mandibular grade II furcation defects.

Dermatofibrosarcoma protuberans with unusual presentation in vulva / Dermatofibrosarcoma protuberans con presentación inusual en vulva

Abstract Gynecological sarcomas are uncommon and their location in the vulva and vagina has an incidence of 5% of all malignant neoplasms of the female genital tract. We present the case of a 54-year-old patient with a diagnosis of dermatofibrosarcoma protuberans in the vulva, an infrequent pathology, with less than 60 cases reported worldwide in this anatomical location. Clinically it has a locally aggressive behavior, due to the proliferation of spindle cells with pleomorphism and frequent figures of mitosis that infiltrate the reticular dermis and subcutaneous cellular tissue, giving rise to tumor lesions of variable size and with high rates of local recurrence. The treatment of first choice is surgical excision of the tumor with Mohs micrographic surgery or other surgical techniques for complete evaluation of the circumferential and deep peripheral margin. However, the identification of carcinogenesis mechanis ms where the chromosomal translocation t (17; 22) (q22; q13) is recognized, forming the COL1A1-PDGFB fusion gene, which participates in stimulating tumor cell proliferation, allowing treatment with tyrosine kinase inhibitors such as imatinib for neoadjuvant therapy of surgically unresectable tumors and local recurrences.

Resumen Los sarcomas ginecológicos son infrecuentes y la localización de estos en vulva y vagina tienen una incidencia del 5% de todas las neoplasias malignas del tracto genital femenino. Presentamos el caso de una paciente de 54 años con diagnóstico de dermatofibrosarcoma protuberans en vulva, una patología infrecuente, con menos de 60 casos reportados a nivel mundial en esta localización anatómica. Clínicamente tiene un comportamiento localmente agresivo, debido a la proliferación de células fusiformes con pleomorfismo y frecuentes figuras de mitosis que infiltran la dermis reticular y tejido celular subcutáneo, dando origen a lesiones tumorales de tamaño variable y con altas tasas de recurrencia local. El tratamiento en primera elección es la escisión quirúrgica del tumor con cirugía micrográfica de Mohs u otras técnicas quirúrgicas para evaluación completa del margen periférico circunferencial y profundo. Sin embargo, la identificación de mecanismos de carcinogénesis donde se reconoce la translocación cromosómica t (17; 22) (q22; q13), formando al gen de fusión COL1A1-PDGFB, el cual participa estimulando la proliferación celular tumoral, ha permitido la utilización de los inhibidores de la tirosina quinasa como el imatinib para la realización de terapia neoadyuvante en casos de tumores irresecables quirúrgicamente y en recurrencias locales.

Effects of Platelet Rich Plasma on Pain in Adult and Elderly Patients with Knee Osteoarthritis: a Systematic Review / Efeitos do Plasma Rico em Plaquetas na Dor em Pacientes Adultos e Idosos com Osteoartrite de Joelho: uma Revisão Sistemática

Platelet-rich plasma (PRP) has been shown to be a promising treatment for osteoarthritis (OA) of the knee in adult and elderly patients, acting in the repair of joint tissues damaged by OA - a degenerative disease that mainly affects articular cartilage. This process of degradation of joint components leads to an increase in the level of pain in these patients. Therefore, this study aimed to evaluate the effectiveness of intra-articular injection of PRP in knee OA in relation to pain. For this systematic review, five databases were used: PubMed, Lilacs, PEDro, Cochrane Library and Google Scholar. Search strategies included the keywords Osteoarthritis, Knee, Platelet-rich plasma, and Pain. In order to be included in this review, the studies had to be of the randomized clinical trial type; use as a sample people between 18 and 80 years old diagnosed with knee OA; use intra-articular PRP treatment compared to placebo; and assess pain intensity. After completion of the selection phase, 5 studies were included in this review. These showed positive results in reducing pain in patients with lower-degeneration OA, and less effective in advanced OA. Furthermore, few side effects were reported after the application of PRP. Therefore, the intra-articular application of PRP can be an important tool for the treatment of knee OA, as it is an effective therapy in decreasing pain levels in patients affected by this pathology. (AU)

O plasma rico em plaquetas (PRP) tem se mostrado um tratamento promissor na osteoartrite (OA) de joelho de pacientes adultos e idosos, atuando no reparo dos tecidos articulares lesados pela OA - doença degenerativa que afeta principalmente a cartilagem articular. Esse processo de degradação dos componentes articulares leva ao aumento do nível de dor nesses pacientes. Sendo assim, este estudo objetivou avaliar a eficácia da injeção intra-articular de PRP na AO de joelho em relação a dor. Para esta revisão sistemática, cinco bases de dados foram utilizadas: PubMed, LILACS, PEDro, Cochrane Library e Google Scholar. As estratégias de busca incluíram as palavras-chave Osteoarthritis, Knee, Platelet-rich plasma e Pain. Para serem incluídos nesta revisão, os estudos deveriam ser do tipo ensaio clínico randomizado; utilizar como amostra pessoas entre 18 e 80 anos diagnosticadas com AO de joelho; utilizar o tratamento com PRP intra articular comparado ao placebo; e avaliar a intensidade da dor. Após o término da fase de seleção, 5 estudos foram incluídos nesta revisão. Estes apresentaram resultados positivos na redução da dor em pacientes com OA de menor grau de degeneração, e menor efetividade na OA avançada. Ademais, poucos efeitos colaterais foram relatados após a aplicação do PRP. Portanto, a aplicação intra-articular do PRP pode ser um importante instrumento para o tratamento da OA de joelhos, visto que é uma terapia eficaz na diminuição dos níveis dor dos pacientes acometidos por tal patologia. (AU)

Plasma rich in growth factors as treatment for a full-thickness macular hole due to macular telangiectasia type 2.

Full-thickness macular hole is a rare complication of macular telangiectasia type 2, and its treatment is still controversial. A patient with a full-thickness macular hole secondary to macular telangiectasia type 2 underwent vitreoretinal surgery with a plasma rich in growth factors membrane in the macular hole. At the sixth month of follow-up, anatomical and functional improvements were achieved, with no adverse effects. Plasma rich in growth factors is a new option, with advantages due to its biological properties that achieves good results in terms of safety and effectiveness in the surgical treatment of macular hole.

Dermatofibrosarcoma protuberans with unusual presentation in vulva.

Gynecological sarcomas are rare and their location in the vulva and vagina has an incidence of 5% of all malignant neoplasms in the female genital tract. We present the case of a 54-year-old patient with a diagnosis of dermatofibrosarcoma protuberans in the vulva, an infrequent pathology with less than 60 cases reported worldwide in this anatomical location. Clinically, it is locally aggressive, due to the proliferation of spindle cells with pleomorphism and frequent mitotic figures infiltrating the reticular dermis and subcutaneous cellular tissue, giving rise to variable size tumors with high local recurrence rates. The first-line treatment is surgical excision of the tumor with Mohs micrographic surgery among other surgical techniques for complete circumferential peripheral and deep margin assessment. However, identification of carcinogenesis mechanisms where the chromosomal translocation t (17; 22) (q22; q13) is recognized, forming the COL1A1-PDGFB fusion gene, which participates in stimulating tumor cell proliferation, allowing treatment with tyrosine kinase inhibitors such as imatinib for neoadjuvant therapy of surgically unresectable tumors and local recurrences.

Los sarcomas ginecológicos son infrecuentes y la localización de estos en vulva y vagina tienen una incidencia del 5% de todas las neoplasias malignas del tracto genital femenino. Presentamos una paciente de 54 años con diagnóstico de dermatofibrosarcoma protuberans en vulva, el cual es una patología infrecuente con menos de 60 casos reportados a nivel mundial en esta localización anatómica. Clínicamente tiene un comportamiento localmente agresivo, debido a la proliferación de células fusiformes con pleomorfismo y frecuentes figuras de mitosis que infiltran la dermis reticular y tejido celular subcutáneo, dando origen a lesiones tumorales de tamaño variable y con altas tasas de recurrencia local. El tratamiento en primera elección es la escisión quirúrgica del tumor con técnicas como cirugía micrográfica de Mohs u otras técnicas quirúrgicas para evaluación completa del margen periférico circunferencial y profundo. Sin embargo, la identificación de mecanismos de carcinogénesis donde se reconoce la translocación cromosómica t (17; 22) (q22; q13), formando al gen de fusión COL1A1-PDGFB el cual participa estimulando la proliferación celular tumoral, ha permitido la utilización de los inhibidores de la tirosina quinasa como el imatinib para la realización de terapia neoadyuvante en casos de tumores irresecables quirúrgicamente y en recurrencias locales.

Gestational and Lactational Iron Deficiency Anemia Impairs Myelination and the Neurovascular Unit in Infant Rats.

Iron deficiency anemia is a prevalent health problem among pregnant women and infants, particularly in the developing countries that causes brain development deficits and poor cognitive outcomes. Since tissue iron depletion may impair myelination and trigger cellular hypoxic signaling affecting blood vessels, we studied myelination and the neurovascular unit (NVU) in infant rats born to mothers fed with an iron deficient (ID) or control diet from embryonic day 5 till weaning. Blood samples and brains of rat pups at postnatal day (PND) 14 and 30 were analyzed. PND 14 ID rats had severe microcytic hypochromic anemia that was almost reversed at PND 30 although hypomyelination and astrocyte immature phenotype in the corpus callosum were significant at that age. In CA1 hippocampal region, PND 14 and PND 30 ID rats showed significant reduced expression of the receptor ß of the platelet-derived growth factor localized in pericytes and associated to aquaporin 4 (AQP4) immunopositive capillaries. Shorter AQP4 + capillaries and reduced AQP4 expression were also evidenced in PND 14 and PND 30 ID rats. In addition, pericyte membrane permeability through large-pore channels was transiently increased in ID rats at PND 14 but not at PND 30, while the blood-brain barrier permeability was not affected. Remarkably, transient increased pericyte permeability found in PND 14 ID rats was not directly related to iron depletion, suggesting the involvement of other iron deficiency anemia-induced mechanisms. In summary, severe ID during gestation and lactation produces persistent hypomyelination and significantly affects hippocampal pericytes and astrocytes in the NVU which may trigger impaired neurovascular function.

Microarray and bioinformatic analysis of conventional ameloblastoma: an observational analysis

Abstract Ameloblastoma is a highly aggressive odontogenic tumor, and its pathogenesis is associated with many participating genes. Objective We aimed to identify and validate new critical genes of conventional ameloblastoma using microarray and bioinformatics analysis. Methodology Gene expression microarray and bioinformatic analysis were performed using CHIP H10KA and DAVID software for enrichment. Protein-protein interactions (PPI) were visualized using STRING-Cytoscape with MCODE plugin, followed by Kaplan-Meier and GEPIA analyses that were used for the candidate's postulation. RT-qPCR and IHC assays were performed to validate the bioinformatic approach. Results 376 upregulated genes were identified. PPI analysis revealed 14 genes that were validated by Kaplan-Meier and GEPIA resulting in PDGFA and IL2RA as candidate genes. The RT-qPCR analysis confirmed their intense expression. Immunohistochemistry analysis showed that PDGFA expression is parenchyma located. Conclusion With bioinformatics methods, we can identify upregulated genes in conventional ameloblastoma, and with RT-qPCR and immunoexpression analysis validate that PDGFA could be a more specific and localized therapeutic target.

Experiencia con el uso de plasma rico en plaquetas autólogo en el tratamiento de las úlceras del pie diabético / Use of platelet rich plasma in the treatment of diabetic foot ulcers. Report of the experience / Experiência com o uso de plasma autólogo rico em plaquetas no tratamento de úlceras do pé diabético

Resumen: Las úlceras de pie diabético (UPD) generan un alto costo para el paciente y el sistema de salud. Una deficiente vascularización, la inhibición de la generación de factores de crecimiento y la migración celular, entre otros factores, influyen en su cronicidad. Los factores de crecimiento derivados de las plaquetas (PDGF) estimulan la quimiotaxis, la neovascularización y la regeneración tisular. Objetivo: evaluar la efectividad de una formulación de plasma rico en plaquetas autólogo (PRP) como una modalidad complementaria para el tratamiento de UPD, valorando su seguridad y efectos adversos. Materiales: estudio observacional. Se aplicó PRP a seis pacientes con UPD crónicas, Wagner II, III, con una media en el retraso de la cicatrización de 94 semanas. Se administró mediante punción y como gel sobre la lesión una vez por semana hasta el cierre de la úlcera o durante12 semanas. Las UPD se analizaron con respecto al área y su progresión mediante Mobile Wound Analyzer (MOWA), software de análisis de imágenes de úlceras. El porcentaje de reducción del área se calculó entre la medición inicial y la semana 12. Como criterio de mejoría se eligió una reducción >70% de la lesión. Resultados: predominó el sexo masculino con una media de edad de 53 años ± 7,3. Luego de 12 semanas de tratamiento, se reportó una reducción media del tamaño de la úlcera de 78%, con dos cicatrizaciones completas y dos en 99%. Un paciente recibió una amputación luego de presentar infección no controlada. Conclusión: la aplicación del PRP mejoró la regeneración de tejidos, acortando la duración de la úlcera, promoviendo su curación sin efectos adversos, al tiempo que eliminó la necesidad de procedimientos de manejo en una úlcera crónica.

Summary: Diabetic foot ulcers imply high expenditure for both patients and the health system. Vascular impairment, growth factor inhibition and cell migration, among other factors, affect the chronicity of the condition. Platelet-derived growth factors (PDGFs) stimulate chemotaxis, neovascularization and tissue regeneration. Objective: to evaluate effectiveness of a formulation of autologous platelet-rich plasma to complement the treatment of diabetic foot ulcer, assessing safety and adverse effects. Material: observational study. Platelet rich plasma was applied to six patients with chronic diabetic foot ulcer Wagner II, III, with an average delayed healing of 94 weeks. PRP gel was administered by puncture technique on the lesion, once a week until the ulcer healed or for 12 weeks. Diabetic foot ulcers were analysed in regards to surface area and progression through the Mobile Wound Analyzer (MOWA) software to analyse ulcer images. The area reduction percentage was calculated comparing the initial measurement and that of week 12. The improvement criteria was defined in a reduction that is >70% of the lesion. Results: men prevailed with an average age of 53 years ± 7.3. After 12 weeks of treatment an average reduction of 78% of the size of the ulcer was reported, two complete healings and two 99% healed. One patient required amputation because of uncontrolled infection. Conclusion: the application of PRP improved tissue regeneration and shortened the duration of the ulcer, promoting healing with no adverse effects, and it likewise eliminated the need for procedures to handle chronic ulcers.

Resumo: As úlceras do pé diabético (UPD) geram alto custo para o paciente e para o sistema de saúde. A vascularização deficiente, a inibição da geração de fatores de crescimento e migração celular, entre outros fatores, influenciam sua cronicidade. Os fatores de crescimento derivados de plaquetas (PDGF) estimulam a quimiotaxia, a neovascularização e a regeneração de tecidos. Objetivo: avaliar a eficácia de uma formulação autóloga de plasma rico em plaquetas (PRP) como modalidade complementar para o tratamento da UPD, avaliando sua segurança e efeitos adversos. Materiais: estudo observacional. O PRP foi aplicado a 6 pacientes com UPD crônica, Wagner II, III, com um atraso médio na cicatrização de 94 semanas. Foi administrado por punção e como gel sobre a lesão uma vez por semana até o fechamento da úlcera ou por 12 semanas. As UPD foram analisadas em relação à área e sua progressão, utilizando o software de análise de imagem de úlcera Mobile Wound Analyzer (MOWA). A redução percentual da área foi calculada entre a medição inicial e a semana 12. Como critério de melhora, optou-se pela redução > 70% da lesão. Resultados: predominou o sexo masculino com média de idade de 53 ± 7,3 anos. Após 12 semanas de tratamento, foi relatada uma redução média no tamanho da úlcera de 78%, sendo 2 com cicatrização completa e 2 em 99%. Um paciente foi amputado após desenvolver infecção descontrolada. Conclusão: a aplicação do PRP melhorou a regeneração tecidual, encurtando o tempo de duração da úlcera, promovendo sua cicatrização sem efeitos adversos, ao mesmo tempo que eliminava a necessidade de procedimentos de manejo na úlcera crônica.

Plasma rich in growth factors as treatment for a full-thickness macular hole due to macular telangiectasia type 2. / Plasma rico en factores de crecimiento como tratamiento para agujero macular de espesor total secundario a telangiectasia macular tipo 2.

Full-thickness macular hole is a rare complication of macular telangiectasia type 2, and its treatment is still controversial. A patient with a full-thickness macular hole secondary to macular telangiectasia type 2 underwent vitreoretinal surgery with a plasma rich in growth factors membrane in the macular hole. At the sixth month of follow-up, anatomical and functional improvements were achieved, with no adverse effects. Plasma rich in growth factors is a new option, with advantages due to its biological properties that achieves good results in terms of safety and effectiveness in the surgical treatment of macular hole.

Effect of platelet-derived growth factor (PDGF-BB) and bone morphogenic protein 2 (BMP-2) transfection of rBMSCs compounded with platelet-rich plasma on adipogenic differentiation

The aim of this study was to inhibit adipogenic differentiation by transfecting two growth factors, platelet-derived growth factor (PDGF-BB) and bone morphogenic protein 2 (BMP-2), into modified rat bone marrow mesenchymal stem cells (rBMSCs) and then compounded with platelet-rich plasma (PRP). To achieve rBMSCs, the osteoporosis model of rats was established, and then the rBMSCs from the rats were isolated and identified. Co-transfection of rBMSCs with PDGF-BB-GFP and BMP-2 and detection of PDGF-BB/BMP-2 expression in transfected BMSCs was assessed by qRT-PCR and western blot, respectively. Moreover, the effect of the two growth factors transfection of rBMSCs on adipogenic differentiation was evaluated by oil red O staining and western blot, respectively. Finally, construction of the two growth factors transfection of rBMSCs compounded with PRP and detection of adipogenic differentiation were assessed by oil red O staining, CCK-8, and western blot, respectively. In vitro studies revealed that the two growth factors transfection of rBMSCs compounded with PRP promoted cell viability and inhibited adipogenic differentiation and could be promising for inhibiting adipogenic differentiation.

Angiotensin-(1-9) prevents vascular remodeling by decreasing vascular smooth muscle cell dedifferentiation through a FoxO1-dependent mechanism.

The renin-angiotensin system, one of the main regulators of vascular function, controls vasoconstriction, inflammation and vascular remodeling. Antagonistic actions of the counter-regulatory renin-angiotensin system, which include vasodilation, anti-proliferative, anti-inflammatory and anti-remodeling effects, have also been described. However, little is known about the direct effects of angiotensin-(1-9), a peptide of the counter-regulatory renin-angiotensin system, on vascular smooth muscle cells. Here, we studied the anti-vascular remodeling effects of angiotensin-(1-9), with special focus on the control of vascular smooth muscle cell phenotype. Angiotensin-(1-9) decreased blood pressure and aorta media thickness in spontaneously hypertensive rats. Reduction of media thickness was associated with decreased vascular smooth muscle cell proliferation. In the A7r5 VSMC cell line and in primary cultures of rat aorta smooth muscle cells, angiotensin-(1-9) did not modify basal proliferation. However, angiotensin-(1-9) inhibited proliferation, migration and contractile protein decrease induced by platelet derived growth factor-BB. Moreover, angiotensin-(1-9) reduced Akt and FoxO1 phosphorylation at 30 min, followed by an increase of total FoxO1 protein content. Angiotensin-(1-9) effects were blocked by the AT2R antagonist PD123319, Akt-Myr overexpression and FoxO1 siRNA. These data suggest that angiotensin-(1-9) inhibits vascular smooth muscle cell dedifferentiation by an AT2R/Akt/FoxO1-dependent mechanism.

Pericyte in Oral Squamous Cell Carcinoma: A Systematic Review.

The microenvironment of oral cancer is highly dynamic and has been proved to affect tumor progression. Pericytes are blood vessels surrounding cells that have recently gained attention for their roles in vascular and cancer biology. The objective of the present study was to survey the scientific literature for conclusive evidence about whether pericytes are part of blood vessels in oral squamous cell carcinoma (OSCC) and their roles in the tumor microenvironment and clinical outcomes. A systematic electronic search was undertaken in Medline Ovid, PubMed, Web of Science, and Scopus. Eligibility criteria were: publications adopting in vivo models of OSCC that included pericyte detection and assessment by pericyte markers (e.g., α-smooth muscle actin, neuron-glial antigen 2 and platelet-derived growth factor receptor-ß). The search yielded seven eligible studies (from 2008 to 2018). The markers most commonly used for pericyte detection were α-smooth muscle actin and neuron-glial antigen 2. The studies reviewed showed the presence of immature vessels exhibiting a reduction of pericyte coverage in OSCC and indicated that anti-cancer therapies could contribute to vessel normalization and pericyte regain. The pericyte population is significantly affected during OSCC development and cancer therapy. While these findings might suggest a role for pericytes in OSCC progression, the limited data available do not allow us to conclude whether they modify the tumor microenvironment and clinical outcome.

Platelet-derived growth factor B restores vascular barrier integrity and diminishes permeability in ovarian hyperstimulation syndrome.

Although advances in the prediction and management of ovarian hyperstimulation syndrome (OHSS) have been introduced, complete prevention is not yet possible. Previously, we and other authors have shown that vascular endothelial growth factor, angiopoietins (ANGPTs) and sphingosine-1-phosphate are involved in OHSS etiology. In addition, we have demonstrated that ovarian protein levels of platelet-derived growth factor (PDGF) ligands -B and -D decrease in an OHSS rat model, whilst PDGFR-ß and ANGPT2 remain unchanged. In the present work, we investigated the role of PDGF-B in OHSS by evaluating ligand protein levels in follicular fluid (FF) from women at risk of developing OHSS and by using an immature rat model of OHSS. We demonstrated that PDGF-B and PDGF-D are lower in FF from women at risk of developing OHSS compared to control patients (P < 0.05). In the OHSS rat model, PDGF-B (0.5 µg/ovary) administration decreased ovarian weight (P < 0.05), reduced serum progesterone (P < 0.05) and lowered the percentage of cysts (P < 0.05), compared to untreated OHSS rats, but had no effect on the proportion of follicles or corpora lutea (CL). PDGF-B treatment also restored the expression of steroidogenic acute regulatory protein (P < 0.05) and P450 cholesterol side-chain cleavage enzyme (P < 0.01) to control levels. In addition, PDGF-B increased the peri-endothelial cell area in CL and cystic structures, and reduced vascular permeability compared to untreated OHSS ovaries. Lastly, PDGF-B increased the levels of junction proteins claudin-5 (P < 0.05), occludin (P < 0.05) and ß-catenin (P < 0.05), while boosting the extracellular deposition of collagen IV surrounding the ovarian vasculature (PP < 0.01), compared to OHSS alone. In conclusion, our findings indicate that PDGF-B could be another crucial mediator in the onset and development of OHSS, which may lead to the development of novel prediction markers and therapeutic strategies.

Uso do hormônio do crescimento associado à fibrina rica em plaquetas e leucócitos injetável (I-PRF) / Use of the growth hormone associated with injectable platelet-rich fibrin (I-PRF)

Objetivo: demonstrar, por meio de uma revisão de literatura, a utilização do hormônio do crescimento (GH) e concentrados plaquetários e sugerir técnica de associação de uso para odontologia em processos de preservação de osso alveolar. Revisão de literatura: enxertos ósseos são uma necessidade na área da saúde, por diversas razões. A utilização de osso autógeno apresenta grande desvantagem em ter um segundo sítio cirúrgico, entretanto, os substitutos ósseos não possuem as características ideais. Assim, existe a busca por alternativas que otimizem a cicatrização e a incorporação dos substitutos ósseos, dentre elas os concentrados sanguíneos, ricos em fatores de crescimento derivados das plaquetas e o hormônio do crescimento. É possível encontrar uma vasta literatura utilizando os concentrados sanguíneos, inclusive utilizando esses como veículos para outras substâncias. Os concentrados sanguíneos são ricos em fatores de crescimento derivados das plaquetas, como fator de crescimento semelhante à insulina (IGF), Fator de crescimento derivado de plaquetas (PDGF) e outros. Além disso, também é possível encontrar, na literatura, o uso tópico de hormônio do crescimento em enxertos ósseos, fraturas e implantes dentários. Entretanto, o GH possui uma meia-vida de 20 minutos, assim, quando utilizado em conjunto com a I-PRF, espera-se um aumento no tempo de ação local. Considerações finais: é possível otimizar os enxertos ósseos utilizando-se L-PRF/I-PRF e hormônio do crescimento. Porém, são necessárias mais pesquisas.(AU)

Objective: this study aims to show through a literature review the use of the growth hormone and platelet concentrates and to suggest an association technique for dentistry use in alveolar bone preservation processes. Literature review: bone grafts are a health requirement for a number of reasons. The use of autogenous bone has the main disadvantage of a second surgical site, while bone substitutes do not present optimal characteristics. Thus, there is a search for alternatives that optimize the healing and incorporation of bone substitutes, which include blood concentrates that are rich in platelet-derived growth factors and the growth hormone. A vast literature can be found on blood concentrates, including their use as vehicles to other substances. Blood concentrates are rich in platelet-derived growth factors such as IGF, PDGF, and others. Moreover, the literature also shows the topical use of the growth hormone in bone grafts, fractures, and dental implants. However, the growth hormone presents a half-life of 20 minutes; therefore, when combined with I-PRF, an increased time in local action is expected. Final considerations: it is possible to optimize bone grafts by using L-PRF/I-PRF and the growth hormone. However, further research is required.(AU)

Adherent muscle connective tissue fibroblasts are phenotypically and biochemically equivalent to stromal fibro/adipogenic progenitors.

Extracellular matrix (ECM) gives structure, support, and is the niche for several cells found in skeletal muscle. ECM is mainly produced by muscle connective tissue (CT) fibroblasts during development and regeneration. Stromal fibroadipogenic progenitors (FAPs) are CT fibroblasts-like mesenchymal progenitors (MPs) with important roles in regeneration and degeneration. Chronic damage restrains the normal regenerative behavior of muscle fibroblasts/FAPs. Thus, the isolation and study of these mesenchymal progenitors are of crucial importance for understanding their behavior and biology. We investigated whether adult muscle CT fibroblasts (hereafter referred to as adherent fibroblasts [aFbs]) cultured via pre-plating strategy belong to a heterogeneous population of FAPs. By combining microscopy, western blot analyses, flow cytometry, and FACS we determined that aFbs isolated from skeletal muscle largely overlap with FAPs. In addition, we used the PDGFRαEGFP mice in order to corroborate our results with EGFP+ FAPs. Moreover, our strategy allows the isolation of activated EGFP+ FAPs from the murine DMD model PDGFRαEGFP; mdx and PDGFRαEGFP denervated mice. Here we report that 1 h 30 min of pre-plating strategy allows the isolation and culture of a highly enriched population of aFbs. These cells are phenotypically and biochemically a FAPs-like population of adherent cells. In addition, aFbs respond in the same fashion as FAPs to Nilotinib, an inducer of FAPs apoptosis. Moreover, flow cytometry characterization of these aFbs suggests that 85% of them express the MP marker PDGFRα, and isolation of aFbs from the PDGFRαEGFP mice suggests that 75% of them show high EGFP expression. Furthermore, TGF-ß1 induces aFbs proliferation, myofibroblast differentiation, and ECM production. We were also able to isolate activated aFbs from skeletal muscle of the DMD mice and from the PDGFRαEGFP mice 2-days after denervation. Our findings suggest that the in vitro pre-plating strategy allows the isolation and culture of a relatively pure aFbs population, which resembles FAPs in vitro.