Elenice Deffune

RESUMO Objetivo: Realizar um estudo de logística de implantação de negócio e viabilização da terapia com colírio de soro autólogo. Métodos: Foram identificados 177 serviços/clínicas/consultórios de oftalmologia na região de Botucatu (SP). Cinco médicos, que aceitaram participar da pesquisa, selecionaram e acompanharam pacientes com indicação para uso do colírio de soro autólogo, seguindo as exigências da análise sorológica para doenças transmissíveis pelo sangue. Os lotes foram personalizados para cada paciente e, na maioria, foram emitidos a cada 3 meses. Resultados: [...] RESUMO Objetivo: Realizar um estudo de logística de implantação de negócio e viabilização da terapia com colírio de soro autólogo. Métodos: Foram identificados 177 serviços/clínicas/consultórios de oftalmologia na região de Botucatu (SP). Cinco médicos, que aceitaram participar da pesquisa, selecionaram e acompanharam pacientes com indicação para uso do colírio de soro autólogo, seguindo as exigências da análise sorológica para doenças transmissíveis pelo sangue. Os lotes foram personalizados para cada paciente e, na maioria, foram emitidos a cada 3 meses. Resultados:

The possibility of chemical contamination is an important issue to consider when designing a cell therapy strategy. Both bisphenol A (BPA) and 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) are among the most environmentally relevant endocrine disrupting chemicals (EDCs, compounds with a high affinity for adipose tissue) recently studied. Adipose‐derived stem cells (ASCs) are mesenchymal stromal cells (MSCs) obtained from adipose tissue widely used in regenerative medicine to prevent and treat diseases in several tissues and organs. Although the experimental use of tissue‐engineered constructs requires careful analysis for approval and implantation, there has been a recent increase in the number of approved clinical trials for this promising strategy. This study aimed to evaluate cell viability, apoptosis, DNA damage, and the adipogenic or osteogenic differentiation potential of rat adipose‐derived stem cells (rASCs) exposed to previously established non‐cytotoxic doses of BPA and TCDD in vitro. Results demonstrated that 10 μM of BPA and 10 nM of TCDD were able to significantly reduce cell viability, while all exposure levels resulted in DNA damage, although did not increase the apoptosis rate. According to the analysis of adipogenic differentiation, 1 μM of BPA induced the significant formation of oil droplets, suggesting an increased adipocyte differentiation, while both 10 μM of BPA and 10 nM of TCDD decreased adipocyte differentiation. Osteogenic differentiation did not differ among the treatments. As such, BPA and TCDD in the concentrations tested can modify important processes in rASCs such as cell viability, adipogenic differentiation, and DNA damage. Together, these findings prove that EDCs play an important role as contaminants, putatively interfering in cell differentiation and thus impairing the therapeutic use of ASCs.

Background: Platelet-rich plasma (PRP) has been used to favor anterior cruciate ligament (ACL) healing after reconstruction surgeries. However, clinical data are still inconclusive and subjective about PRP. Thus, we propose a quantitative method to demonstrate that PRP produced morphological structure changes. Methods: Thirty-four patients undergoing ACL reconstruction surgery were evaluated and divided into control group (sixteen patients) without PRP application and experiment group (eighteen patients) with intraoperative application of PRP. Magnetic resonance imaging (MRI) scans were performed 3 months after surgery. We used Matlab® and machine learning (ML) in Orange Canvas® to texture analysis (TA) features extraction. Experienced radiologists delimited the regions of interest (RoIs) in the T2-weighted images. Sixty-two texture parameters were extracted, including gray-level co-occurrence matrix and gray level run length. We used the algorithms logistic regression (LR), naive Bayes (NB), and stochastic gradient descent (SGD). Results: The accuracy of the classification with NB, LR, and SGD was 83.3%, 75%, 75%, respectively. For the area under the curve, NB, LR, and SGD presented values of 91.7%, 94.4%, 75%, respectively. In clinical evaluations, the groups show similar responses in terms of improvement in pain and increase in the IKDC index (International Knee Documentation Committee) and Lysholm score indices differing only in the assessment of flexion, which presents a significant difference for the group treated with PRP. Conclusions: Here, we demonstrated quantitatively that patients who received PRP presented texture changes when compared to the control group. Thus, our findings suggest that PRP interferes with morphological parameters of the ACL. Trial registration: Protocol no. CAAE 56164316.6.0000.5411.

The immunogenic collagen V (Col V) and the proinflammatory cytokine interleukin (IL)-17 have been implicated in the pathogenesis of multiple autoimmune diseases. Col V is also up-regulated during adipogencsis and can stimulate adipocyte differentiation in vitro. Conditioned medium (CM) generated from adipose-derived mesenchymal stem cells (MSCs) reduces bleomycin (BLM)-induced lung injury in rats, suggesting a crucial role in situ of immunomodulatory factors secreted by MSCs in these beneficial effects. In the present work, we investigated this hypothesis, analyzing levels of plasma inflammatory mediators and inflammatory and fibrotic mediators in the lung tissue of BLM-injured rats after treatment with MSCs and CM. Pulmonary fibrosis was intratracheally induced by BLM. After 10 days, BLM animals were further randomized into subgroups receiving saline, MSCs, or CM intravenously. On days 14 and 21, the animals were euthanized, and the lungs were examined through protein expression of nitric oxide synthase (NOS), IL-17, transforming growth factor-beta (TGF-beta), vascular endothelial growth factor, endothelin-1, and the immunogenic Col V through histological quantitative evaluation and plasma levels of fibrinogen, Von Willebrand factor, and platelet-derived growth factor (PDGF). Rats that had been injected with MSCs and CM showed a significant increase in weight and significant improvements at 14 and 21 days after intravenous injection at both time points of analysis of plasma fibrinogen, PDGF, and Von Willebrand factor and NOS-2 expression, supporting an early anti-inflammatory action, thus reducing TGF-beta and collagen I fibers. In contrast, intravenous injection of CM was able to significantly increase the deposition of Col V fibers and IL-17 on both day 14 and day 21 as compared with the amount observed in rats from the BLM group and MSC groups. In conclusion, this study reinforces previous observations on the therapeutic properties of MSCs and CM and is the first report to demonstrate the association of its actions with immunomodulatory biomarkers on lung tissue. We concluded that adipose-derived stem cells and adipose-derived stem cells-CM modulate an in situ imbalance between collagen I- and Col V-mediated IL-17 immune response, emerging as a promising therapeutic option for recovering from BLM pulmonary fibrosis.

Purpose: To evaluate the influence of mesenchymal stem cells from adipose tissue in the end-to-side neurorrhaphy, focusing in the nerve regeneration and the muscle reinnervation in acute trauma. Methods: 140 animals were randomly divided in seven groups: control, denervated, end-to-side neurorrhaphy between distal stump of common peroneal nerve and tibial nerve (ESN), ESN wrapped in fascia, ESN wrapped in fascia and platelet gel, ESN wrapped in platelet gel, ESN wrapped in fascia and platelet gel within stem cells (without culture) removed from the adipose tissue. Mass measurements of the animal and of cranial tibial muscles, electromyography, walking track analysis tests and histological examinations of the nerves and muscles after 180 days was performed. Results: In the groups where the ESN was performed, the results were always better when compared to the denervated group, showing reinnervation in all ESN groups. The most sensitive methods were walking track and histological analysis. Only the group with stem cells showed values similar to the control group, as well as the functional indices of peroneal nerve and the number of nerve fibers in the peroneal nerve. Conclusions: Stem cells were effective in ESN according with the functional index of the peroneal nerve, evaluated by walking track analysis and the number of nerve fibers in the peroneal nerve.

The cardiovascular disease is the main cause of mortality in the western population and the Peripheral Arterial Disease (PAD) evolves, in large proportion, to the amputation of the affected limb. This study aimed to synthesize blood vessels using scaffolds of rabbit’s Inferior Vena Cava (IVC) and test its interaction with the receiver tissue and test inflammatory responses. Methods: The IVC were obtained from 8 rabbits to decellularization or in natura veins. The descellularized veins (DV) were obtained through protocols of decellularization established previously, using sodium dodecyl sulfate 1% (SDS) and mechanical agitation for 2 hours.12 animals were used to the experiment in vivo (3 animals in each group), being the product implanted in the interescapular dorsal area of each animal. The established groups are: Group 1- in natura allogeneic vein; Group 2- DV-SDS no cells added; Group 3- DV- SDS with 1x105 adipose tissue allogeneic Mesenchymal Stem Cells (MSC) added; Group 4- DV-SDS + autologous MSC. The (MSC) of the autologous receptors were collected 21 days before the implant and expanded in vitro. The explants were analyzed by histomorphological/immunohistochemistry and the peripheral blood was collected in the pre operatory in 1d, 7d, 14d, 30d and 60 day post operatory to dose the inflammatory and anti-inflammatory interleukins. Results: IL-10 and PDGF levels were significantly higher in group 4, which also showed neovascularization and large endothelial reconstruction. We may conclude the existence of an inflammatory response to the use of allogeneic grafts, which is lower when associated with autologous MSC.