UNAM-HIMFG Bacterial Lysate Activates the Immune Response and Inhibits Colonization of Bladder of Balb/c Mice Infected with the Uropathogenic CFT073 Escherichia coli Strain.

Urinary tract infections (UTIs) represent a clinical and epidemiological problem of worldwide impact that affects the economy and the emotional state of the patient. Control of the condition is complicated due to multidrug resistance of pathogens associated with the disease. Considering the difficulty in carrying out effective treatment with antimicrobials, it is necessary to propose alternatives that improve the clinical status of the patients. With this purpose, in a previous study, the safety and immunostimulant capacity of a polyvalent lysate designated UNAM-HIMFG prepared with different bacteria isolated during a prospective study of chronic urinary tract infection (CUTI) was evaluated. In this work, using an animal model, results are presented on the immunostimulant and protective activity of the polyvalent UNAM-HIMFG lysate to define its potential use in the control and treatment of CUTI. Female Balb/c mice were infected through the urethra with Escherichia coli CFT073 (UPEC O6:K2:H1) strain; urine samples were collected before the infection and every week for up to 60 days. Once the animals were colonized, sublingual doses of UNAM-HIMFG lysate were administrated. The colonization of the bladder and kidneys was evaluated by culture, and their alterations were assessed using histopathological analysis. On the other hand, the immunostimulant activity of the compound was analyzed by qPCR of spleen mRNA. Uninfected animals receiving UNAM-HIMFG lysate and infected animals administered with the physiological saline solution were used as controls. During this study, the clinical status and evolution of the animals were evaluated. At ninety-six hours after infection, the presence of CFT073 was identified in the urine of infected animals, and then, sublingual administration of UNAM-HIMFG lysate was started every week for 60 days. The urine culture of mice treated with UNAM-HIMFG lysate showed the presence of bacteria for three weeks post-treatment; in contrast, in the untreated animals, positive cultures were observed until the 60th day of this study. The histological analysis of bladder samples from untreated animals showed the presence of chronic inflammation and bacteria in the submucosa, while tissues from mice treated with UNAM-HIMFG lysate did not show alterations. The same analysis of kidney samples of the two groups (treated and untreated) did not present alterations. Immunostimulant activity assays of UNAM-HIMFG lysate showed overexpression of TNF-α and IL-10. Results suggest that the lysate activates the expression of cytokines that inhibit the growth of inoculated bacteria and control the inflammation responsible for tissue damage. In conclusion, UNAM-HIMFG lysate is effective for the treatment and control of CUTIs without the use of antimicrobials.

Platelet-Rich Plasma and Related Orthobiologics for the Treatment of Equine Musculoskeletal Disorders-A Bibliometric Analysis from 2000 to 2024.

(1) Background: There is increasing interest in the use of platelet-rich plasma and related orthobiologics for the treatment of chronic musculoskeletal disorders in horses; however, there is no information on the bibliometric impact of the literature published in this area. (2) Methods: A bibliometric analysis was performed using the bibliometrix R package by analyzing the documents registered in the WOS and Scopus databases from 2000 to 2024. The included registers were evaluated according to the menu of results from the biblioshiny web app (overview, sources, authors, documents, words, trending topics, clustering, conceptual structure, and social structure). (3) Conclusions: The documents produced were mainly published in Frontiers in Veterinary Science, Journal of Equine Veterinary Science, BMC Veterinary Research, and the American Journal of Veterinary Research). The most productive institutions were Universidad de Caldas, Colorado State University, University of California-Davis, and University of Leipzig, and the most productive countries were the USA, Brazil, and Colombia. Horse, platelet-rich plasma, equine, osteoarthritis, and autologous conditioned serum were the most frequently used keywords. The trending topics in this area are platelet lysates and orthobiologics. The collaboration network of authors, institutions, and countries shows an isolated development of individual author networks with modest collaboration between institutions and countries.

Impact of different formulations of platelet lysate on proliferative and immune profile of equine mesenchymal stromal cells.

Platelet lysate (PL) is investigated as a potential replacement for fetal bovine serum (FBS) in cell culture. However, there is limited research on its impact on the immune profile of equine mesenchymal stromal cells (eMSCs). This study aimed to evaluate the effects of different PL formulations on the proliferative capacity, multipotentiality, and immune profile of equine adipose tissue-derived MSCs (eAD-MSCs). In vitro growth kinetics and trilineage differentiation of eAD-MSCs (n = 7) were assessed under three culture conditions: medium-concentration PL (MPL), high-concentration PL (HPL), and FBS as a control. The immune profile was evaluated by studying the expression of immunogenic receptors such as MHC I, MHC II, and immunomodulatory molecules IL-6, IL-10, and TNF-α, determined by gene expression, surface marker expression, and cytokine quantification. Both PL formulations, pooled from 5 donors, exhibited 3.3 and 6.5-fold higher platelet counts than baseline plasma for MPL and HPL, respectively. Higher concentrations of TGF-ß and PDGF were found in both PL formulations compared to baseline. Furthermore, MPL and HPL subcultures demonstrated proliferative, clonogenic, and multipotent capacities similar to FBS. The immune profile of PL-cultured cells exhibited gene expression levels related to immunogenicity and immunomodulation similar to the reference condition, and the surface antigen presence of MHC II was also similar. However, HPL media exhibited higher IL-6, IL-10, and TNF-α concentrations in the culture supernatant. In conclusion, both PL media contained higher concentrations of growth factors compared to FBS, supporting the in vitro culture of eAD-MSCs with proliferative, clonogenic, and multipotent capacity similar to the reference medium. Nonetheless, PL usage led to a variation in the immunomodulatory cytokine microenvironment, with higher concentrations of IL-6, IL-10, and TNF-α in HPL media compared to MPL and FBS.

Analysis of commercially available snake antivenoms reveals high contents of endotoxins in some products.

As injectable therapeutics, snake antivenoms must meet specifications for endotoxin content. The Limulus amebocyte lysate (LAL) test was used to evaluate the endotoxin content in several commercially available antivenoms released for clinical use. It was found that some products have endotoxin concentrations higher than the accepted limit for these contaminants. These results emphasize the need to include endotoxin determination as part of the routine evaluation of antivenoms by manufacturers and regulatory agencies.

Comparison between Platelet Lysate, Platelet Lysate Serum, and Fetal Bovine Serum as Supplements for Cell Culture, Expansion, and Cryopreservation.

As cell culture supplements, human platelet lysate (PL) and human platelet lysate serum (PLS) are alternatives to fetal bovine serum (FBS) due to FBS-related issues such as ethical concerns, variability between batches, and the possible introduction of xenogenic contaminants. This study compared the composition and efficacy of PL, PLS, and FBS as supplements in the culture and cryopreservation of human dermal fibroblasts, Wharton's jelly-derived mesenchymal stem cells (WJ-MCS), and adipose tissue (AdMSC). Biochemical components, some growth factors, and cytokines present in each of them were analyzed; in addition, the cells were cultured in media supplemented with 5% PL, 5% PLS, and 10% FBS and exposed to different freezing and thawing solutions with the supplements under study. Biochemical parameters were found to be similar in PL and PLS compared to FBS, with some differences in fibrinogen and calcium concentration. Growth factors and cytokines were higher in PL and PLS compared to FBS. Cell proliferation and morphology showed no significant differences between the three culture media. Regarding the cryopreservation and thawing of cells, better results were obtained with PLS and FBS. In conclusion, PL and PLS are an excellent choice to replace the standard supplement of animal origin (FBS) in the media used for the culture and cryopreservation of fibroblasts, WJ-MSC, and AdMSC.

Lisado plaquetario en el tratamiento de la fisura anal crónica y otras indicaciones / Platelet lysate in the treatment of chronic anal fissure and other indications

Introducción: La aplicación del lisado plaquetario alogénico en el tratamiento de la fisura anal crónica es uno de los temas más novedosos y excitantes dentro de las ciencias biomédicas hoy día. Objetivo: Exponer en forma resumida los avances y perspectivas de empleo del lisado plaquetario alogénico en la fisura anal. Métodos: La estrategia de búsqueda abarcó información en diferentes bases de datos como internet y en el buscador google académico, se utilizaron 42 bibliografías seleccionadas para realizar la revisión, 35 publicadas en los últimos cinco años como artículos originales de revisión, monografías y otros documentos científicos especializados en el tema. A partir de la búsqueda se elaboró el presente artículo. Desarrollo: Se hace alusión a la conceptualización de la enfermedad y los pilares terapéuticos, se sustenta la utilización del lisado plaquetario alogénico; los logros alcanzados con su aplicación en el Hospital General Docente Comandante Pinares; del municipio San Cristóbal en la provincia Artemisa en la especialidad; así como a las potencialidades de tan promisorio campo en el presente siglo. Conclusiones: El Lisado plaquetario alogénico en el tratamiento de la fisura anal crónica constituyó una nueva modalidad de tratamiento de la enfermedad(AU)

Introduction: The application of allogeneic platelet lysate in the treatment of chronic anal fissure is one of the most novel and exciting topics within biomedical sciences today. Objective: To summarize the advances and perspectives of the use of allogeneic platelet lysate in anal fissure. Methods: The search strategy included information in different databases such as the internet and the academic google search engine, 42 bibliographies selected for the review were used, 35 published in the last five years as original review articles, monographs and other scientific documents specialized in the subject. The present article was prepared on the basis of the search. Development: Allusion is made to the conceptualization of the disease and the therapeutic pillars, the use of allogeneic platelet lysate is sustained; the achievements reached with its application in the General Teaching Hospital Comandante Pinares; of San Cristóbal municipality in Artemisa province in the specialty; as well as to the potential of such promising field in the present century. Conclusions: Allogeneic platelet lysate in the treatment of chronic anal fissure constituted a new modality of treatment of the disease(AU)

Uso del lisado plaquetario en el tratamiento de la necrolisis tóxica epidérmica: presentación de un caso / Use of platelet lysate in the treatment of epidermal toxic necrolysis: a case presentation

Introducción: En los últimos años se ha producido un extraordinario impulso de la medicina regenerativa. El uso de las plaquetas con este fin, se ha empleado en diferentes especialidades médicas. El lisado plaquetario es una de las alternativas al tratamiento utilizadas en la necrolisis tóxica epidérmica, urgencia dermatológica que alcanza una mortalidad de hasta el 70 por ciento en ocasiones. Objetivo: Describir el uso del lisado de plaquetas como terapia regenerativa en el tratamiento de la necrolisis tóxica epidérmica. Presentación de caso: Paciente de 53 años con antecedentes patológicos de hipertensión arterial, diabetes mellitus, mieloma múltiple e insuficiencia renal crónica, que ingresó con diagnóstico de neumonía adquirida en la comunidad e insuficiencia renal crónica agudizada, para lo que llevó múltiples tratamientos. Comenzó con lesiones en piel eritematosas, no pruriginosas y después ampollares que se desbridan con facilidad, y que están diseminadas por todo el cuerpo con grandes áreas de piel denudada con una mortalidad de más del 90 por ciento. Se diagnosticó una necrolisis tóxica epidérmica, se iniciaron las medidas habituales de tratamiento y se utilizó de forma tópica el lisado plaquetario alogénico. Conclusiones: La aplicación del lisado plaquetario tuvo una respuesta favorable, se observó reepitelización cutánea y mejoría de las lesiones en la necrolisis tóxica epidérmica(AU)

Introduction: In recent years there has been an extraordinary impulse of regenerative medicine, the use of platelets for this purpose has been used in different medical specialties. The most feared of the dermatological complications is toxic epidermal necrolysis, of which up to 4 cases per million inhabitants occur annually, with a mortality that sometimes reaches up to 70 percent Objective: To describe the use of platelet lysate as regenerative therapy in the treatment of epidermal toxic necrolysis. Case presentation: We present a case of a 53-year-old patient with a pathological history of arterial hypertension, diabetes mellitus, multiple myeloma and chronic renal insufficiency, who was admitted with a diagnosis of community acquired pneumonia and acute chronic renal insufficiency, for which multiple treatments. She began with erythematous, non-pruritic, and then bullous skin lesions that are easily debrided into large areas of denuded skin, scattered throughout the body. A toxic epidermal necrolysis was diagnosed, the usual treatment measures were started and the allogeneic platelet lysate was used topically. Conclusions: A favorable response was observed, with cutaneous re-epithelialization of the skin, although the patient's underlying disease and its comorbidities died(AU)

The proteomic and particle composition of human platelet lysate for cell therapy products.

Following health agencies warning, the use of animal origin supplements should be avoided in biological products proposed as therapy in humans. Platelet lysate and several other growth factors sources are alternatives to replace fetal calf serum, the current gold standard in clinical-grade cell culture. However, the platelet supplement's content lacks data due to different production methods. The principle behind these products relays on the lysis of platelets that release several proteins, some of which are contained in heterogeneous granules and coordinate biological functions. This study aims to analyze the composition and reproducibility of a platelet lysate produced with a standardized method, by describing several batches' protein and particle content using proteomics and dynamic light scattering. Proteomics data revealed a diversified protein content, with some related to essential cellular processes such as proliferation, morphogenesis, differentiation, biosynthesis, adhesion, and metabolism. It also detected proteins responsible for activation and binding of transforming growth factor beta, hepatocyte growth factor, and insulin-like growth factor. Total protein, biochemical, and growth factors quantitative data showed consistent and reproducible values across batches. Novel data on two major particle populations is presented, with high dispersion level at 231 ± 96 d.nm and at 30 ± 8 d.nm, possibly being an important way of protein trafficking through the cellular microenvironment. This experimental and descriptive analysis aims to support the content definition and quality criteria of a cell supplement for clinical applications.

PKHB1, a thrombospondin-1 peptide mimic, induces anti-tumor effect through immunogenic cell death induction in breast cancer cells.

Breast cancer is the most commonly diagnosed cancer and the leading cause of cancer death in women worldwide. Recent advances in the field of immuno-oncology demonstrate the beneficial immunostimulatory effects of the induction of immunogenic cell death (ICD). ICD increases tumor infiltration by T cells and is associated with improved prognosis in patients affected by triple negative breast cancer (TNBC) with residual disease. The aim of this study was to evaluate the antitumoral effect of PKHB1, a thrombospondin-1 peptide mimic, against breast cancer cells, and the immunogenicity of the cell death induced by PKHB1 in vitro, ex vivo, and in vivo. Our results showed that PKHB1 induces mitochondrial alterations, ROS production, intracellular Ca2+ accumulation, as well calcium-dependent cell death in breast cancer cells, including triple negative subtypes. PKHB1 has antitumor effect in vivo leading to a reduction of tumor volume and weight and promotes intratumoral CD8 + T cell infiltration. Furthermore, in vitro, PKHB1 induces calreticulin (CALR), HSP70, and HSP90 exposure and release of ATP and HMGB1. Additionally, the killed cells obtained after treatment with PKHB1 (PKHB1-KC) induced dendritic cell maturation, and T cell antitumor responses, ex vivo. Moreover, PKHB1-KC in vivo were able to induce an antitumor response against breast cancer cells in a prophylactic application, whereas in a therapeutic setting, PKHB1-KC induced tumor regression; both applications induced a long-term antitumor response. Altogether our data shows that PKHB1, a thrombospondin-1 peptide mimic, has in vivo antitumor effect and induce immune system activation through immunogenic cell death induction in breast cancer cells.

Recombinant Escherichia coli Cell Lysates as a Low-Cost Alternative for Vaccines Against Veterinary Clostridial Diseases.

This chapter describes a practical, industry-friendly, and efficient vaccine protocol based on the use of Escherichia coli cell fractions (inclusion bodies or cell lysate supernatant) containing the recombinant antigen. This approach was characterized and evaluated in laboratory and farm animals by the seroneutralization assay in mice, thereby showing to be an excellent alternative to induce a protective immune response against clostridial diseases.

Inflammation reduction potential of nanostructured lipid carriers encapsulated with rat's bone marrow cells' lysate / Potencial de redução da inflamação de carreadores lipídicos nanoestruturados encapsulados com lisado de células de medula óssea de ratos

Bone marrow-derived mesenchymal stromal cells (BMSCs) have been used for treating inflammatory disorders. Due to the large size of BMSCs compared to nanoparticles, BMSCs cannot be loaded into the nanoparticles. It is hypothesized that BMSCs lysate loading into the nanocarriers will effectively deliver cellular contents and regulatory elements of BMSCs at the injury site. This study aimed to investigate nanostructured lipid carriers (NLC) loading with BMSCs lysate through basic characterization and morphological analysis. Moreover, this study was mainly designed to investigate the role of NLC loaded BMSCs lysate in reducing inflammation via in-vitro and in-vivoassays. The in-vitro study involves cell viability assays, p53, annexin V and VEGF expression through ELISA and immunocytochemistry, real-time BAX, caspase-3, IL-6, IL-8, TOP2A, PCNA, and Ki-67 gene expression analysis. Additionally, to evaluate in-vivo anti-inflammatory activity, the carrageenan-induced rat paw oedema model was used. In-vitro results showed that NLC loaded BMSCs lysate increased cell viability, decreased apoptosis and pro-inflammatory genes expression and up-regulated angiogenesis and proliferation in H2O2 pre-stimulated cells. Findings of the in-vivo assay also indicated a reduction in rat's paw oedema volume in NLC-loaded BMSCs lysate, and downregulation of BAX, Caspase-3, IL-6, and IL-8 was observed. Enhanced expressions of TOP2A, PCNA, and Ki-67 were obtained. Concluding the results of this study, NLC-loaded BMSCs lysate could reduce inflammation and possibly regenerate damaged tissue mainly via increasing cell viability, angiogenesis and proliferation, and reducing apoptosis and pro-inflammatory cytokines.

Células estromais mesenquimais derivadas da medula óssea (BMSCs) têm sido utilizadas para o tratamento de distúrbios inflamatórios. Devido ao grande tamanho das BMSCs em comparação com as nanopartículas, as BMSCs não podem ser carregadas nas nanopartículas. Supõe-se que o carregamento de lisado de BMSCs no nanocarriers será eficaz na entrega de conteúdos celulares e elementos reguladores de BMSCs no local da lesão. Este estudo teve como objetivo investigar a carga de carreador lipídico nanoestruturado (NLC) com lisado de BMSCs através de caracterização básica e análise morfológica. Além disso, este trabalho foi projetado, principalmente, para investigar o papel do lisado de BMSCs carregado com NLC na redução da inflamação por meio de ensaios anti-inflamatórios in vitro e in vivo. O estudo in vitro envolve ensaios de viabilidade celular, expressão de p53, anexina V e VEGF por ELISA e imunocitoquímica e expressão gênica em tempo real de BAX, caspase-3, IL-6, IL-8, TOP2A, PCNA e Ki-67 . Além disso, para avaliar a atividade anti-inflamatória in vivo,o modelo de edema de pata de rato induzido por carragenina foi utilizado. Os resultados in vitro mostraram que o lisado de BMSCs carregadas com NLC aumentou a viabilidade celular, diminuiu a apoptose e a expressão de genes pró-inflamatórios e aumentou a angiogênese e proliferação em células pré-estimuladas com H2O2. Os achados do ensaio in vivo também indicaram uma redução no volume do edema da pata de rato no lisado de BMSCs carregado com NLC, entretando, foi observada a regulação negativa de BAX, Caspase-3, IL-6 e IL-8. Expressões aumentadas de TOP2A, PCNA e Ki-67 foram obtidas. Assim, concluindo os resultados do estudo, é possível afirmar que o lisado de BMSCs carregado com NLC pode reduzir a inflamação e possivelmente regenerar o tecido danificado principalmente por meio do aumento da viabilidade celular, angiogênese e proliferação e redução da apoptose e citocinas pró-inflamatórias.

Lisado plaquetario autólogo y su efectividad en pacientes con epicondilitis / Autologous platelet lysate and its effectiveness in patients with epicondylitis

Introducción: La epicondilitis constituye uno de los motivos de consulta más frecuentes tanto en la asistencia primaria como especializada y sin duda alguna, es uno de los problemas que tiene mayor repercusión en la persona que la padece. El tratamiento de las epicondilitis constituye un reto para la medicina debido a enormes implicaciones sanitarias, sociolaborales y el dolor e impotencia funcional que provoca. Objetivo: Evaluar la efectividad del lisado plaquetario autólogo como alternativa de tratamiento en pacientes enfermos con epicondilitis. Método: Se realizó un estudio cuasi experimental analítico longitudinal prospectivo en el que se evaluó el uso de lisado plaquetario autólogo como alternativa de tratamiento en pacientes con epicondilitis. El universo estuvo constituido por los pacientes que acudieron a consulta de Ortopedia y traumatología con el diagnóstico de epicondilitis, durante el periodo comprendido entre octubre de 2014 y julio de 2018. La muestra quedo constituida por 80 pacientes que cumplieron con los criterios de inclusión y exclusión. Resultados: El grupo de edad entre 36-56 años y del sexo femenino son los de mayor representación en padecer esta enfermedad. Las infiltraciones de lisado plaquetario autólogo aportan mejores resultados al convencional y se observa la mayor representación de pacientes que tuvieron una remisión total. Las complicaciones fueron mucho más evidentes en el tratamiento convencional. También es relevante el costo-beneficio del tratamiento con lisado plaquetario autólogo. Conclusiones: El tratamiento con lisado plaquetario autólogo puede ser una alternativa para mejorar la calidad de vida de los pacientes con epicondilitis(AU)

Introduction: Epicondylitis is one of the most frequent reasons for attending consultation in both primary and specialized care; while it is undoubtedly one of the problems with the greatest impact on the person who suffers from it. The managment epicondylitis is a challenge for medicine, due to the enormous health-related and social implications, as well as the pain and functional impotence that it causes. Objective: To assess the effectiveness of autologous platelet lysate as a treatment alternative in patients with epicondylitis. Method: A prospective, longitudinal, analytical and quasiexperimental study was carried out, in which the use of autologous platelet lysate as an alternative treatment in patients with epicondylitis was assessed. The universe consisted of patients who attended the orthopedics and traumatology consultation, during the period between October 2014 and July 2018, with a diagnosis of epicondylitis. The sample was made up of eighty patients who met the inclusion criteria; exclusion criteria were also considered. Results: The age group between 36 and 56 years, together with the female sex, are the most represented with respect to suffering from this disease. Infiltrations of autologous platelet lysate provide better outcomes than the conventional one, while greater representation of remitted patients is observed. Complications were much more evident in conventional treatment. The cost-benefit relationship of treatment with autologous platelet lysate is also relevant. Conclusions: Treatment with autologous platelet lysate can be an alternative to improve the quality of life of patients with epicondylitis(AU)

Human platelet lysate - A potent (and overlooked) orthobiologic.

The knowledge of the essential role of platelets in tissue healing is gradually increasing and as regenerative medicine prompts new solutions, platelet-derived bioproducts have been proposed as a potential tool in this field. In orthopaedics and sports medicine, the use of PRP has been rapidly increasing in popularity as patients seek novel non-surgical approaches to acute and chronic musculoskeletal conditions. The concept of having platelets as a secretory organ other than a mere sponge-like coagulation component opens up new frontiers for the use of the platelet secretome. Platelet lysate is a solution saturated by growth factors, proteins, cytokines, and chemokines involved in crucial healing processes and is administered to treat different diseases such as alopecia, oral mucositis, radicular pain, osteoarthritis, and cartilage and tendon disorders. For this purpose, the abundant presence of growth factors and chemokines stored in platelet granules can be naturally released by different strategies, mostly through lyophilization, thrombin activation or ultrasound baths (ultrasonication). As a result, human platelet lysate can be produced and applied as a pure orthobiologic. This review outlines the current knowledge about human platelet lysate as a powerful adjuvant in the orthobiological use for the treatment of musculoskeletal injuries, without however failing to raise some of its most applicable basic science.

Lisado plaquetario homólogo como factor estimulante de la cicatrización en la zona donante de injertos / Homologous Platelet lysate as astimulatinghealing factor for the grafts donor area

Introducción: La búsqueda de alternativas para disminuir el tiempo de cicatrización y de hospitalización constituye uno de los aspectos fundamentales de la investigación actual. Los factores de crecimiento plaquetarios son capaces de potenciar la cicatrización. Objetivo: Determinar los beneficios de la aplicación del lisado de plaquetas homólogo sobre la zona donante del injerto autólogo de piel. Método: Se realizó un estudio longitudinal prospectivo en 20 pacientes tratados en el Hospital Hermanos Ameijeiras entre agosto de 2016 y mayo de 2019, que requirieron de injerto autólogo de piel en las zonas cruentas. Se realizaron dos tomas de injerto en el mismo paciente y región anatómica; una fue tratada con lisado plaquetario (zona de intervención) y otra con tratamiento convencional (zona control). La selección de pacientes fue intencional. Para estudiar las variables cualitativas se utilizaron números absolutos y proporciones y en las cuantitativas medidas de tendencia central y dispersión. Para la comparación de los resultados se aplicó la prueba de Friedman con un nivel de significación p ≤ 0,05. Resultados: La quemadura fue la principal causa de zona cruenta (75 por ciento), el grupo de edad más afectado fue el de mayores de 55 años. El porcentaje de cicatrización fue mayor en el grupo de intervención, con un tiempo de epitelización significativamente menor (p < 0,01), y el dolor en la zona intervenida fue menor. No hubo complicaciones. Conclusiones: Los beneficios encontrados en la zona intervenida con lisado plaquetario fueron significativos con una cicatrización más rápida y menor dolor(AU)

Introduction: The search for treatment alternatives that allow reducing wound healing and hospitalization time are fundamental aspects in research nowadays. Platelet growth factors are capable of enhancing wound healing. Objective: Determine the benefits of applying homologous platelet lysate on the donor area of autologous skin graft. Methods: A prospective longitudinal-section study was conducted in 20 patients with bloody areas that required autologous skin graft at the Hospital Hermanos Ameijeiras between August 2016 and May 2019. Two graft intakes were made in the same patient and anatomical region, one of them treated with platelet lysate (intervention zone) and one with conventional treatment (control zone). Patients selection was intentional. Absolute numbers and proportions were used to study the qualitative variables, and measures of central tendency and dispersion were used in the quantitative variables. To compare de results, the Friedman test was applied, setting a level of significance p < 0,05. Results: The main cause of bloody area was burns (75 percent), the most affected age group was those over 55 years, the healing percentage was greater in the intervention group with a statistically significant shorter epithelization time (p < 0,01) and there was less pain in the intervention zone. There were no complications. Conclusions: The benefits found in the intervened area with platelet lysate were significant with faster healing and less pain(AU)

Polyvalent Bacterial Lysate Protects Against Pneumonia Independently of Neutrophils, IL-17A or Caspase-1 Activation.

Polyvalent bacterial lysates have been in use for decades for prevention and treatment of respiratory infections with reported clinical benefits. However, besides claims of broad immune activation, the mode of action is still a matter of debate. The lysates, formulated with the main bacterial species involved in respiratory infections, are commonly prepared by chemical or mechanical disruption of bacterial cells, what is believed influences the biological activity of the product. Here, we prepared two polyvalent lysates with the same composition but different method of bacterial cell disruption and evaluated their biological activity in a comparative fashion. We found that both bacterial lysates induce NF-kB activation in a MyD88 dependent manner, suggesting they work as TLR agonists. Further, we found that a single intranasal dose of any of the two lysates, is sufficient to protect against pneumococcal pneumonia, suggesting that they exert similar biological activity. We have previously shown that protection against pneumococcal pneumonia can also be induced by prior S. pneumoniae sub lethal infection or therapeutic treatment with a TLR5 agonist. Protection in those cases depends on neutrophil recruitment to the lungs, and can be associated with increased local expression of IL-17A. Here, we show that bacterial lysates exert protection against pneumococcal pneumonia independently of neutrophils, IL-17A or Caspase-1/11 activation, suggesting the existence of redundant mechanisms of protection. Trypsin-treated lysates afford protection to the same extent, suggesting that just small peptides suffice to exert the protective effect or that the molecules responsible for the protective effect are not proteins. Understanding the mechanism of action of bacterial lysates and deciphering the active components shall allow redesigning them with more precisely defined formulations and expanding their range of action.

Human Platelet Lysate Supports Efficient Expansion and Stability of Wharton's Jelly Mesenchymal Stromal Cells via Active Uptake and Release of Soluble Regenerative Factors.

Manufacturing of mesenchymal stromal cell (MSC)-based therapies for regenerative medicine requires the use of suitable supply of growth factors that enhance proliferation, cell stability and potency during cell expansion. Human blood derivatives such as human platelet lysate (hPL) have emerged as a feasible alternative for cell growth supplement. Nevertheless, composition and functional characterization of hPL in the context of cell manufacturing is still under investigation, particularly regarding the content and function of pro-survival and pro-regenerative factors. We performed comparative analyses of hPL, human serum (hS) and fetal bovine serum (FBS) stability and potency to support Wharton's jelly (WJ) MSC production. We demonstrated that hPL displayed low inter-batch variation and unique secretome profile that was not present in hS and FBS. Importantly, hPL-derived factors including PDGF family, EGF, TGF-alpha, angiogenin and RANTES were actively taken up by WJ-MSC to support efficient expansion. Moreover, hPL but not hS or FBS induced secretion of osteoprotegerin, HGF, IL-6 and GRO-alpha by WJ-MSC during the expansion phase. Thus, hPL is a suitable source of factors supporting viability, stability and potency of WJ-MSC and therefore constitutes an essential raw material that in combination with WJ-MSC introduces a great opportunity for the generation of potent regenerative medicine products.

Data on the characterization of a platelet lysate and of the system in which it is included (nanoparticles/hydrogel), intended for the treatment of wounds.

Platelet lysate has attracted attention for different biomedical applications, including biological processes where cellular proliferation and migration have been altered. Spectroscopic properties of a platelet lysate obtained from human platelets were performed in order to be incorporated in polymeric nanoparticles and then into a PluronicⓇ F127 hydrogel, intended for wound healing (more details can be found at https://doi.org/10.1016/j.ejps.2020.105231 [1]). The platelet lysate (PL) was assessed by ultraviolet, infrared and circular dichroism spectroscopy. The developed hydrogel was also analyzed by infrared spectroscopy to evaluate if the PluronicⓇ F127 structure was maintained when the nanoparticles or platelet lysate-loaded nanoparticles were included. The sol-gel transition temperature of the hydrogel was determined through its thermal behavior and by dynamic light scattering.

Activation of BK Channel Contributes to PL-Induced Mesenchymal Stem Cell Migration.

Due to their capacity to proliferate, migrate, and differentiate, mesenchymal stem cells (MSCs) are considered to be good candidates for regenerative medicine applications. The mechanisms underlying proliferation and differentiation of MSCs have been studied. However, much less is known about the mechanisms regulating the migration of MSCs. Platelet lysate (PL), a supplement used to promote cell expansion, has been shown to promote MSCs migration; however, the underlying mechanism are unknown. Here, by using adipose-derived rat MSCs (rMSCs) and the scratch assay in the absence and presence of various BK channels modulators, we evaluated the role of BK channels in mediating the PL-stimulated migration of rMSCs. We found that 5% PL increased rMSCs migration, and this effect was blocked by the addition of the BK channel selective antagonist Iberiotoxin (IBTX). In the absence of PL, the BK channel agonist NS1619, stimulated rMSCs migration to similar level as 5% PL. Addition of both NS1619 and 5% PL resulted in an increase in rMSCs migration, that was higher than when either one was added individually. From whole-cell recordings, it was found that the addition of 5% PL increased the magnitude of BK current density. By using Western blot and flow cytometry, it was found that PL did not affect the expression of BK channels. Together, our results indicate that as shown in other cell types, activation of BK channels by themselves also promote rMSC migration, and show that activation of BK channels contribute to the observed PL-induced increase in migration of rMSC.

Treatment With Platelet Lysate Inhibits Proteases of Synovial Fluid in Equines With Osteoarthritis.

Osteoarthritis (OA) is the most prevalent arthropathy in sport horses. The administration of a platelet lysate (PL) is an alternative method for the treatment of musculoskeletal conditions. The mechanisms by which PL exerts its beneficial effects have not been determined, and less is known about its effect on the activity of the proteolytic enzymes of the synovial fluid of equines with OA. In this work, the effect of the administration of PL to horses with OA was analyzed both clinically and molecularly by determining the levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5), glycosaminoglycans (GAGs), and tissue inhibitor of metalloproteinase 1 (TIMP-1) in synovial fluid. One mL of PL was administered intra-articularly followed by the extraction of synovial fluid on days 0, 10, 30, and 60. Results were evaluated by an analysis of variance for repeated measures. The levels of MMP-9 decreased significantly (P < .05) on day 10 after treatment with PL. A disintegrin and metalloproteinase with thrombospondin motifs 5 decreased significantly on days 10 (P < .05), 30 (P < .05), and 60 (P < .01) after treatment. The levels of synovial TIMP-1 increased significantly on day 30 (P < .001) after treatment. Glycosaminoglycans showed a significant increase on days 10 (P < .05) and 30 (P < .01). A significant decrease was found for MMP-2 on day 10 (P < .01), 30 (P < .01), and 60 (P < .001). In conclusion, the beneficial effects of PL in OA could be attributed to the decreased activity of MMP-2, MMP-9, and ADAMTS-5 and the increased concentration of GAGs and TIMP-1 after the administration of platelet-rich plasma.

Efficient isolation and proliferation of human adipose-derived mesenchymal stromal cells in xeno-free conditions.

Classical methods used for culture of adipose-derived mesenchymal stromal cells (ADSCs) use xenobiotic components, which may present a potential risk for biological contamination and/or elicit immunological reactions. Therefore, the aim of this study was to establish a xeno-free methodology for the isolation and proliferation of human ADSCs (hADSCs). hADSCs were isolated by enzymatic digestion or mechanical dissociation and cultured in the presence of fetal bovine serum or human platelet lysate. Proliferation curves were performed as a function of time from the cell culture and used to calculate the population doubling time. Immunophenotyping and differentiation tests were used to identify and characterize the hADSCs. Human ADSCs isolated and cultured in conventional or xenobiotic-free conditions peaked at different days but achieved similar maximum proliferation. The hADSCs differentiation ability was similar in all groups. The characterization of hADSCs by flow cytometry showed low contamination of the cultures by other cell types. The xenobiotic-free methodology described in this study is a feasible and reproducible alternative for isolation and proliferation of hADSCs. This methodology is in accordance with the recommendations of the National Health Surveillance Agency, which proposes avoidance of xenobiotic products.