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.

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.

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)

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.

Classification systems for platelet-rich plasma.

There is good scientific rationale to support the use of growth factors to promote musculoskeletal tissue regeneration. However, the clinical effectiveness of platelet-rich plasma (PRP) and other blood-derived products has yet to be proven. Characterization and reporting of PRP preparation protocols utilized in clinical trials for the treatment of musculoskeletal disease is highly inconsistent, and the majority of studies do not provide sufficient information to allow the protocols to be reproduced. Furthermore, the reporting of blood-derived products in orthopaedics is limited by the multiple PRP classification systems available, which makes comparison of results between studies challenging. Several attempts have been made to characterize and classify PRP; however, no consensus has been reached, and there is lack of a comprehensive and validated classification. In this annotation, we outline existing systems used to classify preparations of PRP, highlighting their advantages and limitations. There remains a need for standardized universal nomenclature to describe biological therapies, as well as a comprehensive and reproducible classification system for autologous blood-derived products. Cite this article: Bone Joint J 2019;101-B:891-896.

Platelet lysate loaded electrospun scaffolds: Effect of nanofiber types on wound healing.

In healthy individuals, wound healing is a highly efficient process. However, interruptions of normal healing give rise to chronic wounds, characterized by inflammation with impaired angiogenesis and re-epithelialization. The aim of this work was the design and the development of electrospun nanofibrous scaffolds based on sodium alginate (SA) and pullulan (PUL) and loaded with human platelet lysate (PL) intended for skin reparation, to take the advantage of nanofibrous scaffolds (with improved physical structure) and of SA as biopolymer. Two preparation approaches have been used to load PL in the scaffolds: as component of the PUL/SA matrix, to be electrospun, or as coating component, to cover the previously prepared electrospun PUL based membranes. A preformulation study to assess pullulan entanglement concentration and alginate or citric acid critical concentration, to obtain electrospun nanofibers, has been performed. The preparation process allowed to obtain insoluble systems starting from aqueous solutions and these were able to act as scaffolds for tissue engineering with suitable mechanical properties and PL release. PL loading in PUL/SA matrix nanofibers did not substantially modify the nanofiber morphology before crosslinking, while the crosslinking process, in presence of PL, determined less sharp nanofibers probably due to an increase in hydrophilicity caused by PL proteins. On the contrary, the coated nanofibers showed an increase in diameters due to PL loading. The two different approaches affected the fiber dimension and scaffold elasticity, especially for PL loaded systems. Anyhow, these differences were not crucial for fibroblast adhesion and proliferation which were mainly influenced by PL loading. In particular, fibroblasts presented different conformation and orientation mainly due to the presence of PL. This caused a cell random orientation compatible to a fibroblast-to-myofibroblast transition that could enhance wound healing.

Pacientes con retardo de consolidación, infiltrados con lisado plaquetario autólogo / Delayed consolidation patients infiltrated with autologous platelet lysate

Introducción: El retardo de consolidación es una complicación que resulta de un proceso de consolidación ósea anormal. Objetivo: Caracterizar a los pacientes con retardo de consolidación, infiltrados con lisado plaquetario autólogo. Métodos: Se realizó un estudio de casos, observacional, descriptivo, transversal y retrospectivo, en el Hospital General Docente Comandante Pinares, de enero 2008 a diciembre de 2017. Se revisaron 186 historias clínicas y se seleccionaron 80, que cumplían los criterios para el estudio. Se infiltró entre 6 mL de lisado plaquetario autólogo en los focos de fracturas, se evaluó el tiempo de consolidación ósea mediante radiografías seriadas y se registraron las complicaciones. Las variables del estudio fueron: edad, sexo, color de la piel, localizaciones óseas afectadas, sitios anatómicos del hueso, tiempo de consolidación ósea, complicaciones. Se empleó la distribución de frecuencias absoluta y relativa en el análisis de las variables cualitativas y la media aritmética para la variable cuantitativa edad. Para la relación entre variables cualitativas independientes se utilizó la prueba de chi-cuadrado. Resultados: Hubo más frecuencia de retardo de la consolidación en hombres de piel blanca, con edades comprendidas entre 26 y 45 años. Predominó en el tercio inferior de la tibia. El hueso que más tiempo requirió para su consolidación fue la tibia, con más de 16 semanas. No se describen complicaciones asociadas al implante. Conclusiones: El retardo de consolidación continúa siendo una complicación de difícil manejo. Es frecuente en hombres jóvenes y blancos. El lisado plaquetario autólogo es un buen coadyuvante en el tratamiento de esta complicación(AU)

Introduction: The delay of consolidation is a complication that results from an abnormal bone consolidation process. Objective: To characterize patients with delayed consolidation infiltrated with autologous platelet lysate. Methods: An observational, descriptive, cross-sectional and retrospective case study was conducted at Comandante Pinares General Teaching Hospital, from January 2008 to December 2017. One hundred eighty six (186) medical records were reviewed and eighty (80) were selected, since they met the study criteria. 6 mL of autologous platelet lysate was infiltrated in the foci of fractures. Serial radiographs were used to assess the bone healing time and complications were recorded. The variables of the study were age, sex, color of the skin, affected bone locations, anatomical sites of the bone, time of bone consolidation, complications. The distribution of absolute and relative frequencies was used in the analysis of qualitative variables and arithmetic mean for the quantitative age variable. Chi-square test was used for the relationship between independent qualitative variables. Results: The delayed consolidation was more frequency in white men, aged between 26 and 45 years. It predominated in the lower third of the tibia. Tibia was the bone that required more time for consolidation, more than 16 weeks. No complications associated with the implant are described. Conclusions: The delayed consolidation continues being a complication of difficult treatment. It is frequent in young and white men. The autologous platelet lysate is a good adjuvant in the treatment of this complication(AU)

Introduction: Le retard de consolidation est une complication résultant d'un processus de consolidation osseuse anormal.Objectif: Caractériser les patients atteints de retard de consolidation qui sont traités par infiltration de lysat plaquettaire autologue. Méthodes: Une étude observationnelle, descriptive, transversale et rétrospective a été réalisée à l'hôpital général universitaire Comandante Pinares, depuis janvier 2008 jusqu'à décembre 2017. Sur 186 dossiers médicaux révisés, on a sélectionné 80 respectant les critères d'inclusion de l'étude. Six millilitres de lysat plaquettaire autologue ont été injectés dans les foyers des fractures ; le temps de consolidation osseuse a été estimé par des radiographies en série, et les complications ont été enregistrées. On a utilisé des variables telles que l'âge, le sexe, la couleur de la peau, les localisations osseuses affectées, le site anatomique de l'os, le temps de consolidation osseuse, et les complications. On a employé la distribution de fréquences absolue et relative pour l'analyse des variables qualitatives et la moyenne d'âge, tandis que pour la relation entre les variables qualitatives indépendantes on a utilisé le test du chi-carré. Résultats: Le retard de consolidation a été beaucoup plus fréquent chez les hommes blancs entre 26 et 45 ans. Le tiers inférieur du tibia a été touché en prédominance. L'os qui a exigé beaucoup plus de temps de consolidation a été le tibia (plus de seize semaines). Des complications associées à l'implant n'ont pas été décrites. Conclusions: Le retard de consolidation demeure une complication difficile à traiter. Il est fréquemment observé chez les hommes jeunes et blancs. Le lysat plaquettaire autologue est un bon adjuvant dans le traitement de cette complication(AU)

Efectividad del lisado plaquetario en la cicatrización de las fisuras anales / Effectiveness of platelet lysate in anal fissures cicatrization

Introducción: las fisuras anales son un problema de salud importante en la población cubana, incluso con la supresión de los factores desencadenantes, muchas de ellas tienden a la cronicidad; su evolución es tórpida, y es elevado el riesgo de complicación de las mismas. Objetivo: evaluar la cicatrización de las fisuras anales mediante lisado plaquetario en el Hospital General Docente Comandante Pinares de San Cristóbal. Método: se realizó un estudio cuasi experimental en el servicio de cirugía general del Hospital General Docente Comandante Pinares desde enero 2015 hasta junio 2017. Se incluyeron en el estudio todos los pacientes adultos, de ambos sexos, atendidos por el diagnóstico de fisura anal con inadecuada respuesta al tratamiento convencional y sin otras enfermedades de base que impidieran el uso de la terapia regenerativa. A todos los pacientes se les solicitó el consentimiento informado previa explicación de las características del estudio. Resultados: la hemorroidectomía fue el proceder más frecuente asociado a las fisuras anales. El dolor fue el síntoma predominante antes del tratamiento. La reducción del área de la fisura fue evidente en los tratados con lisado. El efecto final en los pacientes tratados con el método fue positivo. Conclusiones: se comprobó que el lisado plaquetario usado para la cicatrización de la fisura anal crónica fue efectivo con un resultado de satisfacción por parte de los pacientes, con un mínimo de reacciones adversas. Puede representar un futuro promisorio en el tratamiento de esta enfermedad(AU)

Introduction: Anal fissures are an important health problem in the Cuban population, even with the suppression of the triggering factors, many of them tend to chronicity; its evolution is torpid, and their risk for complication is high. Objective: To evaluate the healing of anal fissures by platelet lysate in Comandante Pinares General Teaching Hospital in San Cristóbal. Method: A quasi-experimental study was performed in the general surgery service of Comandante Pinares General Teaching Hospital, from January 2015 to June 2017. All adult patients, of both sexes, were attended upon diagnosis of anal fissure and were included in the study. with inadequate response to conventional treatment and without other underlying diseases that could prevent the use of regenerative therapy. All patients were asked for their informed consent prior explanation of the study characteristics. Results: Hemorrhoidectomy was the most frequent procedure associated with anal fissures. Pain was the predominant symptom before treatment. The reduction in the area of ​​the fissure was evident in those patients treated with the lysate. The final effect in patients treated with the method was positive. Conclusions: The usage of the platelet lysate for the healing of chronic anal fissure proved effective, with a result of patient satisfaction, with a minimum of adverse reactions. It may represent a promising future in the treatment of this disease(AU)

Aplicación del lisado de plaquetas en la fistula pilonidal / Application of platelet lysate in intergluteal pilonidal disease

La enfermedad pilonidal o fístula pilonidal es una afección que se presenta a lo largo del pliegue entre los glúteos, desde el hueso sacro hasta el ano. Esta afección involucra los folículos pilosos de la región presacra, los cuales penetran en el tejido celular subcutáneo y forman un quiste alrededor. Se presenta como una o varias fositas o depresiones de diámetro pequeño, puntiforme por la que emergen pelos. Como parte del tratamiento médico se describen medidas locales, la antibióticoterapia de amplio espectro y tratamiento quirúrgico, en el cual se describen técnicas de resección abiertas y cerradas. Se presenta el caso de una adolescente femenina de 16 años de edad, diagnosticada hace año y medio con una fístula pilonidal que requirió tratamiento con vitaminoterapia, medidas locales e intervención quirúrgica en tres ocasiones. En la última de ellas se produjo dehiscencia del sitio quirúrgico, escaso tejido de granulación y pobre cicatrización. Por tal motivo se decidió la aplicación de lisado de plaquetas en toda el área de la fístula a razón de 1 mL en días alternos, durante dos semanas, se continuó una aplicación semanal durante las 4 semanas siguientes hasta que se obtuvo el cierre total. La utilización del lisado de plaquetas favorece el tejido de granulación y la cicatrización en la fistula pilonidal(AU)

Intergluteal pilonidal disease or pilonidal fistula is a condition presenting along the cleft between the buttocks, which runs from the bone at the bottom of the spine (sacrum) to the anus. This condition involves the hair follicles of the presacral region, which penetrates into the subcutaneous tissue and forms a cyst around. It is presented as one or more pits or depressions of small diameter emerging punctuate by hairs. As part of medical treatment local measures, broad-spectrum antibiotic therapy and surgical treatment, which may be open and closed resection techniques, are described. We present a case of a 16- year- old female teenager, diagnosed a year and half ago, with a pilonidal fistula that required surgical treatment in 3 occasions, with local measures and vitamin therapy; dehiscence of the surgical site, poor tissue granulation and poor healing. It was applied platelet lysate throughout the area of the fistula at 1 mL alternate days, for two weeks, infiltration was continued for 4 weeks obtaining the total closure of the same. Therefore the use of platelet lysate promotes granulation tissue and scarring in pilonidal fistula(AU)

Aplicación de las plaquetas en la extrofia vesical / Use of platelet in bladder exstrophy

La extrofia vesical es una anomalía congénita grave del tracto urinario inferior que afecta la vejiga, los huesos pelvianos, la pared abdominal, los genitales externos, el perineo y, en algunos casos, el intestino. Los tratamientos convencionales están basados en técnicas quirúrgicas, para lograr reconstruir la vejiga, los genitales y cerrar el defecto de la pared. Se presenta el caso de una niña de 3 años de edad, en la que el tratamiento quirúrgico se había aplicado en 6 ocasiones sin resultados. Se decidió combinar el tratamiento quirúrgico con la aplicación de medicina regenerativa. Se le aplicó lisado de plaquetas en la pared de la vejiga y los bordes de la pared abdominal a razón de 1 mL semanal, durante 4 semanas. Se logró la regeneración de los tejidos que permitió afrontar los bordes de dichas estructuras y realizar la técnica quirúrgica convencional. Como resultado se logró cierre total de vejiga, uretra y pared abdominal, sin fístulas entre estas estructuras ni al exterior. La utilización del lisado de plaquetas favorece el crecimiento y regeneración de los tejidos que componen el tracto urinario. La cirugía puede ser una solución definitiva, después de haber aplicado los factores de crecimiento plaquetarios, que preparan el tejido en cuanto a calidad y cantidad, favoreciendo el afrontamiento de los bordes, la cicatrización y disminuyendo las complicaciones posquirúrgicas(AU)

Bladder exstrophy is a severe congenital anomaly of lower urinary tract that affects the bladder, pelvic bones, abdominal wall, external genitalia, perineum and in some cases intestine. Conventional treatments are based on surgical techniques, in order to reconstruct bladder, genitals and close wall defect. We present a case of a 3 year old girl, in which surgical treatment was applied 6 times without results. It was decided to combine the surgical treatment with application of regenerative medicine. It was applied platelet lysate in the bladder wall and the edges of the abdominal wall at the rate of 1 mL weekly for 4 weeks, achieving tissue regeneration. It enabled to face the edges of those structures and perform conventional surgical technique. As a result, we achieved a total closure of bladder, urethra and abdominal wall, without fistulas either between these structures or outside. Therefore the use of platelet lysate promotes growth and tissue regeneration comprising the urinary tract, decreasing number of interventions, time exposition structures of the abdominal cavity, and post surgical complications such as fistulas(AU)

Lisado plaquetario y células mononucleares autólogas de la periodontitis crónica. Presentación de un caso / Autologous platelet lysate and mononuclear stem cell of chronic periodontitis. Case presentation

Introducción: El tratamiento de la periodontitis crónica se prolonga por persistencia de la inflamación en tejidos afectados. La medicina regenerativa muestra resultados alentadores. Es la primera vez en Cuba, según bibliografía revisada, que se usa la combinación de lisado plaquetario y células mononucleares autólogas en este tratamiento. Objetivo: Mostrar el efecto de la combinación del lisado plaquetario y células mononucleares autólogas en el tratamiento de la periodontitis. Presentación de un caso: Paciente femenina de 44 años de edad, con diagnóstico de periodontitis, quien desde hace 1 año lleva tratamiento, sin efectuarse procedimientos quirúrgicos por la inflamación persistente. Es remitida a la consulta de periodoncia del Hospital Enrique Cabrera para recibir tratamiento con medicina regenerativa. Se consideró tratar con lisado de plaquetas autólogas, una vez concluido el raspado y alisador radicular, se prefundieron las encías afectadas con el lisado plaquetario. A los 7 días de aplicado el lisado se constatan las encías sin signos clínicos de inflamación, y disminución ostensible del número de dientes afectados y la profundidad o eliminación de las bolsas periodontales y movilidad dentaria. Al mes se trataron quirúrgicamente los defectos óseos con implante de células mononucleares. Al evaluar a los 6 meses, se hallaron de forma variable, signos radiográficos de organización del trabeculado, definición de la cortical y formación ósea en zona de defectos óseos. Conclusiones: El tratamiento combinado del lisado plaquetario autólogo y células mononucleares muestra una evaluación satisfactoria en el tratamiento de la periodontitis crónica y reduce el tiempo de tratamiento(AU)

Introduction: Treatment of chronic periodontitis is prolonged by persistent inflammation in affected tissues. Regenerative medicine shows encouraging results. It is the first time in Cuba, according to the reviewed literature, that the combination of platelet lysate and autologous mononuclear cells is used in this treatment. Objective: to show the effect of the combination of platelet lysate and autologous mononuclear cells in the treatment of periodontitis. Presentation of the case: A 44-year-old female patient diagnosed with periodontitis who had been treated for 1 year without surgical procedures due to persistent inflammation. She is referred to the periodontics consultation of the Dr. Enrique Cabrera General Hospital to receive treatment with regenerative medicine. Treatment with autologous platelet lysate was considered, once the scaling and root planer had been completed, the affected gingivae were prefixed with the platelet lysate. At 7 days after the lysate is applied, the gingiva shows no clinical signs of inflammation, and a noticeable reduction in the number of affected teeth and the depth or elimination of periodontal pockets and tooth mobility. Bone defects with a mononuclear cell implant were surgically treated one month later. When evaluating at 6 months, radiographic signs of trabecular organization, definition of the cortical bone, and bone formation in the area of bone defects were found variably. Conclusions: The combined treatment of autologous platelet lysate and mononuclear cells shows a satisfactory evaluation in the treatment of chronic periodontitis and reduces treatment time(AU)