Protective effects of mesenchymal stromal cell-derived secretome on dermonecrosis induced in rabbits by Loxosceles intermedia spider venom.

Background: Loxoscelism refers to a set of clinical manifestations caused by the bite of spiders from the Loxosceles genus. The classic clinical symptoms are characterized by an intense inflammatory reaction at the bite site followed by local necrosis and can be classified as cutaneous loxoscelism. This cutaneous form presents difficult healing, and the proposed treatments are not specific or effective. This study aimed to evaluate the protective effect of mesenchymal stromal cells-derived secretome on dermonecrosis induced by Loxosceles intermedia spider venom in rabbits. Methods: Sixteen rabbits were distributed into four groups (n = 4). Except for group 1 (G1), which received only PBS, the other three groups (G2, G3, and G4) were initially challenged with 10 µg of L. intermedia venom, diluted in 100 µL of NaCl 0.9%, by intradermic injection in the interscapular region. Thirty minutes after the challenge all groups were treated with secretome, except for group 2. Group 1 (G1-control group) received intradermal injection (ID) of 60 µg of secretome in 0.15 M PBS; Group 2 (G2) received 0.9% NaCl via ID; Group 3 (G3) received 60 µg of secretome, via ID and Group 4 (G4), received 60 µg of secretome by intravenous route. Rabbits were evaluated daily and after 15 days were euthanized, necropsied and skin samples around the necrotic lesions were collected for histological analysis. Results: Rabbits of G1 did not present edema, erythema, hemorrhagic halo, or necrosis. In animals from G2, G3, and G4, edema appeared after 6h. However, minor edema was observed in the animals of G2 and G3. Hemorrhagic halo was observed in animals, six hours and three days after, on G2, G3, and G4. Macroscopically, in G4, only one animal out of four had a lesion that evolved into a dermonecrotic wound. No changes were observed in the skin of the animals of G1, by microscopic evaluation. All animals challenged with L. intermedia venom showed similar alterations, such as necrosis and heterophilic infiltration. However, animals from G4 showed fibroblast activation, early development of connective tissue, neovascularization, and tissue re-epithelialization, indicating a more prominent healing process. Conclusion: These results suggest that secretome from mesenchymal stromal cells cultured in a xeno-free and human component-free culture media can be promising to treat dermonecrosis caused after Loxosceles spiders bite envenoming.

Efectividad del tratamiento con plasma rico en plaquetas y leucocitos en la fisura anal secundaria / Effectiveness of Treating Secondary Anal Fissure with Platelet and Leukocyte-Rich Plasma

Introducción: El plasma rico en plaquetas es un producto derivado de la sangre, rico en péptidos y proteínas de señalización intercelular, así como citoquinas capaces de intervenir en cada una de las etapas de la regeneración de varios tejidos. Objetivo: Evaluar la efectividad del tratamiento con plasma rico en plaquetas y leucocitos en pacientes con fisura anal secundaria que asistieron a la consulta de Coloproctología del Hospital Docente Provincial Oncológico María Curie de Camagüey en el período de enero de 2020 a enero de 2023. Métodos: Se realizó un estudio cuasiexperimental. El universo lo conformaron todos los pacientes que acudieron a consulta en ese período. La muestra no probabilística intencionada la conformaron 130 pacientes adultos con diagnóstico de fisura anal secundaria que recibieron tratamiento con plasma rico en plaquetas y leucocitos. Resultados: El tiempo de cicatrización de la fisura anal permitió corroborar la efectividad del tratamiento; además, la mayoría de los pacientes evolucionaron de forma favorable. El dolor fue la complicación más visible pues presentó significación estadística al establecer la relación entre las variables. Se demostró que el tratamiento con plasma rico en plaqueta y leucocitos en pacientes con fisura anal secundaria es efectivo y seguro en la evaluación final del tratamiento. Conclusiones: Lo expuesto permite considerar que los pacientes tratados obtuvieron buenos resultados al hacer uso del plasma rico en plaquetas y leucocitos. Por consiguiente, tiene un resultado positivo en no mostrar complicaciones y una alta posibilidad de que el paciente tratado evolucione en mejor condición(AU)

Introduction: Platelet-rich plasma is a blood-derived product, rich in peptides and intercellular signaling proteins, as well as cytokines capable of intervening in each of the stages of regeneration of various tissues. Objective: To assess the effectiveness of treatment with platelet- and leucocyte-rich plasma in patients with secondary anal fissure who attended the coloproctology office at Hospital Docente Provincial Oncológico María Curie of Camagüey from January 2020 to January 2023. Methods: A quasiexperimental study was carried out, whose universe consisted of all the patients who came for consultation during that period. The nonprobabilistic purposive sample consisted of 130 adult patients with a diagnosis of secondary anal fissure who received treatment with platelet- and leucocyte-rich plasma. Results: The healing time of the anal fissure allowed corroborating the effectiveness of the treatment; in addition, most of the patients evolved favorably. Pain was the most visible complication since it presented statistical significance when the relationship between the variables were established. Treatment with platelet- and leucocyte-rich plasma in patients with secondary anal fissure proved effective and safe in the final assessment of the treatment. Conclusions: The above allows considering that the treated patients obtained good outcomes when making use of platelet- and leucocyte-rich plasma. Therefore, it has a positive outcome, not showing complications and a high possibility for the treated patient to evolve in a better condition(AU)

Stem cell therapy for erectile dysfunction: a systematic review.

INTRODUCTION: Erectile dysfunction (ED) is a common condition that negatively affects men's quality of life. It can have various causes, including psychological, vascular, and neurologic factors. Existing treatments for ED mainly focus on symptom relief rather than addressing the underlying cause. Stem cells (SCs) have shown potential as a therapeutic approach for ED due to their anti-inflammatory properties. OBJECTIVES: This systematic review aims to assess the current status of trials and determine the potential impact of SCs on male sexual health. METHODS: A comprehensive search strategy was employed to gather relevant articles from 6 electronic databases. The search included articles published until March 2023. The reference lists of articles were manually reviewed to identify additional studies of relevance. The eligibility criteria for inclusion in the analysis focused on clinical trials involving humans that evaluated the safety and efficacy of SC therapy for ED. Exclusion criteria encompassed case reports, case series, abstracts, reviews, and editorials, as well as studies involving animals or SC derivatives. Data extraction was performed via a standardized form with a focus on erectile outcomes. RESULTS: A total of 2847 articles were initially identified; 18 were included in the final analysis. These studies involved 373 patients with ED and various underlying medical conditions. Multiple types of SC were utilized in the treatment of ED: mesenchymal SCs, placental matrix-derived mesenchymal SCs, mesenchymal SC-derived exosomes, adipose-derived SCs, bone marrow-derived mononuclear SCs, and umbilical cord blood SCs. CONCLUSION: SC therapy shows promise as an innovative and safe treatment for organic ED. However, the lack of standardized techniques and controlled groups in many studies hampers the ability to evaluate and compare trials.

Caracterización del tratamiento con plasma rico en plaquetas y leucocitos en la fisura anal secundaria / Characterization of the Treatment with Platelet-Rich Plasma and Leukocytes against Secondary Anal Fissure

Introducción: Las enfermedades benignas de recto son muy frecuentes en la práctica médica habitual en todos los niveles de atención de salud. Dentro de estos procesos se reconoce que la fisura anal es uno de los más comunes. Objetivo: Caracterizar a los pacientes tratados con plasma rico en plaquetas y leucocitos por fisura anal secundaria en la provincia Camagüey en el período comprendido entre septiembre del 2020 y septiembre del 2022. Métodos: Se realizó un estudio cuasiexperimental, cuyo universo lo conformaron los pacientes con el diagnóstico de fisura anal secundaria que asistieron a la consulta de Coloproctología del Hospital Provincial Docente Oncológico María Curie. Se estableció una muestra no probabilística a criterio de los autores conformada por 132 pacientes adultos, quienes recibieron tratamiento con plasma rico en plaquetas y leucocitos por fisura anal secundaria en la provincia Camagüey. Resultados: Con respecto al sexo y a la edad, se observó una mayor frecuencia del grupo de 40 a 49 años y predominó el sexo femenino. El síntoma de mayor frecuencia fue el dolor, mientras que en la etiología prevaleció la proctitis facticia. En la mayoría de los casos estudiados la fisura anal cicatrizó en menos de 15 días y en cuanto a la evaluación final del tratamiento solo la minoría empeoró. Conclusiones: La caracterización de la muestra en estudio permite afirmar que el tratamiento con plasma rico en plaquetas y leucocitos tiene beneficios para los pacientes con fistulas perianales y su calidad de vida(AU)

Introduction: Benign rectal diseases are very frequent in routine medical practice at all levels of health care. Among these processes, anal fissure is recognized as one of the most common. Objective: To characterize patients treated with platelet-rich plasma and leukocytes for secondary anal fissure in Camagüey Province between September 2020 and September 2022. Methods: A quasiexperimental study was carried out, whose universe was made up of patients with the diagnosis of secondary anal fissure who attended the coloproctology office at Hospital Provincial Docente Oncológico María Curie. A nonprobabilistic sample was established according to the authors' criterions, made up of 132 adult patients who received treatment with platelet-rich plasma and leukocytes for secondary anal fissure in Camagüey Province. Results: Regarding sex and age, a higher frequency was observed in the group 40 to 49 years old and the female sex was predominant. The most frequent symptom was pain, while the prevailing etiology was factitious proctitis. In most of the cases under study, the anal fissure healed in less than 15 days and, regarding the final assessment of the treatment, only the minority worsened. Conclusions: The characterization of the sample under study allows to affirm that treatment with platelet-rich plasma and leukocytes has benefits for patients with perianal fistulas and their quality of life(AU)

Addition of synthetic polymer in the freezing solution of mesenchymal stem cells from equine adipose tissue as a future perspective for reducing of DMSO concentration.

The regenerative therapies with stem cells (SC) has been increased by the cryopreservation, permitting cell storage for extended periods. However, the permeating cryoprotectant agents (CPAs) such as dimethylsulfoxide (DMSO) can cause severe adverse effects. Therefore, this study evaluated equine mesenchymal stem cells derived from adipose tissue (eAT-MSCs) in fresh (Control) or after slow freezing (SF) in different freezing solutions (FS). The FS comprise DMSO and non-permeating CPAs [Trehalose (T) and the SuperCool X-1000 (X)] in association or not, totalizing seven different FS: (DMSO; T; X; DMSO+T; DMSO+X; T+X, and DMSO+T+X). Before and after cryopreservation were evaluated, viability, colony forming unit (CFU), and cellular differentiation capacity. After freezing-thawing, the viability of the eAT-MSCs reduced (P< 0.05) in all treatments compared to the control. However, the viability of frozen eAT-MSCs in DMSO (80.3 ± 0.6) was superior (P<0.05) to the other FS. Regarding CFU, no difference (P>0.05) was observed between fresh and frozen cells. After freezing-thawing, the eAT-MSCs showed osteogenic, chondrogenic, and adipogenic lineages differentiation potential. Nonetheless, despite the significative reduction in the osteogenic differentiation capacity between fresh and frozen cells, no differences (P > 0.05) were observed among FS. Furthermore, the number of chondrogenic differentiation cells frozen in DMSO+X solution reduced (P<0.05) comparing to the control, without differ (P>0.05) to the other FS. The adipogenic differentiation did not differ (P>0.05) among treatments. In conclusion, although these findings confirm the success of DMSO to cryopreserve eAT-MSCs, the Super Cool X-1000 could be a promise to reduce the DMSO concentration in a FS.

As terapias regenerativas com células-tronco (CT) têm sido incrementadas pela criopreservação, permitindo o armazenamento celular. No entanto, os agentes crioprotetores (ACPs) penetrantes, como DMSO, podem causar efeitos adversos graves. Portanto, este estudo avaliou células-tronco mesenquimais equinas derivadas de tecido adiposo (CTM-TAe) in natura (Controle) ou após congelamento lento (CL) em diferentes soluções de congelamento (SC). As SCs compreendem DMSO e ACPs não permeáveis [Trealose (T) e o SuperCool X-1000 (X)] associados ou não: (DMSO; T; X; DMSO+T; DMSO+X; T +X e DMSO+T+X). Antes e após a criopreservação foram avaliados, viabilidade, unidade formadora de colônia (UFC) e capacidade de diferenciação celular. Após o congelamento-descongelamento, a viabilidade das CTM-TAe reduziu (P< 0,05) em todos os tratamentos em relação ao controle. Entretanto, a viabilidade das CTM-TAe congeladas em DMSO (80,3 ± 0,6) foi superior (P<0,05) às demais SC. Em relação às UFC, não houve diferença (P>0,05) entre células frescas e congeladas. Após congelamento-descongelamento, as CTM-TAe apresentaram potencial de diferenciação de linhagens osteogênicas, condrogênicas e adipogênicas. No entanto, apesar da redução significativa na capacidade de diferenciação osteogênica entre células frescas e congeladas, não foram observadas diferenças (P > 0,05) entre SCs. Além disso, o número de células de diferenciação condrogênica congeladas em solução de DMSO+X reduziu (P<0,05) em relação ao controle, sem diferir (P>0,05) das demais SCs. A diferenciação adipogênica não diferiu (P>0,05) entre os tratamentos. Em conclusão, embora esses achados confirmem o sucesso do DMSO para criopreservar CTM-TAe, o Super Cool X-1000 pode ser uma promessa para reduzir a concentração de DMSO.

Efectos de la administración local de PTH 1-34 a bajas dosis en un modelo experimental de periodontitis: estudio piloto / Local effects of low dose of PTH 1-34 on experimental periodontitis: pilot study

La periodontitis es una patología inflamatoria que aumenta la resorción de hueso alveolar (HA), pérdida de la inserción dentaria y posible exfoliación. Evaluamos el efecto de la administración intermitente de bajas dosis de parathormona (PTH) 1-34 sobre la recuperación de la masa ósea pérdida en un modelo experimental de periodontitis inducida por una ligadura periodontal (LP) con hilo de algodón alrededor de la pieza dentaria. Las ratas fueron divididas luego de 5 días en instaurada la periodontitis en: CT LP sin trata-miento y PTH LP tratados con 0,2 µg/kg PTH 1-34 subcutánea local, tres veces por semana por 17 días. El control absoluto fue un tercer grupo sin LP (CT). Se estudiaron parámetros antropométricos, bioquímicos e histomosfométricos en tibias y hemimandibulas. La calcemia, fosfatemia, CTX sérico, PTHi y vo-lumen óseo (BV/TV%) de tibias fueron similares en los tres grupos. El BV/TV% del HA fue significativamente menor en PTH LP respecto de CT pero mayor que CT LP (p<0.05). La pérdida ósea de HA porcentual fue significativamente mayor en CT LP (p<0.05). La altura del ligamento periodontal fue significativamente menor en PTH LP que en CT (p<0.05) y mayor respecto de CT LP, sin alcanzar diferencias significativas. Los resultados del presente estudio piloto sugieren que la administración intermitente de PTH en bajas dosis y durante un periodo de tiempo corto disminuye la progresión de la enfermedad periodontal sin generar efectos sistémicos. Como no se logró regenerar totalmente el tejido periodontal se requieren estudios adicionales. (AU)

Periodontitis is an inflammatory chronic disease with high prevalence in adults that induces a progressive alveolar bone (AB) loss leading to tooth loss. Experimental periodontitis can be induced in rats by cotton ligature placement (LP) in the gingival sulcus around the molar teeth. The biofilm accumulation and disruption of the gingival epithelium lead to bone resorption. We investigated whether intermittent administration of a low dose of PTH 1-34 may recover the alveolar bone loss in the experimental periodontitis induced in female Wistar rats. Animals were randomly divided in two groups which were subcutaneously injected with: saline solution (CT LP) or 0,2 µg/kg PTH 1-34 (PTH LP) three times per week during 17 days. Unligated rats were taken as healthy controls (CT). Anthropometric, biochemical and histologic analysis of tibia and hemimandible were done. No differences in serum calcium, phosphorus, CTX, PTHi or subchondral tibia bone volume (BV/TV%) were observed between the three groups. AB BV/TV% was significantly lower in PTH LP than in CT but higher than in CT LP (p<0.05). The highest percentage of AB loss was observed in CT LP. The height of periodontal ligament was lower in PTH LP than in CT (p<0.05) but not significantly higher than CT LP.The increase in AB loss by experimental periodontitis appears to be corrected by the intermittent administration of low doses of PTH without systemic effect. As the recovery of periodontal tissue was only partial, additional studies should be done.

Terapia endodôntica regenerativa: relato de caso clínico / Regenerative endodontic therapy: clinical case report

O presente trabalho, traz um relato de caso clínico em que foi realizada a terapia endodôntica regenerativa (também conhecida como "revascularização pulpar"), em dente permanente de um paciente jovem (11 anos), com história de ter sofrido trauma dentário que resultou em fratura corono-radicular do incisivo central superior direito. A terapia foi realizada na clínica de especialização da Faculdade de Odontologia da Universidade Federal de Minas Gerais (UFMG). O paciente iniciou o tratamento no ano de 2017, mas a terapia regenerativa foi possível de ser concluída apenas em 2022, devido à difícil adesão do paciente ao tratamento. No decorrer dos 5 anos de atendimento, foram realizadas trocas da medicação intracanal (MIC) à base de hidróxido de cálcio. Em 2022, ao retomar o atendimento do paciente, a MIC foi renovada, e na próxima sessão, removida. O sangramento intracanal foi então induzido com auxílio de lima a 2 mm do comprimento de trabalho. Após formação do coágulo sanguíneo, utilizou-se membrana de colágeno tipo 1, para dar apoio à acomodação da barreira de agregado trióxido mineral (MTA). O dente foi então restaurado com resina composta. Após 5 meses, o paciente relatou diminuição da sintomatologia dolorosa, e aguarda-se o retorno deste para avaliação radiográfica.

The present work presents a clinical case report in which regenerative endodontic therapy (also known as "pulp revascularization") was performed on the permanent tooth of a young patient (11 years old), with a history of having suffered dental trauma that resulted in coronal-radicular fracture of the maxillary right central incisor. The therapy was performed at the specialization clinic of the Faculty of Dentistry of the Federal University of Minas Gerais (UFMG). The patient started treatment in 2017, but regenerative therapy was only possible to be completed in 2022, due to the patient's difficult adherence to treatment. During the 5 years of care, intracanal medication (MIC) based on calcium hydroxide was changed. In 2022, upon resuming patient care, the MIC was renewed, and in the next session, removed. Intracanal bleeding was then induced with the aid of a file 2 mm from the working length. After formation of the blood clot, a type 1 collagen membrane was used to support the accommodation of the mineral trioxide aggregate (MTA) barrier. The tooth was then restored with composite resin. After 5 months, the patient reported a decrease in pain symptoms, and he is awaiting his return for radiographic evaluation.

Subluxación sacroilíaca en un equino criollo colombiano / Sacroiliac subluxation in a horse in Colombia

Resumen La subluxación sacroilíaca es una enfermedad claudiocógena de los miembros posteriores que se puede presentar de forma aguda o crónica. La causa de esta patología se debe principalmente a resbalones, tropezones o caídas de los miembros posteriores, el diagnóstico es difícil de realizar debido a la profunda ubicación anatómica de la articulación, métodos como la ecografía y pruebas de estrés entre otros, son los procedimientos más usados en el diagnóstico de la subluxación sacroiliaca. Por tratarse de una alteración que fácilmente puede confundirse con otras condiciones patológicas de la columna, la pelvis y/o de los miembros posteriores, el diagnóstico muchas veces se realiza por eliminación de otras causas de cojera. Los tratamientos convencionales aún se proponen en la literatura, sin embargo, cada vez como opción para el tratamiento de esta patología. El presente reporte describe el caso de un equino criollo que padeció una subluxación sacroiliaca aguda del lado derecho, la cual fue diagnosticada con un examen clínico que incluía ecografía percutánea y transrectal, y tratada con infiltraciones ecoguiadas de corticoides y aspirado de médula ósea. Un mes después de la última terapia, el caballo mostró una mejoría clínica en un 80% a 90% aproximadamente y retoma actividad física controlada.

Abstract Sacroiliac subluxation is a claudiogenic disease of the hind limbs that can present acutely or chronically. The cause of this pathology is mainly due to slipping, tripping, or falling of the hind limbs, the diagnosis is difficult to make due to the deep anatomical location of the joint, methods such as ultrasound and stress tests among others, are the procedures most used in the diagnosis of sacroiliac subluxation. As it is an alteration that can easily be confused with other pathological conditions of the spine, pelvis and / or the hind limbs, the diagnosis is often made by eliminating other causes of lameness. Conventional treatments are still proposed in the literature, however regenerative therapy is increasingly used as an option for the treatment of sacroiliac joint injuries. This report describes the case of a Creole horse that suffered an acute sacroiliac subluxation on the right side, which was diagnosed with a clinical examination that included percutaneous and transrectal ultrasound and treated with ultrasound-guided infiltrations of corticosteroids and cells of the bone marrow of the sternum without processing. One month after the last therapy, the horse showed an 80% to 90% recovery and resumed controlled physical activity.

Alcance de las células madre derivadas de tejido adiposo / Scope of stem cells derived from adipose tissue

Introducción: En el tejido adiposo se han identificado células madre mesenquimales con capacidad autorrenovadora y multipotencial. Mediante digestión enzimática y centrifugado del lipoaspirado se libera una población heterogénea de células denominada fracción vascular estromal, con innumerables potencialidades terapéuticas en el campo de la medicina regenerativa. Objetivo: Actualizar el alcance de las células madre derivadas de tejido adiposo en la terapia regenerativa. Método: Se revisaron 38 artículos entre los años 2000 y 2019 en las bases de datos Scielo, ScienceDirect, Medline y Pubmed. Desarrollo: Las células de la fracción vascular estromal se caracterizan por su capacidad de generar tejido adiposo y vasos sanguíneos, y por la producción de factores de crecimiento que ayudan en la supervivencia de los adipocitos y la formación de una red vascular. El principal mecanismo de acción de las células madre derivadas de tejido adiposo parece deberse a su acción paracrina y a la sinergia con células endoteliales. En el campo de la medicina regenerativa se han utilizado en el tratamiento de cicatrices patológicas y de fibrosis deformantes con impotencia funcional, en las reconstrucciones de secuelas por cáncer y en el cierre precoz de zonas cruentas. Conclusiones: La lipotransferencia es un procedimiento con un mínimo de complicaciones que constituye una de las opciones terapéuticas más empleadas para corregir defectos en los tejidos, debido a que no solo es un medio de relleno, sino que también permite la regeneración y restauración tisular. La presencia de células madre en el tejido adiposo, unido a su accesibilidad, disponibilidad e histocompatibilidad, ha motivado su aplicación cada vez más expandida en la medicina estética, reconstructiva y regenerativa(AU)

Introduction: Mesenchymal stem cells with self-renewing and multipotential capacity have been identified in adipose tissue. By means of enzymatic digestion and centrifugation of the lipoaspirate a heterogeneous population of cells called vascular stromal fraction is released. It has innumerable therapeutic potentialities in the field of regenerative medicine. Objective: To update the scope of stem cells derived from adipose tissue in regenerative therapy. Method: 38 articles published between 2000 and 2019 in the Scielo, ScienceDirect, Medline and Pubmed databases were reviewed. Development: The cells of the vascular stromal fraction are characterized by generating adipose tissue and blood vessels and by the production of growth factors that help in the survival of adipocytes and the formation of a vascular network. The main mechanism of action of stem cells derived from adipose tissue appears to be due to their paracrine action and synergy with endothelial cells. Stem cells derived from adipose tissue have been used in regenerative medicine for the treatment of pathological scars and deforming fibrosis with functional impotence, in the reconstruction of cancer sequelae and in the early closure of bloody areas. Conclusions: Lipotransfer is a procedure with a minimum of complications that constitutes one of the most widely used therapeutic options to correct tissue defects, since it is not only a filling medium, but also allows tissue regeneration and restoration. The presence of stem cells in adipose tissue, together with their accessibility, availability and histocompatibility, has motivated their increasingly widespread application in aesthetic, reconstructive and regenerative medicine(AU)

Caracterización morfológica de las células de sangre de cordón umbilical / Morphological characterization of umbilical cord blood cells

Introducción: El cordón umbilical se ha convertido en un elemento de interés para la medicina regenerativa en los últimos años, pues constituye una fuente importante de células madres y progenitores hematopoyéticos. Objetivo: Caracterizar morfológicamente la sangre del cordón umbilical para terapia regenerativa en recién nacidos del Hospital Universitario Ginecobstétrico Ana Betancourt de Mora. Métodos: Estudio observacional, analítico y transversal realizado en el Centro de Inmunología y Productos Biológicos de Camagüey, entre enero y diciembre de 2017. Se evaluaron 35 muestras de sangre del cordón umbilical obtenidas de recién nacidos, que fueron partos eutócicos y sus madres no tuvieron procesos de enfermedades infecciosas. Resultados: El promedio mayor de células mononucleares correspondió a los linfocitos. En el conteo diferencial los polimorfonucleares neutrófilos ocuparon el primer lugar, seguido de los linfocitos, con medias de 0,50 y 0,46 x 109/L, respectivamente. De las células presentes en el frotis del botón, fueron más frecuente los linfocitos con 0.59 x 109/L; se observó un promedio de monocitos de 0,00-0,07 x 109/L. Conclusiones: La obtención de células mononucleares viables a través de la vena umbilical, constituye una técnica promisoria en las investigaciones biomédicas. Entre las células mononucleares predominaron los linfocitos, tanto en la sangre del cordón umbilical como en el botón celular aislado(AU)

Introduction: In recent years, the umbilical cord has become an element of interest for regenerative medicine, based on its importance as a source of stem cells and hematopoietic progenitors. Objective: To characterize the morphology of umbilical cord blood for regenerative therapy in newborns from Ana Betancourt de Mora Gyneco-obstetric University Hospital. Methods: Observational, analytical and cross-sectional study carried out at the Center of Immunology and Biological Products in Camagüey, between January and December 2017. We evaluated 35 samples of umbilical cord blood obtained from newborns who were eutocic deliveries and whose mothers did not have infectious disease processes. Results: The highest average of mononuclear cells corresponded to lymphocytes. In the differential count, neutrophil polymorphonuclear cells ranked first, followed by lymphocytes, with averages of 0.50x109 and 0.46x109 per liter, respectively. Of the cells present in the button cell smear, lymphocytes were more frequent, with 0.59x109 per L; an average of monocytes was observed, with 0.00-0.07x109 per L was observed. Conclusions: Obtaining viable mononuclear cells through the umbilical vein is a promising technique in biomedical research. Among the mononuclear cells, lymphocytes predominated, both in the cord blood and in the isolated cell button(AU)

Aplicación de la terapia regenerativa en la enfermedad arterial periférica y el pie diabético isquémico / Use of regenerative therapy in the peripheral arterial disease and the ischemic diabetic foot

Introducción: El pie diabético isquémico y la enfermedad arterial periférica, son dos enfermedades que ocasionan dolor por isquemia crítica y ponen en peligro la viabilidad de las extremidades inferiores. Objetivo: Demostrar la efectividad de la terapia regenerativa con plasma rico en plaquetas en pacientes con las enfermedades citadas. Métodos: Estudio longitudinal, prospectivo y aleatorizado realizado en el Servicio de Angiología del Hospital: Julio Trigo López, entre enero 2016-diciembre 2017. Se incluyeron 26 pacientes en dos grupos de estudios, 10 pacientes no diabéticos con claudicación intermitente y 16 pacientes con lesiones isquémicas en el pie del diabético. Se aplicó plasma rico en plaquetas por vía intramuscular en la pierna afectada, en el primer grupo y de forma intra- y perilesional en el pie del diabético isquémico. Resultados: Se obtuvo alivio del dolor y mejoría en más del 70 por ciento de los pacientes en la distancia de claudicación por encima o cerca de los 200 metros. En los pacientes con pie diabético isquémico, alivio del dolor y cierre de las lesiones isquémicas en el 81,3 por ciento, solo 3 pacientes requirieron amputaciones mayores de los miembros inferiores. Conclusiones: La terapia regenerativa resulta efectiva en el salvataje de las extremidades inferiores en los pacientes estudiados, asociada a los procedimientos convencionales empleados en la cirugía revascularizadora; disminuye la amputación mayor de miembros inferiores y es una posibilidad terapéutica más en manos de especialistas entrenados, por su impacto social y en la calidad de vida de pacientes y familiares(AU)

Introduction: Ischemic diabetic foot and peripheral arterial disease are two conditions that cause pain and put in danger the functionality of lower limbs. Objective: To demonstrate the effectiveness of regenerative therapy with platelet-rich plasma in patients with the above mentioned diseases. Methods: Longitudinal, prospective and randomized study that was conducted in the Angiology Service of Julio Trigo López Hospital, from January 2016 to December 2017. There were 26 patients included in 2 groups: 10 non-diabetic patients with intermittent claudication and 16 patients with ischemic lesions in the diabetic foot. Platelet-rich plasma was used instramuscularly in an affected legs of the first group, and intraperilesional and perilesional in the diabetic foot. Results: There was pain relief and improvement in more than 70 percent of patients in the claudication distance higher or near to 200 meters. In the patients presenting ischemic diabetic foot, pain relief and closing of ischemic lesions in the 81,3 percent , just 3 patients required major amputations of lower limbs. Conclusions: Regenerative therapy results effective in the rescue of lower limbs in patients presenting peripheral arterial disease and ischemic diabetic foot. It also reduces major amputations of lower limbs and it is another therapeutic option in the hands of trained specialists due to its social impact and in the life quality of patients and their families(AU)

Changes on the Structural Architecture and Growth Factor Release, and Degradation in Equine Platelet-Rich Fibrin Clots Cultured Over Time.

The aims of the present study were (1) to describe the microscopic and ultrastructural appearance of equine platelet-rich fibrin (PRF) clots and (2) to determine the release and degradation of transforming growth factor beta 1 (TGF-ß1) and insulin-like growth factor type I (IGF-I) from PRF clots incubated over 14 days. Whole blood from six horses was collected into plain tubes and centrifuged at 240 g for 8 minutes. Clots were evaluated by histology and by both transmission and scanning electronic microscopy (TEM and SEM). Growth factor concentrations were measured by ELISA at 48-hour intervals over 14 days and analyzed by one-way repeated-measures ANOVA. Histology showed a clot composed by a fibrin layer and a cellular layer with platelets and leukocytes. Scanning electron microscopy showed the cells trapped by an incipient fibrin network at 1 hour. At day 8, these cells were embedded by an incipient fibrin network. At day 14, the leukocytes and platelet aggregates from the clot were imbibed in an organized web of fibrin fibrils. TEM exhibited platelets with preserved cytoplasm and alpha granules randomly scattered at day 8, and damaged platelets with interrupted cytoplasm and organelle emigration to the periphery at day 14. TGF-ß1 and IGF-I concentrations showed a progressive increase until day 14. TGF-ß1 was released from PRF clots in a gradual and controlled manner, and increasing its concentration for two weeks, which supports TEM findings indicating that platelets began disintegrating by day 14. Furthermore, IGF-I production and release from PRF clots is sustained over time.

Use of platelet-rich plasma in the treatment of desmitis of distal interphalangeal joint collateral ligament in a horse

Background: Platelet-rich plasma (PRP) has been increasingly used to hasten healing of tendonitis and desmitis, allowinghorse athletes to go back to practicing sports in a shorter period of time. However, its use has rarely been reported in thetreatment of desmitis of the collateral ligaments of the distal interphalangeal joint (CL-DIP), which is based on the useof horseshoes and rest for at least six months. This paper describes the use of PRP as an aid in the treatment of CL-DIPin a horse athlete.Case: A male Criollo horse, 450 kg, about 15-year-old, used for paleteada. The animal had a month-long history of lameness, and a visual inspection revealed an increase in volume in the lateral dorsal region of the limb proximal to the coronaryband of the hoof. A specific examination of the locomotive system showed Grade III, Category 5 claudication, which became evident the beginning of the exercise and ceased after a short period. A radiographic evaluation of the distal phalanxrevealed no clinically relevant changes. Therefore, and in view of the location of the increased volume, an ultrasound scanwas performed, which showed an image compatible with an extensive hematoma in the region underlying the periopliccorium, while the CL-DIP presented swelling with large areas of hypoechogenicity. Considering the findings of the image,the physical examination and the history of the horse, desmitis of the collateral ligaments of the distal interphalangeal jointwas diagnosed. The treatment plan consisted of rest, application of a therapeutic horseshoe, and infiltration of the lesionwith platelet-rich plasma (PRP). First, however, warm compresses were applied to reduce the swelling. The PRP wasobtained by collecting whole blood from the external jugular vein, subjecting it to double centrifugation, and separatingthe supernatant blood plasma. The end product was applied to the lesion by ultrasound-guided infiltration. The horseshoewas a model...(AU)

Bone, periodontal and dental pulp regeneration in dentistry: a systematic scoping review

Abstract The aim of presented systematic scoping review was to investigate the actual and future clinical possibilities of regenerative therapies and their ability to regenerate bone, periodontal and pulp with histological confirmation of the nature of formed tissue. Electronic search was conducted using a combination between Keywords and MeSH terms in PubMed, Scopus, ISI-Web of Science and Cochrane library databases up to January 2016. Two reviewers conducted independently the papers judgment. Screened studies were read following the predetermined inclusion criteria. The included studies were evaluated in accordance with Arksey and O'Malley's modified framework. From 1349 papers, 168 completed inclusion criteria. Several characterized and uncharacterized cells used in Cell Therapy have provided bone regeneration, demonstrating bone gain in quantity and quality, even as accelerators for bone and periodontal regeneration. Synthetic and natural scaffolds presented good cell maintenance, however polyglycolid-polylactid presented faster resorption and consequently poor bone gain. The Growth Factor-Mediated Therapy was able to regenerate bone and all features of a periodontal tissue in bone defects. Teeth submitted to Revascularization presented an increase of length and width of root canal. However, formed tissues not seem able to deposit dentin, characterizing a repaired tissue. Both PRP and PRF presented benefits when applied in regenerative therapies as natural scaffolds. Therefore, most studies that applied regenerative therapies have provided promising results being possible to regenerate bone and periodontal tissue with histological confirmation. However, pulp regeneration was not reported. These results should be interpreted with caution due to the short follow-up periods.

Resumo O objetivo da presente Scoping review foi investigar as possibilidades clínicas atuais e futuras das terapias regenerativas e sua capacidade de regenerar tecido ósseo, periodontal e polpar em humanos com confirmação histológica da natureza do tecido formado. Uma busca eletrônica foi realizada utilizando uma combinação entre as palavras-chave e termos MeSH nos bancos de dados PubMed, Scopus, ISI-web of Science e Cochrane library até janeiro de 2016. Dois revisores realizaram de forma independente o julgamento dos documentos. Os estudos selecionados foram lidos seguindo os critérios de inclusão predeterminados. Os estudos incluídos foram avaliados de acordo com a estrutura modificada de Arksey e O'Malley. Dos 1349 artigos, 168 preencheram os critérios de inclusão. Várias células caracterizadas e não caracterizadas promoveram regeneração óssea utilizada em terapias celulares, demonstrando ganho ósseo em quantidade e qualidade, de forma rápida para regeneração óssea e periodontal. Os scaffolds sintéticos e naturais apresentaram boa manutenção celular, no entanto o poliglicol-polilácido apresentou uma reabsorção rápida e, consequentemente, pequeno ganho ósseo. A terapia mediada por fatores de crescimento foi capaz de regenerar tecido ósseo e todas as características de um tecido periodontal. Dentes submetidos à revascularização apresentaram aumento do comprimento e largura do canal radicular. No entanto, os tecidos formados não foram capazes de depositar dentina, caracterizando um tecido reparado. Tanto o PRP quanto o PRF parecem apresentar benefícios quando aplicados em terapias regenerativas sendo um bom scaffold natural. Portanto, a maioria dos estudos que aplicaram terapias regenerativas forneceram resultados promissores sendo possível regenerar tecido ósseo e periodontal com confirmação histológica. No entanto, não foi observada regeneração de polpa dental. Estes resultados devem ser interpretados com cautela.

Mesenchymal Stem Cells for Periodontal Tissue Regeneration in Elderly Patients.

Mesenchymal stem cell (MSC) grafting is a highly promising alternative strategy for periodontal regeneration in periodontitis, which is one of the primary causes of tooth loss in the elderly. However, aging progressively decreases the proliferative and differentiation potential of MSCs and diminishes their regenerative capacity, which represents a limiting factor for their endogenous use in elderly patients. Therefore, tissue regeneration therapy with MSCs in this age group may require a cellular source without the physiological limitations that MSCs exhibit in aging. In this sense, exogenous or allogeneic MSCs could have a better chance of success in regenerating periodontal tissue in elderly patients. This review examines and synthesizes recent data in support of the use of MSCs for periodontal regenerative therapy in patients. Additionally, we analyze the progress of the therapeutic use of exogenous MSCs in humans.

Use of platelet-rich plasma in the treatment of desmitis of distal interphalangeal joint collateral ligament in a horse

Background: Platelet-rich plasma (PRP) has been increasingly used to hasten healing of tendonitis and desmitis, allowinghorse athletes to go back to practicing sports in a shorter period of time. However, its use has rarely been reported in thetreatment of desmitis of the collateral ligaments of the distal interphalangeal joint (CL-DIP), which is based on the useof horseshoes and rest for at least six months. This paper describes the use of PRP as an aid in the treatment of CL-DIPin a horse athlete.Case: A male Criollo horse, 450 kg, about 15-year-old, used for paleteada. The animal had a month-long history of lameness, and a visual inspection revealed an increase in volume in the lateral dorsal region of the limb proximal to the coronaryband of the hoof. A specific examination of the locomotive system showed Grade III, Category 5 claudication, which became evident the beginning of the exercise and ceased after a short period. A radiographic evaluation of the distal phalanxrevealed no clinically relevant changes. Therefore, and in view of the location of the increased volume, an ultrasound scanwas performed, which showed an image compatible with an extensive hematoma in the region underlying the periopliccorium, while the CL-DIP presented swelling with large areas of hypoechogenicity. Considering the findings of the image,the physical examination and the history of the horse, desmitis of the collateral ligaments of the distal interphalangeal jointwas diagnosed. The treatment plan consisted of rest, application of a therapeutic horseshoe, and infiltration of the lesionwith platelet-rich plasma (PRP). First, however, warm compresses were applied to reduce the swelling. The PRP wasobtained by collecting whole blood from the external jugular vein, subjecting it to double centrifugation, and separatingthe supernatant blood plasma. The end product was applied to the lesion by ultrasound-guided infiltration. The horseshoewas a model...

Plasma rico em plaquetas: estudo comparativo entre a dupla centrifugação e o filtro E-PET (Equine Platelet Enhancement Therapy) / Comparative study of double spin and use of E-PET filter (Equine Platelet Enhancement Therapy) to obtain platelet rich plasma in horses

O plasma rico em plaquetas (PRP) consiste em uma alta concentração de plaquetas em um pequeno volume de plasma, sendo, em média, quatro vezes maior que a concentração sérica. O uso de PRP é justificado pela alta concentração de fatores de crescimento presentes em grânulos no interior das plaquetas, que possuem diversas funções como proliferação celular, quimiotaxia, angiogênese e diferenciação celular, que ampliam o poder de reparação tecidual. Há diversos protocolos para obtenção do PRP em equinos descritos na literatura, dentre os quais destacam-se os de dupla centrifugação, os automatizados e os filtros. Há diferenças substanciais no conteúdo do PRP dependendo do seu método de obtenção, principalmente no que se diz respeito à quantidade de leucócitos, plaquetas e concentração de fatores de crescimento. Sendo assim, o objetivo deste estudo foi comparar a utilização do concentrado de plaquetas obtido por protocolo de dupla centrifugação e o obtido pelo filtro E-PET (Equine Platelet Enhancement Therapy), levando-se em consideração a concentração plaquetária e leucocitária final, a quantificação de fatores de crescimento (TGFß e PDGF-BB) e a facilidade de realização entre tais métodos. Utilizou-se nove animais para a obtenção de PRP por dupla centrifugação e através do filtro E-PET, não havendo diferença estatística (p>0,05) entre os métodos de obtenção em relação à concentração plaquetária e leucocitária, entretanto, houve diferença estatística (p=0,002; p=0,004, respectivamente) em relação a concentração de TGFß e PDGF-BB. Dessa forma, concluiu-se que o filtro E-PET mostrou-se um método mais efetivo, sendo possível sua utilização à campo, além de proporcionar uma maior concentração de fatores de crescimento TGFß e PDGF-BB.(AU)

The platelet-rich plasma (PRP) consists of a high concentration of platelets in a small volume of plasma, four times greater (average) than the serum concentration. The use of PRP is justified by the high concentration of growth factors present in granules in the platelets, which have several functions such as cell proliferation, chemotaxis, angiogenesis and differenciation, which extend the power of tissue repair. There are several protocols to obtain PRP in horses described in the literature, among which are highlighted the double centrifugation, automated and filters. There are substantial differences in the PRP content depending on its method of production, especially when it concerns the amount of leukocytes, platelets and concentration of growth factors. Thus, the aim of this study was to compare the use of platelet concentrates obtained by double centrifugation protocol and obtained by the filter E-PET (Equine Platelet Enhancement Therapy) taking into account the platelet and leukocyte final concentration, the quantification of growth factors (TGFß and PDGF-BB) and the facility to do those methods. Nine horses were used to obtain PRP by double centrifugation and through the E-PET filter, with no statistical difference (p>0.05) between the methods relative to the platelet and leukocyte concentration; however, there was statistical difference (p=0.002 and p=0.004 respectively) compared with the concentration of TGFß and PDGF-BB. It was concluded that the E-PET filter proved to be a more effective method possible to use in the field and to provide a higher concentration of growth factors (TGFß and PDGF-BB).(AU)

Plasma rico em plaquetas: estudo comparativo entre a dupla centrifugação e o filtro E-PET (Equine Platelet Enhancement Therapy) / Comparative study of double spin and use of E-PET filter (Equine Platelet Enhancement Therapy) to obtain platelet rich plasma in horses

O plasma rico em plaquetas (PRP) consiste em uma alta concentração de plaquetas em um pequeno volume de plasma, sendo, em média, quatro vezes maior que a concentração sérica. O uso de PRP é justificado pela alta concentração de fatores de crescimento presentes em grânulos no interior das plaquetas, que possuem diversas funções como proliferação celular, quimiotaxia, angiogênese e diferenciação celular, que ampliam o poder de reparação tecidual. Há diversos protocolos para obtenção do PRP em equinos descritos na literatura, dentre os quais destacam-se os de dupla centrifugação, os automatizados e os filtros. Há diferenças substanciais no conteúdo do PRP dependendo do seu método de obtenção, principalmente no que se diz respeito à quantidade de leucócitos, plaquetas e concentração de fatores de crescimento. Sendo assim, o objetivo deste estudo foi comparar a utilização do concentrado de plaquetas obtido por protocolo de dupla centrifugação e o obtido pelo filtro E-PET (Equine Platelet Enhancement Therapy), levando-se em consideração a concentração plaquetária e leucocitária final, a quantificação de fatores de crescimento (TGFß e PDGF-BB) e a facilidade de realização entre tais métodos. Utilizou-se nove animais para a obtenção de PRP por dupla centrifugação e através do filtro E-PET, não havendo diferença estatística (p>0,05) entre os métodos de obtenção em relação à concentração plaquetária e leucocitária, entretanto, houve diferença estatística (p=0,002; p=0,004, respectivamente) em relação a concentração de TGFß e PDGF-BB. Dessa forma, concluiu-se que o filtro E-PET mostrou-se um método mais efetivo, sendo possível sua utilização à campo, além de proporcionar uma maior concentração de fatores de crescimento TGFß e PDGF-BB.(AU)

The platelet-rich plasma (PRP) consists of a high concentration of platelets in a small volume of plasma, four times greater (average) than the serum concentration. The use of PRP is justified by the high concentration of growth factors present in granules in the platelets, which have several functions such as cell proliferation, chemotaxis, angiogenesis and differenciation, which extend the power of tissue repair. There are several protocols to obtain PRP in horses described in the literature, among which are highlighted the double centrifugation, automated and filters. There are substantial differences in the PRP content depending on its method of production, especially when it concerns the amount of leukocytes, platelets and concentration of growth factors. Thus, the aim of this study was to compare the use of platelet concentrates obtained by double centrifugation protocol and obtained by the filter E-PET (Equine Platelet Enhancement Therapy) taking into account the platelet and leukocyte final concentration, the quantification of growth factors (TGFß and PDGF-BB) and the facility to do those methods. Nine horses were used to obtain PRP by double centrifugation and through the E-PET filter, with no statistical difference (p>0.05) between the methods relative to the platelet and leukocyte concentration; however, there was statistical difference (p=0.002 and p=0.004 respectively) compared with the concentration of TGFß and PDGF-BB. It was concluded that the E-PET filter proved to be a more effective method possible to use in the field and to provide a higher concentration of growth factors (TGFß and PDGF-BB).(AU)

Bioestimulación capilar con plasma rico en plaquetas contra la caída del cabello / Capillary biostimulation with platelets rich plasm against alopecia

Se describe el caso clínico de una paciente de 69 años de edad, quien acudió a la consulta de medicina regenerativa por presentar estrés mantenido y pérdida del cabello, para lo cual se decidió aplicar la bioestimulación con plasma rico en plaquetas lisado como terapia regenerativa, con plaquetas alogénicas en forma tópica, puesto que es un tratamiento inocuo y carente de complicaciones. Se obtuvo un incremento de los cabellos, espesamiento de la zona alopécica y la paciente recuperó su apariencia.

The case report of a 69 years patient is described who went to the regenerative medicine service for presenting maintained stress and loss of her hair, for which it was decided to evaluate biostimulation with platelets rich plasm as regenerative therapy for her, with topical allogeneic platelets, since it is an innocuous treatment with no complications. An increment and growing of the hair in the alopecic area was obtained and the patient recovered her look.

Liquid and gel Platelet Rich Plasma as skin healing adjuvant / Plasma Rico em Plaquetas em suas frações líquida e em gel como adjuvantes da cicatrização cutânea

Background: In recent decades, many researches have been conducted on processes involved in tissue repairing, mainly in the development of resources and technology designed to improve the wound healing progress. Platelet rich plasma (PRP) derived from autologous blood is defined as a plasma volume with platelet concentration higher than physiological level. It is an autogenous and low cost source of growth factors, which are essential for tissue regeneration due to their angiogenic, mitogenic, and chemotactic properties. The aim of this study was evaluate two forms of PRP- liquid and gel - regarding their capacity to influence quality and repair time of standardized skin injuries. Materials, Methods & Results: New Zealand healthy rabbits were distributed in three groups (n = 6): control group (CG), liquid platelet rich plasma group (LIQPRP), and gel platelet rich plasma group (GELPRP). Acute skin lesions were inducted in two areas approximately 2 cm close to scapular edge and depth including epidermis, dermis, and hypodermis to external muscular fascia. Animals received treatment according to each group. Injuries were measured with digital pachymeter in two directions: longer length (l) and longer width (w), every two days. Areas and healing rates were calculated. Microscopic analysis samples were collected on days seven and 14 and evaluated through hematoxylin and eosin [...](AU)