Name: mechanical and controlled prp injections in patients affected by androgenetic alopecia
Uploaded: 2018-01-27T14:00:00.000+00:00
Duration: 3 min 22 s
Description: Watch this Scientific Journal Video about Mechanical and Controlled PRP Injections in Patients Affected by Androgenetic Alopecia at JoVE.com

Los autores no tienen ninguna agradecimientos.

El protocolo de estudio cumplió con la declaración de Helsinki, y todos los pacientes proporcionan consentimiento informado antes de participar en el estudio.
1. no activado A-PRP preparación
<ol><li>Cosecha de 55 mL de sangre de una vena periférica del brazo con un tubo de la aguja de la mariposa según el protocolo de recogida de sangre local.</li>
	<li>Agregar 7 mL de solución de ácido citrato dextrosa (ACD) (112 mM de citrato) para el tubo de la sangre como anticoagulante.</li>
	<li>Centrifugar la sangre recogida en el tubo 260 x g por 10 min. Nota: se produjeron 23 mL de PRP A + Plasma pobre en plaquetas autólogo (A-PPP) y 20 mL de suspensión de células de sangre rojas (RBCs).</li>
	<li>Sacar el tubo que contiene A-PRP + A PPP y los glóbulos rojos de la centrifugadora e insertarla en un dispositivo selector de luz de láser.</li>
	<li>Separado A-PRP + A-PPP en una bolsa estéril por succión mecánica realizada por el selector de láser en el cual se coloca el kit. Los glóbulos rojos se eliminan automáticamente por aspiración mecánica. Nota: 14 mL de PPP A manualmente fue cosechado y extraído por la bolsa estéril el kit. 9 mL de PRP A se obtuvo.</li>
</ol>2. no activado A-PRP inyecciones
<ol><li>Dividir el cuero cabelludo de los pacientes afectados por AGA en frontal, parietal, vértice y áreas occipitales. PRECAUCIÓN: No realice anestesia (local o sistémica).</li>
	<li>Realizar inyecciones de PRP A interfollicular (0,20 mL x cm2) por el médico inyector del arma (véase Tabla de materiales) a las zonas del cuero cabelludo.<ol>
<li>En pacientes con pérdida de pelo localizada en las zonas frontal y parietales, realizar las inyecciones de PRP A exclusivamente al área frontal y utilice solución salina como placebo en la zona parietal.</li>
		<li>En pacientes con pérdida de pelo localizada en las áreas parietal y vértice, inyectar A-PRP en la zona parietal y usar solución salina como placebo en la zona de vértice.</li>
	</ol>
</li>
	<li>Realizar un número igual de las inyecciones de PRP A y placebo para cada paciente.</li>
	<li>Realizar interfollicular PRP A las inyecciones a una profundidad de 5 mm uso de la pistola médica del inyector equipada con una aguja de 30G y jeringa de 10 mL.</li>
	<li>Realizar tres sesiones espaciadas 30 días de diferencia.</li>
</ol>

<ol>
	<li>Rousso, D. E., Kim, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+review+of+medical+and+surgical+treatment+options+for+androgenetic+alopecia.">A review of medical and surgical treatment options for androgenetic alopecia.</a> JAMA Facial Plast Surg. 16, 444-450 (2014).</li><li>Schweiger, E. S., Boychenko, O., Bernstein, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Update+on+the+pathogenesis,+genetics+and+medical+treatment+of+patterned+hair+loss.">Update on the pathogenesis, genetics and medical treatment of patterned hair loss.</a> J Drugs Dermatol. 9, 1412-1419 (2010).</li><li>Price, V. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lack+of+efficacy+of+finasteride+in+postmenopausal+women+with+androgenetic+alopecia.">Lack of efficacy of finasteride in postmenopausal women with androgenetic alopecia.</a> J Am Acad Dermatol. 43, 768-776 (2000).</li><li>Imperato-McGinley, J., Guerrero, L., Gautier, T., Peterson, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Steroid+5alpha-reductase+deficiency+in+man:+An+inherited+form+of+male+pseudohermaphroditism.">Steroid 5alpha-reductase deficiency in man: An inherited form of male pseudohermaphroditism.</a> Science. 186, 1213-1215 (1974).</li><li>Drake, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+finasteride+on+scalp+skin+and+serum+androgen+levels+in+men+with+androgenetic+alopecia.">The effects of finasteride on scalp skin and serum androgen levels in men with androgenetic alopecia.</a> J Am Acad Dermatol. 41, 550-554 (1999).</li><li>Van Neste, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Finasteride+increases+anagen+hair+in+men+with+androgenetic+alopecia.">Finasteride increases anagen hair in men with androgenetic alopecia.</a> Br J Dermatol. 143, 804-810 (2000).</li><li>Kaufman, K. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Finasteride+in+the+treatment+of+men+with+androgenetic+alopecia.">Finasteride in the treatment of men with androgenetic alopecia.</a> J. Am Acad Dermatol. 39, 578-589 (1998).</li><li>Afifi, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-level+laser+therapy+as+a+treatment+for+androgenetic+alopecia.">Low-level laser therapy as a treatment for androgenetic alopecia.</a> Lasers Surg Med. 49, 27-39 (2017).</li><li>Uebel, C. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+platelet+plasma+growth+factors+in+male+pattern+baldness+surgery.">The role of platelet plasma growth factors in male pattern baldness surgery.</a> Plast Reconstr Surg. 118, 1458-1466 (2006).</li><li>Cervelli, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+autologous+activated+platelet+rich+plasma+(AA-PRP)+injection+on+pattern+hair+loss:+Clinical+and+histomorphometric+evaluation.">The effect of autologous activated platelet rich plasma (AA-PRP) injection on pattern hair loss: Clinical and histomorphometric evaluation.</a> BioMed Res Int. , 760709 (2014).</li><li>Gkini, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+of+platelet-rich+plasma+injections+in+the+treatment+of+androgenetic+alopecia+through+an+one-year+period.">Study of platelet-rich plasma injections in the treatment of androgenetic alopecia through an one-year period.</a> J Cutan Aesthet Surg. 7, 213-219 (2014).</li><li>Gentile, P., Garcovich, S., Bielli, A., Scioli, M. G., Orlandi, A., Cervelli, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+platelet-rich+plasma+in+hair+regrowth:+A+randomized+placebo-controlled+trial.">The effect of platelet-rich plasma in hair regrowth: A randomized placebo-controlled trial.</a> Stem Cells Transl Med. 4, 1317-1323 (2015).</li><li>Gentile, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+Not-Activated+and+Activated+PRP+in+Hair+Loss+Treatment:+Role+of+Growth+Factor+and+Cytokine+Concentrations+Obtained+by+Different+Collection+Systems.">Evaluation of Not-Activated and Activated PRP in Hair Loss Treatment: Role of Growth Factor and Cytokine Concentrations Obtained by Different Collection Systems.</a> Int J Mol Sci. 18 (2), 14 (2017).</li></ol>

Los autores no tienen nada que revelar.

Los autores demostraron que las inyecciones mecánicas y controladas de PRP A representan opciones de tratamiento viables de AGA, con reducción del dolor durante el procedimiento en comparación con inyección manual realizada por las manos del cirujano. Pacientes tratados con este protocolo fueron encontrados para tener un mayor incremento en la cuenta de cabello y la densidad total de pelo que los pacientes tratados con inyecciones de manual.
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La clínica inyección de desactivado A-PRP se ha convertido en un atractivo recurso para dirigir el crecimiento del cabello. Aquí, el protocolo de la aplicación de PRP A en AGA se divulga, usando inyecciones mecánicas y controladas. Actualmente, sólo oral inhibidor selectivo de la 5-α-reductasa, tópico diaminopirimidilpiperidin-N-ossido (2.0% y 5.0%) y terapia baja del laser (LLLT) están aprobados por la US Food y Drug Administration (FDA) para combatir el MPHL1,2 .
Diaminopirimidilpiperidin-N-ossido 5% espuma también está aprobada por la FDA para FPHL. El inhibidor selectivo de la 5-α-reductasa ha demostrado en gran parte ineficaz en el tratamiento de FPHL3y, dado que la droga puede causar anormalidades en los órganos genitales externos de fetos masculinos, es inadecuada para el uso de las mujeres pre-menopausal4. Por el contrario, el tratamiento diario con 1 mg de inhibidor selectivo de la 5-α-reductasa ha demostrado para reducir niveles de dihidrotestosterona (DHT) en un 70% y promueve crecimiento del pelo del pelo anagen llevando a aumento gradual en el diámetro del cabello y alargamiento de cabello en hombre AGA pacientes5,6, a través de mejoras significativas en la densidad del pelo. El uso de oral inhibidor selectivo de la 5-α-reductasa puede requerir hasta un año de tratamiento y los usuarios pueden desarrollar pérdida de la libido, que puede persistir después de que el medicamento es descontinuado7.
Recientemente, se ha propuesto el uso de la TLBI para estimular el nuevo crecimiento del pelo. Afifi et al. 8 revisado los estudios existentes para determinar si la TLBI fue una terapia eficaz para la AGA. La aplicación tópica de activado plasma rico en plaquetas autólogo (AA-PRP) a folículos cosechadas antes de la implantación ya se ha demostrado para aumentar su tasa de supervivencia de 15% en los pacientes afectados por AGA9, así como la exposición creciente del pelo densidad 3 meses después de la cirugía con la densidad del pelo terminal aumenta en 19% durante ese tiempo de10. Estos resultados fueron confirmados en un estudio siguiente AGA pacientes tratados con PRP activado por calcio en el transcurso de un año11.
Dado los resultados positivos de AA-PRP12 y13 de la A-PRP como un tratamiento de alopecia y la falta de datos para el sistema de clínica de las inyecciones, el objetivo principal de este trabajo es mostrar mecánica y controlada de protocolo de las inyecciones de PRP.
18 pacientes masculinos y 5 femeninos de 21-70 años que exhibió MPHL en la etapa 2 a etapa 5 según la escala de clasificación de Norwood-Hamilton y FPHL en etapa 1 etapa 2 según la escala de clasificación de Ludwig, fueron tratados con mecánica y controlado de las inyecciones de PRP A. AGA diagnósticos fueron hechos por una historia clínica, análisis de trichoscopic, la evaluación clínica con la prueba de tirar pelo negativo, pruebas de laboratorio y análisis de orina.
La etapa de AGA se evaluó usando la escala de Norwood-Hamilton y Ludwig. Todos los participantes incluidos en este estudio fueron evaluados por un médico médico especialista en dermatología y dos expertos de cirujano en cirugía plástica regenerativa y alopecia. Los autores consideran criterios de exclusión localizada y sistémica para todos los pacientes. Criterios sistémicos fueron representados por trastornos de la sangre o plaquetas, terapia anticoagulante o antiagregante, sepsis, inmunosupresión, cáncer, uso de oral inhibidor selectivo de la 5-α-reductasa o antiandrógenos en los 12 meses anteriores.
Criterios de exclusión localizadas fueron representados por el uso de diaminopirimidilpiperidin-N-ossido (2.0% y 5.0%) o los corticosteroides para los tratamientos tópicos en los 12 meses anteriores.

<table><tbody><tr><td>C-PunT system</td><td>Biomed Device, Modena, Italy</td><td>www.biomeddevice.it</td><td>a closed system medical device to obtain autologous concentrated platelet-rich plasma from the blood</td></tr><tr><td>Ultim Gun</td><td>Anti-Aging Medical System</td><td>www.aamedicalsystems.fr</td><td>a gun medical device to perform a mechanical and controlled injections of autologous platelet-rich plasma in a selected area of the scalp</td></tr><tr><td>Trichoscan</td><td>Tricolog GmbH, Freiburg, Germany).</td><td>www.tricholog.de</td><td>trihoScan is a digital&lt;br/&gt; software-supported epiluminescence technique for measuring hair count (number of hairs per 0.65 cm2), hair density (number of hairs per cm2), hair diameter, anagen-to-telogen ratio, and vellus&lt;br/&gt; hair-to-terminal hair ratio.</td></tr><tr><td>sodium citrate ACD-A</td><td>S.A.L.F.</td><td>SAAS2205GPO</td><td>anticoagulant for PRP preparation</td></tr><tr><td>&amp;nbsp;30-Gauge needle&amp;nbsp;</td><td>Becton Dickinson&amp;nbsp;</td><td>Z192368</td><td>needle for injections</td></tr><tr><td>10 mL mL Luer-Lock syringe</td><td>Becton Dickinson&amp;nbsp;</td><td>BD309695</td><td>syringe for PRP collection</td></tr></tbody></table>

mechanical and controlled prp injections in patients affected by androgenetic alopecia

23 pacientes (18 hombres y 5 mujeres) de 21-70 años de edad que muestran pérdida de cabello de patrón masculino (MPHL) en etapa 1 a etapa 5 según la escala de clasificación de Norwood-Hamilton y pérdida de cabello de patrón femenino (FPHL) etapa 1 etapa 2 según lo determinado por el Ludwig escala de clasificación, fueron tratados con el desactivado de plasma rico en plaquetas autólogo (A-PRP). Sangre autóloga (55 mL) se cosechó con citrato de sodio como anticoagulante. A-PRP (23 mL) fue producido para todos los casos mediante un sistema cerrado según el protocolo del servicio de transfusión. Tras centrifugación (260 x g por 10 min) A-PRP fue insertado en un dispositivo selector de luz de láser, y después de la centrifugación, se recolectó 9 mL de A-PRP.
El cuero cabelludo de los pacientes afectados por la alopecia androgenética (AGA) se dividió en cuatro zonas (frontal, parietal, vértice y occipital); no se realizó anestesia local. Inyecciones controladas y mecánicas programadas a una profundidad de 5 mm utilizando una pistola inyector médicos realizaron inyecciones interfollicular A-PRP (0.2 mL x cm2). Sesiones de tratamiento fueron realizadas con un intervalo de 30 días. Se realizaron tres sesiones de tratamiento para cada paciente.
PRP fue inyectado en las áreas relacionadas con el andrógeno del cuero cabelludo afectadas por la pérdida de cabello. Placebo (solución salina normal) se ha cargado en otra jeringa (10 mL) y se inyecta en el lado adyacente de manera similar.

Los parámetros de crecimiento de pelo miden tres meses después de las inyecciones de PRP A tercera por tricoscopic análisis se compararon con las mediciones iniciales y entre el área del tratamiento A-PRP y el área de control, que recibieron inyecciones de solución salina fisiológica como con placebo. Antes del primer tratamiento, al inicio, no hay diferencias estadísticas en la cuenta de cabello (122 ± 10) y la densidad del pelo (218 ± 17) fueron registrados entre el área dirigida y controlada (área 126 ± 9...

Watch this Scientific Journal Video about Mechanical and Controlled PRP Injections in Patients Affected by Androgenetic Alopecia at JoVE.com

Mechanical and Controlled PRP Injections in Patients Affected by Androgenetic Alopecia

Este trabajo pretende describir las inyecciones interfollicular plasma rico en plaquetas autólogo (0,20 mL x cm2) en el cuero cabelludo de los 23 pacientes afectados por la alopecia androgenética a una profundidad de 5 mm utilizando una pistola inyector médica equipada con una aguja de 30 calibre, en tres sesiones.

Inyecciones de PRP mecánica y controlados en pacientes afectados por la Alopecia Androgenética

23 patients (18 male and 5 female) aged 21-70 years who displayed male pattern hair loss (MPHL) in Stage 1 to Stage 5 as determined by the ...

mechanical-controlled-prp-injections-patients-affected-androgenetic

Research

JoVE Journal

Medicine

15.0K vistas.  University of Rome Tor Vergata. Este trabajo pretende describir las inyecciones interfollicular plasma rico en plaquetas autólogo (0,20 mL x cm2) en el cuero cabelludo de los 23 pacientes afectados por la alopecia androgenética a una profundidad de 5 mm utilizando una pistola inyector médica equipada con una aguja de 30 calibre, en tres sesiones.

Video: Mechanical and Controlled PRP Injections in Patients Affected by Androgenetic Alopecia

Isolation of Soluble and Insoluble PrP Oligomers in the Normal Human Brain

The central event in the pathogenesis of prion diseases involves a conversion of the host-encoded cellular prion protein PrPC into its pathogenic isoform PrPSc 1. PrPC is detergent-soluble and sensitive to proteinase K (PK)-digestion, whereas PrPSc forms detergent-insoluble aggregates and is partially resistant to PK2-6. The conversion of PrPC to PrPSc is known to involve a conformational transition of &alpha;-helical to &beta;-sheet structures of the protein. However, the in vivo pathway is still poorly understood. A tentative endogenous PrPSc, intermediate PrP* or "silent prion", has yet to be identified in the uninfected brain7.
Using a combination of biophysical and biochemical approaches, we identified insoluble PrPC aggregates (designated iPrPC) from uninfected mammalian brains and cultured neuronal cells8, 9. Here, we describe detailed procedures of these methods, including ultracentrifugation in detergent buffer, sucrose step gradient sedimentation, size exclusion chromatography, iPrP enrichment by gene 5 protein (g5p) that specifically bind to structurally altered PrP forms10, and PK-treatment. The combination of these approaches isolates not only insoluble PrPSc and PrPC aggregates but also soluble PrPC oligomers from the normal human brain. Since the protocols described here have been used to isolate both PrPSc from infected brains and iPrPC from uninfected brains, they provide us with an opportunity to compare differences in physicochemical features, neurotoxicity, and infectivity between the two isoforms. Such a study will greatly improve our understanding of the infectious proteinaceous pathogens. The physiology and pathophysiology of iPrPC are unclear at present. Notably, in a newly-identified human prion disease termed variably protease-sensitive prionopathy, we found a new PrPSc that shares the immunoreactive behavior and fragmentation with iPrPC 11, 12. Moreover, we recently demonstrated that iPrPC is the main species that interacts with amyloid-&beta; protein in Alzheimer disease13. In the same study, these methods were used to isolate Abeta aggregates and oligomers in Alzheimer's disease13, suggesting their application to non-prion protein aggregates involved in other neurodegenerative disorders.

A new species of cellular prion protein (PrPC) has recently been identified in uninfected human brains using the methods described here. These methods can be used to isolate various PrP species, while some of them are also useful in isolating other misfolded protein aggregates from human brains.

Aislamiento de oligómeros de PrP soluble e insoluble en el cerebro humano normal

The central event in the pathogenesis of prion diseases involves a conversion of the host-encoded cellular prion protein PrPC into its pathogenic isoform ...

Investigación

Medicina

PRP as a New Approach to Prevent Infection: Preparation and In vitro Antimicrobial Properties of PRP

Implant-associated infection is becoming more and more challenging to the healthcare industry worldwide due to increasing antibiotic resistance, transmission of antibiotic resistant bacteria between animals and humans, and the high cost of treating infections.
In this study, we disclose a new strategy that may be effective in preventing implant-associated infection based on the potential antimicrobial properties of platelet-rich plasma (PRP). Due to its well-studied properties for promoting healing, PRP (a biological product) has been increasingly used for clinical applications including orthopaedic surgeries, periodontal and oral surgeries, maxillofacial surgeries, plastic surgeries, sports medicine, etc.
PRP could be an advanced alternative to conventional antibiotic treatments in preventing implant-associated infections. The use of PRP may be advantageous compared to conventional antibiotic treatments since PRP is less likely to induce antibiotic resistance and PRP's antimicrobial and healing-promoting properties may have a synergistic effect on infection prevention. It is well known that pathogens and human cells are racing for implant surfaces, and PRP's properties of promoting healing could improve human cell attachment thereby reducing the odds for infection. In addition, PRP is inherently biocompatible, and safe and free from the risk of transmissible diseases.
For our study, we have selected several clinical bacterial strains that are commonly found in orthopaedic infections and examined whether PRP has in vitro antimicrobial properties against these bacteria. We have prepared PRP using a twice centrifugation approach which allows the same platelet concentration to be obtained for all samples. We have achieved consistent antimicrobial findings and found that PRP has strong in vitro antimicrobial properties against bacteria like methicillin-sensitive and methicillin-resistant Staphylococcus aureus, Group A Streptococcus, and Neisseria gonorrhoeae. Therefore, the use of PRP may have the potential to prevent infection and to reduce the need for costly post-operative treatment of implant-associated infections.

PRP as a New Approach to Prevent Infection: Preparation and In vitro Antimicrobial Properties of PRP

Implant-associated infection is a significant clinical complication. This study describes an approach using platelet-rich plasma (PRP) to prevent implant-associated infections, presents the protocol for preparing PRP with constant platelet concentration, and reports the newly identified antimicrobial properties of PRP and related protocols for examining such antimicrobial properties in vitro.

PRP como un nuevo enfoque para prevenir la infección: Preparación y<em&gt; In vitro</em&gt; Propiedades antimicrobianas de PRP

Implant-associated infection is becoming more and more challenging to the healthcare industry worldwide due to increasing antibiotic resistance, ...

Inmunología e Infección

Characterization of Leukocyte-platelet Rich Fibrin, A Novel Biomaterial

Autologous platelet concentrates represent promising innovative tools in the field of regenerative medicine and have been extensively used in oral surgery. Unlike platelet rich plasma (PRP) that is a gel or a suspension, Leukocyte-Platelet Rich Fibrin (L-PRF) is a solid 3D fibrin membrane generated chair-side from whole blood containing no anti-coagulant. The membrane has a dense three dimensional fibrin matrix with enriched platelets and abundant growth factors. L-PRF is a popular adjunct in surgeries because of its superior handling characteristics as well as its suturability to the wound bed. The goal of the study is to demonstrate generation as well as provide detailed characterization of relevant properties of L-PRF that underlie its clinical success.

Leucocyte-Platelet Rich Fibrin (L-PRF) represents an FDA cleared preparation of autologous platelet concentrates that possesses unique fibrin architecture, enriched platelets and abundant growth factors. Here, we present a protocol for chair-side generation of L-PRF as well as evaluate its mechanical properties including uniaxial testing and suture retention strength testing.

Caracterización de los leucocitos-plaquetas Rich fibrina, una novela de Biomateriales

Autologous platelet concentrates represent promising innovative tools in the field of regenerative medicine and have been extensively used in oral ...

Bioingeniería

Preparing Activated Platelet-Rich Plasma for Culturing Human Adipose-Derived Stem Cells

Activated platelet-rich plasma (PRP) prepared from whole blood via centrifugation demonstrated a proliferation-stimulating effect in several kinds of cultured cells, implying a possible use in regenerative medicine. Here, a double-spin method was used to prepare PRP from whole blood. PRP was further activated by autologous thrombin. The platelet count was measured in the activated PRP and the proliferation-stimulating effect in human adipose-derived stem cells (hASCs) was examined. The resulting platelet count was 11.5-times higher in PRP than in whole blood plasma. The proliferation of hASCs was markedly enhanced by incubation with 1% PRP. The described method can be used to reproducibly prepare PRP with a high concentration of platelets. PRP prepared by this method markedly promotes proliferation of hASCs.

In this report, we describe a double-spin method for preparing activated platelet-rich plasma (PRP). Using autologous thrombin, human adipose-derived stem cells (hASCs) were cultured. Activated PRP was shown to promote proliferation of hASCs.

Activated platelet-rich plasma (PRP) prepared from whole blood via centrifugation demonstrated a proliferation-stimulating effect in several kinds of ...

Biología del Desarrollo

Production of Autologous Platelet-Rich Plasma for Boosting In Vitro Human Fibroblast Expansion

There is currently great clinical interest in the use of autologous fibroblasts for skin repair. In most cases, culture of skin cells in vitro is required. However, cell culture using xenogenic or allogenic culture media has some disadvantages (i.e., risk of infectious agent transmission or slow cell expansion). Here, an autologous culture system is developed for the expansion of human skin fibroblast cells in vitro using a patient&#8217;s own platelet-rich plasma (PRP). Human dermal fibroblasts are isolated from the patient while undergoing abdominoplasty. Cultures are followed for up to 7 days using a medium supplemented with either fetal bovine serum (FBS) or PRP. Blood cell content in PRP preparations, proliferation, and fibroblast differentiation are assessed. This protocol describes the method for obtaining a standardized, non-activated preparation of PRP using a dedicated medical device. The preparation requires only a medical device (CuteCell-PRP) and centrifuge. This device is suitable under sufficient medical practice conditions and is a one-step, apyrogenic, and sterile closed system that requires a single, soft spin centrifugation of 1,500 x g for 5 min. After centrifugation, the blood components are separated, and the platelet-rich plasma is easily collected. This device allows a quick, consistent, and standardized preparation of PRP that can be used as a cell culture supplement for in vitro expansion of human cells. The PRP obtained here contains a 1.5-fold platelet concentration compared to whole blood together, with a preferential removal of red and white blood cells. It is shown that PRP presents a boosting effect in cell proliferation compared to FBS (7.7x) and that fibroblasts are activated upon PRP treatment.

This protocol presents a device that produces PRP to boost the in vitro expansion of cells in a 100% autologous fibroblast culture system.

Producción de plasma autólogo rico en plaquetas para impulsar la expansión in vitro de fibroblastos humanos

There is currently great clinical interest in the use of autologous fibroblasts for skin repair. In most cases, culture of skin cells in vitro is ...

Effects of Allogeneic Platelet-Rich Plasma (PRP) on the Healing Process of Sectioned Achilles Tendons of Rats: A Methodological Description

This article describes the experimental procedures used to observe if PRP can positively affect tendon healing. There are 4 main steps to follow: induce a lesion in the Achilles tendon; prepare PRP and inject it (or the saline solution); remove the tendon; and perform biomechanical, molecular, and histological evaluations. At each step, all the procedures and methods are described in detail, so they can be reproduced easily.
Achilles tendons have been surgically sectioned (removal of a 5-mm long section). Afterwards, PRP or saline solution was injected to study whether PRP has a positive effect on the healing of the tendon. Three groups of 40 animals (a total of 120 rats were used in this study) were subdivided into 2 subgroups: PRP injection group and a saline injection control group. Rats were sacrificed at increasing time points (Group A: 5 days; Group B: 15 days; Group C: 30 days) and tendons were removed. 90 tendons underwent biomechanical testing before performing transcriptomic analysis and the 30 remaining tendons were submitted to histological analysis.

Effects of Allogeneic Platelet-Rich Plasma PRP on the Healing Process of Sectioned Achilles Tendons of Rats: A Methodological Description

This protocol describes the evaluation process of healing tendons in rats that have been injected with allogeneic platelet rich plasma (PRP) or saline solution after removing part of the Achilles tendon. The progress of tendon healing is evaluated at several time points using different types of analyses.

Efectos del trasplante Allogeneic Plasma rico en plaquetas (PRP) en el proceso de cicatrización de los tendones de Aquiles seccionado de las ratas: una descripción metodológica

This article describes the experimental procedures used to observe if PRP can positively affect tendon healing. There are 4 main steps to follow: induce a ...

Preparation, Procedures and Evaluation of Platelet-Rich Plasma Injection in the Treatment of Knee Osteoarthritis

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited effect on the repair of cartilage and on bone regeneration. Platelet-rich plasma (PRP) is an autologous bioactive substance that can repair cartilage injury and accelerate bone regeneration effectively. However, reporting of PRP preparation protocols in clinical studies is highly inconsistent, with the majority of studies providing insufficient information to allow the protocol to be reproduced. We describe a repeatable method of preparing PRP visually, the treatment of KOA using PRP intra-articular injection, and methods of evaluating the outcome. PRP was prepared using manual double centrifugation. The PRP layer was extracted from peripheral blood and used for knee joint cavity injection. Evaluations included assessments of blood platelet concentrations and clinical outcomes. Preparation of PRP by manual centrifugation requires less apparatus and is less costly than plasma filtration or centrifugation using equipment. The centrifugation time of our double centrifugation method was 6 and 5 minutes for the respective centrifugations at forces of 800 and 1400 x g, respectively, to allow for the consistent preparation of standardized PRP. However, a manual method is susceptible to operator error, and PRP batch preparation is not available. Intra-articular injection of PRP proved to be an effective treatment for knee osteoarthritis. The entire treatment procedure took less than 30 minutes, the blood platelet concentration of PRP could be standardized, and treatment was proven to be effective when evaluated by follow-up.

Knee osteoarthritis is frequently seen in the orthopedic department. We introduce in detail the entire knee osteoarthritis treatment process with platelet-rich plasma injection, including preparation, procedures, and evaluation.

Preparación, procedimientos y evaluación de la inyección de Plasma rico en plaquetas en el tratamiento de la osteoartritis de la rodilla

Knee osteoarthritis (KOA) is one of the most frequently encountered diseases in the orthopedic department. Existing non-surgical treatments have a limited ...

A New Technique for Quantitative Analysis of Hair Loss in Mice Using Grayscale Analysis

Alopecia is a common form of hair loss which can occur in many different conditions, including male-pattern hair loss, polycystic ovarian syndrome, and alopecia areata. Alopecia can also occur as a side effect of chemotherapy in cancer patients. In this study, our goal was to develop a consistent and reliable method to quantify hair loss in mice, which will allow investigators to accurately assess and compare new therapeutic approaches for these various forms of alopecia. The method utilizes a standard gel imager to obtain and process images of mice, measuring the light absorption, which occurs in rough proportion to the amount of black (or gray) hair on the mouse. Data that has been quantified in this fashion can then be analyzed using standard statistical techniques (i.e., ANOVA, T-test). This methodology was tested in mouse models of chemotherapy-induced alopecia, alopecia areata and alopecia from waxing. In this report, the detailed protocol is presented for performing these measurements, including validation data from C57BL/6 and C3H/HeJ strains of mice. This new technique offers a number of advantages, including relative simplicity of application, reliance on equipment which is readily available in most research laboratories, and applying an objective, quantitative assessment which is more robust than subjective evaluations. Improvements in quantification of hair growth in mice will improve study of alopecia models and facilitate evaluation of promising new therapies in preclinical studies.

Alopecia is a common form of hair loss which can occur in many different conditions, including as a side-effect of chemotherapy. We have developed a method to quantify hair loss in mice, utilizing a standard gel imager to perform a grayscale analysis, to facilitate study of promising new alopecia therapies.

Una nueva técnica de análisis cuantitativos de la pérdida del pelo en ratones Uso del análisis de escala de grises

Alopecia is a common form of hair loss which can occur in many different conditions, including male-pattern hair loss, polycystic ovarian syndrome, and ...

Biología

Platelet-Rich Plasma Lysate for Treatment of Eye Surface Diseases

Various ocular surface diseases are treated with blood-derived eye drops. Their use has been introduced in clinical practice because of their metabolite and growth factor content, which promotes eye surface regeneration. Blood-based eye drops can be prepared from different sources (i.e., whole blood or platelet apheresis donation), as well as with different protocols (e.g., different dilutions and freeze/thaw cycles). This variability hampers the standardization of clinical protocols and, consequently, the evaluation of their clinical efficacy. Detailing and sharing the methodological procedures may contribute to defining common guidelines. Over the last years, allogenic products have been diffusing as an alternative to the autologous treatments since they guarantee higher efficacy standards; among them, the platelet-rich plasma lysate (PRP-L) eye drops are prepared with simple manufacturing procedures. In the transfusion medicine unit at AUSL-IRCCS di Reggio Emilia, Italy, PRP-L is obtained from platelet-apheresis donation. This product is initially diluted to 0.3 x 109 platelets/mL (starting from an average concentration of 1 x 109 platelets/mL) in 0.9% NaCl. Diluted platelets are frozen/thawed and, subsequently, centrifuged to eliminate debris. The final volume is split into 1.45 mL aliquots and stored at &#8722;80 &#176;C. Before being dispensed to patients, eye drops are tested for sterility. Patients may store platelet lysates at &#8722;15 &#176;C for up to 1 month. The growth factor composition is also assessed from randomly selected aliquots, and the mean values are reported here.

Platelet lysates represent an emerging tool for the treatment of ocular surface diseases. Here, we propose a method for the preparation, dispensation, storage, and characterization of platelet lysate collected from platelet donors.

Lisado plasmático rico en plaquetas para el tratamiento de enfermedades de la superficie ocular

Various ocular surface diseases are treated with blood-derived eye drops. Their use has been introduced in clinical practice because of their metabolite ...

A Platelet-Rich Plasma Injection into the Sectioned Achilles Tendon of a Rat

Source: Greimers, L., et al. <a href="https://app.jove.com/v/55759">Effects of Allogeneic Platelet-Rich Plasma (PRP) on the Healing Process of Sectioned Achilles Tendons of Rats: A Methodological Description.</a> J. Vis. Exp. (2018).
This video demonstrates a technique for injecting platelet-rich plasma (PRP) into the sectioned Achilles tendon of rats. A small segment is excised from the Achilles tendon of a recipient rat, the site is sutured, and the rat is allowed to recover. Concurrently, whole blood is collected from a donor rat and centrifuged to obtain PRP, which is then activated and injected into the sutured site of the recipient rat to induce tendon repair.

Una inyección de plasma rico en plaquetas en el tendón de Aquiles seccionado de una rata

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Animal ...

Inyecciones de PRP mecánica y controlados en pacientes afectados por la Alopecia Androgenética

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