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

Les auteurs n’ont aucuns accusés de réception.

Le protocole d’étude respectées de la déclaration d’Helsinki, et tous les patients si consentement éclairé avant de participer à l’étude.
1. non-activé A-PRP Preparation
<ol><li>Récolter 55 mL de sang autologue d’une veine périphérique du bras à l’aide d’un tube avec aiguille à ailettes selon le protocole de collecte de sang locale.</li>
	<li>Ajouter 7 mL de solution de dextrose (ACD) de citrate acide (112 mM du citrate) dans le tube de sang comme anticoagulant.</li>
	<li>Centrifuger le sang recueilli dans le tube à 260 x g pendant 10 min. Remarque : 23 A-PRP + Plasma pauvre en plaquettes autologue (A-PPP) et 20 mL de suspension de globules rouges (hématies) ont été produites.</li>
	<li>Retirez le tube contenant le PRP-A + A-PPP et globules rouges de la centrifugeuse et l’insérer dans un dispositif de sélecteur de lumière laser.</li>
	<li>A-PRP séparées + A-PPP dans un sachet stérile par aspiration mécanique effectué par le sélecteur de laser dans lequel est placé le kit. Globules rouges sont automatiquement supprimés par aspiration mécanique. Remarque : 14 mL d’A-PPP a été manuellement récoltée et retiré par le sac stérile de la trousse. 9 mL de A-PRP a été obtenue.</li>
</ol>2. les Injections de PRP A non-activé
<ol><li>Diviser le cuir chevelu des patients touchés par la AGA frontal, pariétal, vertex et zones occipitales. ATTENTION : N’effectuez pas d’anesthésie (locale ou systémique).</li>
	<li>Effectuer des injections de PRP A interfolliculaires (0,20 mL x cm2) par l’injecteur médical des armes à feu (voir Table des matières) à certaines zones du cuir chevelu.<ol>
<li>Chez les patients atteints de la perte de cheveux localisée dans les régions frontales et pariétales, effectuer des injections de PRP A exclusivement à la zone frontale et utiliser une solution saline comme placebo dans la région pariétale.</li>
		<li>Chez les patients atteints de la perte de cheveux localisée dans les régions pariétales et vertex, injecter A-PRP dans la région pariétale et utiliser une solution saline comme placebo dans la zone du vertex.</li>
	</ol>
</li>
	<li>Effectuer un nombre égal d’injections A-PRP et le placebo pour chaque patient.</li>
	<li>Effectuer des injections de PRP A interfolliculaires à une profondeur de 5 mm à l’aide du pistolet injecteur médical équipé d’une aiguille de 30G et une seringue de 10 mL.</li>
	<li>Effectuer trois séances espacées de 30 jours d’intervalle.</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>

Les auteurs n’ont rien à divulguer.

Les auteurs ont démontré que les injections mécaniques et contrôlées de la PRP-A représentent des options de traitement AGA viables, avec réduction de la douleur pendant l’intervention par rapport à l’injection manuelle effectuée par les mains du chirurgien. Les patients traités par le présent protocole ont eu des augmentations plus importantes nombre de cheveux et de la densité des cheveux totale que les patients traités avec des injections de manuelles.
<p class="jo...

La clinique injection de PRP A non-activé est devenu une ressource attrayante pour le ciblage de la croissance des cheveux. Ici, le protocole de l’application A-PRP dans AGA est signalé, à l’aide d’injections mécaniques et contrôlées. Actuellement, seulement orale 5-α-réductase inhibiteur sélectif, topique diaminopirimidilpiperidin-N-ossido (2,0 % et 5,0 %) et la thérapie laser à basse altitude (LLLT) sont approuvés par la Food and Drug Administration (FDA) pour lutter contre les EIPRS1,2 .
Diaminopirimidilpiperidin-N-ossido 5 % mousse est également approuvé par la FDA pour FPHL. L’inhibiteur sélectif de la 5-α-réductase a prouvé largement inefficace dans le traitement de FPHL3et, étant donné que le médicament peut provoquer des anomalies dans les organes génitaux externes du fœtus mâles, est impropre à l’usage par les femmes préménopausées4. À l’inverse, un traitement quotidien avec 1 mg d’inhibiteur sélectif de la 5-α-réductase a été montré pour réduire sériques de la dihydrotestostérone (DHT) de 70 % et favorise la croissance des cheveux de cheveu anagène conduisant à une augmentation graduelle de diamètre des cheveux et l’élongation cheveux AGA mâle les patients5,6, grâce à des améliorations significatives dans la densité des cheveux. L’utilisation d’oral inhibiteur sélectif de la 5-α-réductase peut nécessiter jusqu'à un an de traitement et utilisateurs peuvent développer la perte de la libido, qui peut persister après que le médicament est abandonné7.
L’utilisation de LLLT a récemment été proposée pour stimuler la repousse des cheveux. Amal et al. 8 a examiné les études de recherche existants afin de déterminer si le LLLT était un traitement efficace pour l’AGA. L’application topique d’activés plasma autologue de riche en plaquettes (PRP-AA) à follicules récoltés avant l’implantation a déjà été démontrée pour augmenter leur taux de survie de 15 % chez les patients atteints par l’AGA9, ainsi que l’exposition accrue de cheveux densité 3 mois après la chirurgie avec une densité de cheveux terminaux a augmenté de 19 % au cours de cette heure10. Ces résultats ont été confirmés dans une étude suivant AGA patients traités par PRP activés par le calcium au cours d’une année11.
Compte tenu des résultats positifs de l’AA-PRP12 et13 de la PRP-A comme un traitement de l’alopécie et le manque de données pour le système d’injection clinique, l’objectif principal de ce travail est de montrer mécanique et contrôlé le protocole des injections de PRP.
18 mâles et 5 femelles patients âgés de 21 à 70 ans qui affiche indisponibles à l’étape 2 à l’étape 5, tel que déterminé par l’échelle de classification de Norwood-Hamilton et FPHL à l’étape 1 étape 2 tel que déterminé par la grille de classement Ludwig, ont été traités avec mécanique et contrôlé d’injections de PRP-A. AGA diagnostics ont été faites par un antécédents médicaux, l’analyse trichoscopic, l’évaluation clinique avec essai d’arrachement de cheveux négatifs, des tests de laboratoire et analyses d’urine.
La scène de l’AGA a été évaluée à l’aide de l’échelle de Norwood-Hamilton et Ludwig. Tous les participants inclus dans cette étude ont été évalués par un médecin médecin spécialisé en dermatologie et deux spécialistes des chirurgien plasticien Chirurgie régénératrice et alopécie. Les auteurs considèrent que les critères d’exclusion localisées et systémiques pour tous les patients. Critères systémiques étaient représentés par un traitement anticoagulant ou antiagrégant, septicémie, immunosuppression, cancer, troubles sanguins et/ou de plaquettes, utilisation d’oral inhibiteur sélectif de la 5-α-réductase ou anti-androgènes dans les 12 mois précédents.
Critères d’exclusion localisées étaient représentés par l’utilisation de corticostéroïdes ou de diaminopirimidilpiperidin-N-ossido (2,0 % et 5,0 %) pour les traitements topiques dans les 12 mois précédents.

<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 patients (18 hommes et 5 femmes) âgés de 21 à 70 ans qui affichent la calvitie (indisponibles) à l’étape 1 à l’étape 5, tel que déterminé par l’échelle de classification de Norwood-Hamilton et perte de cheveux féminine (FPHL) à l’étape 1 à l’étape 2, tel que déterminé par le Ludwig grille de classement, ont été traités avec non-activé d’autologue plasma riche en plaquettes (PRP-A). Sang autologue (55 mL) a été récolté à l’aide de citrate de sodium comme anticoagulant. A-PRP (23 mL) a été réalisée dans tous les cas à l’aide d’un système fermé selon le protocole de service de transfusion sanguine. Après centrifugation (260 x g pendant 10 min), le PRP-A a été inséré dans un dispositif sélecteur de lumière de laser, et après la centrifugation, 9 mL de A-PRP ont été recueilli.
Le cuir chevelu des patients touchés par l’alopécie androgénétique (AGA) a été divisé en quatre régions (frontal, pariétal, vertex et occipital) ; anesthésie locale n’a pas été effectuée. Des injections de PRP A interfolliculaires (0,2 mL x cm2) ont été réalisées par des injections contrôlées et mécaniques à la demande à une profondeur de 5 mm à l’aide d’un pistolet injecteur médical. Séances de traitement sont effectuées avec un intervalle de 30 jours. Pour chaque patient, trois séances de traitement ont été effectués.
PRP a été injecté dans les domaines liés aux androgènes du cuir chevelu, touchés par la perte de cheveux. Placebo (solution saline normale) a été chargé dans une autre seringue (10 mL) et injectée sur le côté adjacent de manière similaire.

Les paramètres de croissance de cheveux mesuré trois mois après que les injections de PRP A troisième par analyse de tricoscopic ont été comparées avec les mesures de base et entre la zone de traitement A-PRP et la zone de contrôle, qui ont reçu des injections de sérum physiologique comme placebo. Avant le premier traitement, au départ, aucune différence statistique dans nombre de cheveux (122 ± 10) et la densité des cheveux (218 ± 17) ont été signalés entre la zone ciblée et contrôlée (zone 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

Cet article vise à décrire les injections interfolliculaires plasma riche en plaquettes autologue (0,20 mL x cm2) dans le cuir chevelu des 23 patients touchés par l’alopécie androgénétique à une profondeur de 5 mm à l’aide d’un pistolet injecteur médical équipé d’une aiguille de 30 calibre, dans trois sessions.

Injections de PRP mécanique et contrôlée chez les Patients touchés par l’alopécie androgénétique

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 vues.  University of Rome Tor Vergata. Cet article vise à décrire les injections interfolliculaires plasma riche en plaquettes autologue (0,20 mL x cm2) dans le cuir chevelu des 23 patients touchés par l’alopécie androgénétique à une profondeur de 5 mm à l’aide d’un pistolet injecteur médical équipé d’une aiguille de 30 calibre, dans trois sessions.

Vidéo: 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.

Isolement des oligomères de PrP solubles et insolubles dans le cerveau humain 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 ...

Recherche

Médecine

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 comme une nouvelle approche pour prévenir l&#39;infection: Préparation et<em&gt; In vitro</em&gt; Propriétés antimicrobiennes de PRP

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

Immunologie et infection

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.

Caractérisation de leuco-plaquettaire Rich fibrine, un roman biomatériaux

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

Bio-ingénierie

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 ...

Biologie du développement

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.

Production de plasma autologue riche en plaquettes pour stimuler l’expansion in vitro des fibroblastes humains

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.

Effets d’allogéniques Plasma riche en plaquettes (PRP) sur le processus de guérison des Tendons d’Achille sectionné des Rats : une Description méthodologique

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.

Préparation, procédures et l’évaluation de l’Injection de Plasma riche en plaquettes dans le traitement de l’arthrose du genou

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.

Une nouvelle technique pour l&#39;analyse quantitative de la perte de cheveux chez la souris Utilisation de l&#39;analyse des niveaux de gris

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

Biologie

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.

Lysat plasmatique riche en plaquettes pour le traitement des maladies de la surface oculaire

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.

Injection de plasma riche en plaquettes dans le tendon d’Achille sectionné d’un rat

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

Injections de PRP mécanique et contrôlée chez les Patients touchés par l’alopécie androgénétique

Résumé

Explorer plus de vidéos

PLAYLIST