The authors have no acknowledgments.

The study protocol complied with the Declaration of Helsinki, and all patients provided written informed consent before participating in the study.
1. Non-activated A-PRP Preparation
<ol>
	<li>Harvest 55 mL of autologous blood from a peripheral vein of the arm using a tube with butterfly needle according to the local blood collection protocol.</li>
	<li>Add 7 mL of acid citrate dextrose (ACD) solution (112 mM of citrate) to the blood tube as an anticoagulant.</li>
	<li>Centrifuge the blood c...

<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>

The authors demonstrated that mechanical and controlled injections of A-PRP represent viable AGA treatment options, with reduction of pain during the procedure compared with manual injection performed by the hands of the surgeon. Patients treated with this protocol were found to have greater increases in hair count and total hair density than patients treated with manual injections.
The most important aspect to obtain an improve...

The clinical injection of non-activated A-PRP has become an attractive resource for targeting hair growth. Here, the protocol of the A-PRP application in AGA is reported, using mechanical and controlled injections. Currently, only oral selective 5-&#945;-reductase inhibitor, topical diaminopirimidilpiperidin-N-ossido (2.0% and 5.0%), and low-level laser therapy (LLLT) are approved by the US Food and Drug Administration (FDA) to combat MPHL1,2.
Diaminopirimidilpiperidin-N-ossido 5% foam is also approved by the FDA for FPHL. The selective 5-&#945;-reductase inhibitor has proven largely ineffective in treating FPHL3, and, given that the drug may cause abnormalities in the external genitalia of male fetuses, is unsuitable for use by pre-menopausal women4. Conversely, daily treatment with 1 mg of selective 5-&#945;-reductase inhibitor has been shown to reduce serum dihydrotestosterone (DHT) levels by 70% and promotes hair growth of anagen hair leading to gradual increase in hair diameter and hair elongation in male AGA patients5,6, through significant improvements in hair density. The use of oral selective 5-&#945;-reductase inhibitor may require up to one year of treatment and users may develop loss of libido, which may persist after the medication is discontinued7.
Recently, the use of LLLT has been proposed to stimulate hair regrowth. Afifi et al.8 reviewed the existing research studies to determine whether LLLT was an effective therapy for AGA. The topical application of activated autologous platelet-rich plasma (AA-PRP) to harvested follicles prior to implantation has already been shown to increase their survival rate by 15% in patients affected by AGA9, as well as exhibit increased hair density 3 months post-surgery with terminal hair density increasing by 19% during that time10. These findings were confirmed in a study following AGA patients treated with calcium-activated PRP over the course of one year11.
Given the positive results of AA-PRP12 and A-PRP13 as an alopecia treatment, and the lack of data for the clinical injections system, the primary aim of this work is to show mechanical and controlled PRP injections protocol.
18 male and 5 female patients aged 21-70 years who displayed MPHL in Stage 2 to Stage 5 as determined by the Norwood-Hamilton classification scale, and FPHL in Stage 1 to Stage 2 as determined by the Ludwig classification scale, were treated with mechanical and controlled injections of A-PRP. AGA diagnoses were made by a medical history, trichoscopic analysis, clinical evaluation with negative hair pull test, laboratory tests, and urinalysis.
The stage of AGA was evaluated using the Norwood-Hamilton and Ludwig scale. All of the participants included in this study were assessed by a medical doctor specialized in dermatology and two plastic surgeon experts in regenerative plastic surgery and alopecia. The authors considered localized and systemic exclusion criteria for all patients. Systemic criteria were represented by blood and/or platelet disorders, anticoagulant and/or antiaggregation therapy, sepsis, immunosuppression, cancer, use of oral selective 5-&#945;-reductase inhibitor or antiandrogens in the previous 12 months.
Localized exclusion criteria were represented by use of diaminopirimidilpiperidin-N-ossido (2.0% and 5.0%) or corticosteroids for topical treatments in the previous 12 months.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<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 
			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 
			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>&#160;30-Gauge needle&#160;</td>
			<td>Becton Dickinson&#160;</td>
			<td>Z192368</td>
			<td>needle for injections</td>
		</tr>
		<tr>
			<td>10 mL mL Luer-Lock syringe</td>
			<td>Becton Dickinson&#160;</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 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 Norwood-Hamilton classification scale, and female pattern hair loss (FPHL) in Stage 1 to Stage 2 as determined by the Ludwig classification scale, were treated with non-activated autologous platelet-rich plasma (A-PRP). Autologous blood (55 mL) was harvested using sodium citrate as an anticoagulant. A-PRP (23 mL) was produced for all cases using a closed system according to the transfusion service protocol. Following centrifugation (260 x g for 10 min) the A-PRP was inserted in a laser light selector device, and after the centrifugation, 9 mL of A-PRP was collected.
The scalp of the patients affected by androgenetic alopecia (AGA) was divided into four areas (frontal, parietal, vertex, and occipital); local anesthesia was not performed. Interfollicular A-PRP injections (0.2 mL x cm2) were performed by controlled and mechanical injections scheduled at a depth of 5 mm using a medical injector gun. Treatment sessions were performed with a 30-day interval. For each patient, three treatment sessions were performed.
PRP was injected in the androgen-related areas of scalp affected by hair loss. Placebo (normal saline solution) was loaded in another syringe (10 mL) and injected on the adjacent side in a similar fashion.

The hair growth parameters measured three months after the third A-PRP injections by tricoscopic analysis were compared with the baseline measurements and between the A-PRP treatment area and the control area, which received physiological saline injections as placebo. Before the first treatment, at baseline, no statistical differences in hair count (122 &#177; 10) and hair density (218 &#177; 17) were reported between the targeted area and controlled area (126 &#177; 9 baseline hair count controlled area), (225 &#177; 15...

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

This paper aims to describe the interfollicular autologous platelet-rich plasma injections (0.20 mL x cm2) in the scalp of 23 patients affected by androgenetic alopecia at a depth of 5 mm using a medical injector gun equipped with a 30 gauge needle, in three sessions.

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 ...

The aim of this work is to demonstrate the positive effects of mechanical and controlled PRP injection in the scalp of patients affected by androgenic alopecia. First step, autologous not-activated PRP preparation. Now, we can observe our PRP procedure according to the transfusional service.In this case, we can harvest 55 mL of blood from the patients, as you can see in this moment. And after 10 minutes of centrifugation at 1, 200 RPM or 260 g force, we can obtain 23 mL of PRP and PPP, platelet-poor plasma. At this point, we can insert these specials obtained in a light selector device in which it is possible to select only platelet-rich plasma part, as you can see.Second step, injection of autologous not-activated PRP. Now, we can see our mechanical and controlled PRP injection performed by a gun. In this case, we can inject with 10 mL syringe 0.2 mL of autologous not-activated PRP for each centimeter square in the scalp of patients affected by androgenetic alopecia at five millimeter of deep.Now, it is possible to observe a detail of interfollicular PRP injection performed by a gun with 0.2 mL of PRP for each centimeter square. The results of this study showed a great increase of higher count and higher density after three mechanical and controlled PRP injection in the patients affected by androgenetic alopecia, as you can see in this moment. In conclusion, three mechanical and controlled PRP injection performed in the scalp of the patients affected by androgenetic alopecia compared with the old procedure based on manual injection, as you can see in this moment, improve higher count and higher density with reduction of pain during the procedure.Local or systemic anesthesia are not performed.

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

Research

JoVE Journal

Medicine

14.9K Views.  University of Rome Tor Vergata. The aim of this work is to demonstrate the positive effects of mechanical and controlled PRP injection in the scalp of patients affected by androgenic alopecia. First step, autologous not-activated PRP preparation. Now, we can observe our PRP procedure according to the transfusional service.In this case, we can harvest 55 mL of blood from the patients, as you can see in this moment. And after 10 minutes of centrifugation at 1, 200 RPM or 260 g force, we can obtain 23 mL of PRP and PPP,...