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

著者の謝辞があります。

研究プロトコルはヘルシンキ宣言を遵守し、すべての患者が研究に参加する前に書面によるインフォームド コンセントを提供します。
1. 活性化 A PRP 作製
<ol><li>局所血コレクション プロトコルに従って蝶針とチューブを使って腕の末梢静脈から自己血貯 55 mL を収穫します。</li>
	<li>抗凝固剤として血液チューブに酸クエン酸デキスト ロース (ACD) 溶液 (クエン酸の 112 mM) 7 mL を追加します。</li>
	<li>10 分間 260 x g でチューブで遠心血液を収集しました。 注: A PRP + 自家血小板貧しい血しょう (A PPP) 23 mL と 20 mL の懸濁液赤い血球 (赤血球) の作り出されました。</li>
	<li>遠心分離機から A PRP + A PPP と赤血球を含むチューブを取り出して、レーザー光セレクター デバイスに挿入します。</li>
	<li>別の PRP + キットが置かれたレーザー セレクターによって実行される機械の吸引によって滅菌袋には PPP。赤血球は、機械の吸引によって自動的に削除されます。 注: 14 mL A PPP が、手動で収穫、キットから滅菌バッグで削除します。A PRP の 9 mL が得られました。</li>
</ol>2. 活性化 A PRP 注射
<ol><li>前頭部、頭頂、頂点、後頭部に AGA の影響を受ける患者の頭皮を分割します。 注意: は麻酔 (局所または全身) を行わないでください。</li>
	<li>医療インジェクターによって検 A PRP 注射 (cm2x 0.20 mL) を実行銃頭皮の選択された領域を (材料の表を参照してください)。<ol>
<li>前頭葉と頭頂葉の領域にローカライズされた脱毛症、前頭領域に排他的、PRP 注入を行い頭頂領域におけるプラセボとして生理食塩水を使用します。</li>
		<li>頭頂と頂点の地域にローカライズされた脱毛症、頭頂領域では PRP を注入し、頂点の範囲でプラセボとして生理食塩水を使用します。</li>
	</ol>
</li>
	<li>各患者のための PRP とプラセボ注射の等しい数を実行します。</li>
	<li>30 G 針、10 mL 注射器装備医療インジェクター銃を使用して 5 mm の深さで検 A PRP 注射を実行します。</li>
	<li>30 日の間隔の間隔 3 つのセッションを実行します。</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>

著者が明らかに何もありません。

著者は、外科医の手によって実行される手動注入と比較して処理中に痛みを軽減した、実行可能な AGA 治療法の選択肢を表す A PRP の力学と制御の注射を示した。このプロトコルで治療された患者は、手動の注射で治療された患者よりもヘアの数、全体の髪密度が増加して発見されました。
毛髪数、毛髪密度の改善を得るための最も重要な側面は、PRP 作製と注入の技術の中にさまざまなステップの詳細に注意することです。たとえば、良い髪の改善を得るためには、PRP の準備のため輸血サービス規則に従って滅菌クローズド システムを使用して PRP の機械式噴射を実行する非常に便利です。注射を開始する前に、PRP を注射器を銃に、決定的な位置に置くことが重要です。後に、30 G の針を使用する必要は。
前述のよう、PRP; の力学と制御の注射用医療インジェクター銃を使うことが望ましいこれは手動の注射によって生成される主観的なバイアスを最小限に抑えることの付加的な利点を提供します。実際、銃として機械装置の使用が誤ってターゲット地域に異なる深さで PRP のさまざまな量を注入することの確率の減少の付加的な利点です。
銃や機械装置を使用して頭皮のターゲット地域で PRP の力学と制御の注射だけでなくより良い注入 (より厳格な) が可能、また過度の過注入または各 cm 用低注入のリスクを最小限に抑える検討を行った2。
もう一つの重要な技術的なヒント、アッパー サイドからオーバー ラップした注射のリスクを最小限に抑えるため、各スポット間のスペースの 1 cm2を観察側に継続注入を開始します。
場合は遠心分離により得られた PRP の量が非常に低い治療するターゲット地域と比較して、2 回目の血の収穫および遠心分離の新しいキットを使用しての手順を繰り返す必要があります。この方法では、PRP の大きい量が使用されます。
強く臨床目的のためこのような注入を使用しての 2 年以上の経験では、銃や機械装置を使用して PRP の力学と制御インジェクションが PRP 注入療法の MPHL と FPHL の再現可能なモデルであることを示唆しています。同時にこの PRP 準備を使用しての 4 年以上の経験は PRP の準備のためのクローズド システムが PRP プロシージャの安全で再現性の高いモデルであることを示唆しています。
両方組み込むことができます簡単にトレーニング コースに良い教育や美容再生医療や審美的な再生医療の研修生のトレーニング モデル。私たちの経験では技術的に再生手術の初心者いくつかの訓練の後、初めに挑戦得られる PRP 正常に注射することできる場合の 90% でいいえ以降に最小限の痛み患者の頭皮。
アクティブと非アクティブの PRP の準備の多数の方法は文献10、11,12,13に記載されています。商業システムは、最も一般的に使用されるよう臨床、教育、および研究目的。そのシンプルさ、EU 安全マークを使用するため、これらの手順が支持されています。デバイスを使用して同じ PRP コレクション、PRP が特定のシステムを採用するときにアクティブにする必要がないことを示すAA PRP で治療した患者よりもヘアの数、全体の髪密度が増加して、PRP 治療を受けた患者が発見されました。13。
また、PRP の力学と制御の注入の使用は、隣接点から注射の可能性重複を減らすことによって注射時に痛みと主観的なバイアスを削減が。
このモデルの 2 つの主要な制限は、注射のための機械装置を取得する必要性と PRP 作製、治療を遂行するために再生手術スキルの必要です。ここに提示されたモデルのもう一つの潜在的な制限は、プロシージャの時間です。実際には、マニュアルの手順は、機械・制御の注射よりも高速です。機械装置を使用する PRP 注入のプロシージャごとに 25 時間 45 分の平均を報告しました。
PRP を注入する銃は、男性と女性の脱毛の脱毛治療に使用できます。その使用は、特に局所または全身麻酔なし、痛みに敏感な患者に適しています。これらの結果を踏まえ、著者は注射の機械システムの使用に基づいて臨床試験をお勧めします。

非活性化は prp 臨床注入髪の成長をターゲットに魅力的なリソースとなっています。ここでは、機械・制御の注射を使用して AGA では PRP アプリケーションのプロトコルが報告されました。現在、口腔のみ選択的 5 α-還元酵素阻害剤、外用 diaminopirimidilpiperidin-N-ossido (2.0%、5.0%) と低レベル レーザー治療 (LLLT) MPHL1,2と戦うためには、米国食品医薬品局 (FDA) が承認しました。.
Diaminopirimidilpiperidin N ossido 5% 泡また FPHL FDA によって承認されています。選択的 5 α-還元酵素阻害剤は主に FPHL3の治療に効果が実証されてあり、薬男性胎児の外性器の異常を引き起こす可能性があります、4プレ更年期の女性によって使用には適していません。逆に、選択的 5 α-還元酵素阻害剤 1 mg 毎日治療 70% 血中ジヒドロテストステロン (DHT) を減らすために示されているし、髪の毛直径の緩やかな増加と男性の AGA で髪の伸長につながる成長期毛の髪の成長を促進します。患者5,6髪の密度が大幅に向上します。経口選択的 5 α-還元酵素阻害剤の使用は最長 1 年間の治療の必要があります、ユーザー可能性があります薬は中止7後に持続することが性欲の喪失を開発します。
最近では、レーザー装置の使用は、毛髪の再成長を刺激するために提案されている.Afifiら8は、LLLT が AGA の治療をしたかどうかを決定する既存の研究を検討しました。注入前に収穫された毛包を活性化自己血小板血漿 (AA PRP) の局所アプリケーションは、既に 15% 高められた毛の展示だけでなくアガ9、被害を受けた患者の生存率を高めるに示されています。密度はターミナルの髪の密度がその時刻10時 19% 増と術後 3 ヵ月。これらの調査結果は、1 年11のコースでカルシウム活性化 PRP 治療以下の AGA 患者の研究で確認されました。
臨床注入システムの脱毛症治療とデータの欠如として AA PRP12と13 A PRP の肯定的な結果を考えると、この作品の主な目的は機械を PRP 注入プロトコルを制御します。
18 男性と 5 女性患者がノーウッド ハミルトン分類スケールによって定められるようにステージ 5 ステージ 2 の MPHL を表示 21 70 歳を高齢者し、ステージ 1 ステージ 2 の FPHL ルートヴィヒ分類スケールによって決まりますと扱われた機械とA PRP の注入を制御します。AGA の診断は、病歴、trichoscopic 分析、負の髪引っ張りテスト、検査、尿検査と臨床評価が行われました。
AGA のステージは、ノーウッド ハミルトンそして Ludwig のスケールを使用して評価しました。すべてのこの研究に含まれている参加者は医療をおこなった医師専門皮膚科、再生プラスチック手術や脱毛症の 2 つの外科専門家。著者は、すべての患者のローカライズと全身の除外基準を考慮しました。全身の条件は、血液および/または血小板障害、抗凝固および/または antiaggregation 療法、敗血症、免疫抑制、癌、前 12 ヶ月の経口選択的 5 α-還元酵素阻害剤または抗アンドロゲンの使用で表されました。
ローカライズされた除外基準は、前 12 ヶ月間の局所治療のため diaminopirimidilpiperidin-N-ossido (2.0%、5.0%) またはコルチコステロイドの使用によって表されました。

<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 患者 (18 男性と 5 女性) 21-70 歳以上 Ludwig によって決定されるステージ 2 ステージ 1 ステージ 1 ステージ 5 ノーウッド ハミルトン分類スケールによって定められるように男性型脱毛症 (MPHL) と女性型脱毛症 (FPHL) を表示分類スケール、非活性化自己多血小板血漿 (PRP-A) と扱われました。自己血 (55 mL) は、抗凝固剤としてクエン酸ナトリウムを使用して収穫されました。A PRP (23 mL) の場合は輸血サービス プロトコルに従ってクローズド システムを使用して生産されました。遠心分離 (260 x g で 10 分間) に続いて A PRP レーザー光セレクター装置に挿入し、遠心分離後、PRP の 9 mL を採取.
アンドロゲン性脱毛症 (AGA) の影響を受ける患者の頭皮が (前頭部、頭頂、頂点、および後頭); 4 つのエリアに分かれていた局所麻酔は実行されませんでした。検 A PRP 注射 (cm2x 0.2 mL) は、制御と機械的注射医療インジェクター銃を使用して 5 mm の深さでスケジュールによって行われました。30 日間の間隔での治療のセッションを行った。それぞれの患者の 3 つの治療のセッションを行った。
PRP は、頭皮の毛損失の影響を受けるのアンドロゲン関連分野で注入しました。プラセボ (生理食塩水) は別の注射器 (10 mL) 読み込まれ、同様に隣接する側に注入されます。

髪の成長パラメーター測定 3ヶ月後にベースライン測定とは、PRP 治療領域ととして生理食塩水注射を受けたコントロール領域の tricoscopic 解析による 3 A PRP 注射剤を比較しました。プラセボ。ベースラインでの最初の治療の前に毛髪数 (122 ± 10) の統計的な差異と髪の密度 (218 ± 17) は報告されなかったターゲット地域と管理区域 (126 ± 9 ベースライン髪数制御領域)、間 (225 ± 15 ベースライン髪の密度管理区域)。
得られた結果は、機械と制御の PRP と 3 A PRP 注射治療後 3 カ月注射、平均髪カウント増加大幅男性患者における基準値を示すターゲット地域を示した昇格した髪の数 (158 ± 11) (36 ± 3 毛がベースラインと比較して) 髪の密度 (282 ± 20) を合計し、(65 ± 5 髪 cm2ベースラインと比較して)、制御しながらプラセボとして生理食塩水の注射を受けた地域はごくわずか頭髪の変化は、13を数えます。これらの値は治療群と 1% 未満の髪密度の 31% ± 2% の増加を反映してプラセボ群の毛髪密度の増加します。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56406/56406fig1.jpg"> 図 1.30 G 針、10 mL 注射器装備医療インジェクタドンゴンによって検 A PRP 注射。検 A PRP 注射 (cm2x 0.2 mL) は、5 mm の深さで行われました。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56406/56406fig1large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>髪パラメーター</td>
			<td>タイミング</td>
			<td>コントロール エリア (生理食塩水で治療)</td>
			<td>ターゲット地域 (A, prp 治療)</td>
		</tr>
<tr>
<td rowspan="2">髪の密度、 違います。cm2あたり
</td>
			<td>ベースライン</td>
			<td>225 ± 15</td>
			<td>218 ± 17</td>
		</tr>
<tr>
<td>3 ヵ月後 3 番目の治療</td>
			<td>227 ± 16</td>
			<td>282 ± 20</td>
		</tr>
<tr>
<td rowspan="2">髪の数</td>
			<td>ベースライン</td>
			<td>126 ± 9</td>
			<td>122 ± 10</td>
		</tr>
<tr>
<td>3 ヵ月後 3 番目の治療</td>
			<td>127 ± 9</td>
			<td>158 ± 11</td>
		</tr>
<tr>
<td colspan="4">データは、平均 ± 標準誤差として表されます。</td>
		</tr>
</tbody></table>表 1。育毛関連パラメーターはベースラインと 3 番目の治療後 3 カ月で陰性対照ハーフ ヘッド エリア Trichoscan 解析、A PRP 治療群とプラセボによる評価。

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

本稿の目的医療インジェクター ガンを装備した 30 ゲージ針を使用して 5 mm の深さでアンドロゲン性脱毛症の影響を受ける 23 患者の頭皮の検自己血小板血漿注射 (cm2x 0.20 mL) を記述する 3 つのセッション。

アンドロゲン性脱毛症の影響を受ける患者における力学と制御された PRP 注射

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

Universidad San Sebastian

Research

JoVE Journal

Medicine

15.0K ビュー.  University of Rome Tor Vergata. 本稿の目的医療インジェクター ガンを装備した 30 ゲージ針を使用して 5 mm の深さでアンドロゲン性脱毛症の影響を受ける 23 患者の頭皮の検自己血小板血漿注射 (cm2x 0.20 mL) を記述する 3 つのセッション。

ビデオ: 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.

正常なヒト脳内の可溶性および不溶性のPrPオリゴマーの分離

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

医学

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.

感染を防ぐための新しいアプローチとして、PRP：準備と<em&gt;インビトロ</emPRPの&gt;抗菌特性

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

免疫・感染学

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.

白血球、血小板リッチフィブリン、新規生体材料の特性評価

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

生物工学

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 ...

発生生物学

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.

in vitroヒト線維芽細胞増殖を促進するための自己多血小板血漿の産生

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.

ラットの断面アキレス腱の治癒過程に及ぼす同種多血小板血漿 (PRP): 方法論的説明

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.

準備・手順・膝変形性関節症の治療における多血小板血漿注入の評価

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.

グレースケール解析を用いたマウスの抜け毛の定量分析のための新たな手法

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

生物学

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.

眼表面疾患の治療のための多血小板血漿ライセート

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.bdigitaluss.remotexs.co/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.

ラットの切片アキレス腱への多血小板血漿注入

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