作者没有确认。

研究议定书遵守了《赫尔辛基宣言》, 所有病人在参加研究之前都提供了书面的知情同意。
1. 非活化 A-PRP 制剂
<ol><li>根据当地采血协议, 从手臂的外周静脉中采集55毫升自体血液, 用蝶形针管。</li>
	<li>在血液管中加入7毫升柠檬酸盐葡萄糖 (112 毫米柠檬酸) 溶液作为抗凝剂。</li>
	<li>离心机收集的血液在试管中的 260 x g 10 分钟。 注:23 毫升的 a-PRP + 自体血小板不良血浆 (PPP) 和20毫升红细胞悬浮液 (红细胞) 的产生。</li>
	<li>从离心机中取出含有 a-PRP + a-PPP 和红细胞的管子, 并将其插入激光选择器中。</li>
	<li>在无菌袋中分离 a-PRP + a-PPP, 由激光选择器进行机械吸力, 在其中放置试剂盒。红细胞自动去除机械吸力。 注:14 毫升的 A-PPP 被手工收割, 并由无菌袋从试剂盒中取出。获得了9毫升的 A-PRP。</li>
</ol>2. 非活化 A-PRP 注射
<ol><li>将受感染的患者的头皮分为额、顶叶、顶点和枕区。 注意: 不要进行麻醉 (局部或全身)。</li>
	<li>用医用注射器喷枪 (参见材料表) 对头皮的选定区域执行 interfollicular A-PRP 注射 (0.20 毫升 x cm2)。<ol>
<li>在前额和顶叶区域有脱发的患者, 在顶叶区域只进行 PRP 注射, 并使用生理盐水作为安慰剂。</li>
		<li>在脱发患者的顶叶和顶点区域, 注射 A-PRP 在顶骨区, 并使用生理盐水作为安慰剂在顶点地区。</li>
	</ol>
</li>
	<li>为每位患者执行相同数量的 A-PRP 和安慰剂注射。</li>
	<li>使用装有30G 针和10毫升注射器的医用注射器喷枪, 在深度为5毫米处执行 interfollicular a-PRP 注射。</li>
	<li>执行三会话间隔30天分开。</li>
</ol>

<ol><li>Rousso, D. E., Kim, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+review+of+medical+and+surgical+treatment+options+for+androgenetic+alopecia%5BTitle%5D)+AND+%22JAMA+Facial+Plast+Surg%22%5BJournal%5D)">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/pubmed?term=((Update+on+the+pathogenesis%2C+genetics+and+medical+treatment+of+patterned+hair+loss%5BTitle%5D)+AND+%22J+Drugs+Dermatol%22%5BJournal%5D)">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/pubmed?term=((Lack+of+efficacy+of+finasteride+in+postmenopausal+women+with+androgenetic+alopecia%5BTitle%5D)+AND+%22J+Am+Acad+Dermatol%22%5BJournal%5D)">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/pubmed?term=((Steroid+5alpha-reductase+deficiency+in+man%3A+An+inherited+form+of+male+pseudohermaphroditism%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">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/pubmed?term=((The+effects+of+finasteride+on+scalp+skin+and+serum+androgen+levels+in+men+with+androgenetic+alopecia%5BTitle%5D)+AND+%22J+Am+Acad+Dermatol%22%5BJournal%5D)">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/pubmed?term=((Finasteride+increases+anagen+hair+in+men+with+androgenetic+alopecia%5BTitle%5D)+AND+%22Br+J+Dermatol%22%5BJournal%5D)">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/pubmed?term=((Finasteride+in+the+treatment+of+men+with+androgenetic+alopecia%5BTitle%5D)+AND+%22J.+Am+Acad+Dermatol%22%5BJournal%5D)">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/pubmed?term=((Low-level+laser+therapy+as+a+treatment+for+androgenetic+alopecia%5BTitle%5D)+AND+%22Lasers+Surg+Med%22%5BJournal%5D)">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/pubmed?term=((The+role+of+platelet+plasma+growth+factors+in+male+pattern+baldness+surgery%5BTitle%5D)+AND+%22Plast+Reconstr+Surg%22%5BJournal%5D)">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/pubmed?term=((The+effect+of+autologous+activated+platelet+rich+plasma+%28AA-PRP%29+injection+on+pattern+hair+loss%3A+Clinical+and+histomorphometric+evaluation%5BTitle%5D)+AND+%22BioMed+Res+Int%22%5BJournal%5D)">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/pubmed?term=((Study+of+platelet-rich+plasma+injections+in+the+treatment+of+androgenetic+alopecia+through+an+one-year+period%5BTitle%5D)+AND+%22J+Cutan+Aesthet+Surg%22%5BJournal%5D)">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/pubmed?term=((The+effect+of+platelet-rich+plasma+in+hair+regrowth%3A+A+randomized+placebo-controlled+trial%5BTitle%5D)+AND+%22Stem+Cells+Transl+Med%22%5BJournal%5D)">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/pubmed?term=((Evaluation+of+Not-Activated+and+Activated+PRP+in+Hair+Loss+Treatment%3A+Role+of+Growth+Factor+and+Cytokine+Concentrations+Obtained+by+Different+Collection+Systems%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">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>

作者没有什么可透露的。

作者证明, 机械和控制注射的 A-PRP 代表可行的治疗方案, 减少疼痛的过程中, 与手动注射执行的外科医生的手。经本议定书治疗的患者在头发数量和总头发密度上比使用人工注射治疗的患者有更大的增加。
在 PRP 的制备和注射技术的过程中, 要注意各步骤的细节, 才能提高头发的数量和头发的密度。例如, 为了获得良好的头发改善, 这是非常有益的使用一个灭菌封闭系统根据输血服务规则的 prp 准备和执行的机械注射 prp。在开始注射之前, 重要的是将 PRP 放在它的最终位置, 放入枪中。之后, 有必要使用30G 针。
如前所述, 我们更喜欢用医用注射器枪进行机械和控制注射 PRP;这为减少人工注射产生的主观偏差提供了额外的好处。事实上, 使用机械装置作为枪, 有额外的好处, 减少无意中在不同深度向目标区域注射不同数量的 PRP 的可能性。
作者观察到, 机械和控制注射 PRP 在头皮靶区的使用枪或机械设备, 不仅允许更好的注射 (更严格), 但也尽量减少过度注射或低注射的风险, 每 cm2。
另一个重要的技术提示是从上侧开始注射, 并继续向下观察每一个点之间的 1 cm2的空间, 以尽量减少重叠注射的风险。
如果通过离心机获得的 PRP 量与靶区相比非常低, 则可能需要再次重复使用新试剂盒进行采血和离心的过程。这样, 将使用大量的 PRP。
我们的经验超过两年多使用这种注射的临床目的强烈建议, 机械和控制注射 prp 使用枪或机械设备是一个重复的模式 prp 注射液治疗 MPHL 和 FPHL。同时, 我们的经验超过四年的使用这种 prp 的准备, 建议, 封闭系统的 prp 制备是一个安全的和重复的 prp 程序的模式。
这两者都可以很容易地纳入培训课程, 因为他们是良好的教学和培训模式, 为学员的审美再生手术和美容再生医学。在我们的经验中, 虽然在技术上具有挑战性的新手在开始再生手术, 经过一些训练后, 获得的 PRP 可以成功地注射到头皮的病人, 以最小到没有随后的疼痛90% 的情况下。
在文献10、11、12、13中介绍了多种激活和非活化 PRP 制剂的方法。商业系统似乎是最常用的临床, 教学和研究的目的。由于使用简便, 欧盟安全标志, 这些程序受到青睐。经 prp 治疗的患者的头发数量和总发密度比使用相同的 prp 收集设备的 AA-prp 的患者有更大的增加, 这表明在采用特定系统时不需要激活 prp13。
此外, 采用机械和控制注射 PRP 减少了注射从相邻点的可能性重叠的疼痛和主观偏见的注射。
该模型的两个主要局限性是需要再生手术技能, 以进行治疗和 PRP 准备, 并有必要获得一个机械设备注射。本文所提出的模型的另一个潜在缺陷是程序的时间。事实上, 手动程序比机械和控制注射更快。作者报告说, 当使用机械装置时, 每一个 PRP 注射过程均有25-45 分钟的平均值。
注射 PRP 的枪可用于男性和女性脱发的治疗。它的使用特别适用于对疼痛更敏感的病人, 不需要局部或全身麻醉。根据这些结果, 作者建议更多的临床试验的基础上使用的机械系统的注射。

非活化的 A-PRP 的临床注射已成为一个有吸引力的资源, 针对头发生长。在这里, 使用机械和控制注射的方法报告了 A-PRP 应用的协议。目前, 只有口服选择性 5-α-还原酶抑制剂, 局部 diaminopirimidilpiperidin n-ossido (2.0% 和 5.0%) 和低水平激光治疗 (LLLT) 被美国食品和药物管理局 (FDA) 批准, 以打击 MPHL1,2.
Diaminopirimidilpiperidin-n-ossido 5% 泡沫也被 FDA 批准为 FPHL。选择性的 5-α-还原酶抑制剂在治疗 FPHL3方面已经证明是无效的, 而且, 由于药物可能导致男性胎儿外生殖器的异常, 因此不适合绝经前妇女使用4。相反, 每天治疗1毫克选择性 5-α-还原酶抑制剂已表明, 以减少血清睾 (DHT) 的水平 70%, 并促进头发生长的生长期头发导致逐渐增加头发直径和头发伸长的男性遗传患者5,6, 通过显著改善头发密度。口服选择性 5-α-还原酶抑制剂的使用可能需要多达一年的治疗和用户可能会发展失去的性冲动, 这可能会坚持后, 药物停止7。
最近, LLLT 的使用已经被提出来刺激毛发再生。Afifi et al.8回顾了现有的研究研究, 以确定 LLLT 是否是一种有效的治疗方法。局部应用活化的自体富血小板血浆 (AA-PRP) 的收获卵泡在植入之前已经表明, 增加他们的存活率为15% 的患者受感染的9, 以及展览增加头发密度3月后, 后期手术与终端头发密度增加19% 在这段时间内,10。在一年的11过程中, 经钙活化的 PRP 治疗后, 研究证实了这些结果。
鉴于 AA-prp12和 A-prp13作为脱发治疗的积极结果, 以及缺乏临床注射系统的数据, 这项工作的主要目的是显示机械和控制的 prp 注射协议。
18男性和5女性患者21-70 岁, 显示 MPHL 在阶段2至阶段5由诺伍德-汉密尔顿分类规模, 和 FPHL 在阶段1到阶段2由路德维希分类规模确定, 被处理的机械和控制注射的 A-PRP。通过病史、trichoscopic 分析、阴性毛发拉力试验、实验室检查、尿检等方法进行临床诊断。
采用诺伍德-汉密尔顿和路德维希尺度对其进行了评价。这项研究的所有参与者都是由一位皮肤病学专业的医生和再生整形外科和脱发的两名整形外科专家评估的。作者考虑了所有患者的局部性和系统性排斥标准。系统标准由血液和/或血小板紊乱, 抗凝血和/或栓治疗, 脓毒症, 免疫抑制, 癌症, 口服选择性 5-α-还原酶抑制剂或 antiandrogens 在过去12月。
局部排除标准的代表是使用 diaminopirimidilpiperidin n-ossido (2.0% 和 5.0%) 或皮质类固醇的局部治疗在过去的12月。

<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例 (18 男性和5女性) 岁21-70 岁, 显示男性模式脱发 (MPHL) 在阶段1到阶段 5, 由诺伍德-汉密尔顿分类规模, 和女性模式脱发 (FPHL) 在阶段1到阶段2由路德维希确定分类量表, 采用非活化自体富血小板血浆 (A-PRP) 进行治疗。自体血 (55 毫升) 是用枸橼酸钠作为抗凝剂来收割的。根据输血服务协议, 为所有情况下使用封闭式系统生成 PRP (23 毫升)。离心后 (260 x g 为10分钟) 将 a-prp 插入到激光光选择器中, 在离心后, 收集了9毫升的 a-prp。
雄脱发的患者头皮分为四区 (额、顶叶、顶点、枕部);局部麻醉没有执行。Interfollicular a-PRP 注射 (0.2 毫升 x cm2) 是由控制和机械注射计划在深度为5毫米使用医用注射器枪进行。治疗疗程以30天的间隔进行。对每位患者进行三次治疗。
PRP 被注射在雄激素相关的头皮受脱发影响的地区。安慰剂 (生理盐水溶液) 被装在另一个注射器 (10 毫升) 和注射在毗邻边以相似的方式。

用 tricoscopic 分析法对第三次 a-prp 注射后三月的毛发生长参数进行了比较, 并与基线测量和 A-prp 治疗区和控制区之间进行了对比, 接受生理盐水注射安慰.在第一次治疗之前, 在基线, 没有统计的区别在头发计数 (122 ± 10) 和头发密度 (218 ± 17) 在被瞄准的区域和被控制的区域之间 (126 ±9基线头发计数被控制的区域), (225 ±15基线头发密度控制区)。
结果表明, 三月后的第三个 a-prp 注射治疗的机械和控制 A-prp 注射, 平均头发计数增加显着基线值的男性患者。目标区域显示一个高发数 (158 ± 11) (36 ±3毛比基线) 和总头发密度 (282 ± 20) (65 ±5头发 cm2与基线相比), 而接受生理盐水注射安慰剂的受控区域微不足道头发数的变化13。这些值反映了治疗组的头发密度为31% ± 2%, 而安慰剂组的头发密度增加不到1%。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56406/56406fig1.jpg"> 图 1.Interfollicular a-PRP 注射的医疗注射器枪配备了30G 针, 10 毫升注射器.Interfollicular A-PRP 注射 (0.2 毫升 x cm2) 在5毫米的深度进行。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/56406/56406fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>头发参数</td>
			<td>时间</td>
			<td>控制区域 (用生理盐水处理)</td>
			<td>目标区域 (用 PRP 处理)</td>
		</tr>
<tr>
<td rowspan="2">头发密度, 不。每 cm2
</td>
			<td>基线</td>
			<td>225±15</td>
			<td>218±17</td>
		</tr>
<tr>
<td>3月后第三次治疗</td>
			<td>227±16</td>
			<td>282±20</td>
		</tr>
<tr>
<td rowspan="2">头发计数</td>
			<td>基线</td>
			<td>126±9</td>
			<td>122±10</td>
		</tr>
<tr>
<td>3月后第三次治疗</td>
			<td>127±9</td>
			<td>158±11</td>
		</tr>
<tr>
<td colspan="4">数据被显示为平均±标准误差。</td>
		</tr>
</tbody></table>表1。Trichoscan 分析评估的相关毛发生长参数对 A-PRP 治疗和安慰剂阴性控制半头区域在基线和3月在第三治疗以后。

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

本文的目的是描述 interfollicular 自体血小板丰富的血浆注射液 (0.20 毫升 x cm2) 的头皮23例受雄脱发的影响, 在深度5毫米使用医用注射器枪装备30针, 三会议.

雄脱发患者的机械和控制 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 次观看.  University of Rome Tor Vergata. 本文的目的是描述 interfollicular 自体血小板丰富的血浆注射液 (0.20 毫升 x cm2) 的头皮23例受雄脱发的影响, 在深度5毫米使用医用注射器枪装备30针, 三会议.

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

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作为一种新的方法，以防止感染的制备及<em在体外</em&gt;抗菌性能的PRP

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

科研

免疫与感染

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.

<meta charset="utf8"></head><body>制备活化的富血小板血浆，用于培养人脂肪来源的干细胞

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.

制备活化的富血小板血浆，用于培养人脂肪来源的干细胞

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

发育生物学

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.

生产自体富血小板血浆以促进体外人成纤维细胞扩增

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 ...

生物工程

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.

同种异体富血小板血浆 (PRP) 对大鼠分段腱愈合过程的影响: 方法学描述

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 ...

医学

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.

富血小板血浆注射液治疗膝关节骨关节炎的制备、程序及评价

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

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the knees by using different forms of mesenchymal stem cells (MSCs). In Korea, bone marrow concentrates and cord blood-derived stem cells have been approved by the Korean Food and Drug Administration (KFDA) for cartilage regeneration. In addition, an adipose tissue-derived stromal vascular fraction (SVF) has been allowed by the KFDA for joint injections in human patients. Autologous adipose tissue-derived SVF contains extracellular matrix (ECM) in addition to mesenchymal stem cells. ECM excretes various cytokines that, along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, may help MSCs to regenerate cartilage and improve knee functions. In this article, we presented a protocol to improve knee functions by regenerating cartilage-like tissue in human patients with OA. The result of the protocol was first reported in 2011 followed by a few additional publications. The protocol involves liposuction to obtain autologous lipoaspirates that are mixed with collagenase. This lipoaspirates-collagenase mixture is then cut and homogenized to remove large fibrous tissue that may clog up the needle during the injection. Afterwards, the mixture is incubated to obtain adipose tissue-derived SVF. The resulting adipose tissue-derived SVF, containing both adipose tissue-derived MSCs and remnants of ECM, is injected into knees of patients, combined with HA and calcium chloride activated PRP. Included are three cases of patients who were treated with our protocol resulting in improvement of knee pain, swelling, and range of motion along with MRI evidence of hyaline cartilage-like tissue.

Here, we present a protocol to produce an adipose tissue-derived stromal vascular fraction and its application to improve knee functions by regenerating cartilage-like tissue in human patients with osteoarthritis.

制备脂肪组织基质血管部分用于潜在软骨再生的临床协议

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

Isolation of Adipose Derived Regenerative Cells for the Treatment of Erectile Dysfunction Following Radical Prostatectomy

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem cells can be obtained from different tissues and several methods for isolation have been described. The presented method for the isolation of adipose-derived regenerative cells (ADRCs) can be used within many therapeutic areas because the method is a general procedure and, therefore, not limited to erectile dysfunction (ED) therapy. ED is a common and serious side effect to radical prostatectomy (RP) since ED often is not well treated with conventional therapy. Using ADRC&#8217;s as treatment for ED has attracted great interest due to the initial positive results after a single injection of cells into the corpora cavernosum. The method used for the isolation of ADRC&#8217;s is a simple, automated process, that is reproducible and ensures a uniform product. Furthermore, the sterility of the isolated product is ensured because the entire process takes place in a closed system. It is important to minimize the risk of contamination and infection since the stem cells are used for injection in humans. The whole procedure can be done within 2.5-3.5 hours and does not require a classified laboratory which eliminates the need for shipping tissue to an off-site. However, the procedure has some limitations since the minimum amount of drained lipoaspirate for the isolation device to function is 100 g.

Precise disclosure of methods and protocols is crucial for large scale uptake of stem cell therapies. Here, we present a protocol to isolate adipose-derived regenerative cells, used for a single intracavernous injection as treatment of erectile dysfunction (ED) following radical prostatectomy (RP).

分离脂肪衍生的再生细胞用于治疗根治性前列腺切除术后的勃起功能障碍

Stem cells are used in many research areas within regenerative medicine in part because these treatments can be curative rather than symptomatic. Stem ...

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.

新技术脱发的定量分析小鼠使用灰度分析

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.

富血小板血浆裂解物用于治疗眼表疾病

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

雄脱发患者的机械和控制 PRP 注射

摘要

探索更多视频

PRP作为一种新的方法，以防止感染的制备及

制备活化的富血小板血浆，用于培养人脂肪来源的干细胞

制备活化的富血小板血浆，用于培养人脂肪来源的干细胞

生产自体富血小板血浆以促进体外人成纤维细胞扩增

同种异体富血小板血浆 (PRP) 对大鼠分段腱愈合过程的影响: 方法学描述

富血小板血浆注射液治疗膝关节骨关节炎的制备、程序及评价

制备脂肪组织基质血管部分用于潜在软骨再生的临床协议

分离脂肪衍生的再生细胞用于治疗根治性前列腺切除术后的勃起功能障碍

新技术脱发的定量分析小鼠使用灰度分析

富血小板血浆裂解物用于治疗眼表疾病