CN100349556C - 利用自身真皮成纤维细胞修补皮肤和软组织缺损 - Google Patents

利用自身真皮成纤维细胞修补皮肤和软组织缺损 Download PDF

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CN100349556C
CN100349556C CNB96197270XA CN96197270A CN100349556C CN 100349556 C CN100349556 C CN 100349556C CN B96197270X A CNB96197270X A CN B96197270XA CN 96197270 A CN96197270 A CN 96197270A CN 100349556 C CN100349556 C CN 100349556C
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小·W·K·波斯
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Abstract

本申请涉及一种修补受治者皮下组织或真皮缺损的方法。这种方法内容包括从受治者活体样品培养真皮成纤维细胞，然后用这种基本上无免疫原性蛋白的制剂去修补皮下或真皮组织。这种方法可用于修补皱纹，皮肤扩张纹，凹陷性瘢痕，非创伤性皮肤凹陷以及嘴唇增厚美容。

Description

利用自身真皮成纤维细胞修补皮肤和软组织缺损
1.发明领域：
本方法涉及人类受治者皮肤和软组织缺损，包括皱纹。更具体而言，它着重于一种可用于非外科技术中的可增加真皮或皮下组织体积的新材料。通过注射自身细胞悬液，本发明提供了下临近(subadjacent)组织体积长期增加，而不具有现有材料的缺点。
2.发明背景：
将一种材料(即注射剂)注入体内尤其是脸部来获得美容效果可追溯到十九世纪末。例如通过注射石蜡来纠正轮廓缺陷在“一战”前得到短期接受。但是并发症的存在，长期效果不理想使这种做法遭到放弃。可注射的硅氧烷的获得使这一过程从本世纪六十年代起重演。特制的医用硅氧烷溶液，例如Dow Corning MDX 4.4011，在实验的基础上在美国多个获准的测试中心得到使用。对硅氧烷的局部和全身性反应，注射物的迁移，局部组织断裂，已经限制了硅氧烷注射方法的应用。虽然这种方法的最初提议者仍然用有限的受治者来继续他们的实验计划，但这种方法不可能被更多的外科和内科医生接受。参见Matton，G.等发表于1985年Aesthetic Plastic Surgery 9卷133-140页及Spira，M.和Rosen，T.发表于1993年Clin.Plastic Surgery 20卷181-9页的综述。
注射非生物材料的不良效果促使人们尝试利用外源蛋白，尤其是牛胶原来作为注射物。虽然未加工的牛胶原给人体注射免疫原性太强，但如果病人经过预先筛选，排除了那些免疫反应性的病人，用酶降解去除牛胶原氮端或碳端肽，产生的“无末端胶原(atelocollagen)”的材料还是可以限量使用。制备和利用这些产品的描述可见美国专利3,949,073和4,424,208及4,488,911。该产品按35mg/ml及65mg/ml以ZYDERM牌无末端胶原溶液销售。虽然它在世界范围广泛使用-例如在1987年美国有二十万以上的人使用该产品，但在使用ZYDERM时，有90％以上的受治者产生抗牛抗体，大约1-3％的受治者产生明显的免疫并发症。参见Delustro，F.等发表于1987年Plastic and reconstructive Surgery 79卷581页的文章。
溶液态无末端胶原已证明不能完全令人满意，因为该材料在几周到几月的时间里会被受试者从注射部位吸收不被宿主材料替换。虽然有人考虑过用重复注射等方法，但这些方法因为牛无末端胶原产生免疫反应，其成本较高，病人抵制等原因而实际上应用有限。为了克服这些限制，牛无末端胶原可进一步加工：先用戊二醛交联，然后用细筛滤过并剪切。制备和利用这些产品的描述可见美国专利4,582,640和4,642,117。相应产品以ZYPLAST牌的交联牛无末端胶原出售。交联所期望提供的优点-增加对宿主降解的抵抗，也因产品粘性的增加而有所抵销。粘性的增加，尤其是粘性不规则(团块现象)使该产品更难以使用，甚至在某些方面无法使用。举例可见美国专利5,366,498。
更为甚者，一些调查报告称ZYPLAST和ZYDERM相比，注射效果的持久性仅有很少甚至没有增加。它的注射效果的持续时间最多4-6月。参见Matti，B.A.和NIcolle，F.V.发表于1990年Aesthetic PlasticSurgery 14卷227-34页以及Ozgentas，H.E.等发表于1994年Ann PlasticSurgery 33卷171页的文章。大多数施治者的实验结果表明4-6周以后，该注射物吸收很明显。
牛胶原产品的免疫原性限制了人体使用，促使一些人考虑从胎盘中(举例可见美国专利5,002,071和手术样品中(举例可见美国专利4,969,912以及5,332,802)提取人胶原。因为人胶原和牛胶原经受同样的降解过程，需要进一步的交联和其它化学修饰。
完全由受试者样品衍生的人类胶原是可以制做，现以AUTOLOGENTM牌出售。无明显证据表明注射人胶原比牛胶原产生的效果更持久。而且，利用自身的经加工的胶原也局限于整容外科手术的受治者。因为其制备起始材料是这个手术过程中取出的。明显地，虽然自身的经加工的胶原消除了牛胶原的免疫原性，但它和牛胶原一样不能保证长期治疗效果。而且，它局限于动过手术的受治者。
另有人注射了一种明胶粉末和ε-氨基己酸以及受治者的血浆的混合物(称作“FIBERELTM”)，作为无末端胶原的替代品来增加下临近真皮组织。此为多中心试验结果，参见1987年美国皮肤学会杂志，16卷：1155-62页。
FIBRELTM增加软组织的作用看起来部分依赖于诱导瘢痕性，炎性反应。参见Gold，M.H.发表于1994年J.Dermatologic Surg.Oncol.20卷586-90页的文章。FIBRALTM的治疗报告称它们对部分病人有效，举例可见Millikan，L.等发表于1991年J.Dermatologic Surg.Oncol.17卷223-29页等文章。但因有团块现象且效果不持久，FIBRELTM也不理想。FIBRELTM的应用也因伴随注射的不适而受限，另外医生们也发现它制备繁琐。
总而言之，没有一个现有的非生物可注射材料可以完全令人满意地增加近邻真皮和软组织。
注射无末端胶原不能获得长期的治疗效果，使用FIBREL存在很多问题，这促使一些人去获得并注射(移植)活体脂肪组织去增加下临近真皮组织和软组织的体积。把脂肪组织手术去除，然后手术重新植入人体，对于纠正大缺损可起到很好的效果---举例可见McKinney，P.和Pandya.S.发表于1994年Anesthetic Plastic Surgery 18卷383-5页以及Davies，R.E.等发表于1995年Arch of Otolaryngology--Head & NeckSurgery 121：95-100的文章。但是，需要通过注射来放置移植物的修补，其效果注定不容乐观---可见Eresk，R.A.等发表于1991年Plastic&Reconstructive Surgery 87卷219-27页的文章。因此，虽然对其效果有正面报导---举例可见于Hambley，R.M和Carruthers，J.A.发表于1992年J.Derm.Surgery&Oncol.18卷963-8页的文章，该方法的创立者承认，对于大多数开业医生而言，该技术的效果并不令人满意---可见Lewis，C.M.发表于1993年J.Aesthetic Plastic Surgery 17卷109-12页的文章。医生和病人常碰到的问题有：效果不可预测；团块现象---它使这个操作不宜于治疗细小的皱纹；注射后一段时期内局部极度肿胀---这一般持续4-6周。
3.发明概述：
本发明提供了一种将自身真皮成纤维细胞注射到缺损附近的真皮和皮下组织而纠正受治者皮肤美容整容方面缺损的方法。可用这种方法纠正的典型缺损有皮纹(rhytid)，皮肤扩张纹(stretch mark)，凹陷性瘢痕，非创伤性皮肤凹陷，普通粉刺瘢痕，唇发育不良。依照本方法，注射细胞是和受治者组织相容的，它是经过传代在细胞培养系统中扩增而得到的。在一个优选实施方案中，移植的细胞为真皮成纤维细胞，它来自受治者活体样品培养物。
本发明还提供一种方法可使传代的成纤维细胞基本上没有培养基中存在的免疫原性蛋白，以便用于纠正皮肤缺损。该方法一个步骤是将扩增的成纤维细胞在无蛋白培养基中孵育一段时间。
4.发明详述：
本方法部分基于这样的认识：理想的用于增加缺损下临近真皮和皮下组织体积的材料应该是正常情况下存在于真皮组织中的活细胞。该方法还基于这样的认识：通过在注射前几周从受治者得到活体样品，然后培养来可提供大量所需类型自身细胞。该方法进一步还基于这样的认识：经过这样的组织培养扩增，自身细胞会含有一定量的抗原性蛋白，但是只要依照本申请的指导进行，这种蛋白可在注射前去除。
4.1：可注射细胞悬液的获得方法：
本发明可以通过注射任何可扩大培养的未分化的中胚层细胞来进行。因为真皮成纤维细胞易于获得且易于培养，而且是真皮及其下邻近组织正常成分，因此在一个优选实施方案中，用它来注射。
真皮成纤维细胞培养从2×5mm全层活体皮肤样品开始。因为移植外科医师和免疫学家所熟知的同种异型排斥反应，培养的成纤维细胞与宿主组织相容甚为关键。从待修正其皮肤缺损的受治者获得一块活体样品，并从它开始培养成纤维细胞可确保组织相容性一致。
开始培养前，将活体样品在抗生素和抗真菌试剂中多次漂洗，然后小心地去除表皮和皮下脂肪组织----以便得到的培养物基本上没有非成纤维细胞；将真皮用手术刀或手术剪尽量切开。样品切片逐个用镊子置于一个干玻璃培养皿上。待其贴壁5-10分钟后，缓慢加入少量培养液，注意不要吹起贴壁的组织块。孵育24小时后，再添加一些培养基。用T-25培养瓶开始培养时，培养液的起始量是1.5-2.0ml。从活体样品建立细胞系通常需要2-3周，到时候细胞就可以从最初的培养瓶中取出扩增。
在培养的早期阶段，组织块需要保持贴附在培养皿底部。脱离的组织块需要重新植入新培养瓶中。依照这方面专业人员熟知的技术，组织培养物短暂暴露于EDTA-胰蛋白酶可以刺激成纤维细胞的生长。暴露应该短暂到不足以使成纤维细胞从皿底脱离。当培养物建立起来而快达到融合时，成纤维细胞可以采样冻存。冻存宁早勿晚，因为正常成纤维细胞的传代次数是有限的。
这些成纤维细胞可保存在任何适于保存成纤维细胞的培养液中。应用含70％生长液，20％(V/V)胎牛血清，10％二甲基亚砜(DMSO)的培养液效果就不错。融化的细胞可以用来做传代培养，然后用于同一受治者，免除了重新采样品的不便。
任何从活组织检查样品中扩增真皮成纤维细胞培养方法，都可以用于本发明来扩增细胞。为这方面专业人员所熟知的方法，可参见R.I.Freshney主编的《动物细胞培养：实用方法指南》(英国牛津IRL出版社1987年出版)以及R.I.Freshney主编的《动物细胞培养：基本技术手册》(纽约Alan R.Liss公司出版)。这些书做为以参考文献并入该申请。
任何适合做原代成纤维细胞培养的培养液都可应用。大多数情况下，培养液中添加0.5％-20％(V/V)的血清来加快成纤维细胞的生长。血清浓度越高，细胞生长更快。在一个优选实施方案中，血清是胎牛血清，加到终浓度为10％。例如可以用高葡萄糖DMEM作培养液，添加2mM谷氨酰氨，110mg/L的丙酮酸钠，10％(V/V)胎牛血清和抗生素(即形成“完全培养基”)。
细胞可以胰蛋白酶消化后在新培养瓶传代，例如一瓶细胞可以分做三瓶来养。三底T-150型培养瓶有450平方厘米的培养面积，可以用于该发明。一个三底T-150型培养瓶可以接种6×106个细胞，长满可有1.8×107个细胞。通常5-7天后细胞可以长满，这时生长液换成无血清完全培养基；然后细胞孵育在---也就是保持在30-40℃至少6小时，12小时以上更好。最好是在37℃无蛋白培养基中孵育16-18小时。细胞培养在无血清培养液中培养，基本上可以去除细胞所带的衍生于牛血清蛋白的蛋白质。这些蛋白对人体是免疫原性的，如果存在于注射物中，可以产生变态反应。
无血清培养结束后，将细胞从培养瓶中用胰蛋白酶/EDTA消化下来，通过离心重悬彻底洗涤。最终悬于等体积注射用等张盐水中以备注射。六个三底T-150型培养瓶可以长满可以收获108细胞，这最多少可以做成大约1.0毫升悬液。
或者是，只要在配制成悬液的18个小时内被注射，这些细胞被转移到4℃。细胞悬浮于等体积的完全培养基中用于运送，除了缺乏酚红pH指示剂，用受试者血清代替胎牛血清。细胞可在到运送培养基中吸出和注射。
重悬细胞的盐水或者运送用培养基的体积，要求并不严格。它取决于实施者打算注射的细胞数量，待治疗缺损的大小和数目，受治者对治疗效果要求的迫切性等因素。实施者可以让细胞悬液的体积大一些，这样在每个注射部位注射的细胞相应就会少一些。
5.4.2用于获得细胞粘稠悬液的变通方法：
本发明提供了一种制备注射用细胞悬液的变通方法，适用于真皮修复需要大量材料的情况。加厚嘴唇，治疗鼻唇褶和皮下缺损就可遇到这种情况。
变通方法和上面描述的方法在细胞培养到106以前是一样的。在直径100mm的培养皿中，加入2ml受体的血浆和50-100单位(通常是50μl)的自身凝血酶，使其底部形成血浆凝块，处理以后可以形成一个细胞培养面。将悬于3-5ml的1×106个培养成的真皮成纤维细胞接种于凝块表面，在完全培养基中继续培养7天。到7天时，完全培养基换成无血清培养基。在某种操作方法中，以一小时的间隔换两次培养基，然后将细胞在无血清培养基中继续培养14-16小时可以获得满意的效果。在无血清培养基中孵育完成后，凝块可以吸入注射器，随时注射。
在本发明一个变通实施方案中，成纤维细胞不做成悬液，而是以完整凝块形式或者用手术刀切成所需形状来使用。在真皮擦伤时，这种接种有成纤维细胞的凝块表面做为受治者的真皮的覆盖物。
5.4.3将细胞用于受治者
利用这个领域的专业人士使用ZYDERM和ZYPLAST时所用的同样技术，本发明的悬液可以用来治疗皮肤缺损。细胞悬液可以用来替代无末端胶原，而且具有以上开列的优点。关于利用可注射性材料来增厚缺损下真皮和皮下组织的代表性的教程可以见于外科学文献：Gonzales，U.M.发表于1992年Aesthetic Plastic Surgery 16卷231-4页，Nicolle，F.V发表于1985年Aesthetic Plastic Surgery 9卷159-62页以及Pieyre，J.M.发表于1985年Aesthetic Plastic Surgery 9卷153-54页的文章。这些文献的全部内容作为参考文献引入该申请。
本发明的一个方案，可以用来治疗细微的面部皱纹，方法如下：先用酒精处理待治疗区，然后将之牵拉成平面；取一注射器，装满细胞悬液，安上30ga的针头，尽量表浅地将针头刺入该区---夹角的方向并不关键---轻轻加压，进行真皮内注射，直到可以看到局部轻微发白，注射多个地方。
在其它实施方案中，注射物可以放入口周匝肌来治疗唇发育不良，也可以放入皮下组织来治疗深部皮下缺损。
在一个变通方案中，以将真皮打磨到中层或深层来治疗大片粉刺瘢痕。将一块含成纤维细胞的凝块修整后覆盖到剥露的表面，让接种成纤维细胞的一面紧贴暴露的真皮层。所用的凝块然后用外科包裹剂Xeroform，Adaptic或者其它任何非闭合性外科包裹材料覆盖。
5.临床实验总结：
六个有不同真皮缺损的病人用以上方法进行了治疗，诊断是：笑纹(鼻唇褶)，两个病人；口周放射纹，两个病人；眉间皱纹；凹陷性瘢痕；唇发育不良；光化性颊纹。
每个病人先给预前臂测试剂量1.0ml细胞悬液。两个病人发生了轻微的红斑，但没有其它反应的迹象。三周以后，在真皮缺损区进行了总剂量为1.0ml细胞悬液的治疗性注射。再四周以后进行剂量1.0ml的第二次治疗性注射。仅有唇发育不良病人采用第二次注射来治疗同一缺损，而其它病人在每个治疗区仅注射一次。
有的病人有很轻的红斑，没有即刻的全身或局部不良反应，注射后病人立刻就可以工作。每个病人，都立刻可见到疾病状况改善。
伴随注射，病人仅有很轻微的不适。这种不适据报告轻于注射牛无末端胶原。病人对治疗表示满意，渴望更进一步治疗其它缺损。一些对治疗不知情的病人的亲友也反映病人的真皮缺损得到了纠正。虽然触诊可察觉到一些治疗迹象，但是肉眼看不见皮肤治疗的痕迹。
在注射后六个月的实验期间，没有迟发性局部或全身不良反应。没有病人出现包块或不规则和不平整现象。最有意义的是，经过注射后六个月的观察，治疗效果没有减弱。这段时间中，牛无末端胶原预期应该已吸收。相反，一些可以长期随访的病人还可见到治疗效果随时间延长而增加。长期治疗效果的迟发性提示注射的成纤维细胞是代谢活跃的，可以在注射部位产生额外的细胞外基质。
这里所描述的具体方案是作为该发明的某一方面的一个范例而描述的，本发明的范围不局限于此，功能相当的方法和成分也包括在此发明范围内。的确，在此文中所描述的之外的本发明的多种修改对于                            本                          领域技术人员来说，从上面的描述中显而易见的。这些改良是试图包括在所附的权利要求中的。这里所有引用的文献作为参考文献引入该申请。

Claims (11)

1.真皮成纤维细胞在生产化妆改善受试者皮肤的化妆上或审美上缺损的药剂中的应用，其中的药剂包含扩增的自体真皮成纤维细胞，基本上不含有培养基血清衍生的蛋白质。
2.权利要求1的应用，其中该药剂基本上不含有非成纤维细胞。
3.权利要求1或2的应用，其中缺损指：皮肤纹，皮肤扩张纹，凹陷性瘢痕，非创伤性皮肤凹陷或者嘴唇发育不良。
4.权利要求1或2的应用，其中的药剂包含用于注射到受治疗者化妆或审美缺损的下临近组织的悬液。
5.权利要求4的应用，其中悬液还包括足以形成可注射胶的有效量的人纤维蛋白。
6.权利要求5的应用，其中可注射胶包括凝血酶激活的同源血浆。
7.权利要求1或2的应用，其中通过在不含血清的培养基中温孵去除培养基血清衍生的蛋白。
8.权利要求7的应用，其中成纤维细胞在不含血清的培养基中孵育至少6小时。
9.权利要求1或2的应用，其中的成纤维细胞获自下列步骤：
a)将从受治疗者活体样品得到的真皮成纤维细胞在含0.5-20％非人血清的培养基中传代，以提供基本上无脂肪细胞，角质细胞和细胞外基质的成纤维细胞；
b)将传代的真皮成纤维细胞在无血清培养基中于30-40℃孵育至少6小时；以及
c)将孵育的成纤维细胞用蛋白水解酶消化以悬浮成纤维细胞。
10.真皮成纤维细胞在生产可用于在人类受试者中长期化妆改善受治者皮下或真皮组织的手术方法的药剂中的应用，该方法包括：
a)提供经过传代的自身真皮成纤维细胞的、基本上不含有培养基血清衍生的蛋白质的悬液的药剂；
b)确认容易靠增加下临近皮下或真皮组织来改善的化妆上的缺损；
c)把有效体积的药剂注射到下临近组织使其加厚。
11.权利要求1的应用，其中的药剂包含接种真皮成纤维细胞的凝块，这种真皮成纤维细胞来自该受治者，该凝块用于施于普通粉刺后遗症部位，该部位已剥露到中层或深层真皮，这样接种成纤维细胞贴近真皮。
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Families Citing this family (106)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU6662698A (en) * 1997-02-20 1998-09-09 Gregory S. Keller Augmentation and repair of dermal, subcutaneous, and vocal cord tissue defects
US7767452B2 (en) 1997-02-20 2010-08-03 Kleinsek Don A Tissue treatments with adipocyte cells
WO1998040027A1 (en) * 1997-02-20 1998-09-17 Gerigene Medical Corporation Augmentation and repair of dermal, subcutaneous, and vocal cord tissue defects
US6398753B2 (en) 1998-04-03 2002-06-04 Mcdaniel David H. Ultrasound enhancement of percutaneous drug absorption
US6030374A (en) * 1998-05-29 2000-02-29 Mcdaniel; David H. Ultrasound enhancement of percutaneous drug absorption
US8211423B2 (en) * 2006-11-28 2012-07-03 University Of Pittsburgh Muscle derived cells for the treatment of cardiac pathologies and methods of making and using the same
US6866842B1 (en) 1998-05-01 2005-03-15 University Of Pittsburgh Muscle-derived cells (MDCs) for treating muscle-or bone-related injury or dysfunction
US7115417B1 (en) * 1998-05-01 2006-10-03 Chancellor Michael B Soft tissue and bone augmentation and bulking utilizing muscle-derived progenito compositions, and treatments thereof
US6432710B1 (en) * 1998-05-22 2002-08-13 Isolagen Technologies, Inc. Compositions for regenerating tissue that has deteriorated, and methods for using such compositions
WO1999060951A1 (en) * 1998-05-22 1999-12-02 Isolagen Technologies, Inc. Compositions for regenerating tissue that has degenerated, and methods for using such compositions
US7004933B2 (en) 1998-05-29 2006-02-28 Light Bioscience L.L.C. Ultrasound enhancement of percutaneous drug absorption
US6274090B1 (en) * 1998-08-05 2001-08-14 Thermogenesis Corp. Apparatus and method of preparation of stable, long term thrombin from plasma and thrombin formed thereby
US6936044B2 (en) * 1998-11-30 2005-08-30 Light Bioscience, Llc Method and apparatus for the stimulation of hair growth
US6663659B2 (en) 2000-01-13 2003-12-16 Mcdaniel David H. Method and apparatus for the photomodulation of living cells
US6676655B2 (en) 1998-11-30 2004-01-13 Light Bioscience L.L.C. Low intensity light therapy for the manipulation of fibroblast, and fibroblast-derived mammalian cells and collagen
US20060212025A1 (en) * 1998-11-30 2006-09-21 Light Bioscience, Llc Method and apparatus for acne treatment
US6283956B1 (en) 1998-11-30 2001-09-04 David H. McDaniels Reduction, elimination, or stimulation of hair growth
US9192780B2 (en) * 1998-11-30 2015-11-24 L'oreal Low intensity light therapy for treatment of retinal, macular, and visual pathway disorders
US6887260B1 (en) * 1998-11-30 2005-05-03 Light Bioscience, Llc Method and apparatus for acne treatment
US20080267923A2 (en) * 1999-11-05 2008-10-30 Donald Kleinsek Hair undifferentiated cells
US20090074729A2 (en) * 1999-11-05 2009-03-19 Donald Kleinsek Augmentation and repair of spincter defects with cells including fibroblasts
CA2390053A1 (en) * 1999-11-05 2001-05-10 Gerigene Medical Corporation Augmentation and repair of age-related soft tissue defects
US20080286242A2 (en) * 1999-11-05 2008-11-20 Donald Kleinsek Augmentation and repair of spincter defects with cells including mesenchymal cells
US7799325B2 (en) * 1999-11-05 2010-09-21 Kleinsek Donald A Removal of hypertrophic scars
ATE365045T1 (de) 2000-04-14 2007-07-15 Univ Pittsburgh Vermehrung des weich- sowie des knochengewebes anhand von aus muskeln stammenden vorläuferzellen,sowie damit verbundene zusammensetzungen und behandlungsformen
CA2420901A1 (en) 2000-08-08 2002-02-28 Bioamide, Inc. Scaffolds for tissue engineered hair
US20090130066A1 (en) * 2000-11-06 2009-05-21 Gerigene Medical Corporation Augmentation and repair of sphincter defects with cells including muscle cells
CA2438904C (en) * 2001-02-23 2012-09-04 The University Of Pittsburgh Rapid preparation of stem cell matrices for use in tissue and organ treatment and repair
GB0105899D0 (en) * 2001-03-09 2001-04-25 Intercytex Ltd Fibrosis reduction treatment
US7468242B2 (en) 2001-11-05 2008-12-23 Medgenics, Inc. Dermal micro organs, methods and apparatuses for producing and using the same
US8088568B2 (en) 2001-11-05 2012-01-03 Medgentics, Inc. Dermal micro-organs, methods and apparatuses for producing and using the same
US8501396B2 (en) 2001-11-05 2013-08-06 Medgenics Medical Israel Ltd. Dermal micro-organs, methods and apparatuses for producing and using the same
KR100518152B1 (ko) * 2002-05-09 2005-09-29 메디제네스(주) 혈장 또는 혈청을 함유한 창상 치료용 약학 조성물
US20040101959A1 (en) * 2002-11-21 2004-05-27 Olga Marko Treatment of tissue with undifferentiated mesenchymal cells
WO2004092335A2 (en) * 2003-04-10 2004-10-28 Light Bioscience, Inc. Photomodulation methods and devices for regulating cell proliferation and gene expression
EP1617852B1 (en) * 2003-04-25 2010-09-29 The University of Pittsburgh Muscle derived cells (mdcs) for promoting and enhancing nerve repair and regeneration
KR20120102152A (ko) 2003-05-01 2012-09-17 메드제닉스 인코포레이티드 진피 미세 기관, 그를 생산 및 이용하기 위한 방법 및 장치
US7186557B2 (en) * 2003-06-13 2007-03-06 Isolagen Technologies, Inc. Methods of producing neurons
CN1297324C (zh) * 2003-07-25 2007-01-31 吕伟光 组织工程自体复合皮肤及其制备方法
EP1648385B1 (en) * 2003-07-31 2016-05-04 Gentlewaves Llc System and method for the photodynamic treatment of skin
KR20060036933A (ko) * 2003-10-07 2006-05-02 바이오마스터 인코포레이티드 지방-유래 전구세포의 세포 분화
US20070219487A1 (en) * 2003-11-10 2007-09-20 The Cleveland Clinic Foundation Method to Control Ventricular Rate in Atrial Fibrillation Patients
US20070122906A1 (en) * 2003-12-29 2007-05-31 Allan Mishra Method of culturing cells
EP1718366A4 (en) * 2004-02-06 2007-11-21 Daniel Barolet METHOD AND DEVICE FOR TREATING MAMMAL TISSUE
US9526746B2 (en) * 2004-02-13 2016-12-27 Smith & Nephew, Inc. Wound healing composition
KR100680134B1 (ko) * 2004-06-10 2007-02-08 박우삼 무세포 진피를 주사가능하도록 입자 형태로 가공한 필러
KR100614903B1 (ko) * 2004-08-06 2006-08-25 테고사이언스 (주) 케라티노사이트 및/또는 섬유아세포를 함유하는 화장용조성물
WO2006019357A1 (en) 2004-08-16 2006-02-23 Cellresearch Corporation Pte Ltd Isolation of stem/progenitor cells from amniotic membrane of umbilical cord.
FR2874503B1 (fr) * 2004-09-01 2006-11-24 Soc Ind Limousine Dapplication Biologique Silab Procede d'obtention d'un principe actif raffermissant et a visee anti-vergetures, actif obtenu et les fonctionnalites
GB0505202D0 (en) 2005-03-14 2005-04-20 Intercytex Ltd Skin equivalent culture
JP2008542252A (ja) 2005-05-26 2008-11-27 インターサイテックス リミティド 同種真皮繊維芽細胞を用いる組織修復
JP2008545703A (ja) 2005-05-27 2008-12-18 バイアセル インコーポレーティッド 幹細胞を用いた虚血の処置
JP4982865B2 (ja) * 2005-06-29 2012-07-25 国立大学法人名古屋大学 皮膚組織改善材及びその利用
US20070065415A1 (en) * 2005-09-16 2007-03-22 Kleinsek Donald A Compositions and methods for the augmentation and repair of defects in tissue
TW200800240A (en) 2005-11-22 2008-01-01 Aderans Res Inst Inc Hair follicle graft from tissue engineered skin
CN100421731C (zh) * 2006-01-11 2008-10-01 黄靖西 人自体胶原蛋白活细胞注射剂及其制备方法
EP2210635A3 (en) 2006-02-09 2012-11-07 Aderans Research Institute, Inc. Apparatus for delivering fluid and material to a subject
GB2440908A (en) * 2006-05-25 2008-02-20 Intercytex Ltd Tissue Repair and Augmentation
CN100415307C (zh) * 2006-05-30 2008-09-03 中国人民解放军第二军医大学 人成纤维细胞修饰的异体或异种脱细胞真皮替代物
TR200602727A2 (tr) * 2006-05-31 2007-12-24 Tun� T�Ryak� Kemal Cilt defektleri ve yaşlanması tedavisinde kullanılacak bir yöntem ve bir doku maddesi
KR20070122315A (ko) * 2006-06-26 2007-12-31 (주) 에스바이오메딕스 자가 진피 세포 및 히알루론산을 함유하는 주사용 인체연조직 충전제 조성물
KR20070122316A (ko) * 2006-06-26 2007-12-31 (주) 에스바이오메딕스 주사용 인체 연조직 충전제 및 이의 제조 방법
WO2008027984A1 (en) 2006-08-29 2008-03-06 Isolagen Technologies, Inc. Methods for culturing minimally-passaged fibroblasts and uses thereof
US9127084B2 (en) 2006-09-14 2015-09-08 Medgenics Medical Israel Ltd. Long lasting drug formulations
AU2007294858B2 (en) 2006-09-14 2011-05-12 Medgenics Medical Israel, Ltd Long lasting drug formulations
US8454948B2 (en) 2006-09-14 2013-06-04 Medgenics Medical Israel Ltd. Long lasting drug formulations
WO2008069376A1 (en) * 2006-12-07 2008-06-12 Jun-Ho Shin Culture method of fibroblast using autologous serum mixed with placenta extract and composition for skin regeneration using the same
US20100158875A1 (en) 2006-12-18 2010-06-24 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Muscle derived cells for the treatment of gastro-esophageal pathologies and methods of making and using the same
AU2007334331B2 (en) * 2006-12-18 2014-05-29 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Muscle derived cells for the treatment of gastro-esophageal pathologies and methods of making and using the same
KR20090093792A (ko) * 2006-12-28 2009-09-02 고쿠리츠 다이가쿠 호우징 나고야 다이가쿠 피부 조직 개선재 및 그 제조 방법
US8105834B2 (en) * 2007-01-11 2012-01-31 University of Pittsburgh—of the Commonwealth System of Higher Education Muscle derived cells for the treatment of urinary tract pathologies and methods of making and using same
JP4347408B2 (ja) * 2007-04-28 2009-10-21 株式会社エーシーバイオテクノロジーズ 皮膚または歯肉の軟組織の欠損を補修するための組成物，及び自己線維芽細胞の培養方法
US7985537B2 (en) 2007-06-12 2011-07-26 Aderans Research Institute, Inc. Methods for determining the hair follicle inductive properties of a composition
PL2288376T3 (pl) 2008-04-18 2016-07-29 Collplant Ltd Sposoby generowania i wykorzystywania prokolagenu
US9199003B2 (en) * 2008-08-18 2015-12-01 University of Pittsburgh—of the Commonwealth System of Higher Education Bone augmentation utilizing muscle-derived progenitor compositions in biocompatible matrix, and treatments thereof
EP2182055A1 (en) 2008-10-14 2010-05-05 Heinrich-Heine-Universität Düsseldorf Human cord blood derived unrestricted somatic stem cells (USSC)
US20100233138A1 (en) * 2008-11-07 2010-09-16 University Of Pittsburgh - Of The Commonwealth System Of Higher Education Vocal Cord Augmentation Utilizing Muscle-Derived Progenitor Compositions, and Treatments Thereof
FR2948286B1 (fr) * 2009-07-27 2011-08-26 Jean-Noel Thorel Composition injectable associant un agent de comblement et un milieu de croissance des fibroblastes
US8900181B2 (en) 2009-12-18 2014-12-02 Srgi Holdings, Llc Skin treatment and drug delivery device
US8529883B2 (en) 2010-05-07 2013-09-10 Fibrocell Technologies, Inc. Dosage unit formulations of autologous dermal fibroblasts
WO2012000180A1 (zh) * 2010-06-30 2012-01-05 Chen Jinxi 一种皮肤组织细胞聚合物的制备方法及其用途
US11278309B2 (en) 2010-12-17 2022-03-22 Srgi Holdings, Llc Pixel array medical systems, devices and methods
US11103275B2 (en) 2010-12-17 2021-08-31 Srgi Holdings, Llc Pixel array medical systems, devices and methods
US10661063B2 (en) 2010-12-17 2020-05-26 Srgi Holdings, Llc Systems, devices and methods for fractional resection, fractional skin grafting, fractional scar reduction and fractional tattoo removal
US11109887B2 (en) 2013-12-06 2021-09-07 Srgi Holdings, Llc Pixel array medical systems, devices and methods
US11000310B2 (en) 2010-12-17 2021-05-11 Srgi Holdings, Llc Pixel array medical systems, devices and methods
US20160317170A1 (en) 2013-12-06 2016-11-03 Edward KNOWLTON Pixel array medical systems, devices and methods
US10702684B2 (en) 2010-12-17 2020-07-07 Srgi Holdings, Llc Systems, devices and methods for fractional resection, fractional skin grafting, fractional scar reduction and fractional tattoo removal
US10736653B2 (en) 2013-12-06 2020-08-11 Srgi Holdings, Llc Pixel array medical systems, devices and methods
CN103083727A (zh) * 2011-11-07 2013-05-08 北京清美联创干细胞科技有限公司 一种利用自身真皮成纤维细胞和脐带间充质干细胞修补皮肤和软组织缺损的方法
CN102488930A (zh) * 2011-12-15 2012-06-13 无锡市第三人民医院 一种自异体表皮细胞混合悬液的制备方法
US10407665B2 (en) 2012-04-09 2019-09-10 The Usa, As Represented By The Secretary, Dept. Of Health And Human Services Methods for generation of pluripotent and multipotent cells
CA2926322C (en) 2013-10-02 2022-10-18 Srgi Holdings, Llc Pixel array medical devices and methods
ES2827049T3 (es) 2013-10-02 2021-05-19 Srgi Holdings Llc Dispositivos médicos de conjunto de píxeles
US11229452B2 (en) 2013-12-06 2022-01-25 Srgi Holdings, Llc Pixel array medical systems, devices and methods
US11937846B2 (en) 2013-12-06 2024-03-26 Srgi Holdings Llc Pixel array medical systems, devices and methods
US20150301028A1 (en) 2014-04-22 2015-10-22 Q-State Biosciences, Inc. Analysis of compounds for pain and sensory disorders
EP3579767A4 (en) 2014-10-02 2020-11-18 SRGI Holdings, LLC MEDICAL SYSTEMS, DEVICES AND METHODS FOR PIXEL NETWORKS
US10048275B2 (en) 2015-03-13 2018-08-14 Q-State Biosciences, Inc. Cardiotoxicity screening methods
RU2578804C1 (ru) * 2015-04-27 2016-03-27 Екатерина Евгеньевна Фаустова Способ омоложения организма пациента
EP3298448A4 (en) 2015-05-21 2019-03-27 Q-State Biosciences, Inc. OPTOGENETIC MICROSCOPE
US11751903B2 (en) 2015-08-31 2023-09-12 Srgi Holdings, Llc Pixel array medical systems, devices and methods
US11490952B2 (en) 2015-08-31 2022-11-08 Srgi Holdings, Llc Pixel array medical devices and methods
US11564706B2 (en) 2019-10-28 2023-01-31 Srgi Holdings, Llc Pixel array medical systems, devices and methods
WO2018217479A2 (en) 2017-05-11 2018-11-29 Srgi Holdings, Llc Systems, devices and methods for fractional resection, fractional skin grafting, fractional scar reduction and fractional tattoo removal
GB202104130D0 (en) * 2021-03-24 2021-05-05 King S College London Method

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3837669A1 (de) * 1988-11-05 1989-06-01 Nikolaus W Prof Dr Med Klehr Die verwendung koerpereigener fibroblasten und epidermiszellen als wirkstoff-traeger fuer die intrakorporale behandlung und als wirkstoffvermittler fuer die lokalbehandlung
US5002071A (en) * 1987-04-03 1991-03-26 Research Development Foundation Injectable soft tissue augmentation materials from the placenta and their method of manufacture
EP0498992A2 (en) * 1991-02-15 1992-08-19 Nippon Oil Co., Ltd. Serum-free culture medium containing lysozyme
JPH06169761A (ja) * 1992-12-04 1994-06-21 Noevir Co Ltd 動物由来線維芽細胞の無血清培養用培地及び培養方法

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3949073A (en) * 1974-11-18 1976-04-06 The Board Of Trustees Of Leland Stanford Junior University Process for augmenting connective mammalian tissue with in situ polymerizable native collagen solution
US4488911A (en) * 1975-10-22 1984-12-18 Luck Edward E Non-antigenic collagen and articles of manufacture
US4424208A (en) * 1982-01-11 1984-01-03 Collagen Corporation Collagen implant material and method for augmenting soft tissue
US4582640A (en) * 1982-03-08 1986-04-15 Collagen Corporation Injectable cross-linked collagen implant material
US4609551A (en) * 1984-03-20 1986-09-02 Arnold Caplan Process of and material for stimulating growth of cartilage and bony tissue at anatomical sites
US4642117A (en) * 1985-03-22 1987-02-10 Collagen Corporation Mechanically sheared collagen implant material and method
US4969912A (en) * 1988-02-18 1990-11-13 Kelman Charles D Human collagen processing and autoimplant use
GB2220593B (en) * 1988-06-09 1992-04-22 Cyclofil Hydro-cyclone
DE3829183A1 (de) * 1988-08-29 1990-03-08 Wella Ag Feeder-layer aus menschlichen bindegewebszellen, verfahren zu seiner herstellung sowie verfahren zur vermehrung menschlicher epithelzellen und deren verwendung zur restitution der haut
JP2774695B2 (ja) * 1991-03-29 1998-07-09 コラーゲン コーポレイション 顔のしわを処置する装置および方法
WO1998040027A1 (en) * 1997-02-20 1998-09-17 Gerigene Medical Corporation Augmentation and repair of dermal, subcutaneous, and vocal cord tissue defects

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5002071A (en) * 1987-04-03 1991-03-26 Research Development Foundation Injectable soft tissue augmentation materials from the placenta and their method of manufacture
DE3837669A1 (de) * 1988-11-05 1989-06-01 Nikolaus W Prof Dr Med Klehr Die verwendung koerpereigener fibroblasten und epidermiszellen als wirkstoff-traeger fuer die intrakorporale behandlung und als wirkstoffvermittler fuer die lokalbehandlung
EP0498992A2 (en) * 1991-02-15 1992-08-19 Nippon Oil Co., Ltd. Serum-free culture medium containing lysozyme
JPH06169761A (ja) * 1992-12-04 1994-06-21 Noevir Co Ltd 動物由来線維芽細胞の無血清培養用培地及び培養方法

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