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脂肪干细胞复合富血小板血浆对脂肪颗粒移植的影响
Increased Survival of Free Fat Grafts with Adipose-derived Stem Cells and Platelet-rich Plasma in Nude Mice
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摘要: 目的 探讨运用组织工程技术构建的组织工程脂肪对颗粒脂肪移植的影响。 方法 通过酶消化法获取脂肪干细胞(ASCs),采用两步离心法制备富血小板血浆(PRP),运用组织工程技术构建组织工程脂肪。根据移植物的组成不同分为ASCs+PRP+脂肪颗粒、PRP+脂肪颗粒、ASCs+脂肪颗粒及对照组(PBS+脂肪颗粒),分别移植入裸鼠皮下,并于移植后10 d、30 d、60 d及90 d取出移植物,进行大体观察及体积测量,镜下观察组织结构变化。 结果 90 d后ASCs+PRP+脂肪颗粒组移植物质地柔软、颜色淡黄,较其它组别更接近于正常的脂肪组织,且具有更多的存活体积(P<0.05)及更好的组织学形态。 结论 运用ASCs与PRP复合脂肪颗粒能够成功构建组织工程脂肪,是一种新的安全有效的修复颌面部及全身软组织缺损方法,具有较高的应用前景。Abstract: Objecitve To investigate the impact of combined adipose-derived stem cells (ASCs) and platelet-rich plasma (PRP) on the survival of transplanted fat. Methods The ASCs were isolated and cultured from fat tissues by enzyme digestion; and the PRP was prepared by two-step ultracentrifugation. The grafts of fat granules were divided into test groups (ASCs+PRP+fat granules, PRP+fat granules, ASCs+fat granules) and control groups (PBS+fat granules). The grafts were injected into the left and right dorsal subcutaneous areas of nude mice. General observations, volume measurements and microscope examinations were conducted 10 d, 30 d, 60 d and 90 d after transplantation. Results The grafts of the mice in the ASCs+PRP group showed soft structure, with light yellow color and closer to normal adipose tissue compared with those in other groups. Greater survival volumes (P<0.05) and better histology were also observed in the grafts of the mice in the ASCs+PRP group. Conclusion The fat grafts consisting of PRP and ASCs constitute an ideal transplant strategy, which could provide a valuable and needed tool in plastic and reconstructive surgery.
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Keywords:
- Fat transfer /
- Platelet-rich plasma /
- Adip ose-derived stem cells
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患者，女，30岁，因“孕检发现盆腔包块2月”入院。专科查体：宫体3+月孕大；右附件扪及一包块，10 cm×9 cm×6 cm，质中。B超见子宫下段偏右10.3 cm×7.9 cm×9.9 cm囊实混合性占位，形态较规则，边界较清。肿瘤标志物：CA125 46.2 U/mL，HCG 48 357 IU/mL。于孕15+1周入院行右附件包块切除术并保留妊娠。术中见：右卵巢15 cm×12 cm×12 cm，表面光滑，包膜完整。大体检查：灰红卵圆形包块，10 cm×9 cm×3.2 cm，包膜完整，切面灰红、实性、质软。镜下观察(图 1)：富细胞区和水肿区交错分布，可见假小叶结构。小叶间疏松水肿，灶性出血；肿瘤细胞稀疏，短梭或长梭形，胞浆淡染。假小叶内细胞丰富，可见血管外皮细胞瘤样结构；瘤细胞卵圆形、多边形，胞质嗜酸性或空亮，核小，居中或偏位，无核沟，染色质较细腻；核分裂像每10个高倍视野6~7个。免疫组化染色(图 2)显示
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图 1 右附件包块镜下检查A:富细胞区和水肿区交错分布，形成假小叶结构。HE ×40; B:肿瘤细胞间见薄壁小血管；肿瘤细胞圆或卵圆形，胞质嗜酸性或空亮，核小，染色质较细腻，右上角白框内可见细胞核分裂。HE ×400(白框内HE ×400)![]()
图 2 肿瘤组织切片免疫组化染色结果。EnVision ×200A:肿瘤细胞α-inhibin阳性; B:肿瘤细胞Desmin阳性 α-inhibin、calretinin、Vimentin、desmin均(+)，Ki67阳性率30%；PCK、EMA、CD34、CD31、FⅧ、CD117、DOG1和Bcl-2均(-)。病理诊断为右卵巢核分裂活跃的硬化性间质瘤(sclerosing stromal tumors, SSTs)。术后患者恢复良好，并于妊娠40周剖腹产子。新生儿体质量3 400 g，Apgar评分：10-10-10。随访2年无明显复发征象。
讨论 硬化性间质瘤 SSTs非常罕见，占卵巢间质肿瘤的2%~6%。好发于年轻妇女，常见月经失调和骨盆疼痛。本例患者于产检时偶然发现，无明显临床体征，仅血清CA125轻度升高。SSTs大体典型为实性，少数有囊性变，镜下常呈典型的假小叶结构，肿瘤细胞形态多样，大小不均，以圆形、椭圆形为主，夹杂梭形细胞；细胞核呈圆形、卵圆形，居中或偏位，染色质细腻，核分裂像少见。假小叶间为胶原纤维构成的硬化区或由疏松水肿、黏液变性的胶原纤维形成的疏松水肿区，水肿常呈带状。瘤组织内可见较多大小不一的薄壁血管分支。免疫组化：肿瘤细胞常表达Vimentin、desmin和SMA，不同程度表达α-inhibin、calretinin。 2014版WHO女性生殖道肿瘤分类中认为SSTs核分裂罕见；本例镜下形态典型，但每10个高倍视野有6~7个核分裂像，Ki67增殖指数为30%，超出常规普通病例表现。关于核分裂活跃的SSTs可见少数几篇文献报道，已报道的病例大多为良性，预后与普通的SSTs无差异。本例术后定期随访，于孕40周行剖腹产，随访两年未见肿瘤复发。
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[1] Crandall DL, Hausman GJ, Kral JG. A review of the microcirculation of adipose tissue:anatomic, metabolic, and angiogenic perspectives. Microcirculation,1997;4(2):211-232.
[2] Ersek RA. Transplantation of purified autologous fat:a 3-year follow-up is disappointing. Plast Reconstr Surg,1991;87(2):219-227.
[3] Cherubino M, Marra KG. Adipose-derived stem cells for soft tissue reconstruction. Regen Med,2009;4(1):109-117.
[4] Crandall DL, Busler DE, McHendry-Rinde B, et al. Autocrine regulation of human preadipocyte migration by plasminogen activator inhibitor-1. J Clin Endocrinol Metab,2000;85(7):2609-2614.
[5] Hausman GJ, Richardson RL. Adipose tissue angiogenesis. J Anim Sci,2004;82(3):925-934.
[6] Wu CC, Chen WH, Zao B, et al. Regenerative potentials of platelet-rich plasma enhanced by collagen in retrieving pro-inflammatory cytokine-inhibited chondrogenesis. Biomaterials,2011;32(25):5847-5854.
[7] Yoshimura K, Suga H, Eto H. Adipose-derived stem/progenitor cells:roles in adipose tissue remodeling and potential use for soft tissue augmentation. Regen Med,2009;4(2):265-273.
[8] Stosich MS, Moioli EK, Wu JK, et al. Bioengineering strategies to generate vascularized soft tissue grafts with sustained shape. Methods,2009;47(2):116-121.
[9] Butler DL, Goldstein SA, Guilak F. Functional tissue engineering:the role of biomechanics. J Biomech Eng,2000;122(6):570-575.
[10] Gomillion CT, Burg KJ. Stem cells and adipose tissue engineering. Biomaterials,2006;27(36):6052-6063.
[11] Coleman WP. Autologous fat transplantation. Plast Reconstr Surg,1991;88(4):736.
[12] Robiony M, Zorzan E, Polini F, et al. Osteogenesis distraction and platelet-rich plasma:combined use in restoration of severe atrophic mandible. Long-term results. Clin Oral Implants Res,2008;19(11):1202-1210.
[13] Robiony M, Polini F, Costa F, et al. Osteogenesis distraction and platelet-rich plasma for bone restoration of the severely atrophic mandible:preliminary results. J Oral Maxillofac Surg,2002;60(6):630-635.
[14] Nishimura S, Manabe I, Nagasaki M, et al. Adipogenesis in obesity requires close interplay between differentiating adipocytes, stromal cells, and blood vessels. Diabetes,2007;56(6):1517-1526.
[15] Masuda T, Furue M, Matsuda T. Novel strategy for soft tissue augmentation based on transplantation of fragmented omentum and preadipocytes. Tissue Eng,2004;10(11-12):1672-1683.
[16] Shoshani O, Livne E, Armoni M, et al. The effect of interleukin-8 on the viability of injected adipose tissue in nude mice. Plast Reconstr Surg,2005;115(3):853-859.
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