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胡葵葵 戴育成 李剑 袁敬东 李洁 吴琼
【摘要】 目的用表皮干细胞和成纤维细胞作为种子细胞，研制一种增殖能力强具有表皮、真皮的组织工程化人工复合皮肤。方法从幼儿包皮中分离表皮干细胞，用胶原Ⅳ纯化、富集表皮干细胞，接种在3T3细胞滋养层上．将体外传代培养的表皮干细胞和真皮成纤维细胞分别接种在经冷冻干燥及戊二醛交联的J．型胶原基质网架的两侧，在液面下培养2周后，改为气一液界面培养，构建复合皮肤，移植到裸鼠全层皮肤缺损处，以表皮细胞胶原海绵复合皮肤和无细胞接种的胶原海绵膜作为对照。术后进行大体观察，在第7、14、21天取材行组织学、免疫组织化学及电镜观察。结果用表皮干细胞构建的复合人工皮肤增殖能力强．新生的皮肤瘢痕轻，形态满意，创面愈合的速度和质量均优于对照组。结论在胶原海绵上用表皮干细胞构建的复合人工皮肤增殖能力强，较好地解决了种子细胞的老化问题，可望成为一种较为理想的皮肤替代物。
【关键词】 表皮干细胞； 复合人工皮肤； 创面愈合； 组织工程
Repair of fu¨skin Ioss with composite graft of epidermal stem cells in nude mice HU Kui-kui，DAI Yu一cheng，Li Jian ，YUAN Jing—dong，Li Jie，WU Qiong．Center of Plastic，laser and Cosmetic surger一y，Second Affiliated Hospital, Jiangxi Medical Collage , Nanchang 330006，China
【Abstract】
Objective: To constitute a composite skin substitute that can proliferate well with epidermal stem cells and fibroblasts on collagen sponge．
Methods: Epidermal stem cells were selected by rapid attachment to c01lagenⅣfor 10～15 min and cultured on 3T3 feeder layer． Collagen was extracted from rat tail_ The matrix lattice was fabricated by a freeze—dryer and cross—linked with glutaraldehyde． Fibroblasts (1 x 106／cm2) were inoculated on collagen sponge and cultured for 2 weeks prior to inoculation of epidermal stem cells(2×105／cm2) to construct composite skin substitute．The artificial skin was grafted onto full skin 10ss wounds of nude mice．Collagen sponge membrane lacking cell inoculation and a composite skin substitute with keratinocytes and fibroblasts were used as controls． The wounds were observed daily． Tj\issue samples were harvested and examined by means of histology，immunohistochemistry and electron microcopy．
Results：Composite graft with epidermal stem cells proliferated promptly，and achieved a good adherence to wound on the 3rd day of grafting．the mice exhibited less epidermal scar formation and satisfactory contour of the skin．The quality and speed of wound healing in experimental groups were superior to those in the control groups．
Conclusion: Composite graft with epidermal stem cells have potential prospects in application of repairing skin defect with advantage of good proliferation．
【Key words】Epidermal stem cell；Compound artificial skin；Wound healing；Tissue engineerlng
Bonnie C. Carneya,b,h , Mary A. Oliverb , Metecan Erdi c , Liam D. Kirkpatrickb , Stephen P. Tranchinab , Selim Rozyyevd , John W. Keylounb,e , Michele S. Saruwatarid,e , John L. Daristotlef , Lauren T. Moffatta,b,h , Peter Kofinasc , Anthony D. Sandlerd , Jeffrey W. Shuppa,b,g,h,⁎
a Department of Biochemistry and Molecular and Cellular Biology, Georgetown University Medical Center,Washington, DC, USA
b Firefighters’ Burn and Surgical Research Laboratory, MedStar Health Research Institute, Washington, DC, USA
c Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, MD 20742, USA
d Sheikh Zayed Institute for Pediatric Surgical Innovation, Joseph E. Robert Jr. Center for Surgical Care, Children’s National Medical Center, Washington, DC 20010, USA
e Department of Surgery, MedStar Washington Hospital Center and MedStar Georgetown University Hospital, Washington, DC, USA
f David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
g The Burn Center, Department of Surgery, MedStar Washington Hospital Center, Washington, DC, USA h Department of Surgery, Georgetown University School of Medicine, Washington, DC, USA
article info
Keywords: Autologous skin cell suspension Solution-blow spinning polymer Wound dressings Hypertrophic scarring Skin grafting
abstract
Autologous skin cell suspensions (ASCS) can treat burns of varying depths with the advantage of reduced donor site wound burden. The current standard primary dressing for ASCS is a nonabsorbant, non-adherent, perforated film (control) which has limited conformability over heterogeneous wound beds and allows for run-off of the ASCS. To address these concerns, a novel spray-on polymer formulation was tested as a potential primary dressing in porcine deep partial thickness (DPT) and full thickness (FT) wounds. It was Burn wound healing hypothesized that the polymer would perform as well as control dressing when evaluating wound healing and scarring.
DPT or FT wounds were treated with either a spray-on poly(lactic-co-glycolic acid) (PLGA) and poly(lactide-co-caprolactone) (PLCL) formulation or control ASCS dressings. Throughout the experimental time course (to day 50), we found no significant differences between polymer and control wounds in % re-epithelialization, graft-loss, epidermal or dermal thickness, or % dermal cellularity in either model. Pigmentation, erythema, elasticity, and trans-epidermal water loss (TEWL), were not significantly altered between the treatment groups, but differences between healing wounds/scars and un-injured skin were observed. No cytotoxic effect was observed in ASCS incubated with the PLGA and PLCL polymers.
These data suggest that the novel spray-on polymer is a viable option as a primary dressing, with improved ease of application and conformation to irregular wounds. Polymer formulation and application technique should be a subject of future research.
Kara Kallies, M.S.*, Ann M. Rogers, M.D., F.A.C.S., F.A.S.M.B.S., for the American Society for Metabolic and Bariatric Surgery Clinical Issues Committee
Division of Minimally Invasive and Bariatric Surgery, Penn State Health, Hershey, Pennsylvania Received 16 March 2020; accepted 16 March 2020
*Correspondence: Kara Kallies, M.S., Penn State Health, 500 University Drive, Hershey, PA 17033.
E-mail address: 该Email地址已收到反垃圾邮件插件保护。要显示它您需要在浏览器中启用JavaScript。 (K. Kallies).
吴昌炎·，房志强2，江宝华3，陈石海1
(1．广西医科大学附属第一医院整形美容科广西南宁5 30000；2．龙岩华美罗桥医院整形科；3．江苏省昆山市时光医疗美容医院)
[摘要]目的：探讨自体表皮细胞悬液在修复皮肤组织缺损创面的作用。方法：在2 o只大鼠身上分别取邮票大小自体全厚皮片进行消化，分离成单个表皮细胞的悬液；经传代培养后，使用自体表皮细胞悬液分别对2 0只大鼠背部表皮皮肤缺损创面进行覆盖移植，术后观察移植物成活率和植皮区收缩率，同时取移植物进行组织学观察；并与对照组2 o只自体愈合创面进行相互比较。结果：表皮细胞悬液移植后1周渐融合成片，伤1：2愈合；经6周后，伤口平整，轻度瘢痕愈合。而对照组在6周时伤口不平整，呈瘢痕愈合，色素沉着明显。结论：在表皮缺损创面上应用体外培养的自体表皮细胞悬液移植修复可达到重构皮肤的目的。
[关键词]移植：组织缺损；表皮细胞悬液；细胞培养；皮肤重建
[中图分类号]R622
[文献标识码]A
[文章编号]1 008—645 5(2012)12—2201- 03
Application of the Suspension of epidermal cell cultured in tissue defect
WU Chang—yanl，FANG Zhi—qian92,JIANG Bao—hua3,CHEN Shi—hall
(1．Department of Plastic Surgery，Fi rst Affiliated Hospital of Guangxi Medical University，Nanning 530021，Guangxi，China；2 Department of Plastic Surgery，Longyan Huamei Luoqiao Hospital；3 Department of Plastic Surgery，Kunshan Shiguang
Cosmetology Hospital)
Abstract：
Objective To repair the tissue defect by the Suspension of epidermal cell．
Methods a stamp—size of normal skin was harvested in forty rats and epidermal cells suspension was made th rough decomposing，digesting，culturing and re——digesting in lab．Then it Was implanted to the skin defects made on the dorsa of rats．Shrinkage of the graft survival rate was observed after grafting and samples were taken to make routine paraffin sections for microscopica||y examination，then compared with autogenous healing．
Results After Suspension of epidermal cell was cultured for one week．1he wounds healed gradually The surface of healing wounds was smooth with a slight scar three to six weaks later．But the surface of healing wounds of control group was uneven，scar healing and pigmentation was obvious
Conculsion In the skin defect On the application Of in vitro culture wound autologous skin celI suspension liquid transDlantatiOn to repair lhe purpose of lhe reconstruction of the skin．
Key words：transDIantatiOn：tissue defect；suspension Of epidermal cell；cellular culture；reconstruction Of human skin.
