Michalak-Micka Katarzyna, Klar Agnes S, Böttcher-Haberzeth Sophie, Reichmann Ernst, Meuli Martin, Biedermann Thomas
Tissue Biology Research Unit, Department of Surgery, University Children's Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland.
Department of Surgery, University Children's Hospital Zurich, Zurich, Switzerland.
Pediatr Surg Int. 2019 Jan;35(1):121-127. doi: 10.1007/s00383-018-4383-5. Epub 2018 Oct 31.
The use of autologous bio-engineered dermo-epidermal skin substitutes (DESS) yields a pivotal opportunity to cover large skin defects in human patients. These skin grafts consist of both epidermal and dermal compartments necessary for robust and permanent functional wound closure. In this study, we investigated the impact of mesenchymal cells derived from different body site origins on the expression pattern of diverse markers within DESS.
Human keratinocytes were obtained from interfollicular epidermis, and mesenchymal cells were isolated from foreskin, palmar skin, fat tissue, and tonsils. After expansion, epidermal cells were seeded on collagen I hydrogels containing stromal cells. These human DESS were transplanted on the back of immune-incompetent rats. After 3 weeks, transplants were excised and analyzed using immunohistology techniques.
The macroscopic appearance of skin grafts containing tonsil, fat tissue, or palmar derived mesenchymal cells, was similar to substitutes with foreskin derived dermal fibroblasts. All skin grafts had a strong membrane-localized expression of Lingo-1 in the epidermis. Additionally, we observed an intense expression of transglutaminase 5 in upper epidermal cell layers of the skin grafts confirming a proper keratinocyte differentiation. Tropoelastin was localized throughout the dermal compartments and tightly in contact with the dermo-epidermal junction suggesting an advanced maturation of all skin grafts.
Our data implicate that stromal cells derived from tonsil, fat tissue, and palmar skin can assume fibroblast functions supporting keratinocyte proliferation and differentiation. These findings indicate that distinct types of mesenchymal cells can be clinically used for skin engineering purposes.
使用自体生物工程真皮 - 表皮皮肤替代物(DESS)为治疗人类患者的大面积皮肤缺损提供了关键契机。这些皮肤移植片包含表皮和真皮部分,这对于实现稳固且永久性的功能性伤口闭合是必需的。在本研究中,我们调查了源自不同身体部位的间充质细胞对DESS内多种标志物表达模式的影响。
人角质形成细胞取自毛囊间表皮,间充质细胞则从包皮、手掌皮肤、脂肪组织和扁桃体中分离得到。扩增后,将表皮细胞接种到含有基质细胞的I型胶原水凝胶上。这些人DESS被移植到免疫缺陷大鼠的背部。3周后,切除移植片并使用免疫组织学技术进行分析。
含有扁桃体、脂肪组织或手掌来源间充质细胞的皮肤移植片的宏观外观,与含有包皮来源真皮成纤维细胞的替代物相似。所有皮肤移植片的表皮中Lingo - 1均有强烈的膜定位表达。此外,我们在皮肤移植片的上层表皮细胞层中观察到转谷氨酰胺酶5的强烈表达,证实了角质形成细胞的正常分化。原弹性蛋白定位于整个真皮部分,并与真皮 - 表皮交界处紧密接触,表明所有皮肤移植片均已高度成熟。
我们的数据表明,源自扁桃体、脂肪组织和手掌皮肤的基质细胞可发挥成纤维细胞功能,支持角质形成细胞的增殖和分化。这些发现表明,不同类型的间充质细胞可在临床上用于皮肤工程目的。