Institute of Biomedicine, Department of Pharmacology, Biomedicum, University of Helsinki, Finland.
J Biomed Mater Res A. 2010 Nov;95(2):658-64. doi: 10.1002/jbm.a.32881.
Efficient re-epithelialization of skin lesions is dependent on paracrine support from connective tissue fibroblasts. In deep skin defects, the supporting growth factor incentive is lacking. Current methods of keratinocyte transplantation with compromised attachment, spread, and cell proliferation warrant improvement and refinement. We describe here how human keratinocytes can be stimulated by matrix-embedded factors from a novel process of fibroblast activation: nemosis. Interestingly, the unique set of mediators released in this process also plays a key role in normal wound healing. To develop a system for targeted delivery of nemosis-derived paracrine effectors, herein designated as Finectra, we combined them with fibrin to establish a controlled-release gel. Keratinocytes seeded to cover this active matrix showed better adherence, outgrowth, and viability than did cells on control matrix. The matrix incorporating Finectra supported viability of both primary keratinocytes and green fluorescent protein (GFP)-labeled HaCaT cells, as evaluated by MTT assay and persistence of GFP-fluorescence. The fibrin-Finectra matrix promoted migration of keratinocytes to cover a larger area on the matrix, suggesting better wound coverage on transplantation. An inhibitor of EGFR/c-Met receptor tyrosine kinases abolished keratinocyte responses to fibrin-Finectra matrix. This matrix can thus deliver biologically relevant synergistic stimuli to keratinocytes and hasten re-epithelialization.
皮肤损伤的有效再上皮化依赖于结缔组织成纤维细胞的旁分泌支持。在深部皮肤缺损中，缺乏支持生长因子的刺激。目前，角质形成细胞移植的方法存在附着、扩散和细胞增殖能力受损的问题，需要改进和完善。我们在这里描述了如何通过成纤维细胞激活的新过程：Nemosis，刺激人角质形成细胞。有趣的是，在这个过程中释放的独特介质集也在正常伤口愈合中发挥关键作用。为了开发一种用于Nemosis 衍生旁分泌效应物的靶向递药系统，我们将其与纤维蛋白结合，建立了一种控释凝胶。与对照基质相比，接种于该活性基质上的角质形成细胞表现出更好的黏附、生长和活力。通过 MTT 测定和 GFP 荧光的持久性评估，包含 Finectra 的基质支持原代角质形成细胞和 GFP 标记的 HaCaT 细胞的存活。纤维蛋白-Finectra 基质促进角质形成细胞迁移以覆盖基质上更大的区域，提示在移植时更好地覆盖伤口。表皮生长因子受体 (EGFR)/c-Met 受体酪氨酸激酶抑制剂可消除角质形成细胞对纤维蛋白-Finectra 基质的反应。因此，该基质可以向角质形成细胞传递具有生物学相关性的协同刺激物，加速再上皮化。
