Culturing adult human bone marrow stem cells on gelatin scaffold with pNIPAAm as transplanted grafts for skin regeneration

Abstract

Skin tissue engineering is a possible solution for the treatment of extensive skin defect. The ultimate goal of skin tissue engineering is to restore the complete functions of native skin, but until now the structures and functions of skins are only partially restored. By negative immunoselection (CD45 and glycophorin A), we isolated and cultivated adult human bone marrow stem cells (hBMSCs) that are of multilineage differentiation potential. In this study, we first demonstrated that by using gelatin/thermo-sensitive poly N-isopropylacrylamide (pNIPAAm) and the immunocompromised mice model, the hBMSCs possess the differentiation potential of epidermis and the capability of healing skin wounds. The in vitro observations and the results of the scanning electron microscope showed that the hBMSCs can attach and proliferate in the gelatin/thermo-sensitive pNIPAAm. To further monitor the in vivo growth effect of the hBMSCs in the skin-defected nude mice, the green fluorescence protein (GFP) gene was transduced into the hBMSCs by the murine stem cell viral vector. The results showed that the rates of cell growth and wound recovery in the hBMSC-treated group were significantly higher than those in the control group, which was only treated with the gelatin/pNIPAAm (p < 0.01). More importantly, the re-epithelialization markers of human pan-cytokeratin and E-cadherin were significantly increased on day 7, day 14, and day 21 after the hBMSC-scaffold with the pNIPAAM in the mice with skin defects (p < 0.05). Moreover, the stem cell markers of human CD13 and CD105 were gradually decreased during the period of wound healing. In sum, this novel method provides a transferring system for cell therapies and maintains its temperature-sensitive property of easy-peeling by lower-temperature treatment. In addition, the in vitro and in vivo GFP imaging systems provide a new imaging modality for understanding the differentiation process and the effective expression of stem cells in wound healing.

An Overview of Methods for theIn VivoEvaluation of Tissue-Engineered Skin Constructs

“…They investigated the transformation in expression from the human bone marrow stem cells markers CD13 and CD105 to the epithelial cells specific E-cadherin and pancytokeratin. 160 Lamme et al prelabeled fibroblasts with PKH-26 before implantation and later used flow cytometry on a wound-derived single cell suspension to identify the presence of this label in different cell populations. 189 Gene expression levels and mRNA location Construct-assisted wound healing in animal models can also be analyzed by monitoring changes on the gene expression level.…”

Enhanced wound healing effect of canine adipose-derived mesenchymal stem cells with low-level laser therapy in athymic mice

“…However, several studies claim that the effects of stem cell therapy are not dramatic in the absence of scaffolds or stimulators [3,4]. Recently, various scaffolds or growth factors were studied to increase the skin regeneration effect of stem cells [5][6][7].…”

Differentiation of Human Adipose-Derived Stem Cells into Neuron-Like Cells Which Are Compatible with Photocurable Three-Dimensional Scaffolds

“…Since gelatin is derived from denatured collagen and has both negatively and positively charged patches, 25 it carries specific peptide sequences to facilitate cell attachment and proliferation. 26 It is assumed that the hybrid scaffolds contain interconnected pores for easy loading of hADSCs as well as handy incorporation of growth factors and nutrients, thereby allowing hADSC attachment, proliferation, and differentiation into hADSCNLCs both in vitro and in vivo.…”