Applied Materials Today
Induction of dermal fibroblasts into dermal papilla cell-like cells in hydrogel microcapsules for enhanced hair follicle regeneration
Graphical abstract
Introduction
Hair loss is a common skin disorder associated with aging, diseases and medications, along with the full-thickness skin loss during severe wound injuries [1], [2], [3]. Current therapeutic treatments include medication and autologous hair transplantation [1,4]. Therapeutic drugs mainly prevent further hair loss rather than regenerate new hairs [5]. Hair transplant, a surgery that moves healthy hair follicles (HFs) from parts of the scalp to the bald area, is insufficient for patients with severe hair loss owing to the shortage of HFs [6]. With the advances of tissue engineering and regenerative medicine, cell therapy that aims at regenerating HFs emerges as a promising alternative to traditional HFs transplantation [7,8]. Dermal papilla (DP), a cluster of active specialized fibroblasts derived from dermal mesenchyme, is a spheroid structure located at the base of the HFs. DP cells (DPCs) are essential for HFs morphogenesis and regulation of hair cycling in both embryogenesis and postnatal regeneration [9,10]. However, the development of efficient expansion of DPCs with intrinsic features remains a great challenge, as DPCs proliferate slowly in vitro in two-dimensional (2D) cultures and quickly lose their hair-inducing activity during passaging [11,12]. 2D cultivation is unable to recapitulate the in vivo structures of densely packed DPCs within DPs [13]. Three-dimensional (3D) cultures could enable a microenvironment to facilitate HFs growth [14]. In light of such finding, we hypothesize that the 3D cultivation will allow us to obtain adequate DPCs for de novo folliculogenesis.
The prerequisite of 3D expansion of DPCs is the isolation and collection of a large number of DPCs. As DPCs proliferate slowly, the DPC shortage in DPC-based HFs regeneration still remains a bottleneck. The development of alternative sources of DPCs from other cell lines is thus desirable. DPCs are a specialized cell population sharing a same origin as inter-follicular dermal fibroblasts (DFs) and distinguish from the latter cells by their unique characteristics, such as aggregative behavior, distinct gene expression and the ability to induce the formation of new HFs. Fibroblasts are widely present in skin tissue, and can be extracted and expanded following well-established protocols. These advantages make fibroblasts a valuable cell source for cell therapy studies [15,16]. Although human dermal fibroblasts (hDFs) could differentiate into osteoblasts, neurons, cardiomyocytes, and melanocytes [17], [18], [19], [20], little effort has been made to investigate the derivation of functional DPC-like cells from hDFs.
Here we report a 3D cultivation system of alginate-poly-L-lysine-alginate (APA) microcapsules that restore the trichogenecity of DPCs pre-cultured under 2D conditions. Moreover, we for the first time induced hDFs into functional DPC-like cells through APA microencapsulation (Fig. S1). The hDFs in APA microcapsules exhibited the hDPCs features on the molecular level. We further elucidated that the hDFs into hDF-DPCs in APA microcapsules was activated by Wnt signaling pathways. These two strategies, separately or in combination, can supply a large number of safe and functional DPCs and circumvent the bottleneck in DPC-based regenerative strategies for de novo folliculogenesis.
Section snippets
Human Cell sources and isolation
All clinical procedures and protocols were based on informed consent and approved by the Ethics Committee of Central South University Xiangya Hospital. Human hair follicles were manually isolated from the occipital scalp obtained during hair transplant surgery. Primary hDFs were isolated from the circumcised foreskins of healthy human donors. All procedures were approved by Institutional Review Board of Central South University Xiangya Hospital (IRB No. 201611609) in accordance with the
Trichogenecity of cDPCs within APA microcapsules
Hydrogels have found great success in regenerating skin and skin appendage [27,28]. Our prior studies demonstrated that hydrogels can establish robust interaction with endogenous cells to promote complete skin regeneration [29]. In order to create a microenvironment to retain the hair inducing capacity, we developed APA hydrogel microcapsules, and encapsulated cDPCs within APA to establish spheroids (APA-DPCs) (Fig. 1A). ALP activity, which indicates the hair-inducing capacity of DPCs [30], was
Discussion
Many efforts had been made to obtain DPC-like cells from various stem cell lines, such as embryonic stem cells (ESCs), mesenchymal stem cells (MSCs) and inducible pluripotent stem cells (iPSCs) [25,[42], [43], [44]. Traditional processes are time-consuming and quite expensive. The induction of iPSCs into DPC-like cells, for example, requires the differentiation of iPSCs into neural crest (NC) cells, with a yield of approximately 20%-30%. The NC cells then undergo a second induction and
Conclusion
In summary, we first demonstrated that APA-cultivation successfully reinstated the hair-inducing capacity of 2D-cultured DPCs. Moreover, we for the first time differentiated hDFs into functional DPC-like cells through APA microencapsulation. The hDFs in APA microcapsules exhibited the hDPC features on the molecular level. We further elucidated that the hDFs into hDF-DPCs in APA microcapsules was activated by Wnt signaling pathways. It is worth to note that this approach is not limited to hDFs,
Data availability
Data supporting the findings of this study are within this manuscript or available from the corresponding authors upon reasonable request.
CRediT authorship contribution statement
Bei Xie: Conceptualization, Investigation, Formal analysis, Writing - original draft. Mengting Chen: Investigation, Validation, Writing - original draft. Pinghui Ding: Investigation, Validation, Formal analysis. Lei Lei: Funding acquisition, Writing - review & editing. Xing Zhang: Writing - review & editing. Dan Zhu: Methodology. Yujian Zou: Methodology. Zhili Deng: . Guoming Sun: Writing - review & editing. Ji Li: Methodology, Funding acquisition, Project administration, Supervision. Hemin Nie:
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
This work was supported by the National Natural Science Foundation of China (Nos. 31670997, 81773351, 81900982), Hunan Provincial Science and Technology Department (Nos. 2018SK2106, 2017JJ3421), and Changsha Municipal Bureau of Science and Technology (No. kq1901024). The authors declare no competing interest associated with this publication.
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These authors contributed equally to this work.