Tissue Eng Regen Med.  2020 Aug;17(4):525-536. 10.1007/s13770-020-00266-4.

Human Hair Outer Root Sheath Cells and Platelet-Lysis Exosomes Promote Hair Inductivity of Dermal Papilla Cell

• Nilforoushzadeh MA
• Aghdami N
• Taghiabadi E

• KMID: 2505180
• DOI: http://doi.org/10.1007/s13770-020-00266-4

Abstract

BACKGROUND
Hair loss is a prevalent medical problem in both men and women. Maintaining the potential hair inductivityof dermal papilla cells (DPCs) during cell culture is the main factor in hair follicle morphogenesis and regeneration. Thepresent study was conducted to compare the effects of different concentrations of human hair outer root sheath cell(HHORSC)and platelet lysis (PL) exosomes to maintain hair inductivity of the human dermal papilla cells (hDPCs).
METHODS
In this study, hDPCs and HHORSCswere isolated from healthy hair samples. Specific markers of hDPCs (versican,a-SMA) and HHORSCs (K15) were evaluated using flow cytometric and immunocytochemical techniques. The exosomes wereisolated fromHHORSCsand PL with ultracentrifugation technique.Western blot was used to detect specific markers of HHORSCsand PL exosomes. Particle size and distribution of the exosomes were analyzed by NanoSight dynamic light NanoSight DynamicLight Scattering. Different methods such as proliferation test (MTS assay), migration test (Transwell assay) were used to evaluatethe effects of different concentrations of exosomes (2,550,100 lg/ml) derived from HHORSC and PL on hDPCs. Expression ofspecific genes in the hair follicle inductivity, including ALP, versican and a-SMA were also evaluated using real time-PCR.
RESULTS
The flow cytometry of the specific cytoplasmic markers of the hDPCs and HHORSCs showed expression ofversican (77%), a-SMA (55.2%) and K15 (73.2%). The result of particle size and distribution of the exosomes wereanalyzed by NanoSight dynamic light NanoSight Dynamic Light Scattering, which revealed the majority of HHORSC andPL exosomes were 30–150 nm. For 100 lg/ml of HHORSC exosomes, the expressions of ALP, versican and a-SMAproteins respectively increased by a factor of 2.1, 1.7and 1.3 compared to those in the control group.
CONCLUSION
In summary, we applied HHORSC exosomes as a new method to support hair inductivity of dermalpapilla cells and improve the outcome for the treatment of hair loss.

Keyword

Dermal papilla cells; Exosome; Hair inductivity; Hair loss; Hair outer root sheath cell