TMT-Based Quantitative Proteomic Analysis Reveals the Effect of Bone Marrow Derived Mesenchymal Stem Cell on Hair Follicle Regeneration

Hair loss (HL) is a common chronic problem of poorly defined etiology. Herein, we explored the functionality of bone marrow-derived mesenchymal stem cell (BMSC) and conditioned medium (MSC-CM) as regulators of hair follicle proliferation and regeneration, and the mechanistic basis for such activity. BMSC were cultured and identified in vitro through the induction of multilineage differentiation and the use of a CCK-8 kit. The dorsal skin of mice was then injected with BMSC and MSC-CM, and the impact of these injections on hair cycle transition and hair follicle stem cell (HFSC) proliferation was then evaluated via hematoxylin and eosin (H&E) staining and immunofluorescent (IF) staining. We then conducted a tandem mass tags (TMT)-based quantitative proteomic analysis of control mice and mice treated with BMSC or MSC-CM to identify differentially expressed proteins (DEPs) associated with these treatments. Parallel reaction monitoring (PRM) was utilized as a means of verifying our proteomic analysis results. Herein, we found that BMSC and MSC-CM injection resulted in the transition of telogen hair follicles to anagen hair follicles, and we observed the enhanced proliferation of HFSCs positive for Krt15 and Sox9. Our TMT analyses identified 1,060 and 770 DEPs (fold change＞1.2 or＜0.83 and p ＜ 0.05) when comparing the BMSC vs. control and MSC-CM vs. control groups, respectively. Subsequent PRM validation of 14 selected DEPs confirmed these findings, and led to the identification of Stmn1, Ncapd2, Krt25, and Ctps1 as hub DEPs in a protein-protein interaction network. Together, these data suggest that BMSC and MSC-CM treatment can promote the proliferation of HFSCs, thereby facilitating hair follicle regeneration. Our proteomics analyses further indicate that Krt25, Cpm, Stmn1, and Mb may play central roles in hair follicle transition in this context and may represent viable clinical targets for the treatment of HL.

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A Mouse Model for Studying Stem Cell Effects on Regeneration of Hair Follicle Outer Root Sheaths

Open Life Sciences ◽

10.1515/biol-2020-0005 ◽

2020 ◽

Vol 15 (1) ◽

pp. 41-50

Author(s):

Jingxu Guo ◽

Shuwei Li ◽

Hongyang Wang ◽

Tinghui Wu ◽

Zhenhui Wu ◽

...

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Mouse Model ◽

Hair Follicle ◽

Smooth Muscle Actin ◽

Hair Follicles ◽

Alpha Smooth Muscle Actin ◽

Papillary Structure ◽

Glass Membrane

AbstractObjectiveStem cells hold promise for treating hair loss. Here an in vitro mouse model was developed using outer root sheaths (ORSs) isolated from hair follicles for studying stem cell-mediated dermal papillary regeneration.MethodsUnder sterile conditions, structurally intact ORSs were isolated from hair follicles of 3-day-old Kunming mice and incubated in growth medium. Samples were collected daily for 5 days. Stem cell distribution, proliferation, differentiation, and migration were monitored during regeneration.ResultsCell proliferation began at the glass membrane periphery then spread gradually toward the membrane center, with the presence of CD34 and CD200 positive stem cells involved in repair initiation. Next, CD34 positive stem cells migrated down the glass membrane, where some participated in ORS formation, while other CD34 cells and CD200 positive cells migrated to hair follicle centers. Within the hair follicle matrix, stem cells divided, grew, differentiated and caused outward expansion of the glass membrane to form a dermal papillary structure containing alpha-smooth muscle actin. Neutrophils attracted to the wound site phagocytosed bacterial and cell debris to protect regenerating tissue from infection.ConclusionIsolated hair follicle ORSs can regenerate new dermal papillary structures in vitro. Stem cells and neutrophils play important roles in the regeneration process.

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α-hemihydrate calcium sulfate/octacalcium phosphate combined with sodium hyaluronate promotes bone marrow-derived mesenchymal stem cell osteogenesis in vitro and in vivo

Drug Design Development and Therapy ◽

10.2147/dddt.s173289 ◽

2018 ◽

Vol Volume 12 ◽

pp. 3269-3287 ◽

Cited By ~ 1

Author(s):

Changshun Chen ◽

Chen Zhu ◽

Xiang Hu ◽

Qiuli Yu ◽

Qianjin Zheng ◽

...

Keyword(s):

Bone Marrow ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Calcium Sulfate ◽

Sodium Hyaluronate ◽

Octacalcium Phosphate ◽

Osteogenesis In Vitro

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microRNA‐199a counteracts glucocorticoid inhibition of bone marrow mesenchymal stem cell osteogenic differentiation through regulation of Klotho expression in vitro

Cell Biology International ◽

10.1002/cbin.11460 ◽

2020 ◽

Vol 44 (12) ◽

pp. 2532-2540 ◽

Cited By ~ 1

Author(s):

Jinshan Tang ◽

Huaixi Yu ◽

Yunqing Wang ◽

Gang Duan ◽

Bin Wang ◽

...

Keyword(s):

Bone Marrow ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Osteogenic Differentiation

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Canine Bone Marrow Mesenchymal Stem Cell Conditioned Media Affect Bacterial Growth, Biofilm-Associated Staphylococcus aureus and AHL-Dependent Quorum Sensing

Microorganisms ◽

10.3390/microorganisms8101478 ◽

2020 ◽

Vol 8 (10) ◽

pp. 1478 ◽

Cited By ~ 1

Author(s):

Dobroslava Bujňáková ◽

Anna Čuvalová ◽

Milan Čížek ◽

Filip Humenik ◽

Michel Salzet ◽

...

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Bone Marrow ◽

Stem Cell ◽

Quorum Sensing ◽

Mesenchymal Stem Cell ◽

Bacterial Growth ◽

Conditioned Media ◽

Amyloid Β Peptide

The present study investigated the in vitro antibacterial, antibiofilm and anti-Quorum Sensing (anti-QS) activities of canine bone marrow mesenchymal stem cell-conditioned media (cBM MSC CM) containing all secreted factors <30 K, using a disc diffusion test (DDT), spectrophotometric Crystal Violet Assay (SCVA) and Bioluminescence Assay (BA) with QS-reporter Escherichia coli JM109 pSB1142. The results show a sample-specific bacterial growth inhibition (zones varied between 7–30 mm), statistically significant modulation of biofilm-associated Staphylococcus aureus and Escherichia coli bioluminescence (0.391 ± 0.062 in the positive control to the lowest 0.150 ± 0.096 in the experimental group, cf. 11,714 ± 1362 to 7753 ± 700, given as average values of absorbance A550 ± SD versus average values of relative light units to growth RLU/A550 ± SD). The proteomic analysis performed in our previous experiment revealed the presence of several substances with documented antibacterial, antibiofilm and immunomodulatory properties (namely, apolipoprotein B and D; amyloid-β peptide; cathepsin B; protein S100-A4, galectin 3, CLEC3A, granulin, transferrin). This study highlights that cBM MSC CM may represent an important new approach to managing biofilm-associated and QS signal molecule-dependent bacterial infections. To the best of our knowledge, there is no previous documentation of canine BM MSC CM associated with in vitro antibiofilm and anti-QS activity.

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The Potential of a Hair Follicle Mesenchymal Stem Cell-Conditioned Medium for Wound Healing and Hair Follicle Regeneration

Applied Sciences ◽

10.3390/app10082646 ◽

2020 ◽

Vol 10 (8) ◽

pp. 2646

Author(s):

Keng-Liang Ou ◽

Yun-Wen Kuo ◽

Chia-Yu Wu ◽

Bai-Hung Huang ◽

Fang-Tzu Pai ◽

...

Keyword(s):

Wound Healing ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Hair Follicle ◽

Conditioned Medium ◽

Hair Growth ◽

Hair Regeneration ◽

Hacat Keratinocyte ◽

Cell Conditioned Medium

The study elucidated the wound healing and hair regeneration properties of a conditioned medium prepared from the culture of human hair follicle mesenchymal stem cells (HFMSCs). The wound-healing effects of mesenchymal stem cell-conditioned medium (MSC-CM) were tested in vitro using scratch assays co-cultured with HaCaT keratinocyte and monitored through optical microscopy. The cell proliferation of HFMSCs and the HaCaT keratinocyte were observed in the presence of different kinds of drugs including UK5099, sodium L-lactate, lactate dehydrogenase-A, MSC-CM, caffeine, and caffeic acid. The hair regeneration properties were investigated in vivo by administrating the MSC-CM solutions to adult B6 mouse models. For quantification, hematoxylin and eosin staining were performed following euthanasia. In vitro results revealed that MSC-CM promotes dermal cell migrations and enhances proliferation of HFMSCs and HaCaT keratinocytes, demonstrating wound-healing properties. Moreover, when the MSC-CM solutions were applied to the shaved mouse skin, a dark area that expanded overtime was seen. Although no hair growth was found, histological analysis proved that a fat layer thickness increment was found under the mouse’s skin, ultimately projecting the formation of new hair growth. MSC-CM promotes the migration and proliferation of dermal keratinocytes that are beneficial for wound healing and hair growth. It is believed that MSC-CM can potentially serve as the basis of alternative therapeutic applications for wound closure and skin regeneration as well as hair growth stimulation and hair loss prevention in alopecia.

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Tcf12, A Member of Basic Helix-Loop-Helix Transcription Factors, Mediates Bone Marrow Mesenchymal Stem Cell Osteogenic Differentiation In Vitro and In Vivo

Stem Cells ◽

10.1002/stem.2491 ◽

2016 ◽

Vol 35 (2) ◽

pp. 386-397 ◽

Cited By ~ 17

Author(s):

Siqi Yi ◽

Miao Yu ◽

Shuang Yang ◽

Richard J. Miron ◽

Yufeng Zhang

Keyword(s):

Bone Marrow ◽

Stem Cell ◽

Transcription Factors ◽

Mesenchymal Stem Cell ◽

Osteogenic Differentiation ◽

Basic Helix Loop Helix ◽

Helix Loop Helix

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Mechanotransductive Differentiation of Hair Follicle Stem Cells Derived from Aged Eyelid Skin into Corneal Endothelial-Like Cells

Stem Cell Reviews and Reports ◽

10.1007/s12015-021-10249-0 ◽

2021 ◽

Author(s):

Christian Olszewski ◽

Jessika Maassen ◽

Rebecca Guenther ◽

Claudia Skazik-Voogt ◽

Angela Gutermuth

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Hair Follicle ◽

Hair Follicles ◽

Adherent Cell ◽

Promising Alternative ◽

Eyelid Skin ◽

Hair Follicle Stem Cells ◽

Follicle Stem Cells

AbstractCorneal endothelial insufficiency is one of the leading causes of blindness. The main contemporary treatment for corneal blindness is endothelial keratoplasty, which, however, is unsatisfactory as a medical therapy due to the lack of donor corneas and graft rejection. Therefore, autologous stem cell-based corneal endothelial tissue substitutes may be a promising alternative to conventional grafts in the future. To address the age of most patients suffering from corneal endothelial deficiencies, we investigated the presence and potential of hair-derived stem cells from older tissue donors. Our studies revealed the presence of pluripotency- and neural crest-associated markers in tissue sections from blepharoplasty patients aged 50 to 80 years. In vitro outgrowths from eyelid hair follicles on collagen-coated tissue culture plates revealed a weak decrease in stem-cell potency. In contrast, cells within the spheres that spontaneously formed from the adherent cell layer retained full stem-cell potency and could be differentiated into cells of the ecto- meso and endodermal lineages. Although these highly potent hair follicle derived stem cells (HFSC) were only very slightly expandable, they were able to recognize the biomimicry of the Descemet’s-like topography and differentiate into corneal endothelial-like cells. In conclusion, HFSCs derived from epidermal skin of eyelid biopsies are a promising cell source to provide autologous corneal endothelial replacement for any age group of patients. Graphical Abstract

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Emilin-2 is a component of bone marrow extracellular matrix regulating mesenchymal stem cell differentiation and hematopoietic progenitors

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02674-2 ◽

2022 ◽

Vol 13 (1) ◽

Author(s):

Francesco Da Ros ◽

Luca Persano ◽

Dario Bizzotto ◽

Mariagrazia Michieli ◽

Paola Braghetta ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Extracellular Matrix ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Adipogenic Differentiation ◽

Stem Cell Differentiation ◽

Dependent Manner ◽

Hematopoietic Stem

Abstract Background Dissection of mechanisms involved in the regulation of bone marrow microenvironment through cell–cell and cell–matrix contacts is essential for the detailed understanding of processes underlying bone marrow activities both under physiological conditions and in hematologic malignancies. Here we describe Emilin-2 as an abundant extracellular matrix component of bone marrow stroma. Methods Immunodetection of Emilin-2 was performed in bone marrow sections of mice from 30 days to 6 months of age. Emilin-2 expression was monitored in vitro in primary and mesenchymal stem cell lines under undifferentiated and adipogenic conditions. Hematopoietic stem cells and progenitors in bone marrow of 3- to 10-month-old wild-type and Emilin-2 null mice were analyzed by flow cytometry. Results Emilin-2 is deposited in bone marrow extracellular matrix in an age-dependent manner, forming a meshwork that extends from compact bone boundaries to the central trabecular regions. Emilin-2 is expressed and secreted by both primary and immortalized bone marrow mesenchymal stem cells, exerting an inhibitory action in adipogenic differentiation. In vivo Emilin-2 deficiency impairs the frequency of hematopoietic stem/progenitor cells in bone marrow during aging. Conclusion Our data provide new insights in the contribution of bone marrow extracellular matrix microenvironment in the regulation of stem cell niches and hematopoietic progenitor differentiation.

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Graphene Oxide Enables the Reosteogenesis of Previously Contaminated Titanium In Vitro

Journal of Dental Research ◽

10.1177/0022034520913873 ◽

2020 ◽

Vol 99 (8) ◽

pp. 922-929

Author(s):

W. Qin ◽

C. Wang ◽

C. Jiang ◽

J. Sun ◽

C. Yu ◽

...

Keyword(s):

Bone Marrow ◽

Graphene Oxide ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Osteogenic Potential ◽

Implant Surface ◽

Significant Difference ◽

Titanium Surfaces ◽

Group B

The main goal of peri-implantitis treatment is to control infection and arrest bone loss, which requires the removal of polymicrobial biofilms on the implant surface and the reduction of tissue invasion. Additionally, prognosis can be improved if reosseointegration occurs on previously contaminated implants. To evaluate whether graphene oxide (GO) can remove polymicrobial biofilms, biofilms were established on titanium surfaces in vitro and treated with different methods: group B, removed only with brushing; group G, treated with different GO concentrations (64, 128, 256, and 512 μg/mL); group GB, combined treatments of groups B and G; and group C, untreated. Subsequently, to evaluate reosteogenesis on previously contaminated titanium, 4 groups were used: groups C, B, GB-256, and GB-512 (treated with 256 and 512 μg/mL of GO, respectively). Intact clean titanium (IC) was used as a control. Additionally, cell behavior on IC treated with GB-256 (IGB-256) and GB-512 (IGB-512) was compared with that of the GB-256 and GB-512 groups, respectively. The results showed that at high concentrations (≥256 μg/mL), GO eliminated residual bacteria and inhibited biofilm reformation after brushing, whereas neither GO nor brushing alone could achieve this. Bone marrow–derived mesenchymal stem cell viability in groups GB-256 and IC was higher than that in groups GB-512, C, and B ( P < 0.05). No significant difference was found between group GB-256 and group IC ( P > 0.05). Osteogenic differentiation of bone marrow–derived mesenchymal stem cells in group GB-256 was higher than that in groups IC, GB-512, C, and B. No difference was found between groups IGB-256 and IGB-512 and groups GB-256 and GB-512, respectively ( P > 0.05). In conclusion, 256 μg/mL of GO combined with brushing significantly removed polymicrobial biofilms that remained on the previously contaminated titanium surfaces. The bone marrow–derived mesenchymal stem cell osteogenic potential was regained or even enhanced on the titanium surfaces treated this way in vitro, which might provide a new idea for treating peri-implantitis.

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