臺灣博碩士論文加值系統

生物體內的細胞組織持續受到環境中複雜的機械刺激，這些機械刺激影響了細胞增生、分化、遷移，並決定了組織恆定性及修復。於此研究中，我們使用了特殊設計的皮膚張力裝置，並發現給予特定強度及時間的皮膚拉伸會刺激毛囊幹細胞的增生及毛髮再生。而藉由分子及基因實驗的分析，我們發現WNT及BMP-2訊息的平衡及兩階段的巨噬細胞變化扮演了及重要的角色。皮膚拉伸刺激的趨化因子吸引了巨噬細胞，並促進其進行M2極化。而M2巨噬細胞會藉由分泌肝細胞生長因子(HGF)及類胰島素生長因子(IGF-1)以活化毛囊幹細胞並促進毛髮再生。此研究揭示了由機械刺激、化學訊號、到細胞行為及組織反應的階級特性，並替再生醫學及疾病控制開啟了新的方向，即單用機械刺激去調控細胞反應。

Tissues and cells in organism are continuously exposed to complex mechanical cues from the environment. Mechanical stimulations affect cell proliferation, differentiation, and migration as well as determining tissue homeostasis and repair. By using a specially designed skin-stretching device, we discover that hair stem cells proliferate in response to stretch and hair regeneration occurs only when applying proper strain for an appropriate duration. A counterbalance between WNT and BMP-2 and the subsequent two-step mechanism are identified through molecular and genetic analyses. Macrophages are first recruited by chemokines produced by stretch and polarized to M2 phenotype. Growth factors such as HGF and IGF-1, released by M2 macrophages, then activate stem cells and facilitate hair regeneration. A hierarchical control system is revealed, from mechanical and chemical signals to cell behaviors and tissue responses, elucidating avenues of regenerative medicine and disease control by demonstrating the potential to manipulate cellular processes through simple mechanical stimulation.

Acknowledgement 1
English abstract 7
Chinese abstract 8
List of Abbreviations 9
1. Introduction 11
2. Material and Methods 15
3. Results 22
4. Discussion 31
5. Conclusion 35
6. Perspectives 36
7. References 37
8. Figures 41
Figure 1 41
Figure 2 42
Figure 3 44
Figure 4 45
Figure 5 47
Figure 6 48
Figure 7 49
Figure 8 50
Figure 9 51
Figure 10 52
Figure 11 53
Figure 12 54
Figure 13 55
Figure 14 56
Figure 15 57
Figure 16 58
Figure 17 59
Figure 18 60
Figure 19 61
Figure 20 62
Figure 21 63
Figure 22 64
9. Tables 65
Table 1 65
Table 2 67
Table 3 68
10.Publication 69

1. Discher DE, Mooney DJ, Zandstra PW. Growth factors, matrices, and forces combine and control stem cells. Science. 2009;324(5935):1673-7.
2. Guo CL, Harris NC, Wijeratne SS, Frey EW, Kiang CH. Multiscale mechanobiology: mechanics at the molecular, cellular, and tissue levels. Cell Biosci. 2013;3(1):25.
3. Aragona M, Panciera T, Manfrin A, Giulitti S, Michielin F, Elvassore N, et al. A mechanical checkpoint controls multicellular growth through YAP/TAZ regulation by actin-processing factors. Cell. 2013;154(5):1047-59.
4. Huang S, Ingber DE. The structural and mechanical complexity of cell-growth control. Nat Cell Biol. 1999;1(5):E131-8.
5. Sasai Y. Cytosystems dynamics in self-organization of tissue architecture. Nature. 2013;493(7432):318-26.
6. Mammoto T, Ingber DE. Mechanical control of tissue and organ development. Development. 2010;137(9):1407-20.
7. Mammoto T, Mammoto A, Torisawa YS, Tat T, Gibbs A, Derda R, et al. Mechanochemical control of mesenchymal condensation and embryonic tooth organ formation. Dev Cell. 2011;21(4):758-69.
8. Moore KA, Lemischka IR. Stem cells and their niches. Science. 2006;311(5769):1880-5.
9. Chen CC, Chuong CM. Multi-layered environmental regulation on the homeostasis of stem cells: the saga of hair growth and alopecia. J Dermatol Sci. 2012;66(1):3-11.
10. Chen CC, Wang L, Plikus MV, Jiang TX, Murray PJ, Ramos R, et al. Organ-level quorum sensing directs regeneration in hair stem cell populations. Cell. 2015;161(2):277-90.
11. Torkamani N, Rufaut NW, Jones L, Sinclair R. Destruction of the arrector pili muscle and fat infiltration in androgenic alopecia. Br J Dermatol. 2014;170(6):1291-8.
12. Torkamani N, Rufaut NW, Jones L, Sinclair RD. Beyond goosebumps: does the arrector pili muscle have a role in hair loss? Int J Trichology. 2014;6(3):88-94.
13. Yazdabadi A, Whiting D, Rufaut N, Sinclair R. Miniaturized Hairs Maintain Contact with the Arrector Pili Muscle in Alopecia Areata but not in Androgenetic Alopecia: A Model for Reversible Miniaturization and Potential for Hair Regrowth. Int J Trichology. 2012;4(3):154-7.
14. Agha R, Ogawa R, Pietramaggiori G, Orgill DP. A review of the role of mechanical forces in cutaneous wound healing. J Surg Res. 2011;171(2):700-8.
15. Evans ND, Oreffo RO, Healy E, Thurner PJ, Man YH. Epithelial mechanobiology, skin wound healing, and the stem cell niche. J Mech Behav Biomed Mater. 2013;28:397-409.
16. Bouffard NA, Cutroneo KR, Badger GJ, White SL, Buttolph TR, Ehrlich HP, et al. Tissue stretch decreases soluble TGF-beta1 and type-1 procollagen in mouse subcutaneous connective tissue: evidence from ex vivo and in vivo models. J Cell Physiol. 2008;214(2):389-95.
17. Osaka N, Takahashi T, Murakami S, Matsuzawa A, Noguchi T, Fujiwara T, et al. ASK1-dependent recruitment and activation of macrophages induce hair growth in skin wounds. J Cell Biol. 2007;176(7):903-9.
18. Samuelov L, Sprecher E, Tsuruta D, Biro T, Kloepper JE, Paus R. P-cadherin regulates human hair growth and cycling via canonical Wnt signaling and transforming growth factor-beta2. J Invest Dermatol. 2012;132(10):2332-41.
19. Di Loreto C, La Marra F, Mazzon G, Belgrano E, Trombetta C, Cauci S. Immunohistochemical evaluation of androgen receptor and nerve structure density in human prepuce from patients with persistent sexual side effects after finasteride use for androgenetic alopecia. PLoS One. 2014;9(6):e100237.
20. Rossi A, Cantisani C, Melis L, Iorio A, Scali E, Calvieri S. Minoxidil use in dermatology, side effects and recent patents. Recent Pat Inflamm Allergy Drug Discov. 2012;6(2):130-6.
21. Karolchik D, Baertsch R, Diekhans M, Furey TS, Hinrichs A, Lu YT, et al. The UCSC Genome Browser Database. Nucleic Acids Res. 2003;31(1):51-4.
22. Kim D, Pertea G, Trapnell C, Pimentel H, Kelley R, Salzberg SL. TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions. Genome Biol. 2013;14(4):R36.
23. Trapnell C, Hendrickson DG, Sauvageau M, Goff L, Rinn JL, Pachter L. Differential analysis of gene regulation at transcript resolution with RNA-seq. Nat Biotechnol. 2013;31(1):46-53.
24. Huang da W, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 2009;4(1):44-57.
25. Muller-Rover S, Handjiski B, van der Veen C, Eichmuller S, Foitzik K, McKay IA, et al. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages. J Invest Dermatol. 2001;117(1):3-15.
26. Plikus MV, Mayer JA, de la Cruz D, Baker RE, Maini PK, Maxson R, et al. Cyclic dermal BMP signalling regulates stem cell activation during hair regeneration. Nature. 2008;451(7176):340-4.
27. Plikus MV, Widelitz RB, Maxson R, Chuong CM. Analyses of regenerative wave patterns in adult hair follicle populations reveal macro-environmental regulation of stem cell activity. Int J Dev Biol. 2009;53(5-6):857-68.
28. Chen CC, Plikus MV, Tang PC, Widelitz RB, Chuong CM. The Modulatable Stem Cell Niche: Tissue Interactions during Hair and Feather Follicle Regeneration. J Mol Biol. 2016;428(7):1423-40.
29. Greco V, Chen T, Rendl M, Schober M, Pasolli HA, Stokes N, et al. A two-step mechanism for stem cell activation during hair regeneration. Cell Stem Cell. 2009;4(2):155-69.
30. Enshell-Seijffers D, Lindon C, Kashiwagi M, Morgan BA. beta-catenin activity in the dermal papilla regulates morphogenesis and regeneration of hair. Dev Cell. 2010;18(4):633-42.
31. Lowry WE, Blanpain C, Nowak JA, Guasch G, Lewis L, Fuchs E. Defining the impact of beta-catenin/Tcf transactivation on epithelial stem cells. Genes Dev. 2005;19(13):1596-611.
32. Demicheva E, Hecker M, Korff T. Stretch-induced activation of the transcription factor activator protein-1 controls monocyte chemoattractant protein-1 expression during arteriogenesis. Circ Res. 2008;103(5):477-84.
33. Gruden G, Setti G, Hayward A, Sugden D, Duggan S, Burt D, et al. Mechanical stretch induces monocyte chemoattractant activity via an NF-kappaB-dependent monocyte chemoattractant protein-1-mediated pathway in human mesangial cells: inhibition by rosiglitazone. J Am Soc Nephrol. 2005;16(3):688-96.
34. Boniakowski AE, Kimball AS, Jacobs BN, Kunkel SL, Gallagher KA. Macrophage-Mediated Inflammation in Normal and Diabetic Wound Healing. J Immunol. 2017;199(1):17-24.
35. Mills CD. M1 and M2 Macrophages: Oracles of Health and Disease. Crit Rev Immunol. 2012;32(6):463-88.
36. Mills CD, Kincaid K, Alt JM, Heilman MJ, Hill AM. M-1/M-2 macrophages and the Th1/Th2 paradigm. J Immunol. 2000;164(12):6166-73.
37. Mills CD. Anatomy of a discovery: m1 and m2 macrophages. Front Immunol. 2015;6:212.
38. Mills CD, Ley K. M1 and M2 macrophages: the chicken and the egg of immunity. J Innate Immun. 2014;6(6):716-26.
39. Gordon S. Alternative activation of macrophages. Nat Rev Immunol. 2003;3(1):23-35.
40. Kano F, Matsubara K, Ueda M, Hibi H, Yamamoto A. Secreted Ectodomain of Sialic Acid-Binding Ig-Like Lectin-9 and Monocyte Chemoattractant Protein-1 Synergistically Regenerate Transected Rat Peripheral Nerves by Altering Macrophage Polarity. Stem Cells. 2017;35(3):641-53.
41. Das A, Sinha M, Datta S, Abas M, Chaffee S, Sen CK, et al. Monocyte and macrophage plasticity in tissue repair and regeneration. Am J Pathol. 2015;185(10):2596-606.
42. Gordon S, Martinez FO. Alternative activation of macrophages: mechanism and functions. Immunity. 2010;32(5):593-604.
43. Mori T, Agata N, Itoh Y, Inoue-Miyazu M, Mizumura K, Sokabe M, et al. Post-injury stretch promotes recovery in a rat model of muscle damage induced by lengthening contractions. J Physiol Sci. 2017.
44. Song G, Ju Y, Shen X, Luo Q, Shi Y, Qin J. Mechanical stretch promotes proliferation of rat bone marrow mesenchymal stem cells. Colloids Surf B Biointerfaces. 2007;58(2):271-7.
45. Sun L, Qu L, Zhu R, Li H, Xue Y, Liu X, et al. Effects of Mechanical Stretch on Cell Proliferation and Matrix Formation of Mesenchymal Stem Cell and Anterior Cruciate Ligament Fibroblast. Stem Cells Int. 2016;2016:9842075.
46. Yu HS, Kim JJ, Kim HW, Lewis MP, Wall I. Impact of mechanical stretch on the cell behaviors of bone and surrounding tissues. J Tissue Eng. 2016;7:2041731415618342.
47. Eckes B, Zweers MC, Zhang ZG, Hallinger R, Mauch C, Aumailley M, et al. Mechanical tension and integrin alpha 2 beta 1 regulate fibroblast functions. J Investig Dermatol Symp Proc. 2006;11(1):66-72.
48. Kippenberger S, Bernd A, Loitsch S, Guschel M, Muller J, Bereiter-Hahn J, et al. Signaling of mechanical stretch in human keratinocytes via MAP kinases. J Invest Dermatol. 2000;114(3):408-12.
49. Lu D, Liu X, Gao Y, Huo B, Kang Y, Chen J, et al. Asymmetric migration of human keratinocytes under mechanical stretch and cocultured fibroblasts in a wound repair model. PLoS One. 2013;8(9):e74563.
50. Tokuyama E, Nagai Y, Takahashi K, Kimata Y, Naruse K. Mechanical Stretch on Human Skin Equivalents Increases the Epidermal Thickness and Develops the Basement Membrane. PLoS One. 2015;10(11):e0141989.
51. Lee Y, Gil MS, Hong JJ. Histomorphologic changes of hair follicles in human expanded scalp. Plast Reconstr Surg. 2000;105(7):2361-5.
52. Bao C, Wang B, Yang F, Pang J, Jiang Y, Chen L. Blockade of interleukin-7 receptor shapes macrophage alternative activation and promotes functional recovery after spinal cord injury. Neuroscience. 2017.
53. Martinez FO, Helming L, Gordon S. Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol. 2009;27:451-83.
54. ter Horst EN, Hakimzadeh N, van der Laan AM, Krijnen PA, Niessen HW, Piek JJ. Modulators of Macrophage Polarization Influence Healing of the Infarcted Myocardium. Int J Mol Sci. 2015;16(12):29583-91.
55. Xing L, Genhua M, Chunmei S, Liangliang Z, Lei H, Qin J, et al. Role of M2 Macrophages in Sepsis-Induced Acute Kidney Injury. Shock. 2017.
56. Cheng Y, Rong J. Macrophage Polarization as a Therapeutic Target in Myocardial Infarction. Curr Drug Targets. 2017.
57. Wang X, Chen H, Tian R, Zhang Y, Drutskaya MS, Wang C, et al. Macrophages induce AKT/beta-catenin-dependent Lgr5+ stem cell activation and hair follicle regeneration through TNF. Nat Commun. 2017;8:14091.
58. Qi Y, Li M, Xu L, Chang Z, Shu X, Zhou L. Therapeutic role of human hepatocyte growth factor (HGF) in treating hair loss. PeerJ. 2016;4:e2624.
59. El Kasmi KC, Stenmark KR. Contribution of metabolic reprogramming to macrophage plasticity and function. Semin Immunol. 2015;27(4):267-75.
60. He C, Carter AB. The Metabolic Prospective and Redox Regulation of Macrophage Polarization. J Clin Cell Immunol. 2015;6(6).
61. Adair-Kirk TL, Atkinson JJ, Kelley DG, Arch RH, Miner JH, Senior RM. A chemotactic peptide from laminin alpha 5 functions as a regulator of inflammatory immune responses via TNF alpha-mediated signaling. J Immunol. 2005;174(3):1621-9.
62. Hance KA, Tataria M, Ziporin SJ, Lee JK, Thompson RW. Monocyte chemotactic activity in human abdominal aortic aneurysms: role of elastin degradation peptides and the 67-kD cell surface elastin receptor. J Vasc Surg. 2002;35(2):254-61.
63. Adair-Kirk TL, Senior RM. Fragments of extracellular matrix as mediators of inflammation. Int J Biochem Cell Biol. 2008;40(6-7):1101-10.
64. McKee CM, Penno MB, Cowman M, Burdick MD, Strieter RM, Bao C, et al. Hyaluronan (HA) fragments induce chemokine gene expression in alveolar macrophages. The role of HA size and CD44. J Clin Invest. 1996;98(10):2403-13.
65. Jiang A, Bloom O, Ono S, Cui W, Unternaehrer J, Jiang S, et al. Disruption of E-cadherin-mediated adhesion induces a functionally distinct pathway of dendritic cell maturation. Immunity. 2007;27(4):610-24.
66. Ali N, Zirak B, Rodriguez RS, Pauli ML, Truong HA, Lai K, et al. Regulatory T Cells in Skin Facilitate Epithelial Stem Cell Differentiation. Cell. 2017;169(6):1119-29 e11.
67. Maryanovich M, Frenette PS. T-Regulating Hair Follicle Stem Cells. Immunity. 2017;46(6):979-81.