血管生長 (Vascularization) 過程在人體正常的生理和病理發展過程中持續進行， 研究發現血管的形成 (Vascular formation) 與氧氣張力 (Oxygen tension)之間有著密不可分的關係。 然而因為技術的限制，氧氣在空間中的分佈即氧氣梯度 (Oxygen gradient) 對血管發育的影響仍有待進一步的探索。本論文利用微流體技術設計了兩個不同的細胞培養平台，以建構具有氧氣梯度的體外細胞模型，用來探索血管形成過程與環境中氧氣梯度之間的相關性。為了在體外構建類似體內的微觀環境，本論文亦應用水凝膠來形成具有適當的三維環境作為細胞外基質（Extra-cellular matrix, ECM），並為細胞及組織提供必需的生長因子。為了研究微流體細胞培養平台所創造的氧氣梯度，本論文利用頻率域螢光生命週期顯微術 (Frequency domain fluorescence lifetime imaging microscopy, FD-FLIM) 來測量微流體平台中所生成的氧氣梯度。 利用本論文所發展的第一個細胞培養平台，實驗中可以觀察到三維水凝膠內存在的氧氣梯度，可促進血管細胞網路的形成，氧氣梯度亦對血管細胞網路形成的方向有著調節的作用。 此外，在第二個細胞培養平台的實驗中，觀察到氧氣梯度的存在對血管細胞出芽行為扮演關鍵的角色，梯度具有增加血管細胞出芽長度之特性。
本論文所開發具有能夠產生氧氣梯度的微流體細胞培養平台，提供了生醫研究先進的體外模型，可以應用於研究各種類似體內氧氣微環境下的血管細胞網路形成和血管萌芽行為。 更進一步，可以應用在研究與氧氣相關的腫瘤微環境，以更好地探索腫瘤發生及轉移的過程，更具有幫助改善臨床檢驗和藥物開發及篩選的淺力。 另一方面，如此可形成氧氣梯度的微流道細胞培養平台可以進一步加速血管化類器官之開發，期許在未來能應用在類器官晶片的開發上，加速實現個人化臨床藥物篩檢及個人精準治療。

Vascularization processes are ongoing in human bodies throughout normal physiological and pathological development. It has been shown that oxygen tensions can greatly affect vascular formation. However, the influence of the spatial distribution of oxygen (i.e., oxygen gradient) on vascular development is still underexplored. Therefore, two microfluidic platforms are designed in this thesis to explore the relationship between the blood vessel-forming processes and oxygen gradient generations. Furthermore, to construct the human-like microenvironment in vitro, hydrogels are exploited to construct a three-dimensional extracellular matrix (ECM) with proper properties and supply essential growth factors for cell/tissue growth. In order to investigate the oxygen microenvironments inside the devices, the generated oxygen gradients are characterized using frequency-domain fluorescence lifetime microscopy (FD-FLIM) measurements. The results of the first vascular formation studies reveal that the oxygen gradient can be established within the hydrogels in the microfluidic device. The experimental results show that the oxygen gradients can promote three-dimensional vascular cell network formation. In addition, the gradient also plays an important role in regulating the vascular cell network orientation. The second device is exploited to investigate vascular cell sprouting, and the results show that the presence of an oxygen gradient has great effects on increasing vascular cell sprouting.
It is confirmed that the two developed microfluidic platforms can generate oxygen gradients for cell culture applications. The platforms provide powerful in vitro models to investigate both vascular cellular network formations and vascular sprouting behaviors under various in vivo-like oxygen microenvironments. The platforms can be further applied to study the oxygen-related tumor microenvironments to better picture the process of tumorigenesis and metastasis. In addition, the microfluidic platforms capable of generating oxygen gradients can be further exploited to develop vascularized organoids for the development of the advanced organ-on-a-chip that can be used in personalized medicine in the coming future.

口試委員會審定書 #
誌謝 i
中文摘要 ii
ABSTRACT iv
CONTENTS vi
LIST OF FIGURES ix
LIST OF TABLES xiv
Chapter 1 Introduction 1
1.1 Background 1
1.2 Motivation 3
1.3 Specific Aims 5
1.4 Thesis Outline 6
Chapter 2 Literature Review 8
2.1 Vascularization 8
2.2 Oxygen Levels 10
2.3 Extracellular Matrix 14
2.4 Main Growth Factors for Vascularization 16
2.5 Microfluidic Systems 17
Chapter 3 Development of an upside-down microfluidic platform capable of generating oxygen gradient to study influence of oxygen gradient on initial vascular cell network formation 23
3.1 Introduction 23
3.2 Materials and Methods 26
3.2.1 Microfluidic Device Design and Fabrication 26
3.2.2 Cell Culture 28
3.2.3 Three-dimensional (3D) Hydrogel Preparation 29
3.2.4 Oxygen Gradient Generation Method 31
3.2.5 Consistence of Oxygen Levels 32
3.2.6 Numerical Simulation 32
3.2.7 Frequency Domain Fluorescence Lifetime Microscopy System 34
3.2.8 Characterization of Oxygen Gradient 37
3.2.9 Characterization of the Hydrogel Stiffness 39
3.2.10 Cell Viability 39
3.2.11 Immunocytochemistry 40
3.2.12 Proportions of Single Cell Analysis 41
3.2.13 Statistical Analysis 44
3.3 Results and Discussion 44
3.3.1 Oxygen Gradient Profiles in the Microfluidic Platform 44
3.3.2 Cell Viability Analysis 46
3.3.3 Single Cell Population Analysis 52
3.3.4 Cell Network Length and Orientation Analysis 55
3.4 Elasticity of Hydrogel Compositions 61
3.5 Conclusion 63
Chapter 4 Studying the development of vascular endothelial cell sprouting under combinations of oxygen conditions and hydrogel compositions 64
4.1 Introduction 64
4.2 Materials and Methods 66
4.2.1 Microfluidic Platform 66
4.2.2 Cell Culture 68
4.2.3 Three-Dimensional (3D) Matrix 69
4.2.4 Oxygen Gradient Generation Method 70
4.2.5 The Characterization of Oxygen Gradients 72
4.2.6 Assay of Angiogenesis Sprouting 73
4.2.7 Analysis of Angiogenic Sprouting Length 73
4.2.8 Cell Viability Assay 74
4.2.9 Immunocytochemistry Staining 75
4.2.10 Cytokine Array Analysis 76
4.2.11 Atomic Force Microscopy (AFM) Analysis 77
4.2.12 Statistical Analysis 78
4.3 Results and Discussion 79
4.3.1 Oxygen Gradient Profiles in the Angiogenic Sprouting Microfluidic Platform 79
4.3.2 Analysis of Cell Viability 82
4.3.3 HUVECs sprouting in Various Oxygen Microenvironments 83
4.3.4 Sprouting of HUVECs with Co-culture of MRC-5 in the 3D Hydrogel Matrixes 87
4.3.5 Cytokine Array Analysis 91
4.3.6 Analysis of Hydrogel Stiffness 93
Chapter 5 Conclusion and Future Works 100
List of Papers 103
REFERENCE 104

[1] J. Folkman, "Tumor angiogenesis: therapeutic implications," New england journal of medicine, vol. 285, no. 21, pp. 1182-1186, 1971.
[2] J. Folkman, D. M. Long Jr, and F. F. Becker, "Growth and metastasis of tumor in organ culture," Cancer, vol. 16, no. 4, pp. 453-467, 1963.
[3] U. Folkman, R. Kalluri, D. Kufe, R. Pollock, and R. Weichselbaum, "Beginning of angiogenesis research," Holland-frei cancer medicine, vol. 6, 2003.
[4] A. M. Duffy, D. J. Bouchier-Hayes, and J. H. Harmey, "Vascular endothelial growth factor (VEGF) and its role in non-endothelial cells: autocrine signalling by VEGF," in Madame Curie Bioscience Database [Internet]: Landes Bioscience, 2013.
[5] A. Tufro-McReddie, V. Norwood, K. Aylor, S. Botkin, R. Carey, and R. Gomez, "Oxygen regulates vascular endothelial growth factor-mediated vasculogenesis and tubulogenesis," Developmental biology, vol. 183, no. 2, pp. 139-149, 1997.
[6] Q. Zhang, Q. Yan, H. Yang, and W. Wei, "Oxygen sensing and adaptability won the 2019 Nobel Prize in Physiology or medicine," Genes & diseases, vol. 6, no. 4, pp. 328-332, 2019.
[7] S. Ramakrishnan, V. Anand, and S. Roy, "Vascular endothelial growth factor signaling in hypoxia and inflammation," Journal of neuroimmune pharmacology, vol. 9, pp. 142-160, 2014.
[8] Y. M. Salinas-Vera et al., "Three-dimensional 3D culture models in gynecological and breast cancer research," Frontiers in Oncology, vol. 12, p. 826113, 2022.
[9] J. Lomei, "Functional characterization of pro-angiogenic neutrophils," Acta Universitatis Upsaliensis, 2018.
[10] P. Carmeliet and R. K. Jain, "Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases," Nature reviews Drug discovery, vol. 10, no. 6, pp. 417-427, 2011.
[11] C. W. Peak, L. Cross, A. Singh, and A. K. Gaharwar, "Microscale technologies for engineering complex tissue structures," Microscale Technologies for Cell Engineering, pp. 3-25, 2016.
[12] F. Toschi and M. Sega, Flowing matter. Springer Nature, 2019.
[13] D. J. Hicklin and L. M. Ellis, "Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis," Journal of clinical oncology, vol. 23, no. 5, pp. 1011-1027, 2005.
[14] L. C. Goldie, M. K. Nix, and K. K. Hirschi, "Embryonic vasculogenesis and hematopoietic specification," Organogenesis, vol. 4, no. 4, pp. 257-263, 2008.
[15] K. L. Marcelo, L. C. Goldie, and K. K. Hirschi, "Regulation of endothelial cell differentiation and specification," Circulation research, vol. 112, no. 9, pp. 1272-1287, 2013.
[16] S. Kazemi et al., "Differential role of bFGF and VEGF for vasculogenesis," Cellular Physiology and Biochemistry, vol. 12, no. 2-3, pp. 55-62, 2002.
[17] X. Lin et al., "Oxygen-induced cell migration and on-line monitoring biomarkers modulation of cervical cancers on a microfluidic system," Scientific reports, vol. 5, no. 1, p. 9643, 2015.
[18] T. H. Adair and J.-P. Montani, "Angiogenesis," 2011.
[19] A. Schmidt, K. Brixius, and W. Bloch, "Endothelial precursor cell migration during vasculogenesis," Circulation research, vol. 101, no. 2, pp. 125-136, 2007.
[20] C. K. Griffith et al., "Diffusion limits of an in vitro thick prevascularized tissue," Tissue engineering, vol. 11, no. 1-2, pp. 257-266, 2005.
[21] K. Kisler et al., "Pericyte degeneration leads to neurovascular uncoupling and limits oxygen supply to brain," Nature neuroscience, vol. 20, no. 3, pp. 406-416, 2017.
[22] J. C.-Y. Chung and D. Shum-Tim, "Neovascularization in tissue engineering," Cells, vol. 1, no. 4, pp. 1246-1260, 2012.
[23] B. Bourghardt Peebo, "Angiogenesis from a new perspective," Linköping University Electronic Press, 2012.
[24] S. Patan, "Vasculogenesis and angiogenesis as mechanisms of vascular network formation, growth and remodeling," Journal of neuro-oncology, vol. 50, pp. 1-15, 2000.
[25] R. K. Jain, K. Schlenger, M. Hockel, and F. Yuan, "Quantitative angiogenesis assays: progress and problems," Nature medicine, vol. 3, no. 11, pp. 1203-1208, 1997.
[26] W. Cai et al., "Pericytes in brain injury and repair after ischemic stroke," Translational Stroke Research, vol. 8, pp. 107-121, 2017.
[27] M. Murakami and M. Simons, "Fibroblast growth factor regulation of neovascularization," Current opinion in hematology, vol. 15, no. 3, p. 215, 2008.
[28] E. Hadjipanayi and A. F. Schilling, "Regeneration through autologous hypoxia preconditioned plasma," Organogenesis, vol. 10, no. 2, pp. 164-169, 2014.
[29] T. L. Place, F. E. Domann, and A. J. Case, "Limitations of oxygen delivery to cells in culture: An underappreciated problem in basic and translational research," Free Radical Biology and Medicine, vol. 113, pp. 311-322, 2017.
[30] S. S. Kety, "The theory and application of the exchange of inert gas at the lungs and tissues," Pharmacol rev, vol. 3, pp. 1-41, 1951.
[31] A. Krogh, "The supply of oxygen to the tissues and the regulation of the capillary circulation," The Journal of physiology, vol. 52, no. 6, p. 457, 1919.
[32] S. McKeown, "Defining normoxia, physoxia and hypoxia in tumours—implications for treatment response," The British journal of radiology, vol. 87, no. 1035, p. 20130676, 2014.
[33] L. Tiede, E. Cook, B. Morsey, and H. Fox, "Oxygen matters: tissue culture oxygen levels affect mitochondrial function and structure as well as responses to HIV viroproteins," Cell death & disease, vol. 2, no. 12, pp. e246-e246, 2011.
[34] A. Carreau, B. E. Hafny‐Rahbi, A. Matejuk, C. Grillon, and C. Kieda, "Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia," Journal of cellular and molecular medicine, vol. 15, no. 6, pp. 1239-1253, 2011.
[35] M. C. Simon and B. Keith, "The role of oxygen availability in embryonic development and stem cell function," Nature reviews Molecular cell biology, vol. 9, no. 4, pp. 285-296, 2008.
[36] K. E. Hacker, C. M. Lee, and W. K. Rathmell, "VHL type 2B mutations retain VBC complex form and function," PloS one, vol. 3, no. 11, p. e3801, 2008.
[37] L. Østergaard et al., "Microcirculatory dysfunction and tissue oxygenation in critical illness," Acta Anaesthesiologica Scandinavica, vol. 59, no. 10, pp. 1246-1259, 2015.
[38] R. M. Winslow, "Oxygen: the poison is in the dose," Transfusion, vol. 53, no. 2, pp. 424-437, 2013.
[39] M. Ponticos and B. D. Smith, "Extracellular matrix synthesis in vascular disease: hypertension, and atherosclerosis," Journal of biomedical research, vol. 28, no. 1, p. 25, 2014.
[40] J. T. Thorne, T. R. Segal, S. Chang, S. Jorge, J. H. Segars, and P. C. Leppert, "Dynamic reciprocity between cells and their microenvironment in reproduction," Biology of reproduction, vol. 92, no. 1, pp. 25, 1-10, 2015.
[41] R. Jayadev and D. R. Sherwood, "Basement membranes," Current Biology, vol. 27, no. 6, pp. R207-R211, 2017.
[42] C. Bonnans, J. Chou, and Z. Werb, "Remodelling the extracellular matrix in development and disease," Nature reviews Molecular cell biology, vol. 15, no. 12, pp. 786-801, 2014.
[43] H. Järveläinen, A. Sainio, M. Koulu, T. N. Wight, and R. Penttinen, "Extracellular matrix molecules: potential targets in pharmacotherapy," Pharmacological reviews, vol. 61, no. 2, pp. 198-223, 2009.
[44] M. Dietrich et al., "Guiding 3D cell migration in deformed synthetic hydrogel microstructures," Soft Matter, vol. 14, no. 15, pp. 2816-2826, 2018.
[45] M. W. MOSESSON, "Fibrinogen and fibrin structure and functions," Journal of thrombosis and haemostasis, vol. 3, no. 8, pp. 1894-1904, 2005.
[46] J. Arulmoli et al., "Combination scaffolds of salmon fibrin, hyaluronic acid, and laminin for human neural stem cell and vascular tissue engineering," Acta biomaterialia, vol. 43, pp. 122-138, 2016.
[47] O. Moreno-Arotzena, J. G. Meier, C. Del Amo, and J. M. García-Aznar, "Characterization of fibrin and collagen gels for engineering wound healing models," Materials, vol. 8, no. 4, pp. 1636-1651, 2015.
[48] S. G. Zambuto, K. B. Clancy, and B. A. Harley, "A gelatin hydrogel to study endometrial angiogenesis and trophoblast invasion," Interface Focus, vol. 9, no. 5, p. 20190016, 2019.
[49] P. A. Janmey, J. P. Winer, and J. W. Weisel, "Fibrin gels and their clinical and bioengineering applications," Journal of the Royal Society Interface, vol. 6, no. 30, pp. 1-10, 2009.
[50] M. Venkatesan et al., "Recombinant production of growth factors for application in cell culture," Iscience, vol. 25, no. 10, 2022.
[51] M. Geindreau, M. Bruchard, and F. Vegran, "Role of cytokines and chemokines in angiogenesis in a tumor context," Cancers, vol. 14, no. 10, p. 2446, 2022.
[52] Q. Luo et al., "Vasculogenic mimicry in carcinogenesis and clinical applications," Journal of hematology & oncology, vol. 13, pp. 1-15, 2020.
[53] X. Yang et al., "Fibroblast growth factor signaling in the vasculature," Current atherosclerosis reports, vol. 17, pp. 1-11, 2015.
[54] H. Wu, T. Lee, P. Ko, H. Chiang, C. Peng, and Y. Tung, "Review of microfluidic cell culture devices for the control of gaseous microenvironments in vitro," Journal of Micromechanics and Microengineering, vol. 28, no. 4, p. 043001, 2018.
[55] P. F. Davies, "Flow-mediated endothelial mechanotransduction," Physiological reviews, vol. 75, no. 3, pp. 519-560, 1995.
[56] Y. Kamotani et al., "Individually programmable cell stretching microwell arrays actuated by a Braille display," Biomaterials, vol. 29, no. 17, pp. 2646-2655, 2008.
[57] C. Moraes, J.-H. Chen, Y. Sun, and C. A. Simmons, "Microfabricated arrays for high-throughput screening of cellular response to cyclic substrate deformation," Lab on a Chip, vol. 10, no. 2, pp. 227-234, 2010.
[58] C. M. Leung et al., "A guide to the organ-on-a-chip," Nature Reviews Methods Primers, vol. 2, no. 1, p. 33, 2022.
[59] A. P. Vollmer, R. F. Probstein, R. Gilbert, and T. Thorsen, "Development of an integrated microfluidic platform for dynamic oxygen sensing and delivery in a flowing medium," Lab on a Chip, vol. 5, no. 10, pp. 1059-1066, 2005.
[60] M.-C. Kim, R. H. Lam, T. Thorsen, and H. H. Asada, "Mathematical analysis of oxygen transfer through polydimethylsiloxane membrane between double layers of cell culture channel and gas chamber in microfluidic oxygenator," Microfluidics and Nanofluidics, vol. 15, pp. 285-296, 2013.
[61] J. F. Lo, E. Sinkala, and D. T. Eddington, "Oxygen gradients for open well cellular cultures via microfluidic substrates," Lab on a Chip, vol. 10, no. 18, pp. 2394-2401, 2010.
[62] Y.-A. Chen et al., "Generation of oxygen gradients in microfluidic devices for cell culture using spatially confined chemical reactions," Lab on a Chip, vol. 11, no. 21, pp. 3626-3633, 2011.
[63] C.-W. Chang et al., "A polydimethylsiloxane–polycarbonate hybrid microfluidic device capable of generating perpendicular chemical and oxygen gradients for cell culture studies," Lab on a Chip, vol. 14, no. 19, pp. 3762-3772, 2014.
[64] M. D. Brennan, M. L. Rexius-Hall, L. J. Elgass, and D. T. Eddington, "Oxygen control with microfluidics," Lab on a Chip, vol. 14, no. 22, pp. 4305-4318, 2014.
[65] S. Kim, W. Kim, S. Lim, and J. S. Jeon, "Vasculature-on-a-chip for in vitro disease models," Bioengineering, vol. 4, no. 1, p. 8, 2017.
[66] J. Brown, "Vasculogenesis: a crucial player in the resistance of solid tumours to radiotherapy," The British journal of radiology, vol. 87, no. 1035, p. 20130686, 2014.
[67] J. P. Greenfield, W. S. Cobb, and D. Lyden, "Resisting arrest: a switch from angiogenesis to vasculogenesis in recurrent malignant gliomas," The Journal of clinical investigation, vol. 120, no. 3, pp. 663-667, 2010.
[68] R. S. Kerbel, "Tumor angiogenesis," New England Journal of Medicine, vol. 358, no. 19, pp. 2039-2049, 2008.
[69] A. J. Giaccia, M. C. Simon, and R. Johnson, "The biology of hypoxia: the role of oxygen sensing in development, normal function, and disease," Genes & development, vol. 18, no. 18, pp. 2183-2194, 2004.
[70] K. T. Morin and R. T. Tranquillo, "In vitro models of angiogenesis and vasculogenesis in fibrin gel," Experimental cell research, vol. 319, no. 16, pp. 2409-2417, 2013.
[71] M. Skolimowski et al., "Microfluidic dissolved oxygen gradient generator biochip as a useful tool in bacterial biofilm studies," Lab on a Chip, vol. 10, no. 16, pp. 2162-2169, 2010.
[72] H.-M. Wu, T.-A. Lee, P.-L. Ko, W.-H. Liao, T.-H. Hsieh, and Y.-C. Tung, "Widefield frequency domain fluorescence lifetime imaging microscopy (FD-FLIM) for accurate measurement of oxygen gradients within microfluidic devices," Analyst, vol. 144, no. 11, pp. 3494-3504, 2019.
[73] H. H. Hsu, P. L. Ko, H. M. Wu, H. C. Lin, C. K. Wang, and Y. C. Tung, "Study 3D Endothelial Cell Network Formation under Various Oxygen Microenvironment and Hydrogel Composition Combinations Using Upside‐Down Microfluidic Devices," Small, vol. 17, no. 15, p. 2006091, 2021.
[74] Y.-H. Chen, C.-C. Peng, and Y.-C. Tung, "Flip channel: A microfluidic device for uniform-sized embryoid body formation and differentiation," Biomicrofluidics, vol. 9, no. 5, p. 054111, 2015.
[75] D. Jiang, J. Liang, and P. W. Noble, "Hyaluronan as an immune regulator in human diseases," Physiological reviews, vol. 91, no. 1, pp. 221-264, 2011.
[76] B. P. Toole, "Hyaluronan: from extracellular glue to pericellular cue," Nature Reviews Cancer, vol. 4, no. 7, pp. 528-539, 2004.
[77] W. Zhong, P. Urayama, and M.-A. Mycek, "Imaging fluorescence lifetime modulation of a ruthenium-based dye in living cells: the potential for oxygen sensing," Journal of Physics D: Applied Physics, vol. 36, no. 14, p. 1689, 2003.
[78] S. R. Caliari and J. A. Burdick, "A practical guide to hydrogels for cell culture," Nature methods, vol. 13, no. 5, pp. 405-414, 2016.
[79] C.-H. Lin, C.-K. Wang, Y.-A. Chen, C.-C. Peng, W.-H. Liao, and Y.-C. Tung, "Measurement of in-plane elasticity of live cell layers using a pressure sensor embedded microfluidic device," Scientific reports, vol. 6, no. 1, p. 36425, 2016.
[80] C. A. Schneider, W. S. Rasband, and K. W. Eliceiri, "NIH Image to ImageJ: 25 years of image analysis," Nature methods, vol. 9, no. 7, pp. 671-675, 2012.
[81] P. Fraisl, M. Mazzone, T. Schmidt, and P. Carmeliet, "Regulation of angiogenesis by oxygen and metabolism," Developmental cell, vol. 16, no. 2, pp. 167-179, 2009.
[82] H. E. Abaci, R. Truitt, E. Luong, G. Drazer, and S. Gerecht, "Adaptation to oxygen deprivation in cultures of human pluripotent stem cells, endothelial progenitor cells, and umbilical vein endothelial cells," American Journal of Physiology-Cell Physiology, vol. 298, no. 6, pp. C1527-C1537, 2010.
[83] S. Zhao et al., "Melatonin protects against hypoxia/reoxygenation-induced dysfunction of human umbilical vein endothelial cells through inhibiting reactive oxygen species generation," Acta Cardiologica Sinica, vol. 34, no. 5, p. 424, 2018.
[84] R. C. Wang and B. Levine, "Autophagy in cellular growth control," FEBS letters, vol. 584, no. 7, pp. 1417-1426, 2010.
[85] E. Könönen, M. Gursoy, and U. K. Gursoy, "Periodontitis: a multifaceted disease of tooth-supporting tissues," Journal of clinical medicine, vol. 8, no. 8, p. 1135, 2019.
[86] H. Lin, C. Wang, Y. Tung, F. Chiu, and Y. Su, "Increased vasculogenesis of endothelial cells in hyaluronic acid augmented fibrin-based natural hydrogels-from in vitro to in vivo models," Eur Cell Mater, vol. 40, pp. 133-145, 2020.
[87] S. Ghose, S. Biswas, K. Datta, and R. K. Tyagi, "Dynamic Hyaluronan drives liver endothelial cells towards angiogenesis," BMC cancer, vol. 18, no. 1, pp. 1-13, 2018.
[88] W. Shigeeda et al., "Hyaluronic acid enhances cell migration and invasion via the YAP1/TAZ-RHAMM axis in malignant pleural mesothelioma," Oncotarget, vol. 8, no. 55, p. 93729, 2017.
[89] R. M. Simpson et al., "Hyaluronan is crucial for stem cell differentiation into smooth muscle lineage," Stem Cells, vol. 34, no. 5, pp. 1225-1238, 2016.
[90] Z. Zhu, Y.-M. Wang, J. Yang, and X.-S. Luo, "Hyaluronic acid: a versatile biomaterial in tissue engineering," Plast Aesthet Res, vol. 4, no. 219-27, 2017.
[91] P. W. Noble, "Hyaluronan and its catabolic products in tissue injury and repair," Matrix biology, vol. 21, no. 1, pp. 25-29, 2002.
[92] M. Slevin, S. Kumar, and J. Gaffney, "Angiogenic oligosaccharides of hyaluronan induce multiple signaling pathways affecting vascular endothelial cell mitogenic and wound healing responses," Journal of biological chemistry, vol. 277, no. 43, pp. 41046-41059, 2002.
[93] M. Kretschmer, D. Rüdiger, and S. Zahler, "Mechanical aspects of angiogenesis," Cancers, vol. 13, no. 19, p. 4987, 2021.
[94] C. R. Pfeifer, C. M. Alvey, J. Irianto, and D. E. Discher, "Genome variation across cancers scales with tissue stiffness–An invasion-mutation mechanism and implications for immune cell infiltration," Current opinion in systems biology, vol. 2, pp. 103-114, 2017.
[95] J. Pauty et al., "A vascular endothelial growth factor-dependent sprouting angiogenesis assay based on an in vitro human blood vessel model for the study of anti-angiogenic drugs," EBioMedicine, vol. 27, pp. 225-236, 2018.
[96] J. E. Bader, K. Voss, and J. C. Rathmell, "Targeting metabolism to improve the tumor microenvironment for cancer immunotherapy," Molecular cell, vol. 78, no. 6, pp. 1019-1033, 2020.
[97] K. De Bock et al., "Role of PFKFB3-driven glycolysis in vessel sprouting," Cell, vol. 154, no. 3, pp. 651-663, 2013.
[98] F. Bordeleau et al., "Matrix stiffening promotes a tumor vasculature phenotype," Proceedings of the National Academy of Sciences, vol. 114, no. 3, pp. 492-497, 2017.
[99] A. Costa, A. Scholer-Dahirel, and F. Mechta-Grigoriou, "The role of reactive oxygen species and metabolism on cancer cells and their microenvironment," in Seminars in cancer biology, 2014, vol. 25: Elsevier, pp. 23-32.
[100] T. Hashimoto and F. Shibasaki, "Hypoxia-inducible factor as an angiogenic master switch," Frontiers in pediatrics, vol. 3, p. 33, 2015.
[101] B. L. Krock, N. Skuli, and M. C. Simon, "Hypoxia-induced angiogenesis: good and evil," Genes & cancer, vol. 2, no. 12, pp. 1117-1133, 2011.
[102] A. C. Newman, M. N. Nakatsu, W. Chou, P. D. Gershon, and C. C. Hughes, "The requirement for fibroblasts in angiogenesis: fibroblast-derived matrix proteins are essential for endothelial cell lumen formation," Molecular biology of the cell, vol. 22, no. 20, pp. 3791-3800, 2011.
[103] I. Mahmud and D. Liao, "DAXX in cancer: phenomena, processes, mechanisms and regulation," Nucleic acids research, vol. 47, no. 15, pp. 7734-7752, 2019.
[104] S. Germain, C. Monnot, L. Muller, and A. Eichmann, "Hypoxia-driven angiogenesis: role of tip cells and extracellular matrix scaffolding," Current opinion in hematology, vol. 17, no. 3, pp. 245-251, 2010.
[105] R. Edmondson, J. J. Broglie, A. F. Adcock, and L. Yang, "Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors," Assay and drug development technologies, vol. 12, no. 4, pp. 207-218, 2014.
[106] M. Mehling and S. Tay, "Microfluidic cell culture," Current opinion in Biotechnology, vol. 25, pp. 95-102, 2014.
[107] M.-C. Liu et al., "Electrofluidic pressure sensor embedded microfluidic device: a study of endothelial cells under hydrostatic pressure and shear stress combinations," Lab on a Chip, vol. 13, no. 9, pp. 1743-1753, 2013.
[108] H.-H. Hsu, P.-L. Ko, C.-C. Peng, Y.-J. Cheng, H.-M. Wu, and Y.-C. Tung, "Studying sprouting angiogenesis under combination of oxygen gradients and Co-culture of fibroblasts using microfluidic cell culture model," Materials Today Bio, p. 100703, 2023.
[109] M. Dewerchin and P. Carmeliet, "PlGF: a multitasking cytokine with disease-restricted activity," Cold Spring Harbor perspectives in medicine, vol. 2, no. 8, p. a011056, 2012.
[110] H.-C. Shih, T.-A. Lee, H.-M. Wu, P.-L. Ko, W.-H. Liao, and Y.-C. Tung, "Microfluidic collective cell migration assay for study of endothelial cell proliferation and migration under combinations of oxygen gradients, tensions, and drug treatments," Scientific reports, vol. 9, no. 1, p. 8234, 2019.
[111] M. Migdal et al., "Neuropilin-1 is a placenta growth factor-2 receptor," Journal of Biological Chemistry, vol. 273, no. 35, pp. 22272-22278, 1998.
[112] L. Xiang, R. Varshney, N. A. Rashdan, J. H. Shaw, and P. G. Lloyd, "Placenta growth factor and vascular endothelial growth factor a have differential, cell‐type specific patterns of expression in vascular cells," Microcirculation, vol. 21, no. 5, pp. 368-379, 2014.
[113] D. Ribatti, "The discovery of the placental growth factor and its role in angiogenesis: a historical review," Angiogenesis, vol. 11, no. 3, pp. 215-221, 2008.
[114] H. Iwamoto et al., "PlGF-induced VEGFR1-dependent vascular remodeling determines opposing antitumor effects and drug resistance to Dll4-Notch inhibitors," Science Advances, vol. 1, no. 3, p. e1400244, 2015.
[115] C. J. Green et al., "Placenta growth factor gene expression is induced by hypoxia in fibroblasts: a central role for metal transcription factor-1," Cancer research, vol. 61, no. 6, pp. 2696-2703, 2001.
[116] S. De Falco, "The discovery of placenta growth factor and its biological activity," Experimental & molecular medicine, vol. 44, no. 1, pp. 1-9, 2012.
[117] L. Tudisco, A. Orlandi, V. Tarallo, and S. De Falco, "Hypoxia activates placental growth factor expression in lymphatic endothelial cells," Oncotarget, vol. 8, no. 20, p. 32873, 2017.
[118] M. Shibuya, "Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) signaling in angiogenesis: a crucial target for anti-and pro-angiogenic therapies," Genes & cancer, vol. 2, no. 12, pp. 1097-1105, 2011.
[119] N. Rahimi, "VEGFR-1 and VEGFR-2: two non-identical twins with a unique physiognomy," Frontiers in bioscience: a journal and virtual library, vol. 11, p. 818, 2006.
[120] F. Morfoisse, E. Renaud, F. Hantelys, A.-C. Prats, and B. Garmy-Susini, "Role of hypoxia and vascular endothelial growth factors in lymphangiogenesis," Molecular & cellular oncology, vol. 2, no. 4, p. e1024821, 2015.
[121] D. Gazit, Y. Zilberman, G. Turgeman, S. Zhou, and A. Kahn, "Recombinant TGF‐β1 stimulates bone marrow osteoprogenitor cell activity and bone matrix synthesis in osteopenic, old male mice," Journal of Cellular Biochemistry, vol. 73, no. 3, pp. 379-389, 1999.
[122] J. Ma, G. Sanchez-Duffhues, M.-J. Goumans, and P. Ten Dijke, "TGF-β-induced endothelial to mesenchymal transition in disease and tissue engineering," Frontiers in cell and developmental biology, vol. 8, p. 260, 2020.
[123] M. S. Pepper, "Transforming growth factor-beta: vasculogenesis, angiogenesis, and vessel wall integrity," Cytokine & growth factor reviews, vol. 8, no. 1, pp. 21-43, 1997.
[124] J. Massagué, "TGFβ signalling in context," Nature reviews Molecular cell biology, vol. 13, no. 10, pp. 616-630, 2012.
[125] A. B. Roberts et al., "Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro," Proceedings of the National Academy of Sciences, vol. 83, no. 12, pp. 4167-4171, 1986.
[126] J. A. Madri, B. M. Pratt, and A. M. Tucker, "Phenotypic modulation of endothelial cells by transforming growth factor-beta depends upon the composition and organization of the extracellular matrix," The Journal of cell biology, vol. 106, no. 4, pp. 1375-1384, 1988.
[127] E. Y. Yang and H. L. Moses, "Transforming growth factor beta 1-induced changes in cell migration, proliferation, and angiogenesis in the chicken chorioallantoic membrane," The Journal of cell biology, vol. 111, no. 2, pp. 731-741, 1990.
[128] M. J. Pollman, L. Naumovski, and G. H. Gibbons, "Vascular cell apoptosis: cell type–specific modulation by transforming growth factor-β1 in endothelial cells versus smooth muscle cells," Circulation, vol. 99, no. 15, pp. 2019-2026, 1999.
[129] O. Saksela, D. Moscatelli, and D. B. Rifkin, "The opposing effects of basic fibroblast growth factor and transforming growth factor beta on the regulation of plasminogen activator activity in capillary endothelial cells," The Journal of cell biology, vol. 105, no. 2, pp. 957-963, 1987.
[130] M. Pepper, D. Belin, R. Montesano, L. Orci, and J. Vassalli, "Transforming growth factor-beta 1 modulates basic fibroblast growth factor-induced proteolytic and angiogenic properties of endothelial cells in vitro," The Journal of cell biology, vol. 111, no. 2, pp. 743-755, 1990.
[131] H. O. Akman, H. Zhang, M. Siddiqui, W. Solomon, E. L. Smith, and O. A. Batuman, "Response to hypoxia involves transforming growth factor-β2 and Smad proteins in human endothelial cells," Blood, The Journal of the American Society of Hematology, vol. 98, no. 12, pp. 3324-3331, 2001.
[132] F. Soncin, "Angiogenin supports endothelial and fibroblast cell adhesion," Proceedings of the National Academy of Sciences, vol. 89, no. 6, pp. 2232-2236, 1992.
[133] X. Gao and Z. Xu, "Mechanisms of action of angiogenin," Acta biochimica et biophysica Sinica, vol. 40, no. 7, pp. 619-624, 2008.
[134] C. Chen et al., "CXCL5 induces tumor angiogenesis via enhancing the expression of FOXD1 mediated by the AKT/NF-κB pathway in colorectal cancer," Cell death & disease, vol. 10, no. 3, p. 178, 2019.
[135] W. Zhang et al., "CXCL5/CXCR2 axis in tumor microenvironment as potential diagnostic biomarker and therapeutic target," Cancer Communications, vol. 40, no. 2-3, pp. 69-80, 2020.
[136] G. Gopinathan et al., "Interleukin-6 stimulates defective angiogenesis," Cancer research, vol. 75, no. 15, pp. 3098-3107, 2015.
[137] C. O. Crosby and J. Zoldan, "Mimicking the physical cues of the ECM in angiogenic biomaterials," Regenerative biomaterials, vol. 6, no. 2, pp. 61-73, 2019.
[138] D. J. LaValley, M. R. Zanotelli, F. Bordeleau, W. Wang, S. C. Schwager, and C. A. Reinhart-King, "Matrix stiffness enhances VEGFR-2 internalization, signaling, and proliferation in endothelial cells," Convergent Science Physical Oncology, vol. 3, no. 4, p. 044001, 2017.
[139] T. Bertucci, S. Kakarla, D. Kim, and G. Dai, "Differentiating human pluripotent stem cells to vascular endothelial cells for regenerative medicine, tissue engineering, and disease modeling," Vascular Tissue Engineering: Methods and Protocols, pp. 1-12, 2022.
[140] Y. Guo et al., "Matrix stiffness modulates tip cell formation through the p-PXN-Rac1-YAP signaling axis," Bioactive materials, vol. 7, pp. 364-376, 2022.