TY - JOUR
T1 - Designing topographically textured microparticles for induction and modulation of osteogenesis in mesenchymal stem cell engineering
AU - Amer, Mahetab H.
AU - Alvarez-Paino, Marta
AU - McLaren, Jane
AU - Pappalardo, Francesco
AU - Trujillo, Sara
AU - Wong, Jing Qian
AU - Shrestha, Sumana
AU - Abdelrazig, Salah
AU - Stevens, Lee A.
AU - Lee, Jong Bong
AU - Kim, Dong Hyun
AU - González-García, Cristina
AU - Needham, David
AU - Salmerón-Sánchez, Manuel
AU - Shakesheff, Kevin M.
AU - Alexander, Morgan R.
AU - Alexander, Cameron
AU - Rose, Felicity RAJ
N1 - Funding Information:
Funding: This work was supported by the Engineering and Physical Sciences Research Council (EPSRC) Programme Grants for “Next Generation Biomaterials Discovery” [ EP/N006615/1 ]. We also acknowledge financial support from EPSRC Programme Grant “ Engineering growth factor microenvironments - a new therapeutic paradigm for regenerative medicine ” [ EP/P001114/1 ].
Funding Information:
General: We acknowledge Dr. Alison Woodward (University of Nottingham) for assistance with metabolomics, Dr. Robert Markus ( University of Nottingham ) for assistance with confocal microscopy (equipment funded by Biotechnology and Biological Sciences Research Council [ BB/L013827/1 ]), and Dr. Tim Constantin (University of Nottingham) for assistance with PCR. We also thank the Nanoscale and Microscale Research Centre at University of Nottingham for providing access to SEM and XPS instrumentation. Portions of the schematic in Fig. 6 was produced using Servier Medical Art artwork.
Funding Information:
Extracellular metabolite levels can be linked with intracellular metabolism [34]. Metabolomics data analyses suggest that hMSCs are both a producer and target of vitamin D3 metabolites when cultured on dimpled particulate surfaces, which suggests a potential role of vitamin D metabolism in topographically induced differentiation. While some mammalian cells can make their own 1,25-dihydroxyvitamin D (1,25(OH)2D3) and respond to it [69], hMSCs express low levels of 25(OH)D3-1?-hydroxylase and vitamin D receptor [70]. Nevertheless, hydroxy-derivatives of vitamin D3 have been shown to stimulate osteogenesis [70]. Vit D has also been reported to prompt expression of ?v?3 integrin, in turn favouring the formation of focal adhesions and osteogenesis [71]. PGE2 and TGF-?1 levels were shown to be regulated by 1?,25(OH)2D3 in osteoblast-like cells, with effects synergistic with increasing substrate microroughness and absent on smooth surfaces [72]. LPG (20:1(11Z)/0:0), a monoacylglycerophosphoglycerol, was also identified as a significantly increased metabolite when hMSCs were cultured on dimpled microparticles relative to smooth ones. Osteoblastic differentiation requires lipidomic remodeling, driven by polyunsaturated lipids with long acyl chains [73]. Palmitic, stearic and oleic acids are major fatty acid components in human plasma lysophosphatidic acid (LPA) [36], which were revealed to be significantly increased in the metabolic profile of dimpled media samples relative to smooth. LPAs regulate ERK pathways, which are central to cell differentiation [74]. This agrees with the major signaling networks identified by Ingenuity Pathway Analysis (IPA), where ERK, interleukin-1 (IL-1) and JNK pathways are central. Supporting literature reports that integrin-related signaling has a central role in the modulation of stem cell phenotype on nano-topographical surfaces, acting through ERK1/2 and Jnk, with LPA being vital in driving osteogenic differentiation [37]. IPA of canonical signalling suggested the osteo-inductive effect of dimpled microparticles may occur via enhanced activity of iNOS, which is expressed in bone marrow stromal cells and osteoblasts, and is involved in the stimulation of osteoblasts by NO production [75]. Other significantly affected pathways included arginine, which plays an important role in bone healing through the production of nitric oxide [76], and proline, a crucial component of collagen [77]. Carnitine has also been linked to osteogenic differentiation [78]. Ketone body metabolism is a significant contributor to mammalian energy metabolism within extrahepatic tissues when glucose is not readily available [79]. Top upstream regulators identified by IPA included ZC3H10, consistent with its activated state. ZC3H10 is a poorly characterized RNA-binding protein which regulates mitochondrial biogenesis [80], which correlates with the importance of mitochondrial dynamics in osteogenesis [81]. More studies are needed to reveal how it regulates osteogenesis, and further experiments on intracellular metabolites will confirm mechanisms associated with the metabolic adaptation of hMSCs to topographically textured surfaces. The fundamental causes behind these differences in cellular responses presents exciting avenues for future studies.General: We acknowledge Dr. Alison Woodward (University of Nottingham) for assistance with metabolomics, Dr. Robert Markus (University of Nottingham) for assistance with confocal microscopy (equipment funded by Biotechnology and Biological Sciences Research Council [BB/L013827/1]), and Dr. Tim Constantin (University of Nottingham) for assistance with PCR. We also thank the Nanoscale and Microscale Research Centre at University of Nottingham for providing access to SEM and XPS instrumentation. Portions of the schematic in Fig. 6 was produced using Servier Medical Art artwork. Funding: This work was supported by the Engineering and Physical Sciences Research Council (EPSRC) Programme Grants for ?Next Generation Biomaterials Discovery? [EP/N006615/1]. We also acknowledge financial support from EPSRC Programme Grant ?Engineering growth factor microenvironments - a new therapeutic paradigm for regenerative medicine? [EP/P001114/1].
Publisher Copyright:
© 2020 The Authors
PY - 2021/1
Y1 - 2021/1
N2 - Mesenchymal stem cells are the focus of intense research in bone development and regeneration. The potential of microparticles as modulating moieties of osteogenic response by utilizing their architectural features is demonstrated herein. Topographically textured microparticles of varying microscale features are produced by exploiting phase-separation of a readily soluble sacrificial component from polylactic acid. The influence of varying topographical features on primary human mesenchymal stem cell attachment, proliferation and markers of osteogenesis is investigated. In the absence of osteoinductive supplements, cells cultured on textured microparticles exhibit notably increased expression of osteogenic markers relative to conventional smooth microparticles. They also exhibit varying morphological, attachment and proliferation responses. Significantly altered gene expression and metabolic profiles are observed, with varying histological characteristics in vivo. This study highlights how tailoring topographical design offers cell-instructive 3D microenvironments which allow manipulation of stem cell fate by eliciting the desired downstream response without use of exogenous osteoinductive factors.
AB - Mesenchymal stem cells are the focus of intense research in bone development and regeneration. The potential of microparticles as modulating moieties of osteogenic response by utilizing their architectural features is demonstrated herein. Topographically textured microparticles of varying microscale features are produced by exploiting phase-separation of a readily soluble sacrificial component from polylactic acid. The influence of varying topographical features on primary human mesenchymal stem cell attachment, proliferation and markers of osteogenesis is investigated. In the absence of osteoinductive supplements, cells cultured on textured microparticles exhibit notably increased expression of osteogenic markers relative to conventional smooth microparticles. They also exhibit varying morphological, attachment and proliferation responses. Significantly altered gene expression and metabolic profiles are observed, with varying histological characteristics in vivo. This study highlights how tailoring topographical design offers cell-instructive 3D microenvironments which allow manipulation of stem cell fate by eliciting the desired downstream response without use of exogenous osteoinductive factors.
KW - 3D
KW - Mesenchymal stem cells
KW - Microcarriers
KW - Osteogenic differentiation
KW - Osteoinductive
KW - Topography
UR - http://www.scopus.com/inward/record.url?scp=85092910243&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2020.120450
DO - 10.1016/j.biomaterials.2020.120450
M3 - Article
C2 - 33096376
AN - SCOPUS:85092910243
SN - 0142-9612
VL - 266
JO - Biomaterials
JF - Biomaterials
M1 - 120450
ER -