Publications

@article{Owen2024-ih,

title = {β-glycerophosphate, not low magnitude fluid shear stress,  increases osteocytogenesis in the osteoblast-to-osteocyte cell  line IDG-SW3},

author = {Robert Owen and Claudia Wittkowske and Damien Lacroix and Cecile M Perrault and Gwendolen C Reilly},

doi = {10.1080/03008207.2024.2375065},

year  = {2024},

date = {2024-07-01},

urldate = {2024-07-01},

journal = {Connect. Tissue Res.},

volume = {65},

number = {4},

pages = {313\textendash329},

publisher = {Informa UK Limited},

abstract = {AIM: As osteoblasts deposit a mineralized collagen network, a 

 subpopulation of these cells differentiates into osteocytes. 

 Biochemical and mechanical stimuli, particularly fluid shear 

 stress (FSS), are thought to regulate this, but their relative 

 influence remains unclear. Here, we assess both biochemical and 

 mechanical stimuli on long-term bone formation and 

 osteocytogenesis using the osteoblast-osteocyte cell line 

 IDG-SW3. METHODS: Due to the relative novelty and uncommon 

 culture conditions of IDG-SW3 versus other osteoblast-lineage 

 cell lines, effects of temperature and media formulation on 

 matrix deposition and osteocytogenesis were initially 

 characterized. Subsequently, the relative influence of 

 biochemical (β-glycerophosphate (βGP) and ascorbic 

 acid 2-phosphate (AA2P)) and mechanical stimulation on 

 osteocytogenesis was compared, with intermittent application of 

 low magnitude FSS generated by see-saw rocker. RESULTS: 

 βGP and AA2P supplementation were required for 

 mineralization and osteocytogenesis, with 33°C cultures 

 retaining a more osteoblastic phenotype and 37°C cultures 

 undergoing significantly higher osteocytogenesis. βGP 

 concentration positively correlated with calcium deposition, 

 whilst AA2P stimulated alkaline phosphatase (ALP) activity and 

 collagen deposition. We demonstrate that increasing βGP 

 concentration also significantly enhances osteocytogenesis as 

 quantified by the expression of green fluorescent protein linked 

 to Dmp1. Intermittent FSS (~0.06 Pa) rocker had no effect on 

 osteocytogenesis and matrix deposition. CONCLUSIONS: This work 

 demonstrates the suitability and ease with which IDG-SW3 can be 

 utilized in osteocytogenesis studies. IDG-SW3 mineralization was 

 only mediated through biochemical stimuli with no detectable 

 effect of low magnitude FSS. Osteocytogenesis of IDG-SW3 

 primarily occurred in mineralized areas, further demonstrating 

 the role mineralization of the bone extracellular matrix has in 

 osteocyte differentiation.},

keywords = {biomechanics, bone tissue engineering, extracellular matrix, Matrix mineralization, mechanobiology},

pubstate = {published},

tppubtype = {article}

}

@article{Lonsmann2023-pc,

title = {Biomarkers of type IV collagen turnover reflect disease 

 activity in patients with early-stage non-alcoholic fatty liver 

 (NAFL)},

author = {Ida L\onsmann and Jane I Grove and Asma Haider and Philip Kaye and Morten A Karsdal and Diana J Leeming and Guruprasad P Aithal},

year  = {2023},

date = {2023-08-01},

journal = {Biology (Basel)},

volume = {12},

number = {8},

abstract = {BACKGROUND: Identification of progressive liver disease 

 necessitates the finding of novel non-invasive methods to 

 identify and monitor patients in need of early intervention. 

 Investigating patients with early-liver injury may help identify 

 unique biomarkers. Early-liver injury is characterized by 

 remodeling of the hepatocyte basement membrane (BM) of the 

 extracellular matrix. Thus, we quantified biomarkers targeting 

 two distinct neo-epitopes of the major BM collagen, type IV 

 collagen (PRO-C4 and C4M), in patients spanning the non-alcoholic 

 fatty liver disease (NAFLD) spectrum. METHODS: We evaluated 

 PRO-C4 and C4M in a cross-sectional study with 97 patients with 

 NAFLD confirmed on histology. Serological levels of PRO-C4 and 

 C4M were quantified using validated competitive enzyme-linked 

 immunosorbent assays (ELISA). Using the fatty liver inhibition of 

 progression (FLIP) algorithm, we stratified patients into two 

 groups: non-alcoholic fatty liver (NAFL) and non-alcoholic 

 steatohepatitis (NASH). Biomarker levels were investigated in the 

 two groups in patients stratified by the NAFLD activity score 

 (NAS). In both groups, biomarker measurements were analyzed in 

 relation to histological scorings of steatosis, inflammation, 

 ballooning, and fibrosis. RESULTS: Patients had a body mass index 

 (BMI) of 30.9 $±$ 5.6 kg/m2, age of 53 $±$ 13 years and a NAS 

 range of 1-8. Upon stratification by FLIP, the NASH patients had 

 higher platelets, ALT, and AST levels than the NAFL group. Both PRO-C4 (p = 0.0125) and C4M (p = 0.003) increased with increasing 

 NAS solely within the NAFL group; however, a large variability 

 was present in the NASH group. Furthermore, both markers were significantly associated with lobular inflammation (p = 0.020 and p = 0.048) and steatosis (p = 0.004 and p = 0.015) in patients 

 with NAFL. CONCLUSIONS: This study found that type IV collagen 

 turnover increased with the increase in NAS in patients with 

 NAFL; however, this was not the case in patients with NASH. These 

 findings support the assessments of the BM turnover using 

 biomarkers in patients with early-disease development. These 

 biomarkers may be used to track specific processes involved in 

 the early pathobiology of NAFL.},

keywords = {basement membrane, biomarkers, collagen turnover, extracellular matrix, NAFLD},

pubstate = {published},

tppubtype = {article}

}

@article{Ashworth2020-so,

title = {Peptide gels of fully-defined composition and mechanics for 

 probing cell-cell and cell-matrix interactions in vitro},

author = {J C Ashworth and J L Thompson and J R James and C E Slater and S Pijuan-Galit\'{o} and K Lis-Slimak and R J Holley and K A Meade and A Thompson and K P Arkill and M Tassieri and A J Wright and G Farnie and C L R Merry},

year  = {2020},

date = {2020-01-01},

journal = {Matrix Biol.},

volume = {85-86},

pages = {15\textendash33},

publisher = {Elsevier BV},

abstract = {Current materials used for in vitro 3D cell culture are often 

 limited by their poor similarity to human tissue, batch-to-batch 

 variability and complexity of composition and manufacture. Here, 

 we present a ``blank slate'' culture environment based on a 

 self-assembling peptide gel free from matrix motifs. The gel can 

 be customised by incorporating matrix components selected to 

 match the target tissue, with independent control of mechanical 

 properties. Therefore the matrix components are restricted to 

 those specifically added, or those synthesised by encapsulated 

 cells. The flexible 3D culture platform provides full control 

 over biochemical and physical properties, allowing the impact of 

 biochemical composition and tissue mechanics to be separately 

 evaluated in vitro. Here, we demonstrate that the peptide gels 

 support the growth of a range of cells including human induced 

 pluripotent stem cells and human cancer cell lines. Furthermore, 

 we present proof-of-concept that the peptide gels can be used to 

 build disease-relevant models. Controlling the peptide gelator 

 concentration allows peptide gel stiffness to be matched to 

 normal breast (1 kPa), with higher stiffness favouring the 

 viability of breast cancer cells over normal breast cells. In 

 parallel, the peptide gels may be modified with matrix 

 components relevant to human breast, such as collagen I and 

 hyaluronan. The choice and concentration of these additions 

 affect the size, shape and organisation of breast epithelial 

 cell structures formed in co-culture with fibroblasts. This 

 system therefore provides a means of unravelling the individual 

 influences of matrix, mechanical properties and cell-cell 

 interactions in cancer and other diseases.},

keywords = {biomaterials, Cancer, extracellular matrix, Stem cells, Stiffness},

pubstate = {published},

tppubtype = {article}

}