Publications by authors named "Payal Ganguly"

9 Publications

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Electrospun and 3D printed polymeric materials for one-stage critical-size long bone defect regeneration inspired by the Masquelet technique: Recent Advances.

Injury 2022 Feb 16. Epub 2022 Feb 16.

Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, UK; Leeds General Infirmary, Department of Trauma and Orthopaedic Surgery, University of Leeds, Leeds, UK. Electronic address:

Critical-size long bone defects represent one of the major causes of fracture non-union and remain a significant challenge in orthopaedic surgery. Two-stage procedures such as a Masquelet technique demonstrate high level of success however their main disadvantage is the need for a second surgery, which is required to remove the non-resorbable cement spacer and to place the bone graft into the biological chamber formed by the 'induced membrane'. Recent research efforts have therefore been dedicated towards the design, fabrication and testing of resorbable implants that could mimic the biological functions of the cement spacer and the induced membrane. Amongst the various manufacturing techniques used to fabricate these implants, three-dimensional (3D) printing and electrospinning methods have gained a significant momentum due their high-level controllability, scalable processing and relatively low cost. This review aims to present recent advances in the evaluation of electrospun and 3D printed polymeric materials for critical-size, long bone defect reconstruction, emphasizing both their beneficial properties and current limitations. Furthermore, we present and discuss current state-of-the art techniques required for characterisation of the materials' physical, mechanical and biological characteristics. These represent the essential first steps towards the development of personalised implants for single-surgery, large defect reconstruction in weight-bearing bones.
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http://dx.doi.org/10.1016/j.injury.2022.02.036DOI Listing
February 2022

Evaluation of Human Bone Marrow Mesenchymal Stromal Cell (MSC) Functions on a Biomorphic Rattan-Wood-Derived Scaffold: A Comparison between Cultured and Uncultured MSCs.

Bioengineering (Basel) 2021 Dec 21;9(1). Epub 2021 Dec 21.

Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS9 7TF, UK.

The reconstruction of large bone defects requires the use of biocompatible osteoconductive scaffolds. These scaffolds are often loaded with the patient's own bone marrow (BM) cells to facilitate osteoinductivity and biological potency. Scaffolds that are naturally sourced and fabricated through biomorphic transitions of rattan wood (B-HA scaffolds) offer a unique advantage of higher mechanical strength and bioactivity. In this study, we investigated the ability of a biomorphic B-HA scaffold (B-HA) to support the attachment, survival and gene expression profile of human uncultured BM-derived mesenchymal stromal cells (BMSCs, = 6) and culture expanded MSCs (cMSCs, = 7) in comparison to a sintered, porous HA scaffold (S-HA). B-HA scaffolds supported BMSC attachment (average 98%) and their survival up to 4 weeks in culture. Flow cytometry confirmed the phenotype of cMSCs on the scaffolds. Gene expression indicated clear segregation between cMSCs and BMSCs with MSC osteogenesis- and adipogenesis-related genes including and being higher expressed in BMSCs. These data indicated a unique transcriptional signature of BMSCs that was distinct from that of cMSCs regardless of the type of scaffold or time in culture. There was no statistical difference in the expression of osteogenic genes in BMSCs or cMSCs in B-HA compared to S-HA. VEGF release from cMSCs co-cultured with human endothelial cells ( = 4) on B-HA scaffolds suggested significantly higher supernatant concentration with endothelial cells on day 14. This indicated a potential mechanism for providing vasculature to the repair area when such scaffolds are used for treating large bone defects.
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http://dx.doi.org/10.3390/bioengineering9010001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8773042PMC
December 2021

Aging, Bone Marrow and Next-Generation Sequencing (NGS): Recent Advances and Future Perspectives.

Int J Mol Sci 2021 Nov 12;22(22). Epub 2021 Nov 12.

Novogene Europe, 25 Science Park, Milton, Cambridge CB4 0FW, UK.

The aging of bone marrow (BM) remains a very imperative and alluring subject, with an ever-increasing interest among fellow scientists. A considerable amount of progress has been made in this field with the established 'hallmarks of aging' and continued efforts to investigate the age-related changes observed within the BM. Inflammaging is considered as a low-grade state of inflammation associated with aging, and whilst the possible mechanisms by which aging occurs are now largely understood, the processes leading to the underlying changes within aged BM remain elusive. The ability to identify these changes and detect such alterations at the genetic level are key to broadening the knowledgebase of aging BM. Next-generation sequencing (NGS) is an important molecular-level application presenting the ability to not only determine genomic base changes but provide transcriptional profiling (RNA-seq), as well as a high-throughput analysis of DNA-protein interactions (ChIP-seq). Utilising NGS to explore the genetic alterations occurring over the aging process within alterative cell types facilitates the comprehension of the molecular and cellular changes influencing the dynamics of aging BM. Thus, this review prospects the current landscape of BM aging and explores how NGS technology is currently being applied within this ever-expanding field of research.
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http://dx.doi.org/10.3390/ijms222212225DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8620539PMC
November 2021

Bone Marrow Multipotent Mesenchymal Stromal Cells as Autologous Therapy for Osteonecrosis: Effects of Age and Underlying Causes.

Bioengineering (Basel) 2021 May 17;8(5). Epub 2021 May 17.

Leeds Institute of Rheumatic and Musculoskeletal Medicine, School of Medicine, University of Leeds, Leeds LS2 9JT, UK.

Bone marrow (BM) is a reliable source of multipotent mesenchymal stromal cells (MSCs), which have been successfully used for treating osteonecrosis. Considering the functional advantages of BM-MSCs as bone and cartilage reparatory cells and supporting angiogenesis, several donor-related factors are also essential to consider when autologous BM-MSCs are used for such regenerative therapies. Aging is one of several factors contributing to the donor-related variability and found to be associated with a reduction of BM-MSC numbers. However, even within the same age group, other factors affecting MSC quantity and function remain incompletely understood. For patients with osteonecrosis, several underlying factors have been linked to the decrease of the proliferation of BM-MSCs as well as the impairment of their differentiation, migration, angiogenesis-support and immunoregulatory functions. This review discusses the quality and quantity of BM-MSCs in relation to the etiological conditions of osteonecrosis such as sickle cell disease, Gaucher disease, alcohol, corticosteroids, Systemic Lupus Erythematosus, diabetes, chronic renal disease and chemotherapy. A clear understanding of the regenerative potential of BM-MSCs is essential to optimize the cellular therapy of osteonecrosis and other bone damage conditions.
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http://dx.doi.org/10.3390/bioengineering8050069DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8156020PMC
May 2021

Intrinsic Type 1 Interferon (IFN1) Profile of Uncultured Human Bone Marrow CD45CD271 Multipotential Stromal Cells (BM-MSCs): The Impact of Donor Age, Culture Expansion and IFNα and IFNβ Stimulation.

Biomedicines 2020 Jul 15;8(7). Epub 2020 Jul 15.

Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds LS9 7TF, UK.

Skeletal aging is associated with reduced proliferative potential of bone marrow (BM) multipotential stromal cells (MSCs). Recent data suggest the involvement of type 1 interferon (IFN1) signalling in hematopoietic stem cell (HSC) senescence. Considering that BM-HSCs and BM-MSCs share the same BM niche, we investigated IFN1 expression profile in human BM-MSCs in relation to donor age, culture-expansion and IFN1 (α and β) stimulation. Fluorescence-activated cell sorting was used to purify uncultured BM-MSCs from younger (19-41, = 6) and older (59-89, = 6) donors based on the CD45CD271 phenotype, and hematopoietic-lineage cells (BM-HLCs, CD45CD271) were used as controls. Gene expression was analysed using integrated circuits arrays in sorted fractions as well as cultured/stimulated BM-MSCs and Y201/Y202 immortalised cell lines. IFN1 stimulation led to BM-MSC growth arrest and upregulation of many IFN1-stimulated genes (ISGs), with IFNβ demonstrating stronger effects. Uncultured MSCs were characterised by a moderate-level ISG expression similar to Y201 cells. Age-related changes in ISG expression were negligible in BM-MSCs compared to BM-HLCs, and intracellular reactive oxygen species (ROS) levels in BM-MSCs did not significantly correlate with donor age. Antiaging genes Klotho and SIRT6 correlated with more ISGs in BM-MSCs than in BM-HLCs. In patients with osteoarthritis (OA), BM-MSCs expressed considerably lower levels of several ISGs, indicating that their IFN1 signature is affected in a pathological condition. In summary, BM-MSCs possess homeostatic IFN1 gene expression signature in health, which is sensitive to in vitro culture and external IFN1 stimulation. IFN signalling may facilitate in vivo BM-MSC responses to DNA damage and combating senescence and aberrant immune activation.
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http://dx.doi.org/10.3390/biomedicines8070214DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7399891PMC
July 2020

Colony Formation, Migratory, and Differentiation Characteristics of Multipotential Stromal Cells (MSCs) from "Clinically Accessible" Human Periosteum Compared to Donor-Matched Bone Marrow MSCs.

Stem Cells Int 2019 21;2019:6074245. Epub 2019 Nov 21.

Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, UK.

Periosteum is vital for fracture healing, as a highly vascular and multipotential stromal cell- (MSC-) rich tissue. During surgical bone reconstruction, small fragments of periosteum can be "clinically accessible," yet periosteum is currently not ultilised, unlike autologous bone marrow (BM) aspirate. This study is aimed at comparing human periosteum and donor-matched iliac crest BM MSC content and characterising MSCs in terms of colony formation, growth kinetics, phenotype, cell migration patterns, and trilineage differentiation capacity. "Clinically accessible" periosteum had an intact outer fibrous layer, containing CD271+ candidate MSCs located perivasculary; the inner cambium was rarely present. Following enzymatic release of cells, periosteum formed significantly smaller fibroblastic colonies compared to BM (6.1 mm vs. 15.5 mm, = 4, = 0.0006). Periosteal colonies were more homogenous in size (range 2-30 mm vs. 2-54 mm) and on average 2500-fold more frequent (2.0% vs. 0.0008%, = 10, = 0.004) relative to total viable cells. When expanded , similar growth rates up to passage 0 (P0) were seen (1.8 population doublings (PDs) per day (periosteum), 1.6 PDs per day (BM)); however, subsequently BM MSCs proliferated significantly slower by P4 (4.3 PDs per day (periosteum) vs. 9.3 PDs per day (BM), = 9, = 0.02). In early culture, periosteum cells were less migratory at slower speeds than BM cells. Both MSC types exhibited MSC phenotype and trilineage differentiation capacity; however, periosteum MSCs showed significantly lower (2.7-fold) adipogenic potential based on Nile red : DAPI ratios with reduced expression of adipogenesis-related transcripts . Altogether, these data revealed that "clinically accessible" periosteal samples represent a consistently rich source of highly proliferative MSCs compared to donor-matched BM, which importantly show similar osteochondral capacity and lower adipogenic potential. Live cell tracking allowed determination of unique morphological and migration characteristics of periosteal MSCs that can be used for the development of novel bone graft substitutes to be preferentially repopulated by these cells.
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http://dx.doi.org/10.1155/2019/6074245DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6906873PMC
November 2019

The Biological Fitness of Bone Progenitor Cells in Reamer/Irrigator/Aspirator Waste.

J Bone Joint Surg Am 2019 Dec;101(23):2111-2119

Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, United Kingdom.

Background: The biological waste collected during use of the Reamer/Irrigator/Aspirator (RIA; DePuy Synthes) has been described as an abundant source of bone progenitor cells with a comparable osteogenic gene profile to donor-matched iliac crest bone marrow (IC-BM). However, it is not clear whether these RIA-waste (RIA-W) cells are biologically fit. We aimed to evaluate the stress levels and functions of RIA-W progenitor cells.

Methods: Reactive oxygen species (ROS) levels were tested in freshly collected bone progenitor cells (defined as CD45CD271 cells) using flow cytometry. ROS levels induced in these cells by hypoxia and/or oxidative stress as well as by an experimental simulation of the RIA procedure were also measured. Furthermore, the alkaline phosphatase (ALP) expression levels, proliferation, and senescence of culture-expanded RIA-W and IC-BM mesenchymal stromal cells (MSCs) were compared.

Results: RIA-W and donor-matched IC-BM CD45CD271 cells were 97% and 98% viable, but the ROS levels were significantly higher for RIA-W cells than for IC-BM cells (p = 0.0020). Also, ROS induced by hypoxia, oxidative stress, and both were higher for RIA-W cells (p = 0.0312, 0.0156, and 0.0703, respectively). Dilution with saline solution, suction pressure, and irrigation reduced cell viability, with a positive correlation with the ROS level (p = 0.0035). The RIA-W and IC-BM colony-forming cells (average, 96,100 and 11,500, respectively) showed comparable ALP levels. Furthermore, culture-expanded RIA-W and IC-BM MSCs showed comparable ROS levels, ALP levels, susceptibility to death, and proliferation.

Conclusions: Although freshly collected RIA-W bone progenitor cells appeared to be transiently stressed, these cells were as viable as IC-BM cells and present in greater numbers. The proliferation and osteogenesis of both cell types were comparable.

Clinical Relevance: The RIA waste bag contains bone progenitor cells with promising potential for regenerative applications, and should not be wasted.
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http://dx.doi.org/10.2106/JBJS.19.00133DOI Listing
December 2019

The Analysis of Aging in Human Bone Marrow Mesenchymal Stromal Cells Using Colony-Forming Unit-Fibroblast Assay and the CD45CD271 Phenotype.

Stem Cells Int 2019 1;2019:5197983. Epub 2019 Aug 1.

Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, UK.

Uncultured mesenchymal stromal cells (MSCs) are increasingly used in therapies; however, the effects of donor age on their biological characteristics and gene expression remain unclear. The aim of this study was to investigate age-related changes in bone marrow (BM) MSCs following minimal or no culture manipulation. Iliac crest BM was aspirated from 67 healthy donors (19-89 years old) and directly used for the colony-forming unit-fibroblast (CFU-F) assay or CD45CD271 cell enumeration. The colonies were analysed for colony area and integrated density (ID) when grown in standard MSC media or media supplemented with human serum from young (YS) or old (OS) donors. There was a notable age-related decline in the number of MSCs per millilitre of BM aspirate revealed by the CFU-F assay ( = -0.527, < 0.0001) or flow cytometry ( = -0.307, = 0.0116). Compared to young donors (19-40 years old), colony IDs were significantly lower in older donors (61-89 years old), particularly for smaller-sized colonies (42% lower, < 0.01). When cultured in media supplemented with OS, young and old donor MSCs formed colonies with lower IDs, by 21%, < 0.0001, and 27%, < 0.05, respectively, indicating the formation of smaller sparser colonies. No significant differences in the expression of selected adipogenic, osteogenic, stromal, and bone remodelling genes as well as CD295, CD146, CD106, and connexin 43 surface molecules were found in sorted CD45CD271 MSCs from young and old donors ( = 8 donors each). Altogether, these results show similar trends for age-related decline in BM MSC numbers measured by the CFU-F assay and flow cytometry and reveal age-related effects of human serum on MSC colony formation. No significant differences in selected gene expression in uncultured CD45CD271 MSCs suggest that old donor MSCs may not be inferior in regard to their multipotential functions. Due to large donor-to-donor variation in all donor groups, our data indicate that an individual's chronological age is not a reliable predictor of their MSC number or potency.
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http://dx.doi.org/10.1155/2019/5197983DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6701348PMC
August 2019

Age-related Changes in Bone Marrow Mesenchymal Stromal Cells: A Potential Impact on Osteoporosis and Osteoarthritis Development.

Cell Transplant 2017 09;26(9):1520-1529

1 Leeds Institute of Rheumatic and Musculoskeletal Medicine, University of Leeds, Leeds, United Kingdom.

Aging at the cellular level is a complex process resulting from accumulation of various damages leading to functional impairment and a reduced quality of life at the level of the organism. With a rise in the elderly population, the worldwide incidence of osteoporosis (OP) and osteoarthritis (OA) has increased in the past few decades. A decline in the number and "fitness" of mesenchymal stromal cells (MSCs) in the bone marrow (BM) niche has been suggested as one of the factors contributing to bone abnormalities in OP and OA. It is well recognized that MSCs in vitro acquire culture-induced aging features such as gradual telomere shortening, increased numbers of senescent cells, and reduced resistance to oxidative stress as a result of serial population doublings. In contrast, there is only limited evidence that human BM-MSCs "age" similarly in vivo. This review compares the various aspects of in vitro and in vivo MSC aging and suggests how our current knowledge on rejuvenating cultured MSCs could be applied to develop future strategies to target altered bone formation processes in OP and OA.
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http://dx.doi.org/10.1177/0963689717721201DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5680949PMC
September 2017
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