Anna Sorushanova

Abstract Recent data suggest that collagen retains the memory of the tissue that it derives from. With this in mind, collagen (from bovine skin and tendon) sponges were fabricated with different crosslinking densities, and their physicochemical and biological properties were assessed. As the crosslinking density was increased, the resistance to collagenase digestion, denaturation temperature, compressive stress, and compressive modulus was significantly increased and the free amine content, % swelling, and human dermal fibroblast cytocompatibility were significantly reduced. The tendon‐collagen‐derived scaffolds exhibited significantly higher compressive stress and compressive modulus values and induced significantly higher human tenocyte DNA concentration and metabolic activity than the skin‐collagen‐derived scaffolds. In human tenocyte cultures on day 14, the 1 mM 4‐arm polyethylene‐glycol succinimidyl glutarate tendon‐collagen‐derived collagen sponges induced significantly higher collagen type III synthesis (as expected at early stages of physiological tendon healing) and downregulated actin alpha 2 (associated with myofibroblast differentiation) and the skin‐collagen‐derived collagen sponges induced significantly higher collagen type IV synthesis (found primarily at the dermal‐epidermal junction) and upregulated prolyl 4‐hydroxylase subunit alpha‐1 (associated with collagen biosynthesis and constitutes a target for antifibrotic compounds). Our data indicate that the tissue from which collagen is extracted should be considered in the development of medical devices.

Type I collagen

Myofibroblast

Human skin

10.1002/mba2.24

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Non-destructive determination of collagen fibril width in extruded collagen fibres by piezoresponse force microscopy

Biomedical Physics & Engineering Express (2017)

Arwa Bazaid Sabine M. Neumayer Anna Sorushanova Jill Guyonnet Dimitrios I. Zeugolis

Extruded collagen fibres are a promising platform for tissue engineering applications.Ensuring that the functional properties of the engineered tissues possess similar structural properties as native tissues is important for biomedical applications.Advanced imaging tools including scanning electron microscopy (SEM) and atomic force microscopy (AFM) have revealed the structural features of collagen fibrils within such fibres; however, these techniques often require modification steps that can alter the sample in the process.Here, lateral piezoresponse force microscopy (LPFM), which is sensitive to the polar orientation of piezoelectric collagen fibrils, is demonstrated as a promising tool to assess the width of individual fibrils and moreover map their organisation and polar orientation without altering the sample.Within the fibres studied, the collagen fibrils showed a highly anisotropic arrangement with preferred alignment along the length of the fibre.Fibril widths of 74 ± 18 nm and 73 ± 19 nm in untreated and bleached fibres, respectively, were measured from LPFM amplitude images.These values agreed with values from SEM (70 ± 10 nm) and AFM (71 ± 19 nm) measurements that could only be obtained from bleached fibres.

Collagen fibril

Collagen fiber

10.1088/2057-1976/aa85ec

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Citations (6)

Development and Characterisation of Three-Dimensional (3D) Cell Culture Models of Adrenocortical Carcinoma

Endocrinology (2024)

Sarah Feely Nathan Mullen Padraig Donlon Eileen Reidy Ritihaas Surya Challapalli

Adrenocortical carcinoma (ACC) is a rare malignancy of the adrenal cortex that is associated with a poor prognosis. Developing effective treatment options for ACC is challenging owing to the current lack of representative pre-clinical models. This study addressed this limitation by developing and characterising 3D cell cultures incorporating the ACC cell lines, MUC-1, HAC15 and H295R in a type I collagen matrix. ACC tissue samples were analysed by immunohistochemistry to determine the presence of type I collagen in the tumour microenvironment. Cell viability and proliferation were assessed using flow cytometry and confocal microscopy. mRNA expression of steroidogenic enzymes and steroid secretion was analysed by comparing the 3D and monolayer cell culture models. All cells were successfully cultured in a type I collagen matrix, which is highly expressed in the ACC tumour microenvironment and showed optimal viability until day 7. All three models showed increased metabolic and proliferative activity over time. 3D cell cultures were steroidogenic and demonstrated increased resistance to the gold-standard chemotherapy, mitotane compared to monolayer. The use of these models may lead to an improved understanding of disease pathology and provide a better representative platform for testing and screening of potential therapies.

10.1210/endocr/bqae159

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The past, present and future in scaffold-based tendon treatments

Advanced Drug Delivery Reviews (2014)

Alex Lomas Christina N. M. Ryan Anna Sorushanova Naledi Shologu Aikaterini I. Sideri

Scar tissue

10.1016/j.addr.2014.11.022

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Citations (190)

Scaffold and scaffold-free self-assembled systems in regenerative medicine

Biotechnology and Bioengineering (2015)

Dilip Thomas Diana Gaspar Anna Sorushanova Gesmi Milcovich Kyriakos Spanoudes

Self-assembly in tissue engineering refers to the spontaneous chemical or biological association of components to form a distinct functional construct, reminiscent of native tissue. Such self-assembled systems have been widely used to develop platforms for the delivery of therapeutic and/or bioactive molecules and various cell populations. Tissue morphology and functional characteristics have been recapitulated in several self-assembled constructs, designed to incorporate stimuli responsiveness and controlled architecture through spatial confinement or field manipulation. In parallel, owing to substantial functional properties, scaffold-free cell-assembled devices have aided in the development of functional neotissues for various clinical targets. Herein, we discuss recent advancements and future aspirations in scaffold and scaffold-free self-assembled devices for regenerative medicine purposes. Biotechnol. Bioeng. 2016;113: 1155-1163. © 2015 Wiley Periodicals, Inc.

Regenerative Medicine

10.1002/bit.25869

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Citations (39)

Sublethal hyperthermia decreases cellular proliferation and transiently disrupts steroidogenesis in adrenal cells

Endocrine Abstracts (2022)

Nathan Mullen Padraig Donlon Katen Duffy Sarah Feely Kate Warde

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

10.1530/endoabs.81.p644

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12407 Assessing Human Monocyte Migration And Co-culture With Adrenocortical Cancer Cells

Journal of the Endocrine Society (2024)

Cheng William Hong Shawna Feely Ram Challapalli Nathan Mullen Anna Sorushanova

Abstract Disclosure: C. Hong: None. S. Feely: None. R. Challapalli: None. N. Mullen: None. A. Sorushanova: None. C. Hantel: None. M.D. Griffin: None. M.C. Dennedy: None. Adrenocortical carcinoma (ACC) is a lipid, typically steroidogenic cancer which carries a 5-year prognosis of <10%. Therapeutic options are limited including surgery and mitotane – a poorly tolerated and efficacious insecticide. Recent data have demonstrated that the immune environment of adrenocortical carcinoma is deficient in lymphocytes while demonstrating a relative rich monocyte/macrophage population in the tumour microenvironment. We have investigated the gradient for migration of circulating peripheral monocytes to ACC. ACC cells were seeded in the bottom chamber of the transwell system. Monocytes were isolated from human peripheral blood, by negative selection using magnetic beads with high purity (>95%) and quality. Migration of monocytes at 24/48h was evaluated by identifying those remaining in the top chamber, the floating fraction in the bottom chamber and those adherent to ACC cells in the bottom chamber. Analysis of monocyte/macrophages was undertaken by flow cytometry using the following markers: Sytox Blue, CD45, CD14, CD16 and HLA-DR. At 24h, when compared to control conditions, more monocytes had migrated to the ACC cells seeded lower chamber and attached to the bottom. Similar results were observed in migration at 48h. Predominantly Classical monocytes (CD14++CD16-HLA-DR+) migrated to metastatic cancer cells (MUC1), and Non-Classical monocytes (CD14+CD16+HLA-DR+) migrated to primary cancer cells (H295R/ HAC15) at 48h. We then investigated the polarization cytokine profile of migrated monocytes with or without ACC cells at 72 h by multiplex bead-based assay. Migrated monocytes indicated a significant shift toward the M2 phenotype in the presence of H295R or HAC15. The production of the pro-inflammatory factors of IL-6, TNF-alpha, etc. decreased and immunosuppressive factors of IL-10, TARC, etc. increased. MUC1 didn’t increase M2 or M1 cytokine secretion. Moreover, surface markers of migrated monocytes were evaluated at 72h by flow cytometry. When compared to the control group, all 3 ACC cell lines switched monocytes/ macrophages into M2 phenotype, as evidenced by the decrease in the expression of the M1 marker CD86 and the increase in the expression of the M2 markers CD163 and CD206. We demonstrate monocytes migrate and associate with ACC cells: (i) subpopulations differ in transmigration properties in response to ACC cells (ii) monocytes/macrophages exhibit M2 polarization phenotype. Presentation: 6/2/2024

Monocyte

10.1210/jendso/bvae163.2223

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12407 Assessing Human Monocyte Migration And Co-culture With Adrenocortical Cancer Cells

Journal of the Endocrine Society (2024)

Chang‐Ki Hong Shawna Feely Ritihaas Surya Challapalli Nathan Mullen Anna Sorushanova

Abstract Disclosure: C. Hong: None. S. Feely: None. R. Challapalli: None. N. Mullen: None. A. Sorushanova: None. C. Hantel: None. M.D. Griffin: None. M.C. Dennedy: None. Adrenocortical carcinoma (ACC) is a lipid, typically steroidogenic cancer which carries a 5-year prognosis of <10%. Therapeutic options are limited including surgery and mitotane – a poorly tolerated and efficacious insecticide. Recent data have demonstrated that the immune environment of adrenocortical carcinoma is deficient in lymphocytes while demonstrating a relative rich monocyte/macrophage population in the tumour microenvironment. We have investigated the gradient for migration of circulating peripheral monocytes to ACC. ACC cells were seeded in the bottom chamber of the transwell system. Monocytes were isolated from human peripheral blood, by negative selection using magnetic beads with high purity (>95%) and quality. Migration of monocytes at 24/48h was evaluated by identifying those remaining in the top chamber, the floating fraction in the bottom chamber and those adherent to ACC cells in the bottom chamber. Analysis of monocyte/macrophages was undertaken by flow cytometry using the following markers: Sytox Blue, CD45, CD14, CD16 and HLA-DR. At 24h, when compared to control conditions, more monocytes had migrated to the ACC cells seeded lower chamber and attached to the bottom. Similar results were observed in migration at 48h. Predominantly Classical monocytes (CD14++CD16-HLA-DR+) migrated to metastatic cancer cells (MUC1), and Non-Classical monocytes (CD14+CD16+HLA-DR+) migrated to primary cancer cells (H295R/ HAC15) at 48h. We then investigated the polarization cytokine profile of migrated monocytes with or without ACC cells at 72 h by multiplex bead-based assay. Migrated monocytes indicated a significant shift toward the M2 phenotype in the presence of H295R or HAC15. The production of the pro-inflammatory factors of IL-6, TNF-alpha, etc. decreased and immunosuppressive factors of IL-10, TARC, etc. increased. MUC1 didn’t increase M2 or M1 cytokine secretion. Moreover, surface markers of migrated monocytes were evaluated at 72h by flow cytometry. When compared to the control group, all 3 ACC cell lines switched monocytes/ macrophages into M2 phenotype, as evidenced by the decrease in the expression of the M1 marker CD86 and the increase in the expression of the M2 markers CD163 and CD206. We demonstrate monocytes migrate and associate with ACC cells: (i) subpopulations differ in transmigration properties in response to ACC cells (ii) monocytes/macrophages exhibit M2 polarization phenotype. Presentation: 6/2/2024

Monocyte

10.1210/jendso/bvae163.2139

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The Collagen Suprafamily: From Biosynthesis to Advanced Biomaterial Development

Advanced Materials (2018)

Anna Sorushanova Luis M. Delgado Zhuning Wu Naledi Shologu Aniket Kshirsagar

Collagen is the oldest and most abundant extracellular matrix protein that has found many applications in food, cosmetic, pharmaceutical, and biomedical industries. First, an overview of the family of collagens and their respective structures, conformation, and biosynthesis is provided. The advances and shortfalls of various collagen preparations (e.g., mammalian/marine extracted collagen, cell-produced collagens, recombinant collagens, and collagen-like peptides) and crosslinking technologies (e.g., chemical, physical, and biological) are then critically discussed. Subsequently, an array of structural, thermal, mechanical, biochemical, and biological assays is examined, which are developed to analyze and characterize collagenous structures. Lastly, a comprehensive review is provided on how advances in engineering, chemistry, and biology have enabled the development of bioactive, 3D structures (e.g., tissue grafts, biomaterials, cell-assembled tissue equivalents) that closely imitate native supramolecular assemblies and have the capacity to deliver in a localized and sustained manner viable cell populations and/or bioactive/therapeutic molecules. Clearly, collagens have a long history in both evolution and biotechnology and continue to offer both challenges and exciting opportunities in regenerative medicine as nature's biomaterial of choice.

Biomaterial

10.1002/adma.201801651

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Citations (846)

2.15 Collagen: Materials Analysis and Implant Uses ☆

Elsevier eBooks (2017)

Anna Sorushanova João Q. Coentro Abhay Pandit Dimitrios I. Zeugolis Michael Raghunath

Lysyl Oxidase

Type I collagen

Biomaterial

10.1016/b978-0-12-803581-8.10155-9

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