scholarly journals 4485 Silicone Implant Shells Increase the Rate of Proliferation of Patient-Derived BIA-ALCL Cells but Not Primary T Cells in an Engineered Biomimetic Breast Platform

2020 ◽  
Vol 4 (s1) ◽  
pp. 113-113
Author(s):  
Ishani Premaratne ◽  
Matthew Wright ◽  
Mariam Gadjiko ◽  
Daniel Lara ◽  
Arash Samadi ◽  
...  
Keyword(s):  
T Cells ◽  
Cell Count ◽  
Progenitor Cells ◽  
Type I Collagen ◽  
Breast Implant ◽  
Stromal Vascular Fraction ◽  
Large Cell ◽  
Silicone Implant ◽  
Seeding Density ◽  
Type I

OBJECTIVES/GOALS: We use a tissue engineered, biomimetic, 3D model to study the pathogenesis of breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) by comparing the effect of silicone implant shell on proliferation of patient-derived BIA-ALCL to its precursor T cells within the breast microenvironment. METHODS/STUDY POPULATION: Patient-derived breast tissue was processed for component adipocytes, ductal organoids, and stromal vascular fraction. These were suspended within 50 µl of 0.3% type I collagen matrix to which was added 200,000 cells/mL of either patient-derived BIA-ALCL cells or T progenitor cells. These were then plated into 6mm wells. As a control, both BIA-ALCL cells and T progenitor cells were suspended within type I collagen alone at the same seeding density without breast components. Before plating, wells were lined circumferentially with either textured, smooth, or no implant shell. These were 1cm by 2cm pieces dissected from the whole implant. Wells were imaged using confocal microscopy over 8 days. RESULTS/ANTICIPATED RESULTS: Unstimulated T progenitor cell count showed no significant increase in any of the conditions tested. The change in cell count over 8 days was 3.85% in each condition (p = 0.3352). A Tukey’s multiple comparison test comparing each condition revealed no significant increase in cell count over 8 days for all six conditions. Notably, our previous studies have shown proliferation of BIA-ALCL cells to be significantly more robust in the biomimetic platform compared to collagen-only groups, regardless of implant shell type (p < 0.01). BIA-ALCL cells grew nearly 30% faster in textured and smooth shell biomimetic groups compared to biomimetic wells lacking implant shell. DISCUSSION/SIGNIFICANCE OF IMPACT: Towards elucidating BIA-ALCL’s etiopathology, we show that silicone implant shell has a significant effect on proliferation of BIA-ALCL cells, but not their precursor T cells. If breast implant silicone shell is not a sufficient stimulus for T cell proliferation, co-stimulatory factors are required.

Hyperactivation of RAP1 and JAK/STAT Signaling Pathways Contributes to Fibrosis during the Formation of Nasal Capsular Contraction

2021 ◽  
pp. 1-12
Author(s):  
Meng Wu ◽  
Ming Li ◽  
Hong-Ju Xie ◽  
Hong-Wei Liu
Keyword(s):  
Signaling Pathways ◽  
Type I Collagen ◽  
Ex Vivo ◽  
Capsular Contracture ◽  
Silicone Implant ◽  
Type I ◽  
Loss Of Function ◽  
Sequencing Data ◽  
Functional Changes ◽  
Stat Signaling

Silicone implant-based augmentation rhinoplasty or mammoplasty induces capsular contracture, which has been acknowledged as a process that develops an abnormal fibrotic capsule associated with the immune response to allogeneic materials. However, the signaling pathways leading to the nasal fibrosis remain poorly investigated. We aimed to explore the molecular mechanism underlying the pathogenesis of nasal capsular contracture, with a specific research interest in the signaling pathways involved in fibrotic development at the advanced stage of contracture. By examining our recently obtained RNA sequencing data and global gene expression profiling between grade II and grade IV nasal capsular tissues, we found that both the RAP1 and JAK/STAT signaling pathways were hyperactive in the contracted capsules. This was verified on quantitative real-time PCR which demonstrated upregulation of most of the representative component signatures in these pathways. Loss-of-function assays through siRNA-mediated Rap1 silencing and/or small molecule-directed inhibition of JAK/STAT pathway in ex vivo primary nasal fibroblasts caused a series of dramatic behavioral and functional changes, including decreased cell viability, increased apoptosis, reduced secretion of proinflammatory cytokines, and synthesis of type I collagen, compared to control cells, and indicating the essential role of the RAP1 and JAK/STAT signaling pathways in nasal capsular fibrosis. Our results sheds light on targeting downstream signaling pathways for the prevention and therapy of silicone implant-induced nasal capsular contracture.

A Cautionary Tale and Update on Breast Implant–Associated Anaplastic Large Cell Lymphoma (BIA-ALCL)

2020 ◽  
Vol 40 (12) ◽  
pp. 1288-1300 ◽  
Author(s):  
Trina Ghosh ◽  
Eric Duncavage ◽  
Neha Mehta-Shah ◽  
Patricia A McGuire ◽  
Marissa Tenenbaum ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Cell Lymphoma ◽  
Breast Implant ◽  
Large Cell ◽  
World Health ◽  
Multidisciplinary Teams ◽  
Level Of Evidence ◽  
Anaplastic Lymphoma

Abstract Breast implant–associated anaplastic large T-cell lymphoma (BIA-ALCL) was first recognized by the World Health Organization in 2016. The total number of cases worldwide continues to increase, with &gt;800 cases confirmed through a combination of Food and Drug Administration data, verified reports, and registries. To date, 33 deaths have been reported. Typical presentation includes a late seroma containing monoclonal T cells that are CD30 positive and anaplastic lymphoma kinase negative. We present a review of the current literature and report on 3 cases of BIA-ALCL at our institution, which serve to illustrate our approach to diagnosis and management of this disease. In 2 cases, the diagnosis of BIA-ALCL was not initially confirmed due to an incomplete workup but was recognized upon explantation. The seroma fluid was sent for flow cytometry. Initially, the cells were reported as morphologically suspicious for malignancy with phenotypically normal T cells based on standard CD3+ T-cell gating. Subsequent cytology specimens were reported as consistent with recurrent adenocarcinoma. However, upon regating of flow-cytometry data, a population of CD30+, CD3– T cells was noted and the diagnosis of BIA-ALCL was confirmed by immunohistochemical stains of the excised breast capsule specimen. Given the increasing incidence of this disease, as plastic surgeons we must stay informed to order the correct workup to avoid misdiagnosis and be prepared to appropriately refer affected patients to centers with multidisciplinary teams experienced in the management of BIA-ALCL. Level of Evidence: 4

scholarly journals IL-1/TNF-αInflammatory and Anti-Inflammatory Synchronization Affects Gingival Stem/Progenitor Cells’ Regenerative Attributes

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Fan Zhang ◽  
Misi Si ◽  
Huiming Wang ◽  
Mohamed K. Mekhemar ◽  
Christof E. Dörfer ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Osteogenic Differentiation ◽  
Progenitor Cells ◽  
Type I Collagen ◽  
Basic Medium ◽  
Control Group ◽  
Type I ◽  
Alizarin Red ◽  
Cell Numbers ◽  
Anti Inflammatory

Cytokines play major roles in tissue destruction/repair. The present study investigates proliferative and osteogenic differentiation potentials of gingival mesenchymal stem/progenitor cells (G-MSCs), influenced by IL-1/TNF-αinflammatory/anti-inflammatory conditions. Human G-MSCs were isolated, characterized, and cultured in basic medium (control group, M1), in basic medium with IL-1β, TNF-α, and IFN-γ(inflammatory group, M2) and with IL-1ra/TNF-αi added to M2 (anti-inflammatory group, M3). MTT tests at days 1, 3, and 7 and CFU assay at day 12 were conducted. Osteogenic differentiation was analyzed by bone-specific transcription factors (RUNX2), alkaline phosphatase (ALP), type I collagen (Col-I), osteopontin (OPN), and osteonectin (ON) expression at days 1, 3, 7, and 14 and Alizarin red staining at day 14. At day 3, the control group showed the highest cell numbers. At day 7, cell numbers in inflammatory and anti-inflammatory group outnumbered the control group. At day 12, CFUs decreased in the inflammatory and anti-inflammatory groups, with altered cellular morphology. The anti-inflammatory group demonstrated elevated bone-specific transcription factors at 14 days. After 14 days of osteogenic induction, calcified nodules in the anti-inflammatory group were higher compared to control and inflammatory groups. For regeneration, initial inflammatory stimuli appear essential for G-MSCs’ proliferation. With inflammatory persistence, this positive effect perishes and is followed by a short-term stimulatory one on osteogenesis. At this stage, selective anti-inflammatory intervention could boost G-MSCs’ differentiation.

scholarly journals Syndecan-4 Functionalization of Tissue Regeneration Scaffolds Improves Interaction with Endothelial Progenitor Cells

2021 ◽  
Author(s):  
Harleigh Warner ◽  
Yidi Wu ◽  
William D Wagner
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Tissue Regeneration ◽  
Binding Sites ◽  
Type I Collagen ◽  
Type I ◽  
Endothelial Progenitor ◽  
Hybrid Scaffold ◽  
Scaffold Materials

Abstract Key to most implanted cell free scaffolds for tissue regeneration is the ability to sequester and retain undifferentiated mesenchymal stem cells at the repair site. In this report, syndecan-4, a heparan sulfate containing proteoglycan, was investigated as a unique molecule for use in scaffold functionalization. An electrospun hybrid scaffold comprised of poly (glycerol) sebacate (PGS), silk fibroin and type I collagen (PFC) was used as a model scaffold to develop a procedure and test the hypothesis that functionalization would result in increased scaffold binding of endothelial progenitor cells (EPCs). For these studies both Syndecan-4 and stromal derived factor-1α (SDF-1α) were used in functionalization PFC. Syndecan-4 functionalized PFC bound 4.8 fold more SDF-1α compared to nonfunctionalized PFC. Binding was specific as determined by heparin displacement studies. After culture for 7 days, significantly, more EPCs were detected on PFC scaffolds having both syndecan-4 and SDF-1α compared to scaffolds of PFC with only syndecan-4, or PFC adsorbed with SDF-1α, or PFC alone. Taken together, this study demonstrates that EPCs can be bound to and significantly expanded on PFC material through syndecan-4 mediated growth factor binding. Syndecan-4 with a multiplicity of binding sites has the potential to functionalize and expand stem cells on a variety of scaffold materials for use in tissue regeneration.

Key Survival Pathways and Chemotherapeutic Sensitivity of Breast Implant Associated Anaplastic Large Cell Lymphomas,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3739-3739
Author(s):  
Melissa G Lechner ◽  
Connor Church ◽  
Carolina Megiel ◽  
Rikki S Bass ◽  
Garry S Brody ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cell Lines ◽  
Primary Tumor ◽  
Breast Implant ◽  
Large Cell ◽  
Gene Rearrangements ◽  
Tumor Biopsy ◽  
Biopsy Specimens

Abstract Abstract 3739 Primary lymphomas of the breast are very rare (0.2–1.5% of breast malignancies) and the vast majority (95%) is of B-cell origin. Recently, a series of anaplastic large-cell kinase (ALK)-negative, T-cell, anaplastic, non-Hodgkin lymphomas (T-ALCL) have been reported worldwide in patients with saline and silicone breast implants (Lechner et al. Cancer 2010, Lazzeri D et al. Clin Breast Cancer 2011, Carty et al. Plast Reconstr Surg 2011, Popplewell et al. Leuk Lymphoma 2011, Roden et al. Mod Pathol 2008, de Jong et al. JAMA 2008, and others). These cases are striking for their homogeneous clinical presentation and pathology. Histological and cytogenetic analyses of the primary tumor biopsy specimens consistently demonstrates CD30+ EMA(+) PAX5− lymphoma cells with significant chromosomal atypia that lack the NPM-ALK (2;5) translocation, express pan T-cell markers, and show monoclonal TCR gene rearrangements. Increasing evidence suggests that these seroma-associated ALK− ALCLs are a distinct clinicopathologic entity from either systemic ALK− ALCL or cutaneous ALCL, and while the majority of cases are indolent, several patients have died from this disease. In order to understand the nature of this newly emerging clinical entity, we have established three new cell lines, (TLBR-1, TLBR-2, TLBR-3,) from the primary tumor biopsy specimens of three patients diagnosed with breast implant-associated ALCL. Characterization of these pre-clinical models confirmed fidelity to the original tumor biopsy specimens and highlighted unique features that may aid the development of effective treatments for these cancers. Cytogenetics and FISH performed on TLBR-1, -2, and -3 revealed nuclear atypia with partial or complete trisomy (modal chromosome number 47, 76, and 81, respectively) and absence of the NPM-ALK (2;5) translocation. Phenotypic characterization of TLBR cell lines by flow cytometry and immunocytochemistry showed strong positivity for CD30, CD71, pan T cell CD2/5/7, natural killer (CD56), and antigen presentation (HLA-DR, CD80, CD86) markers, and IL-2 receptors (CD25, CD122). Multiplex or consensus sequence PCR confirmed monoclonal TCR gene rearrangements and showed no onco-virus incorporation (EBV, HTLV1/2), respectively, for TLBR-1, -2, and -3. To identify effective therapies for patients with breast-implant associated ALCL, in vitro studies were used to evaluate the mechanisms by which TLBR evades immune detection and promotes cell cycle dysregulation. Gene expression analysis for proto-oncogenes, tumor suppressor genes, and regulators of apoptosis demonstrated significant up-regulation of survivin and down-regulation of pro-apoptotic genes (BID, BAK, BBC3) by all TLBR cell lines relative to healthy donor T cells. Interestingly, TLBR cell lines shared many features with human regulatory T cells, including phenotype (CD4+/8+, CD25+), high FoxP3 expression, up-regulation of immunosuppressive cytokines IL-10 and TGFβ, and suppression of T cell responses to stimuli. Inhibitor studies were used to identify critical signaling pathways in survival and proliferation of breast implant-associated ALCL models TLBR-1, -2, and -3. Data from these studies revealed that inhibition of the JAK/STAT, PI3K/mTOR, and WNT/beta-catenin signaling pathways resulted in a substantial increase in cell death in the TLBR cell lines. Significantly, TLBR-2, which was derived from a fatal breast implant associated ALCL, showed increased resistance to all inhibitors compared with TLBR-1 and TLBR-3, which were derived from more indolent cases. Analysis of protein lysates showed elevated cleaved Notch1 in TLBR-2, compared with cell lines TLBR-1 and -3, which may drive increased cell proliferation and resistance to apoptotic regulators. Since the treatment of these lymphomas has not been studied, we evaluated the sensitivity of the TLBR cell lines to mainstay chemotherapy regimens (CHOP, methotrexate). These studies found that doxorubicin and vinblastine exert potent cytotoxic effects against all three TLBR cell lines, whereas cyclophosphamide was largely ineffective. In summary, TLBR-1, -2, and -3 closely resemble the primary breast implant-associated lymphomas from which they were derived, and as such provide valuable preclinical models to study the cell of origin and biology of this newly emerging clinical entity. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Differential Production of Cartilage ECM in 3D Agarose Constructs by Equine Articular Cartilage Progenitor Cells and Mesenchymal Stromal Cells

2020 ◽  
Vol 21 (19) ◽  
pp. 7071
Author(s):  
Stefanie Schmidt ◽  
Florencia Abinzano ◽  
Anneloes Mensinga ◽  
Jörg Teßmar ◽  
Jürgen Groll ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Progenitor Cells ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Type I Collagen ◽  
Type Ii Collagen ◽  
Cell Types ◽  
Type I ◽  
Cartilage Engineering ◽  
Hypoxic Conditions

Identification of articular cartilage progenitor cells (ACPCs) has opened up new opportunities for cartilage repair. These cells may be used as alternatives for or in combination with mesenchymal stromal cells (MSCs) in cartilage engineering. However, their potential needs to be further investigated, since only a few studies have compared ACPCs and MSCs when cultured in hydrogels. Therefore, in this study, we compared chondrogenic differentiation of equine ACPCs and MSCs in agarose constructs as monocultures and as zonally layered co-cultures under both normoxic and hypoxic conditions. ACPCs and MSCs exhibited distinctly differential production of the cartilaginous extracellular matrix (ECM). For ACPC constructs, markedly higher glycosaminoglycan (GAG) contents were determined by histological and quantitative biochemical evaluation, both in normoxia and hypoxia. Differential GAG production was also reflected in layered co-culture constructs. For both cell types, similar staining for type II collagen was detected. However, distinctly weaker staining for undesired type I collagen was observed in the ACPC constructs. For ACPCs, only very low alkaline phosphatase (ALP) activity, a marker of terminal differentiation, was determined, in stark contrast to what was found for MSCs. This study underscores the potential of ACPCs as a promising cell source for cartilage engineering.

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