Cell and protein arrays have demonstrated remarkable utility in the high-throughput evaluation of biological responses; however, they lack the complexity of native tissue and organs. Here, we describe tissue extracellular matrix (ECM) arrays for screening biological outputs and systems analysis. We spotted processed tissue ECM particles as two-dimensional arrays or incorporated them with cells to generate three-dimensional cell-matrix microtissue arrays. We then investigated the response of human stem, cancer, and immune cells to tissue ECM arrays originating from 11 different tissues, and validated the 2D and 3D arrays as representative of the in vivo microenvironment through quantitative analysis of tissue-specific cellular responses, including matrix production, adhesion and proliferation, and morphological changes following culture. The biological outputs correlated with tissue proteomics, and network analysis identified several proteins linked to cell function. Our methodology enables broad screening of ECMs to connect tissue-specific composition with biological activity, providing a new resource for biomaterials research and translation.
Micronized porcine urinary bladder matrix (UBM) is an extracellular matrix biomaterial that has immunomodulatory and pro-regenerative properties. The objective of this study was to assess the ability of UBM to alter disease progression in a mouse model of post-traumatic osteoarthritis (OA). Ten-week-old wild-type C57BL/6 male mice underwent anterior cruciate ligament transection (ACLT) to induce OA. Two weeks after ACLT, UBM (50 mg/mL) or saline was injected into the mouse joint. At 4 and 8 weeks post-ACLT, cartilage integrity was assessed using OARSI scoring of histology, pain was evaluated, and joints were harvested for quantitative RT-PCR analysis of cartilage-specific and inflammatory gene expression. UBM-treated animals showed improved cartilage integrity at 4 and 8 weeks and reduced pain at 4 weeks compared to saline-injected mice. Animals injected with UBM expressed higher levels of genes encoding structural cartilage proteins, such as collagen2α1 and aggrecan, as well as anti-inflammatory cytokines, including interleukins 10 and 4. UBM decreased cartilage degeneration in the murine ACLT model of OA, which may be due to reduced inflammation in the joint and maintenance of high expression levels of proteoglycans.
Purpose: Osteoarthritis (OA) is a degenerative joint disease in which synergistic interactions between synovial fluid lubricants and the cartilage surface are lost. We designed a strategy to bind HA to the cartilage surface via a polymer-peptide binding system, mimicking the function of lubricin on the healthy cartilage surface. This system contains an HA binding peptide and a collagen binding peptide linked by poly(ethylene glycol) (PEG) to target and concentrate HA at the cartilage surface. This technology enhanced HA retention in vivo and cartilage lubrication in vitro.In the current study, the polymer-peptide technology was applied to a small animal model of post traumatic OA. A synthetic peptide (HABP1), RHAMM mimetic peptide (HABP2), and another synthetic peptide (HABP3), each conjugated to 1kDa peg-COLBP and also to an 8-arm pegcolbp, were studied to determine which one had the greatest efficacy reducing OA progression in vivo and in binding HA in vitro. Methods: The anterior cruciate ligament (ACL) of each mouse was transected and OA was allowed to develop over two weeks at which time saline controls or polymer-peptides were injected intra-articularly. Cartilage integrity was assessed using OARSI scoring of histology at 4 weeks post ACLT. Additionally, joints were harvested for quantitative PCR (qPCR) of structural cartilage genes and inflammatory markers at 4 weeks post ACLT. Pain and function was monitored at 2 and 4 weeks post ACLT by hot plate and incapacitance testing, which are functional assessments of sensitivity and pain. In vitro, quartz crystal microbalance with dissipation monitoring (QCM-D) was used to compare binding of different HABP formulations to HA. Results: The average OARSI score was reduced to varying degrees in each treatment group. Among peptides, the highest concentration of each peptide possible was tested. This resulted in a reduction from the average saline control score of 3.28e1.33 for HABP1-peg-colbp, 1.33 for HABP2-peg-colbp, 1.66 for HABP3-peg-colbp, 1.83 for HABP1-8 arm peg-colbp, and 0.833 for HABP2-8 arm peg-colbp (n ¼ 3, HABP2-8 arm peg-colbp was significant at p < 0.05). These preliminary results showed that all peptide treated groups had more intense proteoglycan staining and intact articular cartilage than saline controls; more animals will be added to select treatments based on these preliminary OARSI scores and on in vitro binding outcomes. These OARSI results tended to correlate with the functional pain tests, in which the HABP1-peg-colbp, HABP2-peg-colbp, and HABP2-8 arm pegcolbp injected mice displayed a significant reduction of pain on the hot plate test (p < 0.05). HABP2-peg-colbp and HABP2-8 arm peg-colbp also reduced pain in the incapacitance test (p < 0.05). qPCR analysis was performed 4 weeks post ACLT and fold changes calculated with respect to unoperated, age matched controls. This revealed that, compared to saline controls, HABP1-peg-colbp injected animals expressed significantly lower levels of the matrix metalloproteinase MMP13 (on average 1.16 fold ...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.