Hydrogels composed of hydrophilic polymers such as polyethylene glycol and alginate have been used as scaffolds for various tissue engineering applications. This chapter describes procedures for encapsulation of cells in hydrogels and subsequently characterizing the extracellular matrix (ECM) production by those cells using biochemical assays, gene expression analysis, and histology. In particular, the biochemical assays described here are used to quantify collagen, glycosaminoglycan (GAG), and DNA content in each scaffold. The methods for analyzing the level of gene expression of specific ECM molecules such as collagen I, collagen II, and aggrecan are also described. Finally, included are protocols for histological methods used to analyze overall matrix production and GAG synthesis via hematoxylin and eosin staining and Safranin-O, respectively. These methods can be modified so that other scaffolds apart from hydrogels can be used.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1007/978-1-59745-413-1_23 | DOI Listing |
Int J Biol Macromol
January 2025
Second Department of Cardiovascular Medicine, Shengjing Hospital Affiliated to China Medical University, No. 36, Sanhao Street, Heping District, Shenyang 110004, Liaoning Province, China. Electronic address:
This work optimized proteoglycan-degrading enzymes through targeted mutagenesis to enhance their interaction with the tumor microenvironment in Renal Cell Carcinoma (RCC). A comprehensive mutagenesis approach identified 60 key mutations significantly improving enzymatic activity, stability, and structural integrity. When compared to Wild Type (WT) enzyme, a remarkable increase in specific activity by 35 % (p < 0.
View Article and Find Full Text PDFFront Bioeng Biotechnol
January 2025
Department of Agricultural and Environmental Sciences - Production, Landscape, Agroenergy, Università degli Studi di Milano, Milan, Italy.
Accelerating the genetic selection to obtain animals more resilient to climate changes, and with a lower environmental impact, would greatly benefit by a substantial shortening of the generation interval. One way to achieve this goal is to generate male gametes directly from embryos. However, spermatogenesis is a complex biological process that, at present, can be partially reproduced only in the mouse.
View Article and Find Full Text PDFImmunology
January 2025
Department of Clinical Immunology, Jawaharlal Institute of Post-Graduate Medical Education and Research (JIPMER), Puducherry, India.
Mechanisms contributing to non-response to treatment in lupus nephritis (LN) are unclear. We characterised the transcriptome of paired peripheral blood mononuclear cells (PBMCs) and renal tissues in LN before and after cyclophosphamide (CYC) treatment and identified markers that predicted treatment response. Total RNA isolated from paired PBMCs (n = 32) and renal tissues (n = 25) of 16 proliferative LN before CYC treatment, 6 months post-treatment, and during renal flare, was sequenced on Illumina Novaseq-6000 platform.
View Article and Find Full Text PDFMatrix Biol Plus
February 2025
Department of Biomedical Engineering, University of Cincinnati, Cincinnati, OH 45221, USA.
Schwann cells (SCs) hold key roles in axonal function and maintenance in the peripheral nervous system (PNS) and are a critical component to the regeneration process following trauma. Following PNS trauma, SCs respond to both physical and chemical signals to modify phenotype and assist in the regeneration of damaged axons and extracellular matrix (ECM). There is currently a lack of knowledge regarding the SC response to dynamic, temporal changes in the ECM brought on by swelling and the development of scar tissue as part of the body's wound-healing process.
View Article and Find Full Text PDFMater Today Bio
February 2025
Anhui University of Chinese Medicine, Hefei, 230012, China.
The therapeutic effect of immune checkpoint inhibitors (ICIs) in triple-negative breast cancer (TNBC) is unsatisfactory. The immune "cold" microenvironment caused by tumor-associated fibroblasts (TAFs) has an adverse effect on the antitumor response. Therefore, in this study, mixed cell membrane-coated porous magnetic nanoparticles (PMNPs) were constructed to deliver salvianolic acid B (SAB) to induce an antitumor immune response, facilitating the transition from a "cold" to a "hot" tumor and ultimately enhancing the therapeutic efficacy of immune checkpoint inhibitors.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!