Development of a chitosan‐based tissue‐engineered renal proximal tubule conduit

Abstract

Renal proximal tubule cells (RPTCs) are responsible for glomerular filtration and maintenance of water/electrolyte balance. To regenerate a proximal tubule, sufficient cell numbers and normal cell function are requisite. Collagen has been routinely used as a substrate for culturing human RPTCs (HRPTCs); however, the role of biomaterials has not been thoroughly explored. In this study, RPTCs retrieved from human nephrectomy/nephroureterectomy specimens were cultivated on chitosan as a substrate in serum-free condition for up to 150 days. HRPTCs could maintain a typical epithelial morphology and the specific differentiation feature of transporting epithelia after such long-term culture. As compared with HRPTCs cultivated on collagen, those cultivated on chitosan showed more dome formation, higher Na -K ATPase expression, lower vimentin expression, and lower transepithelial electrical resistance, indicating that HRPTCs cultivated on chitosan presented better differentiation status and would be more functional with better active transportation. Thus, the current study indicates greater scope for the use of chitosan as a biomaterial for preparing a HRPTC-coated chitosan conduit, which might be useful for the scaffold design of tissue-engineered proximal tubules. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 9-20, 2018.

Preclinical evaluation of melanocyte transplantation by chitosan‐based melanocyte spheroid patch to skin prepared by controlled sunburn blistering

“…Chitosan membrane was prepared as described . Briefly, chitosan (C‐3646, Sigma) was dissolved in 4% acetic acid at the concentration of 15 mg/mL (W/V).…”

“…A sterilized Teflon ring of a 15‐mm diameter was placed on top of each membranes in the well to prevent them from floating. The wells were sterilized in 70% alcohol overnight and rinsed extensively with phosphate buffered saline (PBS) before cell seeding …”

“…Chitosan membrane was prepared as described. 14,26 Briefly, chitosan (C-3646, Sigma) was dissolved in 4% acetic acid at the concentration of 15 mg/mL (W/V). Chitosan solution was added into a glass dish and left dried at 508C to form a thin dry membrane.…”

“…The wells were sterilized in 70% alcohol overnight and rinsed extensively with phosphate buffered saline (PBS) before cell seeding. 26 Cell and culture The experiment involving human cells was approved by our Institutional Review Board and informed consent was obtained from patients undergoing routine skin surgery or circumcision before the skin samples were collected. The skin specimen was treated with 0.2% protease (Sigma) solution at 48C overnight, and then the dermis was removed gently.…”

Chitin and Chitosan Derivatives as Biomaterial Resources for Biological and Biomedical Applications

“…For instance, chitosan nanoplex effectively covered siRNA for knocking down PDGF-B and PDGFR-beta [ 100 ]. Chitosan was also used as biomaterials in combination with collagen for culturing human renal proximal tubular cells [ 101 ]. The potential beneficial effects of chitosan oligosaccharide on kidney diseases have been also reviewed elsewhere [ 102 ].…”