A high molecular weight hyaluronic acid biphasic dispersion as potential therapeutics for interstitial cystitis

Abstract

Interstitial cystitis (IC) is a progressive bladder disease characterized by increased urothelial permeability, inflammation of the bladder with abdominal pain. While there is no consensus on the etiology of the disease, it was believed that restoring the barrier between urinary solutes and (GAG) urothelium would interrupt the progression of this disease. Currently, several treatment options include intravesical delivery of hyaluronic acid (HA) and/or chondroitin sulfate solutions, through a catheter to restore the urothelial barrier, but have shown limited success in preclinical, clinical trials. Herein we report for the first time successful engineering and characterization of biphasic system developed by combining cross‐linked hyaluronic acid and naïve HA solution to decrease inflammation and permeability in an in vitro model of interstitial cystitis. The cross‐linking of HA was performed by 4‐arm‐polyethyeleneamine chemistry. The HA formulations were tested for their viscoelastic properties and the effects on cell metabolism, inflammatory markers, and permeability. Our study demonstrates the therapeutic effects of different ratios of the biphasic system and reports their ability to increase the barrier effect by decreasing the permeability and alteration of cell metabolism with respect to relative controls. Restoring the barrier by using biphasic system of HA therapy may be a promising approach to IC.

Comprehensive transcriptome profiling of urothelial cells following TNFα stimulation in an in vitro interstitial cystitis/bladder pain syndrome model

“…As normal human bladder urothelial cells are difficult to obtain for ethical reasons, while cell lines are widely accessible, our in vitro model consisted of the human non-invasive bladder cancer urothelial cell line RT4. In contrast to the more commonly used muscle-invasive bladder cancer urothelial cell line T24 in previously published in vitro IC/BPS models (18,20,21,23), RT4 cells isolated from grade I transitional cell carcinoma better reflect normal urothelial cells, especially with respect to differentiation markers (24, 25), as well as growth and migration characteristics (26). However, to increase the credibility of our results, we also confirmed our main findings from RT4 cells in normal porcine bladder urothelial (NPU) cells, which are more accessible and share most structural, molecular, and physiological features with normal human bladder urothelial cells (27-29).…”

“…Furthermore, TNFa signaling pathway has been determined as significantly enriched in the bladders of IC/BPS patients, specifically in those with Hunner's lesions (17). It is therefore not surprising that published in vitro models of bladder urothelial cells frequently use TNFa to mimic an inflammatory milieu found in IC/BPS (18)(19)(20)(21)(22)(23). However, the exact effects of TNFa on global gene expression and alterations of cellular processes in urothelial cells are still not yet entirely known.…”

“…Hence, we here determined the overall effect of TNFa on the transcriptomic profile of urothelial cells using RNA sequencing (RNA seq) and validated our result using qPCR, ELISA, Western blots, and immunofluorescence. The concentration of TNFa (20 ng/ml) that was used to stimulate urothelial cells in the current study was selected based on previously published in vitro studies (18)(19)(20)(21)(22)(23), viability assays and the ability to elicit a maximum measurable inflammatory response with the methods used that reached statistical significance. The ultimate goal of the in vitro model was to recapitulate the most important pathological processes and pathways, found enriched in the bladders of IC/BPS patients, and identify new potential therapeutic targets.…”

Hyaluronic Acid: A Key Ingredient in the Therapy of Inflammation

“…The biphasic system in the proportion of 1:1 does not increase the levels of IL-6, IL-8 or MCP-1 at concentrations of 1 and 3 mg/mL. In these conditions, the biphasic system decreases the permeability of the cell monolayer (assessed by the value of transepithelial electrical resistance (TEER)), making it a promising approach for IC treatment [169].…”

A Systematic Review of Therapeutic Approaches Used in Experimental Models of Interstitial Cystitis/Bladder Pain Syndrome

“…In contrast, the follow-up study in 2020 revealed that none of the commercially available GAG formulations containing HA or HA with CS were able to attenuate the TNFα-induced production of IL8 and IL6, the expression of GAG synthesis enzymes, or markers of tissue remodeling and pain [ 34 ]. Later on, Rooney et al also reported on a newly developed biphasic system combining cross-linked and native HA in a 1:1 ratio that was able to reduce permeability, while at the same time did not alter the production of proinflammatory cytokines in HTB2 cells [ 35 ]. The significant recovery in various cystometric parameters following HA treatment was shown in vivo in H 2 O 2 -induced IC in female Wistar rats.…”