Synergistic Effects of N-Acetylcysteine and Mesenchymal Stem Cell in a Lipopolysaccharide-Induced Interstitial Cystitis Rat Model

Abstract

The purpose of this study was to reduce the amount of stem cells used in treating preclinical interstitial cystitis (IC model) by investigating the synergistic effects of multipotent mesenchymal stem cells (M-MSCs; human embryonic stem cell-derived) and N-acetylcysteine (NAC). Eight-week-old female Sprague-Dawley rats were divided into seven groups, i.e., sham (n = 10), lipopolysaccharide/protamine sulfate (LPS/PS; n = 10), LPS/PS + NAC (n = 10), LPS/PS with 25K MSC (n = 10), LPS/PS with 50K MSC (n = 10) LPS/PS + 25K MSC + NAC (n = 10), and LPS/PS + 50K MSC + NAC (n = 10). To induce the IC rat model, protamine sulfate (10 mg, 45 min) and LPS (750 µg, 30 min) were instilled once a week for five consecutive weeks via a transurethral PE-50 catheter. Phosphate-buffered saline (PBS) was used in the sham group. One week after the final instillation, M-MSCs with two suboptimal dosages (i.e., 2.5 or 5.0 × 10 4 cells) were directly transplanted into the outer-layer of the bladder. Simultaneously, 200 mg/kg of NAC or PBS was intraperitoneally injected daily for five days. The therapeutic outcome was evaluated one week after M-MSC or PBS injection by awake cystometry and histological analysis. Functionally, LPS/PS insult led to irregular micturition, decreased intercontraction intervals, and decreased micturition volume. Both monotherapy and combination therapy significantly increased contraction intervals, increased urination volume, and reduced the residual volume, thereby improving the urination parameters compared to those of the LPS group. In particular, a combination of NAC dramatically reduced the amount of M-MSCs used for significant restoration in histological damage, including inflammation and apoptosis. Both M-MSCs and NAC-based therapy had a beneficial effect on improving voiding dysfunction, regenerating denudated urothelium, and relieving tissue inflammation in the LPS-induced IC/BPS rat model. The combination of M-MSC and NAC was superior to MSC or NAC monotherapy, with therapeutic efficacy that was comparable to that of previously optimized cell dosage (1000K) without compromised therapeutic efficacy.

Clinical Remission Using Personalized Low-Dose Intravenous Infusions of N-acetylcysteine with Minimal Toxicities for Interstitial Cystitis/Bladder Pain Syndrome

“…Based on experimental data and successful treatment of several chronic inflammation-related diseases [ 17 ], we personalized the dose and schedule of low-dose NAC by monitoring symptoms and plasma cytokines before and after infusions. Recent studies using NAC to treat liposaccharide-induced interstitial cystitis in rats demonstrated positive results [ 25 , 26 ]. Intraperitoneal NAC injections in rat subjects reduced inflammation, reversed the progression of fibrosis, and restored the integrity of the urothelium, resulting in improvement of abnormal voiding symptoms [ 25 ].…”

Functional enhancement strategies for immunomodulation of mesenchymal stem cells and their therapeutic application

“…Recently, Hu et al have demonstrated the synergistic effect of BM-MSC and botulinum toxin type A to treat hypertrophic scars, downregulating related mRNA and protein levels including alpha-smooth muscle actin (α-SMA) [73]. Compared to both monotherapies, combined treatment with N-acetylcysteine (NAC) and MSC exerts better therapeutic effects on the resolution of inflammation or apoptosis in the interstitial cystitis experimental model [74].…”

A Preclinical Study of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells for Treating Detrusor Underactivity by Chronic Bladder Ischemia

“…The contractile response to 80 mM KCl (P9333; Sigma-Aldrich, St. Louis, MO, USA), EFS (electrical field stimulation, 1, 2, 4, 8, 16, and 32 Hz), 1 mM ATP (A2383; Sigma-Aldrich), or carbachol concentration (PHR1511; Sigma-Aldrich; 1 nM to 1 mM) were recorded as previously described [12,19]. An electrical pulse (1 millisecond pulse width and 80 V in bath) was delivered using a stimulator (D-7806; Hugo Sachs Elektronik, Germany) for 5 s at increasing frequencies (1,2,4,8,16, and 32 Hz), with 5 min intervals between electrical field stimulations.…”

“…MSCs can been isolated from adult or fetal tissues [5], or they can be also derived from pluripotent stem cells (PSCs) such as embryonic stem cells (ESCs) and induced PSCs (iPSCs) [6][7][8]. The beneficial effects of MSC therapy include the generation of various preclinical bladder dysfunction models and reports of positive patient outcomes from clinical trials [9][10][11][12][13][14][15][16][17].…”