Implanted Muscle-Derived Stem Cells Ameliorate Erectile Dysfunction in a Rat Model of Type 2 Diabetes, but Their Repair Capacity Is Impaired by Their Prior Exposure to the Diabetic Milieu

Abstract

Introduction Muscle derived stem cells (MDSC) and other stem cells implanted into the penile corpora cavernosa ameliorate erectile dysfunction in type 1 diabetic (T1D) rat models by replenishing the lost corporal smooth muscle cells (SMC) and reducing fibrosis. However, no conclusive data on this question in T2/D/obesity models is available. Aim. We studied whether: a) MDSC from T2D Obese Zucker (OZ) rats at an early stage of diabetes (ED-SC), counteract corporal veno-occlusive dysfunction (CVOD) and corporal SMC loss/lipofibrosis when implanted in the OZ rats at a late stage of diabetes; b) MDSC from these late diabetes OZ rats (LD-SC) differ from ED-SC in their gene transcriptional phenotype and repair capacity. Methods and Outcomes ED-SC and LD-SC were compared by DNA microarray assays, and ED-SC were incubated in vitro under high glucose conditions (ED-HG-SC). These three MDSC types were injected into the corpora cavernosa of late diabetes OZ rats (OZ/ED, OZ/LD, and OZ/ED-HG rats respectively). Untreated OZ (OZ/UT) and non-diabetic Lean Zucker (LZ/UT) rats were controls. Two months later, rats were subjected to cavernosometry and the penile shaft and corporal tissues were subjected to histopathology and DNA microarray assays. Results Implanted ED-SC and ED-HG-SC, improved CVOD, counteracted corporal SMC/collagen decrease and fat infiltration in long-term T2D rats, and upregulated nNOS and eNOS. LD-SC acquired an inflammatory/profibrotic/oxidative/dyslipidemic transcriptional phenotype, and failed to repair the corporal tissue. Conclusions MDSC from pre-diabetic rats injected into the corpora cavernosa of long-term diabetic T2D rats improve CVOD and the underlying histopathology. In contrast, MDSC from long-term uncontrolled diabetic T2D rats, are imprinted by the hyperglycemic/dyslipidemic milieu with a noxious phenotype associated with an impaired tissue repair capacity. Diabetes-impacted stem cells may lack tissue repair efficacy as autografts, and should either be reprogrammed in vitro, or substituted by stem cells from allogenic non-diabetic sources.

Muscle Derived Stem Cells Stimulate Muscle Myofiber Repair and Counteract Fat Infiltration in a Diabetic Mouse Model of Critical Limb Ischemia

“…MDSC implanted into the muscle immediately after UFAL prevented mortality. Supplementation of the MDSC treatment with molsidomine, but not molsidomine alone [ 38 ], improved limb preservation. However, the possibility of self-mutilation complicates the interpretation of leg loss.…”

“…The homogenate was clarified by centrifugation at 16,000 g for 5 minutes, following which 20 μg protein samples were electophoresed through Bio-Rad Mini-Protean TGX 4–15% gels and transferred to PVDF membranes. Blots were probed with primary antibodies: (a) Oct 4 (anti-rabbit Oct 4, polyclonal, 1:500; BioVision, Inc., Milpitas, California, U.S.A.) [ 12 ]; (b) ASMA (anti-human ASMA, monoclonal, 1:1,000; Calbiochem, Billerica, Massachusetts, USA) [ 12 ]; (c) Calponin I (anti-mouse calponin 1, monoclonal, 1:500; Santa Cruz Biotechnology, Dallas, Texas, U.S.A) [ 12 ]; (d) VEGF (anti-mouse VEGF, monoclonal, 1:500; Santa Cruz Biotechnology) [ 43 , 44 ]; (e) Von Willebrand (anti-mouse Von Willebrand, polyclonal, 1:500; Abcam, Cambridge, Massachusetts, U.S.A.) [ 45 ]; (f) CD31 (anti-mouse CD31, monoclonal, 1:1000, Abcam) [ 46 ]; (g) BDNF (anti-mouse BDNF, rabbit monoclonal, 1:1000; Abcam) [ 38 ]; (h) MHC (fast) (anti-mouse MHC-II, monoclonal, 1:5000; Abcam) [ 12 ]; (i) myostatin (anti-mouse myostatin, rabbit polyclonal 1:1,000; Chemicon International Inc, Billerica, Massachusetts, U.S.A.) [ 12 , 47 ]; (j) follistatin (anti-mouse follistatin, rabbit polyclonal, 1:1000, Abcam) [ 48 ]; and (k) GAPDH (anti-mouse GAPDH, monoclonal, 1:5,000, Chemicon International).…”

Low‐intensity extracorporeal shock wave therapy promotes myogenesis through PERK/ATF4 pathway

“…The satellite cells reside between the basal lamina and the plasma membrane of skeletal muscle fibers. Most previous studies have shown that MDSCs can be isolated from hind limb . There is no previous report about the urethral striated muscle stem cells.…”

“…Most previous studies have shown that MDSCs can be isolated from hind limb. 17 There is no previous report about the urethral striated muscle stem cells. In this study, the MDSCs were harvested from the rat urethra.…”

Effects of Vitamin D Restricted Diet Administered during Perinatal and Postnatal Periods on the Penis of Wistar Rats

“…Although the rat penis presents these differences when compared to the penis of humans, several studies consider a suitable model to investigate the morphological changes resulting from pathological conditions [ 29 , 30 ].…”