Korean J Urol.  2015 Apr;56(4):280-287. 10.4111/kju.2015.56.4.280.

Use of nanoparticles to monitor human mesenchymal stem cells transplanted into penile cavernosum of rats with erectile dysfunction

• Kim JH
• Lee HJ
• Doo SH
• Yang WJ
• Choi D
• Kim JH
• Won JH
• Song YS

• KMID: 2155303
• DOI: http://doi.org/10.4111/kju.2015.56.4.280

Abstract

PURPOSE
This study was performed to examine the treatment of erectile dysfunction by use of superparamagnetic iron oxide nanoparticles-labeled human mesenchymal stem cells (SPION-MSCs) transplanted into the cavernous nerve injured cavernosa of rats as monitored by molecular magnetic resonance imaging (MRI).
MATERIALS AND METHODS
Eight-week-old male Sprague-Dawley rats were divided into three groups of 10 rats each: group 1, sham operation; group 2, cavernous nerve injury; group 3, SPION-MSC treatment after cavernous nerve injury. Immediately after the cavernous nerve injury in group 3, SPION-MSCs were injected into the cavernous nerve injured cavernosa. Serial T2-weighted MRI was done immediately after injection and at 2 and 4 weeks. Erectile response was assessed by cavernous nerve stimulation at 2 and 4 weeks.
RESULTS
Prussian blue staining of SPION-MSCs revealed abundant uptake of SPION in the cytoplasm. After injection of 1x10(6) SPION-MSCs into the cavernosa of rats, T2-weighted MRI showed a clear hypointense signal induced by the injection. The presence of SPION in the corpora cavernosa was confirmed with Prussian blue staining. At 2 and 4 weeks, rats with cavernous nerve injury had significantly lower erectile function than did rats without cavernous nerve injury (p<0.05). The group transplanted with SPION-MSCs showed higher erectile function than did the group without SPION-MSCs (p<0.05). The presence of SPION-MSCs for up to 4 weeks was confirmed by MRI imaging and Prussian blue staining in the corpus cavernosa.
CONCLUSIONS
Transplanted SPION-MSCs existed for up to 4 weeks in the cavernous nerve injured cavernosa of rats. Erectile dysfunction recovered and could be monitored by MRI.

Keyword

Erectile dysfunction; Magnetic resonance imaging; Mesenchymal stem cell transplantation; Nanoparticles

MeSH Terms

Animals
Contrast Media/pharmacology
Dextrans/*pharmacology
Disease Models, Animal
Drug Delivery Systems/methods
*Erectile Dysfunction/diagnosis/etiology/therapy
Magnetic Resonance Imaging/methods
*Magnetite Nanoparticles
Male
Mesenchymal Stem Cell Transplantation/*methods
Monitoring, Physiologic/methods
Penis/*innervation
*Peripheral Nerve Injuries/complications/diagnosis/physiopathology/therapy
Rats
Suspensions
Treatment Outcome
Contrast Media
Dextrans
Magnetite Nanoparticles
Suspensions

Figure

• 

  Fig. 1 Prussian blue staining of human mesenchymal stem cells with or without superparamagnetic iron oxide nanoparticles labeling. (A) Prussian blue-stained unlabeled human mesenchymal stem cells (×400). (B) Prussian blue-stained labeled human mesenchymal stem cells (×400). Note the abundant iron particles (blue dots) in the cytoplasm of the cells (arrow).

• 

  Fig. 2 (A) Schematic view of in vivo MRI according to the concentration of SPION-labeled hMSCs on the back muscle of rats (105 SPION-hMSCs, 5×105 SPION-hMSCs, 106 SPION-hMSCs, 104 SPION-hMSCs, 5×104 SPION-hMSCs, 106 hMSCs/mL). MRI showed a clear hypointense signal at all concentrations greater than 1×105 SPION-hMSCs/mL. (B) In vivo MR image according to the duration SPION-labeled human MSCs on the back muscle of rats. MRI showed a clear hypointense signal until 48 days after transplantation. MRI, magnetic resonance imaging; SPION, superparamagnetic iron oxide nanoparticles; hMSCs, human mesenchymal stem cells.

• 

  Fig. 3 In vivo MRI of magnetically labeled hMSCs. (A) Before transplantation in rat. (B) Immediately after transplantation in rat. (C) At 1 week after transplantation in rat. (D) At 2 weeks after transplantation in rat. (E) At 4 weeks after transplantation in rat. (F) At 6 weeks after transplantation in rat. (G) At 12 weeks after transplantation in rat. (H) Schematic drawing of glans penis, cavernosum, urethra, bladder, and transplanted hMSCs. Arrow shows the decrease of MR signal intensity. The areas of decreased MR signal intensity in the in the penile cavernosum were confined locally. G, glans penis; C, cavernosum; U, urethra; B, bladder; MRI, magnetic resonance imaging; hMSCs human mesenchymal stem cells.

• 

  Fig. 4 Histologic findings of magnetically labeled human mesenchymal stem cells in rat corpus cavernosum after transplantation by use of Prussian blue staining. (A-C) Two weeks after transplantation (A: ×40; B: ×100; C: ×400). (D-F) Four weeks after transplantation (D: ×40; E: ×100; F: ×400). Intracytoplasmic SPION particles (blue dots) are clearly visible with Prussian blue staining (arrows). SPION, superparamagnetic iron oxide nanoparticles.

• 

  Fig. 5 Recovery of erection after 1-V electrical stimulation. ICP/MAP significantly decreased and recovered at 2 and 4 weeks after SPION-hMSCs transplantation. At 2 weeks after transplantation, sham vs. SPION-hMSCs noninjection, p<0.05, sham vs. SPION-hMSCs injection, p<0.05. At 4 weeks, after transplantation, sham vs. SPION-hMSCs noninjection, p<0.05, sham vs. SPION-hMSCs injection, p<0.05. Sham, sham operation; CNI, cavernous nerve injury; CNI+MSCs, mesenchymal stem cell transplantation after cavernous nerve injury; SPION, superparamagnetic iron oxide nanoparticles; hMSCs, human mesenchymal stem cells; ICP/MAP, ratio of maximal intracavernosal pressure to mean arterial pressure.

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