Mujun Lu

Penile and Scrotal Paget’s disease: 130 Chinese patients with long‐term follow‐up

To report a retrospective series of 130 Chinese patients with penoscrotal extramammary Paget’s diseases (EMPD), with a long‐term follow‐up, and thus improve the diagnosis and therapy of this disease.

Time-dependent bladder tissue regeneration using bilayer bladder acellular matrix graft-silk fibroin scaffolds in a rat bladder augmentation model.

With advances in tissue engineering, various synthetic and natural biomaterials have been widely used in tissue regeneration of the urinary bladder in rat models. However, reconstructive procedures remain insufficient due to the lack of appropriate scaffolding, which should provide a waterproof barrier function and support the needs of various cell types. To address these problems, we have developed a bilayer scaffold comprising a porous network (silk fibroin [SF]) and an underlying natural acellular matrix (bladder acellular matrix graft [BAMG]) and evaluated its feasibility and potential for bladder regeneration in a rat bladder augmentation model. Histological (hematoxylin and eosin and Massons trichrome staining) and immunohistochemical analyses demonstrated that the bilayer BAMG-SF scaffold promoted smooth muscle, blood vessel, and nerve regeneration in a time-dependent manner. At 12weeks after implantation, bladders reconstructed with the BAMG-SF matrix displayed superior structural and functional properties without significant local tissue responses or systemic toxicity. These results demonstrated that the bilayer BAMG-SF scaffold may be a promising scaffold with good biocompatibility for bladder regeneration in the rat bladder augmentation model.

Two-stage urethroplasty is a better choice for proximal hypospadias with severe chordee after urethral plate transection: a single-center experience

It is still debatable whether single- or two-stage urethroplasty is a more suitable technique for treating hypospadias with severe chordee after urethral plate transection. This retrospective study evaluated these two techniques. A total of 66 patients of proximal hypospadias with severe chordee were divided into two groups according to the techniques they underwent: 32 and 34 patients underwent single-stage (Duckett) or two-stage urethroplasty, respectively. Median ages at presentation were 7.5 years and 11.0 years in single-stage and two-stage repair groups, respectively. Median follow-ups were 28.5 months (20−60 months) and 35 months (18−60 months) in the single-stage and two-stage groups, respectively. The meatus of the neourethra was located at the top of the glans in all patients. No recurrence of chordee was found during follow-up, and all patients or parents were satisfied with the penile length and appearance. Complications were encountered in eight patients in both groups, with no statistically significant differences between the two techniques. The late complication rate of stricture was higher after the single-stage procedure (18.75% vs 0%). The complication rate after single-stage repairs was significantly lower in the prepubescent subgroup (10.52%) than in the postpubescent cohort (46.15%). These results indicate that the urethral plate transection effectively corrects severe chordee associated with proximal hypospadias during the intermediate follow-up period. Considering the higher rate of stricture after single-stage urethroplasty, two-stage urethroplasty is recommended for proximal hypospadias with severe chordee after urethral plate transection.

Remodeling of buccal mucosa by bladder microenvironment.

OBJECTIVES The optimal cell source for bladder mucosa reconstruction is in question. This study explored the feasibility of phenotype transformation of oral mucosa towards bladder urothelium by transplanting the oral mucosa into bladder local microenvironment. METHODS Porcine oral mucosa grafts were transplanted into autologous bladder mucosa defects, and the specific marker expressions of both oral epithelium and bladder urothelium were examined by immunohistochemistry and RT-PCR at 3, 6, and 12 months to detect a potential phenotype transformation. RESULTS The grafts could retain the phenotype and structure of oral mucosa within 3 months with positive expression of cytokeratin 14 (CK 14, a specific marker of oral epithelium) and negative expression of uroplakin II (UPII, a urothelium-specific marker). However, after 6 months of transplantation, the grafts expressed UPII at both protein and mRNA levels and the phenotype persisted at 12 months postsurgery. All the grafts continuously retained the positive expression of CK 14 at all time points. CONCLUSIONS These findings, for the first time, revealed the transdifferential potential of oral keratinocytes toward urothelial cells and the important role of bladder local microenvironment for remodeling oral mucosa epithelium. These results support the hypothesis that oral keratinocytes can serve as a potential cell source for reconstructing bladder mucosa.

Adipose-derived stem cells-seeded bladder acellular matrix graft-silk fibroin enhances bladder reconstruction in a rat model

The unfavourable clinical outcomes of host cell-seeded scaffolds for bladder augmentation warrant improved bioactive biomaterials. This study aimed to examine the feasibility of adipose-derived stem cells (ASCs)-seeded bilayer bladder acellular matrix graft (BAMG)-silk fibroin (SF) scaffold in enhancing bladder reconstruction. Sprague Dawley rats were randomly divided into three groups: the BAMG-SF-ASCs group, the acellular BAMG-SF group and the cystotomy group. The BAMG-SF-ASCs group was sampled at 2, 4 and 12 weeks, and compared with the other groups at 12 weeks. In the BAMG-SF-ASCs group, the normal bladder contour was reformed similar to that in the cystotomy group, with abundant urothelium and smooth muscle regeneration, as well as a suitable scaffold degradation speed, and trivial fibrosis and inflammation. The ASCs seeded in BAMG-SF were maintained in the regenerated region during the 12-week experimental period and significantly enhanced the vessel density, nerve regeneration and bladder function compared with acellular BAMG-SF. In addition, the BAMG-SF-ASCs group presented elevated levels of SDF-1α, VEGF and their receptors, with an obvious increase in ERK 1/2 phosphorylation. BAMG-SF is a promising biomaterial for ASCs seeding to facilitate bladder augmentation and demonstrated an enhanced angiogenic potential possibly related to the SDF-1α/CXCR4 pathway via ERK 1/2 activation.

Estrogen Receptor Alpha (ERα)-Associated Fibroblasts Promote Cell Growth in Prostate Cancer

Estrogen receptor (ER) is expressed in cancer-associated fibroblasts (CAFs) in the stromal compartment of cancerous prostate. However, the effect of ERα in CAF cells on prostate cancer (PCa) cell growth remains unclear. We used lentiviral transduction to stably express ERα in CAF cells isolated from transgenic adenocarcinoma of the mouse prostate model. MTT and 3D colony-formation assays demonstrated that conditioned medium from ERα-expressing CAF cells (CAF-ERα+) promoted cell proliferation and colony growth of various PCa cell lines, such as PC3, LNCaP, 22RV1, and C4-2. We further confirmed the in vitro data by orthotopically co-implanting 22RV1, transfected with firefly luciferase, and CAF-ERα+ cells in vivo using mouse model. Mice co-implanted with CAF-ERα+ exhibited stronger luciferase signals and bigger tumor size compared to animals co-implanted with CAF that do not express ER. Our results demonstrate that ER expressed in CAF might play a pro-proliferative role in PCa.

Intraperitoneal incubation of bladder acellular matrix grafts improves bladder smooth muscle regeneration via neovascularization

The objective of this study was to investigate whether intraperitoneal incubation improves the regenerative capacity of bladder acellular matrix grafts (BAMGs) in a rat model of bladder augmentation. After 2 weeks of incubation in the peritoneum of male rats, BAMG flaps with vascular pedicles were harvested for autologous bladder augmentation. As the control, BAMGs were directly used for bladder augmentation without intraperitoneal incubation. Histological analyses of the incubated BAMGs demonstrated extensive cell growth and vasculature in homogeneous collagen bundles. The cells were positive for vimentin and negative for α-smooth muscle actin and pan-cytokeratin AE1/AE3. Cystography revealed smoother contours of the augmented bladders in the incubated group at 4 and 12 weeks postoperatively. However, the bladder capacity was not significantly different between the two groups. In both groups, the entire urothelium regenerated well without obvious differences. At both time points, compared with the control group, increased numbers of smooth muscle cells (SMCs) and blood vessels were found in the incubated group. At 12 weeks, the SMCs in the incubated group were more similar to those in the native smooth muscle fiber bundles of the bladder. Taken together, our results demonstrated that BAMGs preincubated in the peritoneum promote the regeneration of bladder smooth muscle via neovascularization in a rat bladder augmentation model.

Reconstruction of Major Scrotal Defects by Anterolateral Thigh Flap

To summarize the indications and applications of anterolateral thigh flaps (ALTP) in reparation and reconstruction of acute scrotum skin deficiency. We report our experience of treating three patients for scrotal reconstruction using the ALTP method from May of 2007 to December of 2010. The flap was completely islanded, tunneled beneath the fascial septa below the rectus femoris, and placed into the defect. The resulting reconstruction provided a tension-free, cosmetically appealing scrotum with complete testicular coverage. There were no perioperative or postoperative complications. Spermatogenesis (sperm count levels), and sexual function (IIEF) were not changed significantly. ALTP represent an excellent reconstruction option for patients with contaminated perineum. ALTP is available for replacement of the avulsed portion of the scrotal skin resulting in satisfactory shape and color of the penis and scrotum.

A smart bilayered scaffold supporting keratinocytes and muscle cells in micro/nano-scale for urethral reconstruction

Rationale: In urethral tissue engineering, the currently available reconstructive procedures are insufficient due to a lack of appropriate scaffolds that would support the needs of various cell types. To address this problem, we developed a bilayer scaffold comprising a microporous network of silk fibroin (SF) and a nanoporous bacterial cellulose (BC) scaffold and evaluated its feasibility and potential for long-segment urethral regeneration in a dog model. Methods: The freeze-drying and self-assembling method was used to fabricate the bilayer scaffold by stationary cultivation G. xylinus using SF scaffold as a template. The surface morphology, porosity and mechanical properties of all prepared SF-BC scaffolds were characterized using Scanning electron microscopy (SEM), microcomputed tomography and universal testing machine. To further investigate the suitability of the bilayer scaffolds for tissue engineering applications, biocompatibility was assessed using an MTT assay. The cell distribution, viability and morphology were evaluated by seeding epithelial cells and muscle cells on the scaffolds, using the 3D laser scanning confocal microscopy, and SEM. The effects of urethral reconstruction with SF-BC bilayer scaffold was evaluated in dog urethral defect models. Results: Scanning electron microscopy revealed that SF-BC scaffold had a clear bilayer structure. The SF-BC bilayer scaffold is highly porous with a porosity of 85%. The average pore diameter of the porous layer in the bilayer SF-BC composites was 210.2±117.8 μm. Cultures established with lingual keratinocytes and lingual muscle cells confirmed the suitability of the SF-BC structures to support cell adhesion and proliferation. In addition, SEM demonstrated the ability of cells to attach to scaffold surfaces and the biocompatibility of the matrices with cells. At 3 months after implantation, urethra reconstructed with the SF-BC scaffold seeded with keratinocytes and muscle cells displayed superior structure compared to those with only SF-BC scaffold. Principal Conclusion: These results demonstrate that the bilayer SF-BC scaffold may be a promising biomaterial with good biocompatibility for urethral regeneration and could be used for numerous other types of hollow-organ tissue engineering grafts, including vascular, bladder, ureteral, bowel, and intestinal.

Adipose-derived stem-cell-implanted poly(ϵ-caprolactone)/chitosan scaffold improves bladder regeneration in a rat model

AIM The study investigated the feasibility of seeding adipose-derived stem cells (ASCs) onto a poly(ϵ-caprolactone)/chitosan (PCL/CS) scaffold for bladder reconstruction using a rat model of bladder augmentation. MATERIALS & METHODS In the experimental group, the autologous ASCs were seeded onto the PCL/CS scaffold for bladder augmentation. An unseeded scaffold was used for bladder augmentation as control group. The sham group was also set. RESULT 8 weeks after implantation, more densely smooth muscles were detected in the experimental group with a larger bladder capacity and more intensive blood vessels. Immunofluorescence staining demonstrated that some of the smooth muscle cells were transdifferentiated from the ASCs. CONCLUSION Our findings indicated that ASC-seeded PCL/CS may be a potential scaffold for bladder tissue engineering.