Factors secreted from dental pulp stem cells show multifaceted benefits for treating experimental temporomandibular joint osteoarthritis.

OBJECTIVE: Temporomandibular joint osteoarthritis (TMJOA) is a degenerative disease characterized by progressive cartilage degeneration, abnormal bone remodeling, and chronic pain. In this study, we aimed to investigate effective therapies to reverse or suppress TMJOA progression. DESIGN: To this end, we performed intravenous administration of serum free conditioned media from human exfoliated deciduous teeth stem cells (SHED-CM) into a mechanical-stress induced murine TMJOA model. RESULTS: SHED-CM administration markedly suppressed temporal muscle inflammation, and improved bone integrity and surface smoothness of the destroyed condylar cartilage. Moreover, SHED-CM treatment decreased the number of IL-1ß, iNOS, and MMP-13 expressing chondrocytes, whereas it specifically increased PCNA-positive cells in the multipotent polymorphic cell layer. Notably, the numbers of TdT-mediated dUTP nick end labeling (TUNEL)-positive apoptotic chondrocytes in the SHED-CM treated condyles were significantly lower than in those treated with DMEM, whereas the proteoglycan positive area was restored to a level similar to that of the sham treated group, demonstrating that SHED-CM treatment regenerated the mechanical-stress injured condylar cartilage and subchondral bone. Secretome analysis revealed that SHED-CM contained multiple therapeutic factors that act in osteochondral regeneration. CONCLUSIONS: Our data demonstrated that SHED-CM treatment promoted the regeneration and repair of mechanical-stress induced mouse TMJOA. Our observations suggest that SHED-CM has potential to be a potent tissue-regenerating therapeutic agent for patients with severe TMJOA.

Extracellular Vesicles of Stem Cells to Prevent BRONJ.

Extracellular vesicles (EVs), several tens to hundreds of nanometers in size, are vesicles secreted by cells for intercellular communication. EVs released from mesenchymal stem cells (MSC-EVs) have the potential to treat multiple diseases. This study aimed to determine the effects of MSC-EVs on bisphosphonate-related osteonecrosis of the jaw (BRONJ), whose pathogenesis and treatment are not yet established. To this end, zoledronic acid (ZOL) was administered to bone marrow cells and fibroblasts in vitro. In vivo, a BRONJ model was produced by administering ZOL to rats and extracting teeth. Each MSC-EV-treated and nontreated group was compared histologically and molecularly. In vitro, the nontreated group showed an increased number of ß-galactosidase-positive cells and expression of senescence-associated genes p21, pRB and senescence-related inflammatory cytokines. Conversely, MSC-EV administration decreased the number of senescent cells and expression levels of p21, pRB and inflammatory cytokines. In vivo, in the nontreated group, the socket was partially uncovered by the oral epithelium, leaving an exposed bone. Conversely, in the MSC-EV-treated group, the socket was healed. Besides, in the nontreated group, ß-galactosidase-positive cells existed in the socket and colocalized with the CD90 and periostin-positive cells. However, there were few ß-galactosidase-positive cells in the MSC-EV-treated group. Furthermore, gene expression of stem cell markers Bmi1 and Hmga2 and the vascular endothelial marker VEGF was significantly increased in the MSC-EV-treated group, compared with that in the nontreated group. These results indicate that MSC-EVs prevent ZOL-induced senescence in stem cells, osteoblasts, and fibroblasts and reduce inflammatory cytokines. Furthermore, administration of MSC-EVs prevented senescence of cells involved in wound healing and the spread of chronic inflammation around senescent cells, thereby promoting angiogenesis and bone regeneration and preventing BRONJ.

Alveolar cleft osteoplasty using tissue-engineered osteogenic material.

The use of tissue-engineered osteogenic material comprising platelet-rich plasma and autologous mesenchymal stem cells isolated, expanded and induced to osteogenic potential in bone augmentation procedures as a replacement for autologous bone grafts, offers predictable results with minimal donor-site morbidity. This material was applied for an alveolar cleft osteoplasty of a 9-year-old female patient. Serial computed tomograms showed the regenerated bone extending from the cleft walls after 3 months and bridging the cleft after 6 months, with 79.1% of the grafted region after 9 months at the time when the canine and lateral incisor in the affected side erupted in the reconstructed alveolar ridge.

Initiation-stage enhancement by uracil of N-ethyl-N-hydroxyethylnitrosamine-induction of kidney carcinogenesis in rats.

The present study was conducted to clarify the effects of N-ethyl-N-hydroxyethylnitrosamine (EHEN) and uracil in combination on renal carcinogenesis in female F344 rats. Group 1 animals (n = 30) received a 3-week simultaneous administration of 0.05% EHEN and 3% uracil, and group 2 (n = 26) 0.05% EHEN with a lower 1.5% dose of uracil. Group 3 (n = 30) was treated with 0.05% EHEN alone and group 4 (n = 28) received only 3% uracil for 3 weeks. In all the above four groups, the rats were given basal diet and water without chemical addition for a 48-week period after the 3-week treatment period. Group 5 (n = 33) received no chemicals for the entire 52 weeks. At the end of week 52, renal adenocarcinomas were found in 53%, 31% and 13% of the rats in groups 1, 2, and 3, respectively. The present study thus demonstrated that simultaneous administration of uracil enhances the occurrence of EHEN-induced renal adenocarcinomas in rats.

Comparison of the effectiveness of placebo and alpha-blocker therapy for the treatment of idiopathic oligozoospermia.

OBJECTIVE: To determine whether alpha-blocker (bunazosin) improves fertility and/or semen parameters DESIGN: Placebo-controlled, double-blind clinical study. SETTING: Nagoya University Hospital Andrology Clinic, Nagoya, Japan. PATIENTS: Thirty-four men with sperm density between 5 and 20 x 10(6) sperm/mL, normal serum gonadotropins and T, and a fertile partner were enrolled in this study. INTERVENTIONS: After a 3-month control period, patients randomly were prescribed bunazosin 2 mg/d or a placebo, two tablets per day, for 6 months. Semen and blood samples were collected before and after therapy. Semen parameters, serum gonadotropins, T, PRL, and E2 were evaluated before and after therapy. RESULTS: The pregnancy rate (PR) in the alpha-blocker group was 25%, compared with 6.7% in the placebo group. There was no statistical difference in the PR between groups. The alpha-blocker group had significantly higher levels of sperm density and total motile sperm count. There were no differences between the placebo and alpha-blocker groups in seminal volume, the percentage of motile sperm, and normal morphology or hormone levels. CONCLUSIONS: The authors conclude that alpha-blocker is a useful drug in the treatment of idiopathic moderate oligozoospermic men.

New treatment of idiopathic severe oligozoospermia with mast cell blocker: results of a single-blind study.

OBJECTIVE: To determine whether mast cell blocker (tranilast) improves fertility and/or semen parameters in severe oligozoospermia. DESIGN: Placebo-controlled single-blind clinical study. SETTING: Nagoya University Hospital Andrology Clinic, Nagoya, Japan. PATIENTS: Fifty men with sperm density < 5 x 10(6) sperm/mL, normal serum gonadotropins, and T, and a fertile partner were enrolled in this study. INTERVENTIONS: Patients were prescribed randomly 300 mg/d tranilast or a placebo, three tablets per day, for 3 months. Semen and blood samples were collected before and after therapy. Semen parameters, serum gonadotropins, T, and PRL were evaluated before and after therapy. RESULTS: The pregnancy rate (PR) in the mast cell blocker group was 28.6% compared with 0% in the placebo group. There was a statistical difference in the PR between groups. The mast cell blocker group had significantly higher levels of sperm density, sperm motility, and total motile sperm count. There were no differences between the mast cell blocker and placebo groups in seminal volume and normal sperm morphology. CONCLUSIONS: The authors conclude that mast cell blocker is clinically useful for the treatment of idiopathic severe oligozoospermic men.

Modified osteosynthesis for condylar neck fractures in atrophic mandibles.

A fixation system combining both plate and lag screw osteosynthesis for condylar neck fractures of the mandible is described. The currently available device is adapted in that the lag screw is inserted in the lateral cortical bone of the condylar segment instead of the cancellous bone alone.

A case of unilateral giant hydronephrosis with renal insufficiency.

Giant hydronephrosis, which is a symptomless enlargement of the abdomen, is an extremely rare condition in adults, with most cases being discovered in young-to-middle-aged people. We report a case of adult giant hydronephrosis accompanied by renal insufficiency, in which surgical repair was not performed because of the patient's poor cardiac function. Nephrostomy was performed and was followed with replacement of the nephrostomy tube every 2 weeks. For diagnostic purposes, ultrasonography and computerized tomography were preferable. A supplementary antegrade and retrograde pyelography was valuable in locating the stenosis.

Effect of albumin on proluminal movement of 3H-androgen into seminiferous and epididymal tubules and androgen binding in the interstitium of the testis and epididymis after perifusion with fluid containing albumin.

Effect of albumin on proluminal androgen movement from the peritubular space to the intratubular fluids of the adult rat testis and epididymis was examined by using in vivo microperifusion and subsequent micro-puncture of the seminiferous tubules and caput epididymal tubules. Tubules were perfused with four different fluids: (1) Minimum Essential Medium (MEM) containing 3H-testosterone and 14C-polyethyleneglycol (PEG) alone; (2) MEM + 8 mg/ml Bovine Serum Albumin (BSA) containing the same radiolabeled compounds as above; (3) MEM + 80 mg/ml albumin containing the same radiolabeled compounds as above; and (4) Testosterone-free rat serum containing the same radiolabeled compounds as above. Bound 3H-androgens in the interstitial fluids of the testis and epididymis after one-hour perifusion with the four different fluids above were measured by charcoal assay. In the testis, proluminal 3H-androgen movement was not significantly altered by addition of albumin to the perifusion fluid (p = 0.08). Bound 3H-androgens in the interstitial fluid after perifusion were significantly increased with increasing albumin concentrations in the perifusion fluid. In the caput epididymis, proluminal 3H-androgen movement was significantly decreased with increasing albumin concentration in the perifusion fluid. Bound 3H-androgens in the interstitial fluid after perifusion were significantly increased with increasing albumin concentrations in the perifusion fluid (p < 0.05). These findings suggest that proluminal transepithelial movement of 3H-androgens in the reproductive tract could be influenced by the presence of albumin, androgen-binding protein or some other binding protein in the peritubular space.

A comparison of transurethral resection of the prostate and medical treatment for the patient with moderate symptoms of benign prostatic hyperplasia.

Transurethral resection of the prostate is the most common surgical treatment for benign prostatic hyperplasia. We conducted a prospective randomized clinical trial to compare this surgery with medical treatment in men with moderate symptoms of benign prostatic hyperplasia. Of 98 men over the age of 54 years who were screened between June 1993 and June 1995, 53 were studied (25 in the surgery group and 28 in the medication group). Patients' symptoms and the degree to which they were bothered by urinary difficulties were measured with standardized questionnaires and medical evaluations. The men randomly assigned to the surgery group underwent surgery within 2 weeks after the assignment. Surgery was not associated with an impotence or urinary incontinence. The follow-up period was 1 year. Surgery was significantly associated with improvement in residual urinary volume and peak flow rate; and also in the scores for urinary difficulties, sexual performance and interference with activities of daily living (P < 0.001 for all comparisons). We concluded that for patients with moderate symptoms of benign prostatic hyperplasia, surgery is more effective than medication in improving genitourinary symptoms and enhancing the quality of life. Thus, medication as treatment should be reserved for patients who are less bothered by urinary difficulty or who wish to delay surgery.

The incidence of antisperm antibodies in patients with seminal tract obstructions.

Antisperm antibodies are one of the important factors in male infertility caused by vasal obstructions. To investigate the incidence of serum antisperm antibodies in patients with various kinds of seminal tract obstructions, we retrospectively analyzed the sera of 60 patients using the indirect immunobead test (IBT) to find serum antisperm antibodies. Immunoglobulin (Ig)G and IgA class antisperm antibodies were positive in 55% and 18% of those patients with a vasal obstruction caused by inguinal herniorrhaphy and in 60% and 20% of vasectomized patients respectively; whereas these antibodies were positive in 13% and 0% of those patients with an epididymal obstruction of unknown etiology and in 8% and 3% of those patients with congenital bilateral absence of the vas deferens (CBAVD). The incidence of antisperm antibodies was significantly higher in patients with a vasal obstruction than in those with an epididymal obstruction (p < 0.001).

Effects of varicocelectomy on testis volume and semen parameters in adolescents: a randomized prospective study.

Because the treatment of a varicocele in adolescents remains controversial, we undertook a prospective study in 51 male adolescents, aged 15-21 years, who were referred for a varicocele. Twenty-nine patients were treated with high retroperitoneal ligation of the left spermatic vein, and 22 were untreated. Eighteen healthy adolescent volunteers without a varicocele were also assessed. Testicular volume was measured using an orchiometer, and semen analyses were carried out at referral and after 1 year of follow-up. In addition, serum luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone levels were estimated by radioimmunoassay. Both patient groups had significantly smaller testis volumes than the controls at the outset. In the follow-up, the treated patients had testis volumes similar to those of the controls. Both testes increased significantly in volume after treatment. Although semen parameters were comparable in all groups initially, sperm concentration increased significantly after treatment of the varicocele. We concluded that varicocele treatment in adolescents leads to increased testis volume and a higher sperm concentration; however, whether early treatment will improve testicular function remains to be elucidated.

Antibiotic and ejaculation treatments improve resolution rate of leukocytospermia in infertile men with prostatitis.

Leukocytospermia has been associated with notable adverse effects on semen parameters and sperm function. The present study was undertaken to identify men with leukocytospermia and prostatitis in an infertility population and assess the effects of various treatments. One million white blood cells (WBC)/ml semen was defined as leukocytospermia. An expressed prostatic fluid was analyzed for the presence of white blood cells. The presence of more than 10 WBCs/high power field on expressed prostatic secretion was needed for the diagnosis of prostatitis. Those men who had more than one million WBCs in their semen and more than 10 WBCs/high power field on expressed prostatic secretion were enrolled in this study. Of two hundred sixty-three men screened for the presence of leukocytospermia and prostatitis, forty-eight men met both criteria. They were blindly and randomly assigned to one of three groups. Group 1 received no treatment. Group 2 were treated with trimethoprim 80 mg-sulfamethoxazole 400 mg (TMP-SMX) orally twice per day for one month. Group 3 were treated not only with the same antibiotic regimen as group 2 but also were instructed to ejaculate frequently (at least once every three days) for one month. Significant resolution of leukocytospermia occurred in the order of patient groups 3 > 2 > 1 at one month. The resolution rate of leukocytospermia of each group was 76% in group 3, 56% in group 2 and 6.7% in group 1. The rates in groups 2 and 3 were significantly higher than that in group 1 (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Intraluminal content is required for the maintenance of antigrade proluminal movement of 3H-androgens into rat caput epididymal tubules.

The present study was undertaken to determine whether or not antigrade, proluminal 3H-androgen movement in the caput epididymis occurs in the absence of native lumen content. A single tubule was perfused with artificial caput fluid containing no androgen-binding protein for 30 min and subsequently tubules were perifused with Minimum Essential Medium perifusion fluid containing 26.7 microCi/ml 3H-testosterone and 1.3 microCi/ml 14C-polyethyleneglycol for 1 h. Radioactivity of isotopes in perifusion and intraluminal fluids was determined at 1 h after sustaining perifusion, and the percentage of peritubular isotopes appearing in the intraluminal fluid was determined. Net entry of 3H-androgen into the epididymal tubules in the presence of native intraluminal content was approximately 323%. In contrast, intraluminal 3H-androgen concentrations in the epididymal fluid in the absence of native lumen content were significantly reduced, to 100% of those in the peritubular fluid. Antigrade, proluminal movement of 3H-androgen in the caput epididymis does not occur in the absence of native lumen content. Androgen-binding protein in the epididymal lumen may be required to maintain uphill proluminal movement of 3H-androgen into the tubules.

Effects of protein synthesis inhibitor and antimicrotubular agent on transepithelial movement of 3H-androgens in the rat caput epididymis.

The effects of protein synthesis inhibition and disassembly of microtubules in the epididymal epithelia on proluminal movement of 3H-androgens were investigated by using in vivo microperifusion of 3H-testosterone and subsequent micropuncture to obtain peritubular and intraluminal fluids of caput epididymal tubules. Cycloheximide (100 micrograms/ml) was used as protein synthesis inhibitor. Nocodazole (3 micrograms/ml) was used to depolymerize microtubules in the cell. The perifusion fluid was Minimum Essential Medium containing 26.7 microCi/ml 3H-testosterone and 1.3 microCi/ml 14C-polyethyleneglycol (14C-PEG), or the same fluid supplemented with cycloheximide or nocodazole. Radioactivity of 3H-androgen and 14C-PEG in perifusion and intraluminal fluids was determined at one hour after initiation of the sustaining perifusion, and the percentage of radioactivity of 3H-androgen and 14C-PEG appearing in the intraluminal fluid to that in the peritubular fluid was determined. Proluminal movement of 3H-androgens into the caput epididymal tubules in the control rats was 323.4 +/- 73.2%. This value was significantly reduced to 121.8 +/- 13% by addition of cycloheximide to the perifusion fluid (p < 0.01). Transepithelial movement of 3H-androgen in the caput epididymis was significantly decreased to 86.6 +/- 5.3% by exposure of the epididymal tubules to nocodazole (p < 0.01). Inhibition of protein synthesis and disassembly of microtubules in the epididymal epithelial cells completely eliminated antigrade proluminal movement of 3H-androgen into the tubules. Study of the incorporation of 35S-Methionine into epididymal tissue protein revealed significant reduction of the quantity of radiolabeled proteins in the perifused tissue with fluid containing cycloheximide (p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Surgical outcome of microscopic vasectomy reversal: an analysis of 30 cases.

The results of 30 consecutive microscopic vasovasostomy procedures performed at a single institution over a five-year period between 1991 and 1995 were reviewed. When the surgical outcomes of patients who were operated on less than five years after their vasectomy were compared with the outcomes of those patients who received a vasovasostomy more than five years after their vasectomy, decreases in technical success rates were observed as measured by appearance of sperm in ejaculate (56% vs. 36%), biologic recovery as measured by mean sperm counts (56 million vs. 35 million) and mean progressive sperm motility (44% vs. 21%), along with a decrease in clinical success, as measured by overall pregnancy rates (50% vs. 7%, p < 0.05). Therefore, a microscopic vasovasostomy within 5 years of a vasectomy is a favorable procedure for vasectomy reversal.

A case of sex reversal syndrome with sex-determining region (XX male).

We examined a 32-year-old man with a 4-year history of infertility. The man's sex life, male hair pattern, and penis were normal, and he had no history of erection problems. Left and right testicular volumes were 2 ml and 3 ml, respectively. Semen analysis showed no sperm. The endocrine panel revealed increased serum luteinizing hormone and follicle-stimulating hormone levels, and a normal serum testosterone level. A testicular biopsy demonstrated that both Leydig cell and Sertoli cell hyperplasia were present, and that no germ cells were found in the tubules. A chromosome analysis done on the peripheral blood lymphocytes revealed a karyotype of 46, XX. We identified the sex-determining region, Y, by polymerase chain reaction using Y-specific probes in this patient. The diagnosis was XX male.

Serum inhibin levels in normal men and men with idiopathic infertility.

In vitro studies have shown that the Sertoli cell is the primary source of inhibin in the male. To assess the regulation of inhibin production we measured serum inhibin, FSH, LH and testosterone in 21 normal men and 104 men with various testicular disorders resulting in infertility. The infertile men were subdivided into groups on the basis of their mean sperm count, FSH and LH levels. The mean serum concentrations of inhibin in the normal men were 602 +/- 29 U/L and were significantly decreased in those groups with severe oligozoospermia (p < 0.01) or azoospermia (p < 0.01; p < 0.001). The FSH concentrations correlated inversely with serum inhibin concentrations (p < 0.001) in azoospermic men. Azoospermic men with high FSH had significantly lower inhibin and testosterone levels when compared with normal men (p < 0.01; p < 0.001). Serum FSH concentrations were significantly increased in azoospermic men (p < 0.001). Our present results of serum concentrations of inhibin correlating inversely with those of FSH levels suggest that measurement of inhibin may be a useful circulating marker of Sertoli cell function.

The effect of varicocelectomy on testicular volume in infertile patients with varicoceles.

The presence of a varicocele has been associated with decrease in testicular volume both in adult and pediatric patients. However, a study on the effect of varicocelectomy on testicular volume in the adult varicocele patient has never been reported. We determined the testicular volume before and after varicocelectomy in 52 patients with varicocele. A significant increase in testicular volume with the right side showing a greater increase was observed (p < 0.05). No greater increase in testicular volume was noted among the patients who achieved a pregnancy with their spouses. In addition, the age of the patient did not correlate with the observed change in testicular volume. These results suggest that varicocele is clearly related to decreased testicular volume and that varicocelectomy can increase bilateral testis volume resulting in improved testicular function.

Does epididymal length in men with congenital bilateral absence of the vas deferens have a correlation with the fertilization rate of epididymal sperm retrieved by micropuncture technique?

To investigate whether the variable length of the epididymis in men with congenital bilateral absence of the vas deferens (CBAVD) might have a correlation with fertilization and pregnancy rates in in vitro fertilization (IVF) treatment, we conducted a retrospective study involving a total of 60 CBAVD patients. All patients in this study had epididymal micropuncture in conjunction with perivascular nerve stimulation as part of the IVF program at the Nagoya University Hospital Reproduction Center. The patients were classified into 3 groups: group I consisted of patients having only a proximal portion of the caput epididymidis with a length of between 0.5 and 1.9 cm, group II consisted of patients with the caput and a portion of the corpus epididymidis with a length of between 2.0 and 4.0 cm, and group III consisted of patients with the caput, corpus and cauda of the epididymis measuring more than 4.0 cm. There were no differences in the sperm count, progression and normal morphology among the 3 groups; however, the motility was progressively higher in patients with a longer epididymis (12% in group 1, 18% in group II and 31% in group III). It was evident that group III showed the highest fertilization and pregnancy rate per patient (23% and 28%, respectively) among the 3 groups. This study shows that epididymal sperm from CBAVD patients with a longer epididymis have a higher IVF rate than CBAVD patients with a shorter epididymis.