AMPK/autophagy-mediated alleviation of tendinopathy by IL-38: A novel strategy for the treatment of obesity-related tendinopathy.

The effect of interleukin-38 (IL-38), a recently identified member of the IL-1 family with potential applications in various inflammation-related conditions, on ER stress has not been explored. Furthermore, its role in obesity-associated tendinopathy has not been investigated. In this study, human primary tenocytes were treated with palmitate (200 or 400 µM) and palmitate plus IL-38 (0-50 ng/mL) for 24 h. Western blotting was used to assess ER stress and tendinopathogenic markers in tenocytes. Monodansylcadaverine (MDC) staining was used to evaluate autophagosomes. Apoptosis was determined by cell viability assays, caspase 3 activity assays and TUNEL assays. Cell migration was evaluated by a cell scratch assay. Small interfering (si) RNA transfection was used for target gene silencing. Treatment of tenocytes with IL-38 attenuated apoptosis, restored the balance between MMPs and TIMP-1, and alleviated ER stress under palmitate conditions. IL-38 treatment enhanced AMPK phosphorylation and promoted the expression of autophagy markers related to LC3 conversion, p62 degradation, and autophagosome formation in cultured tenocytes. The effects of IL-38 on ER stress, apoptosis, and MMP-9, MMP-13, and TIMP-1 expression in palmitate-treated tenocytes were abrogated by AMPK siRNA or 3-methyladenine (3MA). These results suggest that IL-38 alleviates ER stress through the AMPK/autophagy pathway, thereby reducing apoptosis and preventing extracellular matrix (ECM) degradation in tenocytes under hyperlipidemic conditions. This study provides a promising therapeutic avenue for treating obesity-related tendinopathy using an endogenous compound such as IL-38.

Reinforcement of Positive Electrode-Electrolyte Interface without Using Electrolyte Additives Through Thermoelectrochemical Oxidation of LiPF6 for Lithium Secondary Batteries.

Owing to the limited electrochemical stability window of carbonate electrolytes, the initial formation of a solid electrolyte interphase and surface film on the negative and positive electrode surfaces by the decomposition of the electrolyte component is inevitable for the operation of lithium secondary batteries. The deposited film on the surface of the active material is vital for reducing further electrochemical side reactions at the surface; hence, the manipulation of this formation process is necessary for the appropriate operation of the assembled battery system. In this study, the thermal decomposition of LiPF6 salt is used as a surface passivation agent, which is autocatalytically formed during high-temperature storage. The thermally formed difluorophosphoric acid is subsequently oxidized on the partially charged high-Ni positive electrode surface, which improves the cycleability of lithium metal cells via phosphorus- and fluorine-based surface film formation. Moreover, the improvement in the high-temperature cycleability is demonstrated by controlling the formation process in the lithium-ion pouch cell with a short period of high-temperature storage before battery usage.

Interface Engineering with Nonsacrificial Perfluorinated-Anion Additives for Boosting the Kinetics of Lithium-Ion Batteries.

Though lithium-ion batteries (LIBs) have seen a meteoric rise in worldwide deployment over the last decade, they should be further advanced in constant demand of higher rate capability and wider temperature adaptability. A solid electrolyte interphase (SEI) is the essential part of LIBs, determining the charge-discharge performance and degradation behavior. Herein, improvement of the SEI properties is achieved by regulating the electrochemical double layer structure with a nonsacrificial electrolyte additive, that is, lithium nonafluoro-1-butanesulfonate. The anion adsorption of the additive affects the decomposition behavior of other additive and solvent species, and the generated SEI at the graphite electrode becomes thinner and more uniform, leading to decreased impedance and finally resulting in improved energy efficiency, power capability, and fast charging performance of the graphite/NCM811 cell. Furthermore, the low-temperature cycleability at -20 °C is considerably enhanced with no dendritic Li metal deposition at the negative electrode surface. A mechanistic study on the interfacial phenomena and the effect is carried out by using various theoretical and experimental methods, such as density functional theory calculations, electrochemical quartz crystal microbalance, and transmission electron microscopy. Consequently, the approach of SEI modification with the nonsacrificial electrolyte additive can be one of the effective ways to advance LIB technology in future.

Interphasial Engineering via Individual Moiety Functionalized Organosilane Single-Molecule for Extreme Quick Rechargeable SiO/NCM811 Lithium-Ion Batteries.

The individual moiety-functionalized organosilane single molecule, that is, 1,1,1,5,5,5-hexamethyl-3-[(trimethylsilyl)oxy]-3-vinyltrisiloxane (TMSV), is investigated as an electrolyte additive for a less charge-consuming and viscoelastic solid electrolyte interphase (SEI) forming agent, finally accomplishing extremely quick (6 min) rechargeable SiO/NCM811 lithium-ion batteries. The moiety of the vinyl group serves with a poly(ethylene oxide)-like viscoelastic SEI film on the SiO electrode, which provides a physicochemically stable interphase during long-term cycling. The increase of DC-iR due to electrolyte decomposition on the continuously exposed SiO surface with cycling is inhibited by the alternated SEI composition. Degradation of bulk electrolyte solution caused by thermal decomposition of the LiPF6 salt is also suppressed by the trimethylsilyl moiety in the TMSV additive, which scavenges HF. Owing to the multifunctionality of TMSV, the cycle performance of laminated pouch full cells comprising high-nickel-contented NCM811 positive electrode and SiO-enriched negative electrode is significantly improved at both room and elevated temperatures. Furthermore, the 6 min quick recharging cycle performance is also enhanced by the TMSV additive.

Associations of Earphone Use with Tinnitus and Anxiety/Depression.

OBJECTIVE: This study aimed to investigate the association of earphone use with audiologic and psychologic factors. MATERIALS AND METHODS: Korea National Health and Nutrition Examination Survey 2010-2012 data were collected for participants aged ≥12 years old with earphone use ≥1 hour/day. They were matched to a control group for age, sex, income, and education level. The relationship between earphone use and the hearing thresholds of pure-tone audiometry, tinnitus, and psychologic factors such as depression and anxiety, and other quality of life variables was analyzed using multiple logistic regression tests with complex sampling. RESULTS: Among the participants, 22.9% (449/1955) of earphone users and 18.1% (355/1600) of control participants had tinnitus (P < 0.001). Earphone users showed 1.27-times higher odds for tinnitus (95% confidence interval [CI] = 1.09-1.50, P = 0.003). Moreover, 6.5% (128/1955) of earphone users and 5.0% (97/1600) of control participants had anxiety and depressive symptoms (P = 0.033). Earphone users showed 1.32-times higher odds for anxiety and depressive symptoms (95% CI = 1.14-1.52, P = 0.040). Nevertheless, the hearing thresholds were comparable between earphone users and control participants. CONCLUSION: Earphone use was associated with tinnitus and anxiety or depressive symptoms.

Single-sided Deafness Leads to Changes in Vesicular Synaptic Transporters and Matrix Metalloproteinase 9 in the Primary Auditory Cortex.

Changes in perineuronal nets (PNNs) after hearing loss were described in previous studies. The present study aimed to examine how single-sided deafness (SSD) affects the expression of excitatory and inhibitory synaptic transporters and PNNs in the primary auditory cortex (A1). Sprague-Dawley rats (8-week-old females, n = 30) were divided into three groups: (1) the SSD 2-week group (n = 10), (2) the SSD 4-week group (n = 10), and (3) the 4-week control group (n = 10). The expression levels of vesicular glutamate transporter 1 (VGLUT1), VGLUT2, vesicular GABA transporter (VGAT), and genes related to PNNs were measured using quantitative reverse transcription-polymerase chain reaction. The A1 was immunostained for VGLUT1, glutamate acid decarboxylase (GAD) 67, neurocan, aggrecan, brevican, and Wisteria floribunda agglutinin (WFA). The expression levels of VGLUT1, VGLUT2, and VGAT were elevated in the A1 on the ipsilateral side in the SSD groups compared with those in the control groups. Aggrecan expression was elevated in the A1 on the contralateral side in the SSD 2-week group. The SSD groups had elevated expression levels of metalloproteinase (MMP) 9 on the contralateral side. The presynaptic glutamatergic and GABAergic transporters were increased in the A1 on the ipsilateral side after induction of SSD. Changes in the cortical auditory nervous system accompanied changes in the PNNs and their degradation enzymes MMP9 and MMP14.

Noise exposure alters MMP9 and brevican expression in the rat primary auditory cortex.

BACKGROUND: This study aimed to investigate the changes in molecules related to perineuronal nets (PNNs) and synaptic transporters in the primary auditory cortices of rats with noise-induced hearing loss. Female Sprague-Dawley rats at postnatal day 7 were divided into the noise and control groups. Four hours of 115 dB SPL white noise was delivered for 10 days to the noise group. Thirty days after noise exposure, the primary auditory cortex and the inferior colliculus were harvested. The expression levels of vesicular glutamatergic transporter (VGLUT)1, VGLUT2, vesicular GABA transporter (VGAT), glutamate decarboxylase (GAD)67, brevican, aggrecan, MMP9, and MMP14 were evaluated using real-time reverse transcription polymerase chain reaction or western blot. An immunofluorescence assay was conducted to assess parvalbumin (PV), Wisteria floribunda agglutinin (WFA), and brevican. The immune-positive cells were counted in the primary auditory cortex. RESULTS: The expression level of VGLUT1 in the primary auditory cortex was decreased in the noise group. The expression level of VGLUT2 in the inferior colliculus was elevated in the noise group. The expression levels of brevican and PV + WFA in the primary auditory cortex were decreased in the noise group. The expression level of MMP9 in the primary auditory cortex was increased in the noise group. CONCLUSION: Noise-induced hearing loss during the precritical period impacted PNN expression in the primary auditory cortex. Increased MMP9 expression may have contributed to the decrease in brevican expression. These changes were accompanied by the attenuation of glutamatergic synaptic transporters.

Tauroursodeoxycholic acid attenuates cisplatin-induced hearing loss in rats.

Tauroursodeoxycholic acid (TUDCA) has been reported to be protective against apoptosis and oxidative stress in various cell types. A few studies have demonstrated otoprotective effects of TUDCA in mouse models. This study investigated the otoprotective effects of TUDCA in cisplatin (CXP)-induced hearing-loss rats. Eight-week-old female Sprague-Dawley rats were used. The CXP group received intraperitoneal injection of CXP at a dose of 5 mg/kg from day 1 to day 3. The CXP + TUDCA group received an intraperitoneal injection of 5 mg/kg CXP and 100 mg/kg TUDCA from day 1 to day 3. The mRNA expression levels of heme oxygenase 1 (HO1) and superoxide dismutase 2 (SOD2) were measured, and the protein levels of caspase 3, cleaved caspase 3, and aryl hydrocarbon receptor (AhR) were evaluated. The CXP group demonstrated higher mean auditory brainstem responses (ABR) thresholds than the control group. The mean ABR threshold shifts were lower in the CXP + TUDCA group than in the CXP group. The CXP group showed elevated HO1 and SOD2 mRNA expression levels compared to the control group, but these changes were reversed in the CXP + TUDCA group. Compared to the levels in the control group, caspase 3, cleaved caspase 3, and AhR levels were higher in the CXP group, but the increase in cleaved caspase-3 was attenuated in the CXP + TUDCA group. The cochlea showed a higher number of spiral ganglion cells and outer hair cells in the CXP + TUDCA group than in the CXP group. TUDCA reduced CXP-induced hearing loss in adult rats. The HO1-mediated antioxidative effects attenuated apoptosis in the cochlea, but AhR activation was not reversed.

WT1 Measurable Residual Disease Assay in Patients With Acute Myeloid Leukemia Who Underwent Allogeneic Hematopoietic Stem Cell Transplantation: Optimal Time Points, Thresholds, and Candidates.

The absence of relevant guidelines for Wilms tumor 1 (WT1) gene quantification as a measurable residual disease (MRD) assessment for patients with acute myeloid leukemia (AML) undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) has limited the widespread use in practice. We investigated optimal time points, thresholds, and candidates for the bone marrow WT1 MRD assay in 425 consecutive patients with AML who underwent allo-HSCT. WT1 expression kinetics before allo-HSCT and at 1 or 3 months after allo-HSCT were determined by real-time PCR using the European LeukemiaNet (ELN) normalized method. Relapsed patients had significantly higher WT1 levels before allo-HSCT and at 3 months after allo-HSCT. The best time point for the WT1 MRD assay was before allo-HSCT by the receiver operating characteristic curve. Among various thresholds, 250 copies recommended from ELN researchers were mostly predictive of post-transplant relapse. In multivariate analysis, WT1 MRD positivity independently predicted relapse, resulting in inferior survival. In subgroup analyses, pretransplant WT1 MRD positivity was predictive of post-transplant relapse in the intermediate group, whereas WT1 MRD positivity occurred at 3 months after allo-HSCT in favorable and adverse risk groups. Among MRD-positive patients before allo-HSCT, all patients who were MRD positive at 3 months relapsed within 6 months. The WT1 MRD assay before allo-HSCT or 3 months after allo-HSCT is useful for predicting post-transplant relapse with a different significance in each risk group by time points, showing the benefit of multiple tests over time. Such monitoring is particularly available in patients with AML without specific molecular targets.

Large Cell Neuroendocrine Carcinoma of the Colon With Carcinomatosis Peritonei.

Colorectal large-cell neuroendocrine carcinomas (NECs) are extremely rare and have very poor prognosis compared to adenocarcinomas. A 74-year-old man presented with abdominal pain, diarrhea and hematochezia. The histopathologic report of colonoscopic biopsy performed at a local clinic was a poorly differentiated carcinoma. An abdominopelvic computed scan revealed irregularly enhanced wall thickening at the sigmoid colon with regional fat stranding and lymphnode enlargement. He underwent a laparoscopic high anterior resection with selective peritonectomy for peritoneal carcinomatosis, intraoperative peritoneal irrigation chemotherapy, and early postoperative intraperitoneal chemotherapy for 5 days. The tumor had a high proliferation rate (mitotic count > 50/10 HPFs and 90% of the Ki-67 index) and lymph-node metastases had occurred. On immunohistochemistry, the tumor cells expressed CD56 and synaptophysin. Large-cell NEC was confirmed. Systemic chemotherapy with cisplatin/etoposide was done. The patient is still alive after 3 years with no evidence of recurrence.

Is surgical excision necessary for the treatment of Granulomatous lobular mastitis?

BACKGROUND: We aimed to investigate the role of surgical excision in treating granulomatous lobular mastitis. METHODS: We performed a retrospective chart review of patients with granulomatous lobular mastitis treated from March 2008 to March 2014. We analyzed clinical features and therapeutic modalities and compared the patient outcomes based on treatment. RESULTS: During the study period, a total of 34 patients were diagnosed with granulomatous lobular mastitis and treated. Initial treatments included wide excision (18), oral steroids after incision and drainage (14), and antibiotic therapy (2). The patients receiving only antibiotic therapy showed no improvement after 1 month and wide excision was then performed. Wide excision resulted in nine case of delayed wound healing with fistula. These patients were treated with oral steroids for 1.5-5 months, with subsequent improvement. Overall, 11 out of 20 patients who had underwent wide excision showed improvement without additional treatment. Fourteen patients who had initially received oral steroids for 1 to 6 months (average, 2.8 months) after incision and drainage showed complete remission. During the median follow-up period with 45.5 months (range, 22-98 months), six patients (17.6%) experienced recurrence. Wide excision group experienced recurrence in five (25%) and steroid and drainage group experienced recurrence in one (7.1%). All six recurrences responded to additional steroid therapy for average 3.5 months. Most wide excision group left extensive breast scarring with deformation that was not in steroid and drainage group. CONCLUSIONS: Wide excision resulted high recurrence than steroid and drainage group and left extensive scarring. Steroid therapy with or without abscess drainage may be the first choice of treatment for majority cases with granulomatous lobular mastitis.

The True Deep Femoral Artery Aneurysm: A Case Report.

A 55-year-old man with a palpable pulsatile mass and pain in his left thigh was presented to us. He had no history of trauma in his left leg, interventions, operation, or medical diseases, including cardiac valve disease, endocarditis, and systemic infection. The size of the aneurysm was 10 cm×7 cm with a mural thrombus in ultrasonography and multidetector computer tomography. There was no evidence of other aneurysms or occlusive lesions in the other arteries. The aneurysm was resected without a vascular reconstruction of the deep femoral artery. The patient's symptom improved rapidly. The patient had an uneventful postoperative recovery without complications. We report a case of true deep femoral artery aneurysm, which was successfully treated with resection of an aneurysm without a vascular reconstruction.

Comparison of outcomes of surgeon-performed intraoperative ultrasonography-guided wire localization and preoperative wire localization in nonpalpable breast cancer patients undergoing breast-conserving surgery: A retrospective cohort study.

This study aimed to determine the efficacy of intraoperative ultrasonography-guided wire localization guided breast-conserving surgery (BCS) for nonpalpable breast cancer and compare it to conventional preoperative wire localization (PWL) guided surgery.We retrospectively analyzed the medical charts of 214 consecutive nonpalpable breast cancer patients who underwent BCS using intraoperative ultrasonography-guided wire localization by a surgeon (IUWLS) and PWL, between April 2013 and March 2017. Positive surgical margins, reexcision rates, and resection volumes were investigated.Of the total cohort, 124 patients underwent BCS with IUWLS and 90 patients with PWL. The following did not differ between the IUWLS and PWL groups: positive margin status, re-excision rate, conversion rate, permanent positive margin status, reoperation rate, median optimal resection volume (ORV), median total resection volume (TRV), and median closest tumor-free margin. Rather, median (range) widest tumor-free margin was significantly smaller in the IUWLS group (9 mm [5-12]) than in the PWL group (14 mm [9-20]; P = .003]). Median (range) calculated resection ratio (CRR) was significantly lower in the IUWLS group (1.67 [0.87-9.38]) than in the PWL group (4.83 [1.63-21.04]; P = .02).In nonpalpable breast cancer patients undergoing BCS, IUWLS showed positive resection margins and reexcision rates equivalent to those of the conventional PWL method. Additionally, excision volume and widest tumor-free margin were smaller with IUWLS, confirming that healthy breast tissue is less likely to be resected with this method. Our results suggest that IUWLS offers an excellent alternative to PWL, while avoiding PWL-induced patient discomfort.

Intensive Long Distance Running as a Possible Cause of Multiple Splanchnic Arterial Aneurysms: A Case Report.

This is a case report that suggests the possible association between multiple splanchnic arterial aneurysms and long-distance running. The clinical features of one patient admitted at Chungbuk National University Hospital for treatment of multiple splanchnic arterial aneurysms were reviewed. A 54-year-old man had a recurrent, intermittent and epigastric pain for 2 months. There was no abnormality in gastroscopy and colonoscopy. An abdominal computed tomography angiography documented calcified superior mesenteric artery (SMA) and splenic artery aneurysms. The patient had a history of recreational long-distance running for over 10 years. His average running time per week was more than 10 hours. There was no evidence of systemic arteritis, connective tissue disorder or infectious process that may have caused the aneurysms. He did not take any drugs. The SMA aneurysm was opened, and the aneurysmal segment of SMA was replaced with a vein graft. The splenic aneurysm was observed. The patient recovered without any sequelae.

Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton's jelly for treating bone defects.

Alternative sources of mesenchymal stem cells (MSCs) for replacing bone marrow (BM) have been extensively investigated in the field of bone tissue engineering. The purpose of this study was to compare the osteogenic potential of canine MSCs derived from adipose tissue (AT), BM, umbilical cord blood (UCB), and Wharton's jelly (WJ) using in vitro culture techniques and in vivo orthotopic implantation assays. After canine MSCs were isolated from various tissues, the proliferation and osteogenic potential along with vascular endothelial growth factor (VEGF) production were measured and compared in vitro. For the in vivo assay, MSCs derived from each type of tissue were mixed with ß-tricalcium phosphate and implanted into segmental bone defects in dogs. Among the different types of MSCs, AT-MSCs had a higher proliferation potential and BM-MSCs produced the most VEGF. AT-MSCs and UCB-MSCs showed greater in vitro osteogenic potential compared to the other cells. Radiographic and histological analyses showed that all tested MSCs had similar osteogenic capacities, and the level of new bone formation was much higher with implants containing MSCs than cell-free implants. These results indicate that AT-MSCs, UCB-MSCs, and WJ-MSCs can potentially be used in place of BM-MSCs for clinical bone engineering procedures.

Comparison of mesenchymal stem cells derived from fat, bone marrow, Wharton's jelly, and umbilical cord blood for treating spinal cord injuries in dogs.

Previous animal studies have shown that transplantation of mesenchymal stem cells (MSCs) into spinal cord lesions enhances axonal regeneration and promotes functional recovery. We isolated the MSCs derived from fat, bone marrow, Wharton's jelly and umbilical cord blood (UCB) positive for MSC markers and negative for hematopoietic cell markers. Their effects on the regeneration of injured canine spinal cords were compared. Spinal cord injury was induced by balloon catheter compression. Dogs with injured spinal cords were treated with only matrigel or matrigel mixed with each type of MSCs. Olby and modified Tarlov scores, immunohistochemistry, ELISA and Western blot analysis were used to evaluate the therapeutic effects. The different MSC groups showed significant improvements in locomotion at 8 weeks after transplantation (P<0.05). This recovery was accompanied by increased numbers of surviving neuron and neurofilament-positive fibers in the lesion site. Compared to the control, the lesion sizes were smaller, and fewer microglia and reactive astrocytes were found in the spinal cord epicenter of all MSC groups. Although there were no significant differences in functional recovery among the MSCs groups, UCB-derived MSCs (UCSCs) induced more nerve regeneration and anti-inflammation activity (P<0.05). Transplanted MSCs survived for 8 weeks and reduced IL-6 and COX-2 levels, which may have promoted neuronal regeneration in the spinal cord. Our data suggest that transplantation of MSCs promotes functional recovery after SCI. Furthermore, application of UCSCs led to more nerve regeneration, neuroprotection and less inflammation compared to other MSCs.

Treatment of a recurrent rectourethral fistula by using transanal rectal flap advancement and fibrin glue: a case report.

Rectourethral fistulas (RUFs) in adults are rare and could result from complicated trauma, and prostatic or rectal surgery. RUFs have been treated initially by using primary repair and omental interposition with or without a colostomy during surgery. Recurrent RUFs require complex surgery, such as a low rectal resection and coloanal anastomosis, an interposition flap of the datos muscle or gracilis muscle, and others. Recently, transanal rectal flap advancement and fibrin glue injection have provided an effective occlusion of RUFs. However, no reports about this technique exist for cases of recurrent RUFs. We report a case of a recurrent RUF successfully repaired by using transanal rectal flap advancement combined with fibrin glue injection into the fistula tract. The postoperative course was uneventful without complications. At the 1-year follow-up, no complications such as urethral stricture or recurrence existed, and voiding was normal without anal incontinence.

Effect of acellular dermal matrix as a delivery carrier of adipose-derived mesenchymal stem cells on bone regeneration.

The purpose of this study is to evaluate the effect of acellular dermal matrix (ADM) as a delivery carrier of adipose-derived mesenchymal stem cells (ASCs) on bone regeneration in athymic murine calvarial bone defect. Paired-critical size defects in nude rat skull were made. The right-side defects received ASCs/ADM or only ADM, whereas the left-side defect was not treated. In 3D images, new bone formation in the ASCs/ADM group was apparent at 4 wk, but in the ADM group at 8 wk. At 4 and 8 wk, bone mineral density and tissue volume in rats that received ASCs/ADM were significantly greater than rats that received ADM and control groups. Histological examination revealed that the defect was repaired by bone in the ASCs/ADM group, whereas only minimal bone island with fibrous connection was observed in the control group. In histomorphometric analysis, the total healing score in the ASCs/ADM group at 4 wk was significantly higher than the ADM and negative control group, whereas the score of 8 wk was similar between the ASCs/ADM and ADM group. ASCs/ADM implants promote new bone formation more rapidly than ADM only or no treatment. ADM seeded with ASCs may be potentially useful as a future biomaterial option in bone implants.

Functional recovery after spinal cord injury in dogs treated with a combination of Matrigel and neural-induced adipose-derived mesenchymal Stem cells.

BACKGROUND AIMS: Previous studies have reported that scaffold or cell-based transplantation may improve functional recovery following spinal cord injury (SCI), but these results were based on neuronal regeneration and cell replacement. In this study, we investigated whether a combination of Matrigel and neural-induced mesenchymal stem cells (NMSC) improved hindlimb function in dogs with SCI, and what mechanisms were involved. METHODS: We pre-differentiated canine adipose-derived mesenchymal stem cells into NMSC. A total of 12 dogs subjected to SCI procedures were assigned to one of the following three transplantation treatment groups: phosphate-buffered saline (PBS); Matrigel; or Matrigel seeded with NMSC. Treatment occurred 1 week after SCI. Basso, Beattie and Bresnahan (B.B.B.) and Tarlov scores, histopathology, immunofluorescence staining and Western blot analysis were used to evaluate the treatment effects. RESULTS: Compared with dogs administered PBS or Matrigel alone, dogs treated with Matrigel + NMSC showed significantly better functional recovery 8 weeks after transplantation. Histology and immunochemical analysis revealed that the combination of Matrigel + NMSC reduced fibrosis from secondary injury processes and improved neuronal regeneration more than the other treatments. In addition, the combination of Matrigel + NMSC decreased the expression of inflammation and/or astrogliosis markers. Increased expressions of intracellular molecules related to neuronal extension, neuronal markers and neurotrophic factors were also found in the Matrigel + NMSC group. However, the expression of nestin as a neural stem cell marker was increased with Matrigel alone. CONCLUSIONS: The combination of Matrigel + NMSC produced beneficial effects in dogs with regard to functional recovery following SCI through enhancement of anti-inflammation, anti-astrogliosis, neuronal extension and neuronal regeneration effects.

Effect of matrigel on the osteogenic potential of canine adipose tissue-derived mesenchymal stem cells.

Adipose tissue-derived mesenchymal stem cells (Ad-MSCs) are a promising source of cells for bone tissue engineering. Matrigel is a basement membrane extract containing multiple extracellular components. This mixture may promote the osteogenic differentiation of MSCs and provide a more appropriate microenvironment for transplanted cells. Here, we investigated the effect of Matrigel on the osteogenic potential of Ad-MSCs. Canine Ad-MSCs were cultured in 2D and 3D matrices and implanted into subcutaneous pouches of dogs either with or without Matrigel. Culture mineralization, cell adhesion efficiency, cell proliferation, osteoid matrix production and alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase activities were quantified and compared. Ad-MSCs grown in 2D cultures with Matrigel showed higher levels of calcium deposition and ALP activity than those grown in the absence of Matrigel under osteogenic conditions. In 3D cultures, the cells cultivated with Matrigel showed greater attachment, proliferation and osteogenic differentiation than those grown without Matrigel. In vivo, Ad-MSCs implanted with Matrigel showed higher osteogenic potential than those without Matrigel. In conclusion, these data suggest that the use of Matrigel can increase the osteogenic potential of canine Ad-MSCs.