Effectiveness of novel oral hygiene instruction avoiding inattentional blindness using an application for unique plaque control record calculation: A randomized clinical trial.

OBJECTIVE: This study was performed to estimate the effectiveness of novel oral hygiene instruction (OHI) focusing on areas with deep periodontal pockets for reduction of periodontal inflammation. BACKGROUND DATA DISCUSSING THE PRESENT STATUS OF THE FIELD: Because stained areas on the plaque chart do not always correspond to the areas with deep periodontal pockets, conventional OHI based on O'Leary's plaque control record (PCR) often provides guidance inconsistent with the target area. METHODS: This randomized clinical trial involved two groups: (1) OHI based on the PCR limited in deep pocket sites (novel OHI group) and (2) OHI based on O'Leary's PCR (conventional OHI group). The unique PCR (aggressive target for PCR [agPCR]; only counting the plaque-stained areas with PD at ≥4 mm sites) for the novel OHI was calculate by dedicated expression program. The probing depth (PD), bleeding on probing (BOP), and periodontal inflamed surface area (PISA) were obtained at the baseline and 5 to 6 months later. RESULTS: The approximation curve with PISA before and after instruction indicated that the PISA converged to a lower value after instruction in the novel OHI group. The approximation curve with the improvement rate of the PISA and agPCR showed a positive correlation in the novel OHI group but no correlation in the conventional OHI group. CONCLUSION: Control of inflammation was more effective in the novel OHI group. These results suggest that this novel OHI technique using our developed application could be used as a strategy to improve the effectiveness of brushing instruction.

Factors influencing the continuation of home blood pressure measurement in community-dwelling older adults: the NOSE study.

OBJECTIVE: This study aimed to identify the factors influencing home blood pressure measurement (HBPM) continuation in community-dwelling older adults. METHODS: A longitudinal analysis used the NOSE study intervention group datasets. The participants were encouraged HBPM with self-monitoring devices provided to them twice in the morning and twice in the evening. Every 7-day interval from the HBPM start date was defined as 1 week, and the number of HBPMs per week was counted. The first week in which the number of HBPMs was zero was defined as the week in which HBPM was discontinued. Participants who did not experienced discontinuation until the end of the observation period were considered complete survivors in the survival time analysis. RESULTS: Data from 437 participants were included in the analysis. Of these, 120 (27.5%) discontinued HBPM. In univariate analysis, factors significantly associated with HBPM discontinuation included exercise habits [hazard ratio per one unit 0.47; 95% confidence interval (CI) 0.31-0.69], social participation (hazard ratio 0.65; 95% CI 0.42-0.99), MoCA-J score (hazard ratio 0.94; 95% CI 0.90-0.98), and frailty (hazard ratio 5.20; 95% CI 2.87-9.43). In multivariate analysis, factors significantly associated with HBPM discontinuation included sex (hazard ratio 0.55; 95% CI 0.32-0.95; ref. = female individuals), smoking history (hazard ratio 1.69; 95% CI 1.02-2.80), exercise habits (hazard ratio 0.51; 95% CI 0.30-0.85), MoCA-J score (hazard ratio 0.93; 95% CI 0.88-0.98), and frailty (hazard ratio 3.31; 95% CI 1.50-7.29). CONCLUSION: Among community-dwelling older adults, female sex, smoking history, lack of exercise, cognitive decline, and frailty were identified as factors influencing HBPM discontinuation.

Unusual imaging appearance of cemental tear in the maxillary first molar on cone-beam computed tomography: A case report.

A cemental tear (CeT) is a definitive clinical entity and its radiographic appearance is well known in single-rooted teeth. However, the imaging features of CeT in multi-rooted teeth have not been clarified. We report a case of CeT which arose in the maxillary first molar and exhibited an unusual appearance in cone-beam computed tomography images. The torn structure was verified as cementum by micro-computed tomography and histological analysis. The hypercementosis, most likely induced by occlusal force, might have been torn from the root by a stronger occlusal force caused by the mandibular implant. An unusual bridging structure was created between the two buccal roots. These features may occur in multi-rooted teeth with long-standing deep pockets and abscesses that are resistant to treatment.

Development and characterization of islet-derived mesenchymal stem cells from clinical grade neonatal porcine cryopreserved islets.

BACKGROUND: Porcine tissues display a great potential as donor tissues in xenotransplantation, including cell therapy. Cryopreserving clinical grade porcine tissue and using it as a source for establishing therapeutic cells should be advantageous for transportation and scheduled manufacturing of MSCs. Of note, we previously performed encapsulated porcine islet transplantation for the treatment of unstable type 1 diabetes mellitus in the clinical setting. It has been reported that co-transplantation of islets and Mesenchymal stem cells (MSCs) enhanced efficacy. We assume that co-transplantation of porcine islets and porcine islet-derived MSCs could improve the efficacy of clinical islet xenotransplantation. METHODS: MSCs were established from fresh and cryopreserved non-clinical grade neonatal porcine islets and bone marrow (termed non-clinical grade npISLET-MSCs and npBM-MSCs, respectively), as well as from cryopreserved clinical grade neonatal porcine islets (termed clinical grade npISLET-MSCs). Subsequently, the cell proliferation rate and diameter, surface marker expression, adipogenesis, osteogenesis, and colony-forming efficiency of the MSCs were assessed. RESULTS: Cell proliferation rate and diameter did not differ between clinical grade and non-clinical grade npISLET-MSCs. However, non-clinical grade npBM-MSCs were significantly shorter and smaller than both npISLET-MSCs (p < 0.05). MSC markers (CD29, CD44, and CD90) were strongly expressed in clinical grade npISLET-MSCs and non-clinical grade npISLET-MSCs and npBM-MSCs. The expression of MSC-negative markers CD31, CD34, and SLA-DR was low in all MSCs. Clinical grade npISLET-MSCs derived from adipose and osteoid tissues were positive for Oil Red and alkaline phosphatase staining. The results of colony-forming assay were not significantly different between clinical grade npISLET-MSCs and non-clinical grade npBM-MSCs. CONCLUSION: The method described herein was successful in of developing clinical grade npISLET-MSCs from cryopreserved islets. Cryopreserved clinical grade porcine islets could be an excellent stable source of MSCs for cell therapy.

Suppression of subgingival bacteria by antimicrobial photodynamic therapy using transgingival irradiation: A randomized clinical trial.

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) is an effective method for eradicating bacteria in periodontal therapy. Standard aPDT requires the insertion of a laser tip into a periodontal pocket, in which the direction of irradiation is limited. Therefore, we devised an aPDT method that uses a transgingival near-infrared wavelength and indocyanine green-encapsulated and chitosan-coated nanoparticles as a photosensitizer. METHODS: Forty patients undergoing supportive periodontal therapy, who had a single root tooth with a pocket of 5 mm or deeper, were used as subjects. In the test group, aPDT was performed by laser irradiation from outside the gingiva using photosensitizer nanoparticles. In the control group, pseudo aPDT without photosensitizer was performed by transgingival irradiation. Subgingival plaque was sampled from inside the pocket before, immediately after, and 1 week after treatment, and evaluated by colony counting and real-time polymerase chain reaction. RESULTS: There were no significant differences in age, sex, periodontal pocket depth, and bleeding on probing between the test and control groups. Compared with the colony count before treatment, the count in the test group was significantly reduced immediately after treatment. The number of patients with colony reduction to ≤50% and ≤10% was significantly higher in the test group than in the control group. None of the participants reported pain, although one participant reported discomfort. CONCLUSION: As a bacterial control method for residual pockets in patients undergoing supportive periodontal therapy, transgingival aPDT is a promising treatment strategy that is not generally accompanied by pain or discomfort.

Ebi3 knockout aggravates experimental periodontitis via Th17 polarization.

AIM: To investigate the role of Ebi3-related cytokines (i.e., interleukin [IL]-35 and/or IL-27) in experimental periodontitis using Ebi3 knockout (KO) mice. MATERIALS AND METHODS: The maxillary right second molar teeth of Ebi3 KO mice and C57BL/6 mice were tied with a silk ligature to induce periodontitis. Three days after ligation, gingival tissues were collected for gene expression analyses. Five days after ligation, the maxillae were removed for haematoxylin and eosin staining and immunohistochemistry. Seven days after ligation, the maxillae were removed for micro-computed tomography. RESULTS: The ligated side of Ebi3 KO mice showed intense alveolar bone resorption, which was substantially more pronounced than in wild-type (WT) mice. IL-17A expression was significantly higher in the gingiva of the ligated side of Ebi3 KO mice compared with WT mice. IL-10 expression was significantly lower in Ebi3 KO mice than in WT mice. The ligature-induced alveolar bone resorption in Ebi3 KO mice that received recombinant IL-35 injection was significantly less compared with that in Ebi3 KO mice that received control injection. CONCLUSIONS: Together, these findings suggest that Th17 cells exacerbate experimental periodontitis in mice lacking Ebi3 and that IL-35 may play a critical role in inhibiting periodontal tissue destruction.

Relationship between oxygen partial pressure and inhibition of cell aggregation of human adipose tissue-derived mesenchymal stem cells stored in cell preservation solutions.

Introduction: This study investigated the storage conditions under which cell aggregation occurs and the conditions that inhibit cell aggregation when human adipose tissue-derived mesenchymal stem cells (hADSCs) are stored in lactated Ringer's solution (LR) supplemented with 3% trehalose and 5% dextran 40 (LR-3T-5D). Methods: We first examined the effects of storage temperature and time on the aggregation and viability of hADSCs stored in LR and LR-3T-5D. The cells were stored at 5 °C or 25 °C for various times up to 24 h. We then evaluated the effects of storage volume (250-2,000 µL), cell density (2.5-20 × 105 cells/mL), and nitrogen gas replacement on aggregation, oxygen partial pressure (pO2), and viability of hADSCs stored for 24 h at 25 °C in LR-3T-5D. Results: When stored in LR-3T-5D, viability did not change under either condition compared with pre-storage, but the cell aggregation rate increased significantly with storage at 25 °C for 24 h (p<0.001). In LR, the aggregation rate did not change under either condition, but cell viability decreased significantly after 24 h at both 5 °C and 25 °C (p < 0.05). The cell aggregation rates and pO2 tended to decrease with increasing solution volume and cell density. Nitrogen gas replacement significantly decreased the cell aggregation rate and pO2 (p < 0.05). However, there were no differences in viability among cells stored under conditions of different storage volumes, densities, and nitrogen gas replacement. Conclusions: Aggregation of cells after storage at 25 °C in LR-3T-5D may be suppressed by increasing the storage volume and cell density as well as by incorporating nitrogen replacement, which lowers the pO2 in the solution.

Non-Native Structures of Apomyoglobin and Apoleghemoglobin in Folding Intermediates Related to the Protein Misfolding.

Protein folding is essential for a polypeptide chain to acquire its proper structure and function. Globins are a superfamily of ubiquitous heme-binding α-helical proteins whose function is principally to regulate oxygen homoeostasis. In this review, we explore the hierarchical helical formation in the globin proteins apomyoglobin and leghemoglobin, and we discuss the existence of non-native and misfolded structures occurring during the course of folding to its native state. This review summarizes the research aimed at characterizing and comparing the equilibrium and kinetic intermediates, as well as delineating the complete folding pathway at a molecular level, in order to answer the following questions: "What is the mechanism of misfolding via a folding intermediate? Does the non-native structure stabilize the contemporary intermediate structure? Does the non-native structure induce slower folding?" The role of the non-native structures in the folding intermediate related to misfolding is also discussed.

Characterization of three γ-glutamyltranspeptidases from Pseudomonas aeruginosa PAO1.

The Pseudomonas aeruginosa strain, PAO1, has three putative γ-glutamyltranspeptidase (GGT) genes: ggtI, ggtII, and ggtIII. In this study, the expression of each of these genes in P. aeruginosa PAO1 was analyzed, and the properties of the corresponding GGT proteins were investigated. This is the first report on biochemical characterization of GGT paralogs from Pseudomonas species. The crude extracts prepared from P. aeruginosa PAO1 exhibited hydrolysis and transpeptidation activities of 17.3 and 65.0 mU/mg, respectively, and the transcription of each gene to mRNA was confirmed by RT-PCR. All genes were cloned, and the expression plasmids constructed were introduced into an Escherichia coli expression system. Enzyme activity of the expressed protein of ggtI (PaGGTI) was not detected in the system, while the enzyme activities of the expressed proteins derived from ggtII and ggtIII (PaGGTII and PaGGTIII, respectively) were detected. However, the enzyme activity of PaGGTII was very low and easily decreased. PaGGTII with C-terminal his-tag (PaGGTII25aa) showed increased activity and stability, and the purified enzyme consisted of a large subunit of 40 kDa and a small subunit of 28 kDa. PaGGTIII consisted of a large subunit of 37 kDa and a small subunit of 24 kDa. The maximum hydrolysis and transpeptidation activities of PaGGTII25aa were obtained at 40ºC-50ºC, and the maximum hydrolysis and transpeptidation activities of PaGGTIII were obtained at 50ºC-60ºC. These enzymes retained approximately 80% of their hydrolysis and transpeptidation activities after incubation at 50ºC for 10 min, reflecting good stability. Both PaGGTII25aa and PaGGTIII showed higher activities of hydrolysis and transpeptidation in the alkali range than in the acidic range. However, they were highly stable at a wide pH range (5-10.5).

Increased expression of angiopoietin-like protein 4 regulates matrix metalloproteinase-13 expression in Porphyromonas gingivalis lipopolysaccharides-stimulated gingival fibroblasts and ligature-induced experimental periodontitis.

BACKGROUND: Angiopoietin-like protein 4 (ANGPTL4) is produced in chronic or acute inflammation. Although ANGPTL4 increases in the periodontal ligament fibroblasts during hypoxia, the involvement and role of ANGPTL4 in periodontitis have not been elucidated. OBJECTIVE: In this study, we investigated whether ligature-induced experimental periodontitis and/or Porphyromonas gingivalis lipopolysaccharides (Pg-LPS) would upregulate ANGPTL4 expression and whether ANGPTL4 would somehow involve in the expression of matrix metalloproteinases (MMPs) which are key molecules in the process of periodontal tissue destruction. METHODS: Experimental periodontitis was induced in 6-week-old male Sprague-Dawley rats by placing a nylon suture around the neck of the maxillary second molar. Two weeks after the induction of periodontitis, the periodontal tissue was excised and analyzed by histological/immunohistochemical staining and gene expression analyses. Human gingival fibroblasts (hGFs) were stimulated with Pg-LPS. The gene expression of ANGPTLs and receptors involved in ANGPTL4 recognition were observed. We also confirmed the changes in gene expression of MMPs upon stimulation with human ANGPTL4. Furthermore, we downregulated ANGPTL4 expression by short interfering RNA in hGFs and investigated the effect of Pg-LPS on MMP production. RESULTS: Induction of periodontitis significantly increased the expression of ANGPTL4 in the gingiva. Pg-LPS significantly increased the gene and protein expression of ANGPTL4 in hGFs but not the gene expression of other ANGPTLs or ANGPTL receptors. Recombinant human ANGPTL4 significantly increased MMP13 gene expression in hGFs. We also confirmed that MMP13 expression was increased in the gingiva during experimental periodontitis. Pg-LPS induced MMP13 gene expression in hGFs. These results suggest the pivotal role of ANGPTL4 in periodontitis. CONCLUSION: Periodontitis increases ANGPTL4 expression in the gingiva, further suggesting that increased ANGPTL4 may be a factor involved in enhancing MMP13 expression.

Porphyromonas gingivalis Fimbriae Induce Osteoclastogenesis via Toll-like Receptors in RAW264 Cells.

The effect of Mfa1 fimbriae of Porphyromonas gingivalis on the progression of bone resorption remains unclear, especially compared with another fimbriae, FimA. We investigated the effect of Mfa1 on osteoclastogenesis together with FimA. We also investigated the role of Toll-like receptors (TLRs) in Mfa1 recognition during osteoclast differentiation. Receptor activator of nuclear factor κß ligand (RANKL)-prestimulated RAW264 cells were used to examine the effects of purified Mfa1 fimbriae. The number of osteoclasts was examined by tartrate-resistant acid phosphate (TRAP) staining, osteoclast activation was investigated by bone resorption assays, and gene expression of differentiation markers was examined by quantitative real-time PCR. Transfection of Tlr2 and Tlr4 siRNAs into RAW264 cells was also employed and their role in Mfa1 recognition was investigated. Mfa1 effectively induced the formation of TRAP-positive multinucleated cells and activated osteoclasts. Mfa1 also increased gene expression of Acp5, Mmp9, and Ctsk in RANKL-prestimulated RAW264 cells compared with the control. The osteoclastogenesis induced by Mfa1 was significantly decreased in cells transfected with Tlr2 or Tlr4 siRNAs compared with control siRNA. Our results revealed the role of Mfa1 fimbriae in osteoclastogenesis that may contribute to the partial elucidation of the mechanisms of periodontal disease progression and the development of new therapeutic strategies.

Risk factors for insufficient reduction after short-segment posterior fixation for thoracolumbar burst fractures: Does the interval from injury onset to surgery affect reduction of fractured vertebrae?

BACKGROUND: Many surgeons have encountered patients who could not immediately undergo surgery to treat spinal fractures because they had associated injuries and/or because a complete diagnosis was delayed. For such patients, practitioners might assume that delays could mean that the eventual reduction would be insufficient. However, no report covered risk factors for insufficient reduction of fractured vertebra including duration from injury onset to surgery. The purpose of this study is to investigate the risk factors for insufficient reduction after short-segment fixation of thoracolumbar burst fractures. METHODS: Our multicenter study included 253 patients who sustained a single thoracolumbar burst fracture and underwent short-segment fixation. We measured the local vertebral body angle (VBA) on roentgenograms, before and after surgery, and then calculated the reduction angle and reduction rate of the fractured vertebra by using the following formula: [Formula: see text] A multiple logistical regression analysis was performed to identify risk factors for insufficient reduction. The factors that we evaluated were age, gender, affected spine level, time elapsed from injury to surgery, inclusion of vertebroplasty with surgery, load-sharing score (LSS), AO classification (type A or B), preoperative VBA, and the ratio of canal compromise before surgery. RESULTS: There were 140 male and 113 female patients, with an average age of 43 years, and the mean time elapsed between injury and surgery was 3.8 days. The mean reduction angle was 12°, and the mean reduction rate was 76%. The mean LSS was 6.4 points. Multiple linear regression analysis revealed that a higher LSS, a larger preoperative VBA, a younger age, and being female disposed patients to having a larger reduction angle and reduction rate. The time elapsed from injury to surgery had no relation to the quality of fracture reduction in the acute period. CONCLUSIONS: Our findings indicate that if there is no neurologic deficit, we might not need to hurry surgical reduction of fractured vertebrae in the acute phase.

Next-Generation Examination, Diagnosis, and Personalized Medicine in Periodontal Disease.

Periodontal disease, a major cause of tooth loss, is an infectious disease caused by bacteria with the additional aspect of being a noncommunicable disease closely related to lifestyle. Tissue destruction based on chronic inflammation is influenced by host and environmental factors. The treatment of periodontal disease varies according to the condition of each individual patient. Although guidelines provide standardized treatment, optimization is difficult because of the wide range of treatment options and variations in the ideas and skills of the treating practitioner. The new medical concepts of "precision medicine" and "personalized medicine" can provide more predictive treatment than conventional methods by stratifying patients in detail and prescribing treatment methods accordingly. This requires a new diagnostic system that integrates information on individual patient backgrounds (biomarkers, genetics, environment, and lifestyle) with conventional medical examination information. Currently, various biomarkers and other new examination indices are being investigated, and studies on periodontal disease-related genes and the complexity of oral bacteria are underway. This review discusses the possibilities and future challenges of precision periodontics and describes the new generation of laboratory methods and advanced periodontal disease treatment approaches as the basis for this new field.

Study protocol for periodontal tissue regeneration with a mixture of autologous adipose-derived stem cells and platelet rich plasma: A multicenter, randomized, open-label clinical trial.

Introduction: Adipose-derived stem cells (ASCs) secrete various growth factors to promote wound healing and to regenerate various tissues, such as bone, cartilage, and fat tissue. Subcutaneous adipose tissue is a considerable cell source in clinical practice and can be collected relatively easily and safely under local anesthesia. Moreover, platelet-rich plasma (PRP), a plasma component containing many platelets purified by centrifuging the collected blood, also promotes wound healing. PRP can be easily gelled and is therefore attracting attention as a scaffolding material for transplanted cells. The usefulness of a mixture of ASCs and PRP for periodontal tissue regeneration has been in vitro demonstrated in our previous study. The aim of this study is to present the protocol of translation of tissue regeneration with ASCs and PRP into practical use, evaluating its efficacy. Methods: This study is a multicenter, randomized, open-label comparative clinical trial. Fifteen patients will be randomly assigned to the treatment with mixture of ASCs and PRP or enamel matrix derivate administration into periodontal tissue defects. Increase in height of new alveolar bone in the transplanted area will be evaluated. The evaluation will be performed using dental radiographs after 36 weeks of transplantation. Occurrence of adverse events will be evaluated as secondary outcome. Results: This clinical study was initiated after meeting the regulations to be complied with, including ethical review and regulatory notifications. Conclusions: If effective, this cell therapy using autologous mesenchymal stem cells can represent a useful medical technology for regeneration of periodontal defects.

Longitudinal study on the effect of keratinized mucosal augmentation surrounding dental implants in preventing peri-implant bone loss.

Background: Dental implant therapy is a well-established method of prosthetic rehabilitation of missing teeth. To maintain the health of the surrounding tissue, management of risk factors/indicators and daily maintenance are important. It still remains controversial whether a certain amount of keratinized mucosal width is essential for maintaining the health of peri-implant tissue. The purpose of this multicenter retrospective study was to assess the correlation between bone loss around dental implant and the amount of keratinized tissue width. Methods: A total of 1,644 implants were evaluated. Data was collected about participants' general and dental history, as well as implant details. Bone resorption around implant was calculated from intra-oral radiographs taken after 1 year and more than 3 years of function. Implants were classified into three groups; received free gingival graft or apically repositioned flap surgery for increasing the keratinized mucosa ≥2 mm width (group A), keratinized mucosa width ≥2 mm (group B), and keratinized mucosa width <2 mm (group C). These data were analyzed by propensity score analysis and a generalized linear regression analysis was performed to compare the bone resorption among groups. Results: Mean functional time was 55.8 months (SD = 20.5) in group A, 67.6 months (SD = 28.1) in group B, and 74.5 months (SD = 32.9) in group C. Mean bone resorption of groups A, B, and C were 0.08 mm (SD = 0.40), 0.18 mm (SD = 0.66), and 0.44 mm (SD = 0.40). Groups A and B had significantly lower bone resorption than group C. Conclusion: The results in this study show the importance of keratinized mucosa in maintaining the peri-implant bone. Our findings also suggest that mucosal transplantation is useful, as opposed to narrowing of the keratinized mucosa.

Decoding an Amino Acid Sequence to Extract Information on Protein Folding.

Protein folding is a complicated phenomenon including various time scales (µs to several s), and various structural indices are required to analyze it. The methodologies used to study this phenomenon also have a wide variety and employ various experimental and computational techniques. Thus, a simple speculation does not serve to understand the folding mechanism of a protein. In the present review, we discuss the recent studies conducted by the author and their colleagues to decode amino acid sequences to obtain information on protein folding. We investigate globin-like proteins, ferredoxin-like fold proteins, IgG-like beta-sandwich fold proteins, lysozyme-like fold proteins and ß-trefoil-like fold proteins. Our techniques are based on statistics relating to the inter-residue average distance, and our studies performed so far indicate that the information obtained from these analyses includes data on the protein folding mechanism. The relationships between our results and the actual protein folding phenomena are also discussed.

Short-term preservation of porcine zygotes at ambient temperature using a chemically defined medium.

We aimed to develop a simple method for the short-term preservation of in vitro-produced porcine zygotes at 25°C for up to 2 days. Firstly, we evaluated the efficiency of three storage solutions to preserve porcine zygotes at 25°C for 24 h. Two of these were commercially available defined media for cell storage (Cell-W and Cell-S), and the third was fetal bovine serum (FBS). Thereafter, we examined the effects of storing the zygotes in the Cell-W solution for 24-72 h at 25°C. The Cell-W solution was the most efficient for 24 h storage of porcine zygotes at 25°C, with no apparent effects on blastocyst quality. However, short-term storage of porcine zygotes for 24 h reduced the blastocyst formation rate compared with the fresh control group. As storage duration increased from 24 to 48 or 72 h, blastocyst formation rates were significantly decreased from 11.3% to 4.4% and 0%, respectively. In conclusion, during zygote storage, the developmental competence to the blastocyst stage decreased with time. Storage of zygotes in chemically defined media did not affect blastocyst quality, but the storage in 100% serum had an adverse effect on developing embryos causing apoptosis.

Viability and developmental potential of porcine blastocysts preserved for short term in a chemically defined medium at ambient temperature.

This study developed an efficient method for liquid storage of in vitro-derived porcine blastocysts at ambient temperature for 24 hr. We evaluated the effects of new chemically defined media (cell wash and preservation solution, Cellstor® -W [Cell-W] and cell suspension and preservation solution, Cellstor® -S [Cell-S]) for short-term storage. In the first experiment, in vitro-derived blastocyst were stored at 25ºC for 24 hr in Cell-W solution, Cell-S solution and pig embryo culture (PBM) medium. There were no differences in the rates of survival and development of stored blastocysts between the Cell-S and Cell-W solutions, but the total cell number of embryos that survived after storage in Cell-S solution was significantly higher (p < .05) than that in the Cell-W solution. In the second experiment, Cell-S solution was used to store the in vitro-derived blastocysts at 20°C, 25°C and 30°C. Storage at 20°C resulted in a significant decrease in the rates of survival and development of stored blastocysts compared to storage at 25°C or 30°C. No differences in survival and development rates were observed between storage at 25°C and 30°C, but the damage to the embryo quality after storage and culture was significantly lower at 25°C than at 30°C. In the third experiment, Cell-S solution was supplemented with ß-mercaptoethanol and curcumin, either alone or in combination, as antioxidant agents. Although the supplementation with curcumin did not improve survival, it significantly increased the development rate of stored blastocysts compared with the control blastocysts stored without antioxidants. In conclusion, when porcine blastocysts were stored at 25°C for 24 hr, a Cell-S solution may be effective for maintaining the survival and development of in vitro embryos.