Evaluation of plasma membrane integrity of frozen-thawed goat spermatozoa with or without seminal plasma

The aim of the present work was to evaluate plasma membrane integrity, motility, vigor and morphology of fresh and frozen goat spermatozoa with or without seminal plasma. Semen samples were diluted in Tris solution, before and after thawing, with a combination of carboxifluorescein diacetate and propidium iodide. The results showed differences (P < 0.01) for motility and minor defects in the presence or absence of seminal plasma, for both fresh and frozen samples. Periods of collection had a significant effect on motility, probably due to changes in the photoperiod. Plasma membrane integrity was significantly reduced by the freezing process, whether seminal plasma was present or absent. In conclusion, removal of seminal plasma decreased motility and vigor rates in frozen samples. The photoperiod probably decreased the testosterone level, contributing negatively to the high percentage of sperm abnormalities, mainly damaged membranes. The use of fluorescent probes allowed a better estimation of the percentage of functional cells, instead of only estimating the percentage of motile cells or morphology defects. © 2001 Elsevier Science B.V. All rights reserved.

Graft incorporation and implant osseointegration following the use of autologous and fresh-frozen allogeneic block bone grafts for lateral ridge augmentation

Objectives: To compare autogenous bone (AT) and fresh-frozen allogeneic bone (AL) in terms of histomorphometrical graft incorporation and implant osseointegration after grafting for lateral ridge augmentation in humans. Materials and methods: Thirty-four patients were treated with either AL (20 patients) or AT (14 patients) onlay grafts. During implant installation surgery 6 months after grafting, cylindrical biopsies were harvested perpendicularly to the lateral aspect of the augmented alveolar ridge. Additionally, titanium mini-implants were installed in the grafted regions, also perpendicularly to the ridge; these were biopsied during second-stage surgery. Histological/histomorphometric analysis was performed using decalcified and non-decalcified sections. Results: Histological analysis revealed areas of necrotic bone (NcB) occasionally in contact with or completely engulfed by newly formed vital bone (VB) in both AT and AL groups (55.9 ± 27.6 vs. 43.1 ± 20.3, respectively; P = 0.19). Statistically significant larger amounts of VB (27.6 ± 17.5 vs. 8.4 ± 4.9, respectively; P = 0.0002) and less soft connective tissue (ST) (16.4 ± 15.6 vs. 48.4 ± 18.1, respectively; P ≤ 0.0001) were seen for AT compared with AL. No significant differences were observed between the groups regarding both bone-to-implant contact (BIC) and the bone area between implant threads (BA) on the mini-implant biopsies. Conclusion: Allogeneic bone block grafts may be an option in cases where a limited amount of augmentation is needed, and the future implant can be expected confined within the inner aspect of the bone block. However, the clinical impact of the relatively poor graft incorporation on the long-term performance of oral implants placed in AL grafts remains obscure. © 2013 John Wiley & Sons A/S.

Fate of autologous and fresh-frozen allogeneic block bone grafts used for ridge augmentation. A CBCT-based analysis

Objectives: To evaluate dimensional changes in autologous (AT) and fresh-frozen allogeneic (AL) block bone grafts 6 months after alveolar ridge augmentation. Material and methods: Twenty-six partially or totally edentulous patients treated either with fresh-frozen AL bone or AT bone onlay block grafts prior to implant placement (13 patients in each group), were included in this analysis. Patients received CBCT (i-CAT Classic) examinations prior to surgery and 14 days and 6 months after grafting. Differences in alveolar ridge area among the various observation times were evaluated by planimetric measurements on two-dimensional CBCT images of the grafted regions. Nineteen grafted blocks from each group were evaluated. Results: Significant increase in alveolar ridge dimensions, allowing implant placement, was obtained with both types of grafts 6 months after grafting; no significant differences in alveolar ridge area were observed between the groups at the various observation times. However, graft resorption in the AL group was significantly larger compared to that in the AT group at 6 months. Conclusions: Larger bone graft resorption was seen in patients treated with fresh-frozen AL bone than in those treated with AT bone 6 months following alveolar ridge augmentation. © 2011 John Wiley & Sons A/S.

Evaluation of Sperm Kinetics and Plasma Membrane Integrity of Frozen Equine Semen in Different Storage Volumes and Freezing Conditions

In the present study, different freezing systems (Styrofoam box and Mini Digitcool ZH 400) and storage volumes (0.5- and 0.25-mL straws) were compared with regard to sperm kinetics and plasma membrane integrity of frozen and thawed semen. For that, three ejaculates from four animals were frozen in Styrofoam box and Mini Digitcool ZH 400 machine. The 0.5-mL straws were thawed at 46°C for 20 seconds, and the 0.25-mL straws were thawed at 46°C for 12 seconds. Statistical analysis was performed using program R of descriptive analysis box plot, followed by analysis of variance using PROC MIXED of SAS 9.1 package. Variances of 5% were considered as different. There was no interaction between the straw sizes and volumes; however, statistical differences were observed between the semen storage volumes. The 0.5-mL straws had higher total motility (%), progressive motility (%), average path velocity (μm/s), straight-line velocity (μm/s), curvilinear velocity (μm/s), and rapid sperm percentage (%) than the 0.25-mL straws. However, plasma membrane integrity analysis did not differ between the two straws. Thus, it is possible to conclude that equine sperm cryopreserved in 0.5-mL straws has better sperm kinetics than when stored in 0.25-mL straws. Additionally, it is possible to conclude that automated systems that enable faster freezing rates result in a seminal quality that is similar to the one obtained by the conventional system using Styrofoam boxes. © 2013 Elsevier Inc.

Is peripheral blood cell balanced altered by the use of fresh frozen bone block allografts in lateral maxillary ridge augmentation?

Background: The relationship between the immune response and red and white blood cell homeostasis is cited in literature, but no studies regarding the balance of these cell populations following maxillary bone-graft surgeries can be found. Aim: The aim of this study was to evaluate the possible impairments in the blood cell balance following fresh-frozen allogeneic bone-graft augmentation procedures in patients who needed maxillary reconstruction prior to implants. Material and Methods: From 33 patients elected to onlay bone grafting procedures, 20 were treated with fresh-frozen bone allografts and 13 with autologous bone grafts. Five blood samples were collected from each patient in a 6-month period (baseline: 14, 30, 90, and 180 days postsurgery), and the hematological parameters (erythrogram, leukogram, and platelets count) were accessed. Results: All evaluated parameters were within the reference values accepted as normal, and significant differences were found for the eosinophils count when comparing the treatments (30 days, p=.035) and when comparing different periods of evaluation (allograft-treated group, baseline×180 days, p≤.05 and 90×180 days, p≤.01; autograft-treated group, 30×90 days, p≤.05 and 30×180 days, p≤.05). Conclusions: Both autologous and fresh-frozen allogeneic bone grafts did not cause any impairment in the red and white blood cell balance, based on quantitative hemogram analysis, in patients subjected to maxillary reconstruction. © 2011 Wiley Periodicals, Inc.

Changes during chilled storage of whole tilapia and short-term frozen storage of tilapia fillets

This study evaluated the physicochemical changes in Nile tilapia (n = 82, 373.71 ± 61.91 g) refrigerated for up to 92 h and in the frozen fillets. The tilapias were captured with nets, slaughtered by ice and water shock (1:1) in a temperature of approximately 2°C for 30 min, and stored refrigerated at 4°C in polystyrene boxes containing ice. The fish were filleted, and filets were weighed and frozen. The drip loss and protein were determined after 23 days of frozen storage. After 4 h of storage, all fish were in full rigor mortis. The pH of the muscles decreased for up to 45 h of the storage period. The fillets obtained from tilapia stored for more than 72 h lost more weight and protein. Thus, the filleting or processing of tilapia should be done before 72 h of cold storage, since deterioration of the fish starts to occur after this period. Copyright © Taylor & Francis Group, LLC.

Healing of fresh frozen bone allograft with or without platelet-rich plasma: A histologic and histometric study in rats

Purpose: This study histomorphometrically analyzed the effect of autogenous platelet-rich plasma (PRP) on healing of fresh frozen bone allograft (FFBA) in bony defects in rat calvaria. Materials and Methods: A 5mm-diameter defect was created in the calvarium of 30 rats. Animals were divided into three groups: C (defect was filled by blood clot only), FFBA (defect was filled with 0.01mL of FFBA), and FFBA/PRP (defect was filled with 0.01mL of FFBA combined with 100μL of PRP). All animals were euthanized at 30 days postoperatively. Histomorphometry and histology analyses were performed. Data were statistically analyzed (analysis of variance, Tukey, p<.05). Results: FFBA had a statistically smaller new bone area than groups FFBA/PRP and C. No statistically significant differences were observed between groups FFBA and FFBA/PRP with regard to remaining bone graft particle area. Conclusion: It can be concluded that (1) PRP improved the incorporation of FFBA, increasing the amount of new bone formed; (2) PRP has not influenced the resorption of nonviable particles of the FFBA; and (3) presence of remaining FFBA particles might have accounted for the smaller amount of new bone observed in group FFBA when compared with control group. © 2011 Wiley Periodicals, Inc.

Clinical similarities and histological diversity comparing fresh frozen onlay bone blocks allografts and autografts in human maxillary reconstruction

Background: In the absence of autologous bone for harvesting, fresh-frozen bone allografts turned into an alternative for bone reconstruction procedures. Purpose: The purpose of this study was to make a histological analysis of fresh-frozen onlay bone allografts (ALs), compared with autografts, in patients who needed maxillary reconstruction prior to dental implants placement. Materials and Methods: Twelve patients with bone deficiencies (width inferior to 4mm) in the sites where the implants were planned were enrolled in the study. From these, six were elected to be treated with autogenous (AT) bone grafts and six with fresh-frozen bone AL. This last group included the patients who had absence of a convenient amount of bone in donor sites. Each patient received from one to six graft blocks, totalling to 12 ATs and 17 ALs. Seven months after grafting procedures, biopsies of the grafts were made using 2-mm internal diameter trephine burs, and processed for histological analysis. One biopsy was retrieved from each patient. Results: Clinically, all grafts were found to be firm in consistency and well-incorporated to the receptor bed. Histological analysis showed a large amount of necrotic bone surrounded by few spots of new-formed bone in the AL group, suggesting low rate of graft remodeling. In the AT group, an advanced stage of bone remodeling was seen. Conclusions: Human fresh-frozen bone block AL showed clinical compatibility for grafting procedures, although associated to slow remodeling process. Further studies are needed to define, at long term, the remodeling process chronology the clinical longitudinal results for fresh-frozen bone AL. Copyright © 2013 Wiley Periodicals, Inc.

Immunological aspects of fresh-frozen allogeneic bone grafting for lateral ridge augmentation

Objectives: To present some immunological aspects of fresh-frozen allogeneic bone grafting for lateral bone augmentation, based on the quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α in patients sera. Material and methods: Thirty-three partially or totally edentulous patients received fresh-frozen allogeneic bone (AL - 20 patients) or autologous bone onlay block grafts (AT - 13 patients) prior to oral implant placement. Blood samples were collected from each patient at various time-points during a 6 month-period (baseline, 14, 30, 90 and 180 days postoperatively). Quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α was performed by enzyme linked immunosorbent assay (ELISA). Results: For all evaluated markers and at all evaluated periods, inter-group comparisons showed no statistically significant differences between the groups, while the observed values were within normal levels. For AL-treated patients, intra-group evaluation showed statistically significant increase of TNF-α from baseline to 90 (P < 0.001) and 180 (P < 0.01) days, and from 14 to 90 (P < 0.01) and 180 (P < 0.05) days. IFN- γ showed intercalated results, with a decrease from baseline to 14 days (P < 0.05), and increase from 14 to 90 days (P < 0.001) and 180 (P < 0.05) days. No differences between the periods of evaluation were found for the AT group. Conclusions: AL grafting for lateral bone augmentation, similar to AT grafting, does not seem to challenge the immune system significantly. © 2012 John Wiley & Sons A/S.

Effect of platelet-rich plasma on bone healing of fresh frozen bone allograft in mandibular defects: a histomorphometric study in dogs

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