Objective: To provide reference for clinical application of liquid plasmatrix, and to investigate the optimal centrifugation time of liquid plasmatrix prepared by horizontal centrifugation for soft tissue regeneration from the aspects of mechanical properties, biological properties, and the effect of promoting soft tissue regeneration. Methods: Venous blood was collected from 6 healthy volunteers [3 males and 3 females, aged (26±2) years, with informed consent] who volunteered to donate blood at School of Stomatology, Wuhan University from September to November 2021. The collected venous blood was centrifuged at 500 ×g for 3, 5, 8 and 12 min to obtain liquid plasmatrix. The volume, weight, solidification time, and mechanical properties of liquid plasmatrix prepared at different centrifugation time were measured and recorded (the sample size at each time point was 3). The microstructure of different groups of liquid plasmatrix clot was observed by scanning electron microscope (SEM). The rheological properties of each group of liquid plasmatrix clot were measured by rheological test. The number and concentration of cells in the whole blood group and in each liquid plasmatrix group were measured using complete blood count test. The distribution of cells in the liquid plasmatrix clots was observed by hematoxylin-eosin staining. The effect of control group (Dulbecco's modified Eagle's medium containing 20% fetal bovine serum) and liquid plasmatrix clot exudates in 3, 5, 8, 12 min group (the sample size at each time point was 3) on gingival fibroblast migration was detected by cell migration method. Finally, the effects of control group and liquid plasmatrix clot exudates on the morphology of gingival fibroblasts were observed by fluorescence microscope. Results: The volume of liquid plasmatrix in 3, 5, 8 and 12 min group were approximately (2.47±0.12), (2.67±0.12), (3.53±0.12) and (3.73±0.12) ml, respectively. The weight of liquid plasmatrix in 3, 5, 8 and 12 min group were approximately (0.35±0.01), (0.46±0.02), (0.88±0.06) and (1.03±0.01) g, respectively. The maximum tensile force of liquid plasmatrix clots in 3, 5, 8 and 12 min group were (0.55±0.03), (0.56±0.03), (1.31±0.05) and (1.38±0.02) N, respectively. SEM results showed that the fibers inside the liquid plasmatrix clot became denser with increased centrifugation time. Compared with other groups, the concentrations of leukocytes, neutrophils, monocytes and lymphocytes in 8 min group were the highest, and the distribution of cell was more even. Compared with other groups, the efficiency of stimulating gingival fibroblast migration in 8 min group was the best (1.60±0.01). Fluorescence staining test showed that the liquid plasmatrix clot exudates could make gingival fibroblasts more stretched compared with control group. Conclusions: The present study shows that liquid plasmatrix prepared by centrifugation with 500 ×g centrifugal force for 8 min has higher concentration of viable cells and the ability to promote the migration of gingival fibroblasts.
Cell Movement ; Centrifugation/methods* ; Female ; Fibroblasts ; Gingiva ; Humans ; Male ; Wound Healing

Platelet-rich fibrin (PRF) has been widely used owing to its ability to stimulate tissue regeneration. To date, few studies have described the antibacterial properties of PRF. Previously, PRF prepared by horizontal centrifugation (H-PRF) was shown to contain more immune cells than leukocyte- and platelet-rich fibrin (L-PRF). This study aimed to compare the antimicrobial effects of PRFs against Staphylococcus aureus and Escherichia coli in vitro and to determine whether the antibacterial effects correlated with the number of immune cells. Blood samples were obtained from eight healthy donors to prepare L-PRF and H-PRF. The sizes and weights of L-PRF and H-PRF were first evaluated, and their antibacterial effects against S. aureus and E. coli were then tested in vitro using the inhibition ring and plate-counting test methods. Flow-cytometric analysis of the cell components of L-PRF and H-PRF was also performed. No significant differences in size or weight were observed between the L-PRF and H-PRF groups. The H-PRF group contained more leukocytes than the L-PRF group. While both PRFs had notable antimicrobial activity against S. aureus and E. coli, H-PRF demonstrated a significantly better antibacterial effect than L-PRF. Furthermore, the antimicrobial ability of the PRF solid was less efficient than that of wet PRF. In conclusion, H-PRF exhibited better antibacterial activity than L-PRF, which might have been attributed to having more immune cells.
Anti-Bacterial Agents/pharmacology* ; Anti-Infective Agents ; Centrifugation ; Escherichia coli ; Leukocytes ; Platelet-Rich Fibrin ; Staphylococcus aureus

centrifugation was performed to isolate motile sperm. Samples were incubated with different concentrations (0, 1, 10, 100, and 1,000 ng/mL) of RA. The non-exposed group (0 ng/mL) was defined as the control group. Samples were analyzed for motility at different time points (0, 60, 150, 240, and 300 minutes) and for vitality and oxidation reduction potential (ORP) (at 0, 240, and 300 minutes). Sperm motility was assessed manually and motion kinetic parameters were recorded by computer aided semen analysis.RESULTS: RA at any tested concentration significantly increased sperm motility compared to the control in a time-dependent manner with a maximum increase after 240 minutes. Motion kinetic parameters were not comparable. For sperm vitality, supplementation with RA significantly maintained survival at higher levels, while non-treated sperm gradually died. These higher levels of vitality were maintained with rising RA concentrations of up to 1,000 ng/mL. A non-significant trend of increased ORP was observed in all study groups.CONCLUSIONS: RA increases sperm motility and maintains vitality at any concentration tested. Therefore, RA might be utilized to improve sperm quality in asthenozoospermic specimens. However, further investigation is ongoing to evaluate the effect of RA on other sperm parameters.]]>
Centrifugation, Density Gradient ; Humans ; In Vitro Techniques ; Methylphenidate ; Oxidation-Reduction ; Oxidative Stress ; Semen ; Semen Analysis ; Sperm Motility ; Spermatozoa ; Tissue Donors ; World Health Organization

BACKGROUND: A major complication of peritoneal dialysis (PD) is peritonitis, and bacterial culture of PD effluent in a blood culture bottle is the preferred technique for diagnosis of peritonitis. In this study, we compared dialysate inoculation and culture using the BacT/AlerT® Fastidious Antimicrobial Neutralization Plus blood culture bottles (FAN Plus; bioMérieux, France) to the conventional centrifugation culture method.METHODS: A total of 170 PD effluents were simultaneously processed by the conventional centrifugation culture method and by culture using FAN Plus media with two different inoculation procedures: inoculation after centrifugation and direct bedside inoculation.RESULTS: Of the 52 cultures that were positive on at least one of the culture methods, 27 samples were positive on conventional centrifugation. However, 46 samples showed growth following inoculation into the FAN Plus media after centrifugation, and 47 samples were positive on the direct FAN Plus inoculation method. Using the case definition for PD peritonitis to classify samples, sensitivity of the conventional method was 50.0% (95% CI, 33.7–66.3%), whereas the sensitivity of the FAN Plus media was 78.9% (95% CI, 62.2–89.9%) by inoculation after centrifugation and 86.8% (95% CI, 71.1–95.1%) by direct inoculation. Use of both inoculation methods with FAN Plus media resulted in 92.1% sensitivity (95% CI, 89.2–99.9%).CONCLUSION: Culture using FAN Plus media demonstrated a superior bacterial recovery rate to the conventional centrifugation culture method. A combination of the two inoculation methods with FAN Plus media is recommended for the best diagnostic yield, while direct inoculation alone can be useful due to its simplicity and cost-effectiveness.
Centrifugation ; Culture Media ; Diagnosis ; Methods ; Peritoneal Dialysis ; Peritonitis

OBJECTIVE: How to increase the long-term retention rate of autologous fat grafting has been widely discussed. This study aimed to evaluate long-term fat graft retention rates for the most widely used fat processing methods in the area of facial esthetic surgery, including centrifugation, filtration, and sedimentation, using three-dimensional (3D) imaging. DATA SOURCES: PubMed, Embase, Wiley/Cochrane Library, and Web of Science databases were comprehensively searched from inception to July 2018 according to the guidelines of the American Society of Plastic Surgeons Fat Graft Task Force Assessment Methodology. STUDY SELECTION: Articles were screened using predetermined inclusion and exclusion criteria. Data collected included patient characteristics, follow-up devices, fat grafting techniques, and clinical outcomes. Patient cohorts were pooled, and fat graft retention rates were calculated. Complications were summarized according to different clinical characteristics. RESULTS: Of 77 articles, 10 clinical studies met the inclusion criteria and reported quantified measurement outcomes with 3D imaging which provide precise volumetric data with approximately 2% standard deviation compared to real volumes. Data of 515 patients were included. Fat grafting retention varied from 21% to 82%. We found filtration and centrifugation techniques could result in better retention outcomes. However, retention varied within each processing technique, with no significant difference among the 3 techniques. Twenty-two complications were reported among 515 patients, including donor-site hematoma (1 case), mild post-operative erythema (2 cases), mild volumetric asymmetries (2 cases), chronic edema (2 cases), overcorrection (2 cases), skin irregularity (6 cases), and headache or dysesthesia (7 cases). CONCLUSIONS Filtration and centrifugation techniques may result in better fat grafting retention outcomes than gravity sedimentation; however, more accurate statistical evidence is needed. Controversies continue to exist with respect to the performance of the different fat-processing techniques in fat graft retention.
Adipocytes ; cytology ; Adipose Tissue ; cytology ; Centrifugation ; methods ; Filtration ; methods ; Humans ; Imaging, Three-Dimensional ; methods

BACKGROUND: Extraction of cell-free DNA (cfDNA) is a key step for determining the quality of cfDNA-related molecular diagnostics. We evaluated the effect of sample containers and sample storage conditions on cfDNA extraction. METHODS: The cfDNA extraction using the MagMAX Cell-Free DNA Isolation Kit from five healthy controls and five lung cancer patients was evaluated according to the type of sample container and storage conditions: K2-EDTA container, <1, 6, 24, and 48 hr storage at 4℃ after immediate plasma separation; and Cell-Free DNA BCT container, <1, 3, 7, and 14 days stored at room temperature. Mutation analysis of EGFR exons 18–21 was performed. To assess the effect of a delay in centrifugation, EDTA whole blood samples from five healthy individuals were stored at 4℃ for 6, 12, and 24 hr before plasma separation. RESULTS: There was no significant difference in the amount and nucleic acid size of cfDNA in both controls and patients with cancer when EDTA plasma was stored at 4℃ up to 48 hr. The amount and size of cfDNA in the BCT container were not different up to 7 days; however, the 14-day sample showed an increase in cfDNA concentration due to genomic DNA contamination. EGFR mutations were detected on EDTA containers up to 48 hr and with BCT containers up to 14 days. When EDTA whole blood was stored at 4℃ and plasma separation was delayed, the cfDNA concentration increased from 24 hr. CONCLUSIONS: The cfDNA extraction was affected by the sample containers and storage conditions.
Biopsy ; Centrifugation ; DNA Contamination ; DNA ; Edetic Acid ; Exons ; Humans ; Lung Neoplasms ; Pathology, Molecular ; Plasma

OBJECTIVE: Density gradient centrifugation (DGC) is frequently used to isolate high-motility fractions of spermatozoa. We compared the efficacy of four DGC media in terms of the percentage of morphologically normal spermatozoa, DNA fragmentation level, and hyaluronic acid (HA) binding ability. METHODS: Thirty men with a total motile spermatozoa count >80 million participated. Semen samples were divided into four aliquots, which were processed using PureSperm, PureCeption, Sidney, and SpermGrad media, respectively. The DNA fragmentation level was measured using the Halosperm assay kit and HA binding ability was measured using the HBA assay kit. RESULTS: The mean percentage of morphologically normal spermatozoa was significantly enhanced after DGC using all four media (10.3%, 9.9%, 9.8%, and 10.7%, respectively; p<0.05 for each when compared with 6.9% in raw semen). The DNA fragmentation level was significantly reduced after DGC using PureSperm, PureCeption, and SpermGrad media (6.0%, 6.5%, and 4.9%, respectively; p<0.05 for each when compared with 11.2% in raw semen), but not after DGC using Sidney media (8.5%, p>0.05). HA binding ability did not change after DGC using any of the four media. CONCLUSION: The four media were equally effective for obtaining a sperm fraction with highly motile, morphologically normal sperm. PureSperm, PureCeption, and SpermGrad media were equally effective for acquiring a sperm fraction with less DNA fragmentation.
Centrifugation, Density Gradient ; DNA Fragmentation ; DNA ; Humans ; Hyaluronic Acid ; Male ; Semen ; Spermatozoa

OBJECTIVES: This study aimed to investigate the effects of chronic restraint stress on the osseointegration of titanium implants. MATERIALS AND METHODS: Twenty adult male Wistar albino rats were used in the study. After surgical insertion of titanium implants into the metaphyseal part of the tibial bone, rats were randomly divided into two groups: a control group (CNT group) and an experimental restraint stress group (RS group). In the CNT group, titanium implants were inserted surgically, and rats received no further treatment during the 47-day experimental period. In the RS group, restraint stress was applied for 3 hours per day for 45 days, beginning 2 days after implant surgery. Weight of the rats was measured prior to surgery and at the end of the study to analyze the effects of stress. At the end of the experimental period, rats were euthanized, and implants and surrounding bone tissues were used for undecalcified histological analysis. Serum cortisol levels were assessed in cardiac blood samples from the rats following centrifugation. RESULTS: Average weight of rats in the RS group was lower than that of rats in the CNT group after the experimental protocol had been completed (P<0.05). Further, serum cortisol levels were higher in the RS group than in the CNT group (P<0.05). There were no significant differences in bone-implant connection levels between the two groups (P>0.05). CONCLUSION: The data analyzed in this study suggest that chronic restraint stress did not adversely affect rats during a 45-day osseointegration period.
Adult ; Animals ; Bone and Bones ; Centrifugation ; Humans ; Hydrocortisone ; Male ; Osseointegration ; Rats ; Tibia ; Titanium

OBJECTIVES: This study examined the effects of plasma-rich growth factors (PRGF) on accelerating bone regeneration/repair in fresh extraction sockets, and determined the quality and quantity of bone by assessing the bone density using cone-beam computed tomography (CBCT). MATERIALS AND METHODS: Twenty patients, who had undergone bilateral extractions, were included in this study. In one extraction socket, PRGF was used and covered with an autologous fibrin plug. Nothing was used in the opposite side extraction socket. Thirteen weeks post extraction, the level of bone regeneration was evaluated on both sides with CBCT. RESULTS: At the end of the study, the mean bone density according to the Hounsfield units (HU) in the control group and PRGF group was 500.05 HU (type III bone type) and 647.95 HU (type II bone type), respectively. CONCLUSION: This study recommends the use of PRGF in post extraction sites to accelerate the rate of bone regeneration and improve the quality of regenerated bone. The technique to process PRGF was simple compared to previously mentioned techniques used for platelet-rich plasma (PRP) preparation. PRP preparation requires a two-cycle centrifugation procedure, leading to a longer processing time.
Bone Density ; Bone Regeneration ; Centrifugation ; Cone-Beam Computed Tomography ; Dental Implants ; Fibrin ; Humans ; Intercellular Signaling Peptides and Proteins ; Plasma ; Platelet-Rich Plasma

OBJECTIVE: The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST).METHODS: Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern (“a”–“g”) were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed.RESULTS: The HOST examinations showed that >93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type “d”, 98.67% in type “g”, and 98.17% in type “f” spermatozoa.CONCLUSION: We found that the type “d” spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.
Centrifugation, Density Gradient ; Chromatin ; DNA Fragmentation ; DNA ; Humans ; Infertility ; Semen ; Semen Preservation ; Sperm Head ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Tail ; Tissue Donors