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Objective:To observe the clinical efficacy of autologous platelet rich gel (APG) in the treatment of type 2 diabetic foot (DF) patients and the effect of APG on the expression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in peripheral blood mononuclear cells (PBMCs).Methods:A total of 62 patients with DF admitted to the Affiliated Hospital of Kangda College of Nanjing Medical University from February 2021 to May 2022 were randomly divided into a control group (30 cases) and an observation group (32 cases) using a random number table method. The control group received ultrasound debridement and dressing change treatment, while the observation group received ultrasound debridement combined with APG treatment. After 6 weeks of treatment, the effective rate, transcutaneous oxygen partial pressure (TcPO 2), and serum tumor necrosis factor- α (TNF-α), interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), hypoxia inducible factor α (HIF-1 α)and the level of MALAT1 expression in PBMCs of the two groups of patients were observed. The Pearson correlation analysis was used to investigate the relationship between the expression change of MALAT (△ MALAT1) and the total effective rate of treatment. Results:The total effective rate of the observation group was higher than that of the control group [93.75%(30/32) vs 73.33%(22/30), P<0.05]. After treatment, the systolic blood pressure (SBP), diastolic blood pressure (DBP), cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), fasting blood glucose (FPG), glycosylated hemoglobin (HbA 1c), urinary microalbumin/creatinine (UACR), uric acid (UA), white blood cells (WBC), TNF- α and IL-6 of both groups had decreased compared to before; HIF-1 α, VEGF and MALAT1 increased compared to before treatment (all P<0.05); After treatment, there was a statistically significant difference in UA, HIF-1α, VEGF, and MALAT1 between the observation group and the control group (all P<0.05). Pearson correlation analysis showed that Δ MALAT1 in DF patients was negatively correlated with TNF -α ( r=-0.61, P=0.02), IL-6 ( r=-0.52, P=0.04), WBC ( r=-0.53, P=0.03), and positively correlated with VEGF ( r=0.58, P=0.03) and HIF-1α ( r=0.54, P=0.03). The total effective rate of DF treatment was higher in the high change group of△ MALAT [88.37%(38/43) vs 73.68%(14/19), P<0.05]. There was no statistically significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:APG can significantly upregulate the expression of MALAT, improve wound tissue blood perfusion, wound angiogenesis, and inflammatory response, promote ulcer healing, and changes in MALAT expression can help determine the prognosis of DF.
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Objective@#To explore the effects of calcium gluconate and thrombin on the formation of platelet-rich gel (PRG) and the release of bioactive substances in human platelet-rich plasma (PRP) and the clinical significance.@*Methods@#Six healthy blood donors who met the inclusion criteria were recruited in our unit from May to August in 2016. Platelet samples of each donor were collected for preparation of PRP. (1) PRP in the volume of 10 mL was collected from each donor and divided into thrombin activation group (TA, added with 0.5 mL thrombin solution in dose of 100 U/mL) and calcium gluconate activation group (CGA, added with 0.5 mL calcium gluconate solution in dose of 100 g/L) according to the random number table, with 5 mL PRP in each group. Then the PRP of the two groups was activated in water bath at 37 ℃ for 1 h. The formation time of PRG was recorded, and the formation situation of PRG was observed within 1 hour of activation. After being activated for 1 h, one part of PRG was collected to observe the distribution of fibrous protein with HE staining, and another part of PRG was collected to observe platelet ultrastructure under transmission electron microscope (TEM). After being activated for 1 h, the supernatant was collected to determine the content of transforming growth factor β1, platelet-derived growth factor BB (PDGF-BB), vascular endothelial growth factor, basic fibroblast growth factor (bFGF), epidermal growth factor, and insulin-like growth factorⅠby enzyme-linked immunosorbent assay. (2) Another 10 mL PRP from each donor was collected and grouped as above, and the platelet suspension was obtained after two times of centrifugation and resuspension with phosphate buffered saline, respectively. And then they were treated with corresponding activator for 1 h as that in experiment (1). Nanoparticle tracking analyzer was used to detect the concentrations of microvesicles with different diameters and total microvesicles derived from platelet. Data were processed with t test.@*Results@#(1) The formation time of PRG in group TA was (228±40) s, and the PRG volume reached the maximum at this moment. The PRG volume shrunk to the minimum after 30 minutes of activation. The formation time of PRG in group CGA was (690±71) s, and the PRG volume reached the maximum at this moment. After 55 minutes of activation, the PRG volume shrunk to the minimum. The formation time of PRG in group TA was obviously shorter than that in group CGA (t=15.17, P<0.01). (2) HE staining showed that after 1 hour of activation, the red-stained area of fibrous protein in PRG of group TA was large and densely distributed, while that of group CGA was small and loosely distributed. TEM revealed that after 1 hour of activation, the platelets in PRG of group TA were fragmented, while lysing platelet structure, lysing α granule structure, intact α granule structure, and intact dense body structure were observed in PRG of group CGA. (3) The content of PDGF-BB released by PRP in group TA was (7.4±0.8) ng/mL, which was obviously higher than that in group CGA [(4.9±0.5) ng/mL, t=5.41, P<0.01]. The content of bFGF released by PRP in group CGA was (960±151) pg/mL, which was significantly higher than that in group TA [(384±56) pg/mL, t=8.75, P<0.01]. The content of the other 4 growth factors released by PRP in the two groups was close (with t values from 0.11 to 1.97, P values above 0.05). (4) The concentrations of total microvesicles, microvesicles with diameter more than 100 nm, and exosomes with diameter less than or equal to 100 nm derived from platelet in group CGA were (165.8±15.1)×108/mL, (142.4±12.3)×108/mL, and (23.4±2.9)×108/mL respectively, which were significantly higher than those in group TA [(24.7±4.6)×108/mL, (22.6±4.0)×108/mL, and (2.1±0.7)×108/mL, with t values from 17.36 to 22.66, P values below 0.01].@*Conclusions@#Calcium gluconate can slowly activate PRP, resulting in slowly shrunk PRG with high content of bFGF and high concentration of microvesicles, which is suitable for repairing articular cavity and sinus tract wound. Thrombin can rapidly activate PRP, resulting in quickly shrunk PRG with high content of PDGF-BB and a certain concentration of microvesicles, which is suitable for repairing acute trauma.
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Objective To evaluate the effectiveness of autologous platelet-rich gel (APG)in the repair of diabetic foot ulcers.Methods This study was a single-center,prospective,randomized controlled trial.A total of 60 patients with diabetic foot ulcers (Wagner grade 2 - 3 )were randomly divided into autologous APG group (treatment group)and recombinant bovine basic fibroblast growth factor gel group (control group).After 8 weeks, we compared wound healing rate and wound healing time at five levels (overall ulcer,superficial ulcer,sinus ulcer, Wagner 2 and Wagner 3)in the two groups.Results In APG treatment group and control group,the healing rate of overall sample ulcer (93.33% vs .63.33%,P =0.005),sinus ulcer (84.62% vs .36.36%,P =0.033),Wagner 3 (81.82% vs .30%,P =0.030)differed significantly,but did not significantly differ in superficial ulcer (100%vs .78.95%,P =0.106)or Wagner 2 (100% vs .80%,P =0.106).Ulcer healing time was 31 d vs .41.5 d,23 d vs .32 d,32 d vs .56 d,25 d vs .32 d,38 d vs .56 d,with significant difference between the two groups (P <0.05). Conclusion Autologous platelet-rich gel can effectively increase the curative rate of diabetic foot and shorten healing time.
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Objective Preparing platelet rich gel through two-times centrifugal technique and co-culturing chondrocytes with PRG, then observing the proliferation and gene expression of chondrocytes, in order to provide a favorable way to prepare tissue engineering cartilage. Methods Centrifugating venous blood of rabbit through two-times centrifugal technique to obtain platelet rich plasma( PRP) ,then detecting the concentration of various growth factor in PRP. Admixing PRP with chondrocytes of rabbit and activating them with activator. After co-culti-vation,the proliferation of chondrocytes through MTT method and expression of ACAN,CollagenⅡand SOX-9 through realtime-PCR were ob-served,and compared with common cultured chondrocytes. Results The concentrations of PDGF-AB,TGF-β1,IGF-1 and VEGF in PRG were significantly higher than those in blood(P<0. 05). After co-cultivation, the proliferation rate of chondrocytes and the expression of ACAN,Collagen Ⅱ and SOX-9 were significantly higher than that of common cultured chondrocytes(P<0. 05). Conclusion Co-culturing chondrocytes with PRG is able to promote the proliferation and gene expression of chondrocytes. We considered that it is a excellent method to construct tissue engineering cartilage.
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Cartilage is vulnerable to traumatic injury and unable to facilitate a satisfactory healing response due to its poor vascularity and inability to access mesenchymal stem cells.Continuous defects in the joint surfaces cause pain,swelling,and mechanical symptoms that result in functional impairment and limitation of athletic participation.Commonly used repair techniques include marrow stimulation,structural osteo-articular autografts or chondrocyte implantation.Platelet-rich plasma (PRP) is a concentrate extract of platelets from autologous blood,which is rich in growth factors and other cytokines and provides local environment for tissue regeneration and lends a possible option for the stimulation and acceleration of cartilage regeneration.This review gives summarization on the current state of the use of PRP for cartilage regeneration.
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Objective To compare the safety and effectiveness between treatments with autologous platelet gel (APG) versus standard care for treating refractory diabetic dermal ulcers.Methods The 46 patients with proved nonhealing diabetic dermal ulcers were enrolled. Eligible for the study were patients with grade II/III ulcers according to Wagner, lasting for at least 2 weeks and with no signs of infection at recruitment.Patients were given their informed consent document and randomly assigned to two groups: standard care (ST, n=23) or standard care plus topic application of APG (APG, n=23) for twelve weeks.The treatment of blood glucose, blood pressure and lipids was optimized and the empiric antibiotic treatment was further adjusted according to the results of culture and sensitivity testing in all patients. APG treatment consisted of wound dressing with APG, followed by topical washing and cleaning. The APG was then covered with vaseline gauze for 72 hours, after which the ulcers were treated by standard care. Participants were seen thrice a week, twice a week, or at weekly intervals. Twelve weeks observation was set as the end point.Results The would of APG group were improved in 22 patients with ulcers healed completely and 1 case with would area reduced. In the ST group, 13 ulcers were healed, 6 worsened and 4 with would area reduced. The cumulative rate of ulcer healing was 95.7% in the APG group versus 56.5% in the ST group (P=0.002). The total effective rate in APG vs ST group was 100.0% vs 73.9% (P=0.009). By Kaplan-Meier analysis,the time-to-healing of ulcer and time-to-lutation of sinus were significantly different between two groups (log-rank, P=0.006, 0.000, respectively). No treatment-related adverse events were observed. Conclusions Treatment with APG in addition to standard care results in a significantly faster and better healing for a refractory diabetic dermal ulcer and does not raise any safety concerns. So APG treatment can be a valuable and effective aid in the management of diabetic foot ulcers.