Objective To explore the effect of the platelet-rich plasma (PRP) on proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells (MSCs) in China goat in vitro. Methods MSCs from the bone marrow of China goat were cultured. The third passage of MSCs were treated with PRP in the PRP group (the experimental group), but the cells were cultured with only the fetal calf serum (FCS) in the FCS group (the control group). The morphology and proliferation of the cells were observed by an inverted phase contrast microscope. The effect of PRP on proliferation of MSCs was examined by the MTT assay at 2,4,6 and 8 days. Furthermore, MSCs were cultured withdexamethasone（DEX）or PRP; alkaline phosphatase (ALP) and the calcium stainingwere used to evaluate the effect of DEX or PRP on osteogenic differatiation of MSCs at 18 days. The results from the PRP group were compared with those from the FCS group. Results The time for the MSCs confluence in the PRP group was earlier than that in the FCS group when observed under the inverted phase contrast microscope. The MTT assay showed that at 2, 4, 6 and 8 days the mean absorbance values were 0.252±0.026, 0.747±0.042, 1.173±0.067, and 1.242±0.056 in the PRP group, but 0.137±0.019, 0.436±0.052, 0.939±0.036, and 1.105±0.070 in the FCS group. The mean absorbance value was significantly higher in the PRP group than in the FCS group at each observation time (P＜0.01). Compared with the FCS group, the positive-ALP cells and the calcium deposition were decreased in the PRP group; however, DEX could increase boththe number of the positiveALP cells and the calcium deposition. Conclusion The PRP can promote proliferation of the MSCs of China goats in vitro but inhibit osteogenic differentiation.
Objective To study the mechanism of compound of calcium phosphate(TCP) and platelet-rich plasma(PRP) in the treatment of femoral head necrosis.Methods The left femoral heads of 48 New Zealand white rabbits were frozen by liquid nitrogen as to make themodel of femoral head necrosis.Twenty-four rabbits were randomly chosen as theexperimental group and their femoral heads were filled with TCP/PRP. The other 24 rabbits were used as the control group and their femoral heads were filled only with TCP. They were sacrificed at 2, 4,8,12 weeks after operation. The specimens were examined with X-ray and histological study.Results At 2 weeks after operation,there was no significant difference in femoral headdensity between the two groups. Four weeks after operation, femoral head density decreased in both groups, while it decreased more in the control group. At 8,12 weeks after operation, the density of the femoral heads in both groups increased, and it was higher in the experimental group. Histology examination showed thatthere was no difference between the two groups 2 weeks after operation. The head became flat at 4 weeks. Control group had more defects. At 4,8,12 weeks, more repairs were observed in the experimental group than that in the control group. The amount and maturity of osteogenesis in experimental group were much more greaterthan those in control group．Bone histomorphometry showed that the volum of thetrabecular was larger in the experimental group (36.65%±7.22%,38.29%±4.28%，39.24%±3.42%) than that of control group(P＜0.05). Conclusion TCP/PRP does not only provide osteoblasts scaffold, butalso promotes bone formation and the head repair. TCP/PRP is a good biomaterialfor the treatment of femur head necrosis.
OBJECTIVE: To study the effect of platelet-rich plasma in the repair of bone defect. METHODS: Segmental bone defects of 1 cm were created in the mid-upper part of bilateral radius of 24 New Zealand white rabbits. One side was randomly chosen as the experimental side, which was filled with artificial bone with platelet-rich plasma (PRP). The other side filled with artificial bone without PRP as the control. After 2, 4, 8 and 12 weeks of implantation, the gross, radiological, histological observations, and computer graphic analysis were performed to investigate the bone healing of the defect in both sides. RESULTS: Two weeks after operation, new bone and fibrous tissue formation in both the experimental and the control sides were observed only in the areas adjacent to the cut ends of the host bone, but the amount of new tissue in the experimental side was much more than that in the control side. In the 4th and 8th weeks, the surface of the artificial bone was covered with a large amount of new bones, the artificial bone was bridged tightly with the host bone by callus in the experimental side, while new bone was limited mainly in the cut ends and was less mature in the control side. In the 12th weeks, bone defects were entirely healed in the experimental side, which were covered completely with cortical bone, while new bone formation was only observed in the ends of artificial bone and there were not continuous bone callus on the surface in the control side. CONCLUSION: Artificial bone with PRP is effective in the repair of segmental bone defects, and PRP could improve the healing of bone defect.
ObjectiveTo explore the clinical efficacy of the ultrasound-guided intra-articular injection of platelet-rich plasma (PRP) in the treatment of patients with different stages of knee osteoarthritis.MethodsWe retrospectively analyzed the clinical characteristics and X-ray data of patients with knee osteoarthritis who received ultrasound-guided intra-articular injection of PRP in the Department of Rehabilitation Medicine at Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University between May 2018 and June 2019. The patients were grouped according to the Kellgren & Lawrence Classification (K&L 0, Ⅰ, Ⅱ, Ⅲ, and Ⅳ). All the patients received four injections with a one-week interval. The Visual Analogue Scale (VAS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) were used to evaluate the clinical efficacy before the injection, and 3 and 6 months after the injection. Adverse reactions were recorded.ResultsA total of 102 patients were included without any grade 0 cases. There were 20 patients in K&L Ⅰ group, 37 in Ⅱ group, 31 in Ⅲ group, and 14 in Ⅳ group. No adverse event was reported. Significant differences of VAS scores and WOMAC index were observed in Ⅰ, Ⅱ and Ⅲ groups at the 3rd and 6th month follow-up (P<0.05). VAS and WOMAC scores of the three groups at the 3rd and 6th month after the treatment were significantly improved compared with those before the treatment (P<0.05). There was no significant difference in VAS score at the 3rd or 6th month after the treatment three groups (P>0.05). For K&L Ⅰ group, there was no statistically significant difference in WOMAC score at the 3rd or 6th month after the treatment (P>0.05). However, the WOMAC scores at the 3rd month after the treatment were better than those at the 6th month in K&L Ⅱ and Ⅲ groups (P<0.05). There was no significant time-depended changes in VAS score or WOMAC score in K&L Ⅳ group (P>0.05).ConclusionThe ultrasound-guided intra-articular PRP injection is safe and effective for pain relief and function improvement in patients with knee osteoarthritis at the early and middle stage.
Objective To calculate the recovery rate and enrichment factor and to analyse the correlation by measuring the concentrations of platelets, leukocyte, and growth factors in platelet-rich plasma (PRP) so as to evaluate the feasibil ity and stabil ity of a set of PRP preparation. Methods The peripheral blood (40 mL) was collected from 30 volunteers accorded with the inclusion criteria, and then 4 mL PRP was prepared using the package produced by Shandong Weigao Group Medical Polymer Company Limited. Automatic hematology analyzer was used to count the concentrations of platelets and leukocyte in whole blood and PRP. The enrichment factor and recovery rate of platelets or leukocyte were calculated; the platelet and leukocyte concentrations of male and female volunteers were measured, respectively. The concentrations of platelet-derived growth factor (PDGF), transforming growth factor β (TGF-β), and vascular endothel ial growth factor (VEGF) were assayed by ELISA. Results The platelet concentrations of whole blood and PRP were (131.40 ± 29.44) × 109/L and (819.47 ± 136.32) × 109/L, respectively, showing significant difference (t=—27.020, P=0.000). The recovery rate of platelets was 60.85% ± 8.97%, and the enrichment factor was 6.40 ± 1.06. The leukocyte concentrations of whole blood and PRP were (5.57 ± 1.91) × 1012/L and (32.20 ± 10.42) × 1012/L, respectively, showing significant difference (t=—13.780, P=0.000). The recovery rate of leukocyte was 58.30% ± 19.24%, and the enrichment factor was 6.10 ± 1.93. The concentrations of platelets and leukocyte in PRP were positively correlated with the platelet concentration (r=0.652, P=0.000) and leukocyte concentration (r=0.460, P=0.011) in whole blood. The concentrations of platelet and leukocyte in PRP between male and female were not significantly different (P gt; 0.05). The concentrations of PDGF, TGF-β, and VEGF in PRP were (698.15 ± 64.48), (681.36 ± 65.90), and (1 071.55 ± 106.04) ng/ mL,which were (5.67 ± 1.18), (6.99 ± 0.61), and (5.74 ± 0.83) times higher than those in the whole blood, respectively. PDGF concentration (r=0.832, P=0.020), TGF-β concentration (r=0.835, P=0.019), and VEGF concentration (r=0.824, P=0.023) in PRP were positively correlated with platelet concentration of PRP. Conclusion PRP with high concentrations of platelets, white blood cells and growth factors can be prepared stably by this package.
Objective To examine an effect of the locally-used platelet derived growth factor-BB (PDGF-BB) on the healing of the medial collateral ligament (MCL) in the knee joints of rats. Methods Forty-eight rats were equally randomly divided into 2 groups: the experimental group (group A) and the control group(group B). MCL of all the rats were ruptured to establish the wound models. In group A, 5 μg of PDGF-BB was locally injected in the wound of each rat and then the wound was sutured; but in group B, the wound was only sutured. After 2 weeks, histological evaluations were performed to determine whether PDGF-BB could promote the healing of MCL. Results There were significantly more fibroblasts formed during the ligament healing process in group A than in group B (213.44±15.32 vs. 180.42±12.78, Plt;0.01). The fibroblasts were more mature andmore regularlyarranged in group A than in group B. The type, content, and crosslink of the collagen were improved to a greater extent in group A than in group B (Plt;0.01). Conclusion PDGF can promote the healing of the injured ligament.
To investigate the role of platelet-activating factor (PAF), neutrophils in ischemia-reperfusion-induced liver injury and their possible mechanism, PAF and the degree of neutrophil infiltration in liver tissue and the preventive effects of PAF antagonist kadsurenone were evaluated in this study by means of a partial liver ischemia model, in which it was induced by clamping only left and median lobes of the liver without causing intestinal congestion. The present study was undertaken to find out the mechanism of liver ischemia-reperfusion injury and preventive effect of kadsurenone. The results indicate that in early stage of reperfusion liver injury possibly caused by the generation of free radicals, declined of autioxidant defence and increased Ca２＋ influx, and in the later stage of reperfusion injury was mainly mediated by accumulation of PAF in the liver, which elicits the release of polymorphonuclear leukocytes induced toxical free radical, endothelial damage, microcirculatory collapse. The authors conclude that the effectiveness of antagonist kadsurenone in protecting against ischemiareperfusioninduced liver injury is due not only to their action in preventing the direct effects of PAF, but also to their ability to inhibit both PAF priming and PAF dependent feedback processes, thus preventing escalation of auto generated inflammatory damage.
ObjectiveTo explore the clinical efficacy and safety of ultrasound-guided intra-articular injection of platelet-rich plasma (PRP) in the treatment of avascular necrosis of the femoral head.MethodsWe retrospectively collected and analyzed the clinical characteristics, imaging data, and clinical outcomes of patients with femoral head necrosis who received ultrasound-guided intra-articular PRP injection in the Department of Rehabilitation Medicine of Sun Yat-sen Memorial Hospital, Sun Yat-sen University between June 2019 and June 2020. All the patients received 4 injections at one-week intervals. The Visual Analogue Scale (VAS), Western Ontario and McMaster University Osteoarthritis Index (WOMAC), and Harris Hip Joint Function Scale (HHS) were evaluated before treatment and 1 month, 3 months, and 6 months after the first injections. Adverse events were recorded. The normally distributed data were presented as mean±standard deviation, and analyzed by one-way repeated measures analysis of variance; the non-normally distributed data were presented as median (lower quartile, upper quartile), and analyzed by Friedman test.ResultsA total of 29 patients were included. According to the Association Research Circulation Osseous classification standard, 2 patients were classified as stageⅠ, 11 as stageⅡ, 11 as stage Ⅲ, and 5 as stage Ⅳ. Before treatment and 1 month, 3 months, and 6 months after treatment, the VAS scores were 7.0 (5.5, 8.0), 4.0 (3.0, 5.0), 3.0 (2.0, 3.0), and 3.0 (2.0, 5.0), respectively, the WOMAC scores were 39.27±11.70, 28.34±8.08, 22.82±6.09, and 24.13±7.55, respectively, and the HHS were 46.0 (40.0, 64.0), 71.0 (57.5, 75.0), 78.0 (68.0, 80.5), and 78.0 (64.0, 80.0), respectively. The time effects in VAS (χ2=65.423, P<0.001), WOMAC (F=46.710, P<0.001), and HHS (χ2=66.347, P<0.001) were all statistically significant. There were significant differences in each index between the values 1 month, 3 months, and 6 months after treatment and those before treatment respectively, and there was also a significant difference in each index between the value 1 month after treatment and that 3 months after treatment (P<0.05). There was no significant difference in any indicator between the value 6 months after treatment and that 3 months after treatment (P>0.05). Significant difference was shown between the value 6 months after treatment and that 1 month after treatment in WOMAC (P=0.016), but not in VAS or HHS (P>0.05). No obvious adverse event was reported during the follow-up period.ConclusionsUltrasound-guided intra-articular PRP injection can effectively alleviate the pain and improve the hip joint function of patients with femoral head necrosis for at least 6 months. However, randomized controlled studies with a larger sample size and longer-term follow-up are needed in the future to confirm the efficacy and safety of PRP injection in femoral head necrosis.
The authors observed the progressive changes of the platelet activity in 25 cases of acute cholangitis severe type(ASCT)within 2 weeks of pre-and-post-operation.The results revealed that there are signficant changes of the platelet activity in ASCT.The levels of the platelet activity were proportional to the severity of disease and degree of biliary duct obstruction.Persistency of abnormality of platelet activity may predict the worse of disease and bad prognosis.The results are useful to understand the complex changes of pathophysiology in ACST,to the mechanism of multiple organic failure.
Objective To investigate the ability of plateletrich plasma(PRP) combined with cells and artificial bone in accelerating the repair of bone defect. Methods The marrow stromal stem cells (MSCs) of rabbit were cultured and induced into the osteoblast-like cells in vitro. PRP was produced with low-density twice centrifugations. Forty-eight New Zealand rabbits were made 1.2 cm bilateral radius defect models and divided into 4 groups averagely at random: group A(left:PRP／MSCs／β-tricalcium phosphate(β-TCP), right: MSCs／β-TCP), group B (left:autoradius, right: PRP／MSCs／β-TCP)； group C (left:autoradius,right: MSCs／β-TCP), and group D(left:PRP／β-TCP; right:β-TCP). At 2, 6 and 12 weeks after operation， the repair of bone defect was evaluated by the generalobservation, histology, biomechanics and histomorphology. Results There was a stable platelet concentration in PRP and it was about 5.45±0.23 times of whole blood. In the aspect of bone bridge and conture of the defects, at 2 and 6 weeks, PRP／MSCs／β-TCP and MSCs／β-TCP displayed asimilar outcome and were less than auto in general sample and X-ray;at12 weeks，PRP／MSCs／β-TCP was similar to autoradius and better than MSCs／β-TCP.in the aspect of quantity and quality of bone formation，histology showed that PRP／MSCs／β-TCP and autoradius were better than MSCs／β-TCP（P＜0.05），and there was nosignificantdifference between PRP／MSCs／β-TCP and autoradius（P＞0.05）. At 2 and 6 weeks，there was no significant difference between PRP／β-TCP and β-TCP（P＞0.05）。At 12 weeks，PRP／β-TCP was better than β-TCP（P＜0.05）. In the aspect of intensity f bone formation，at 6 and 12 weeks，PRP／MSCs／β-TCP and autoradiuswere better than MSCs／β-TCP（P＜0.05）. At 6 weeks，autoradius was better than PRP／MSCs／β-TCP（P＜0.05）. At 12 weeks，there was no significant difference between PRP／MSCs／β-TCP and auto（P＞0.05）. PRP／TCP and β-TCP had no significant difference at 12 weeks（P＞0.05）. Conclusion PRP／MSCs／β-TCP demonstrated excellent ability of forming bone in experiment. PRP was most likely to accelerate the repair of bone defect through increasing the activity of proliferation and differentiationof MSCs and osteoblasts.