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.
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.
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 investigate the effects of platelet-derived growth factor(PDGF) on the expression of α-smooth muscle actin(α-SMA) of cultured human retinal pigment epithelium cells(RPE). Methods Cultured human RPE cells of the 4-6 th passages were divided into two groups: Delbecco′s modified Eagle′s medium (DMEM) and 2%DMEM (20 g/L foeta calf serum+DMEM). PDGF (0,1,50 ng/ml) was added to medium.The expression of α-SMA was detected and quantitatively analyzed by image process of immunofluorescence.Results PDGF stimulated the expression of α-SMA of human RPE cells.In group of DMEM, The rate of RPE of α-SMA expression was 40%-50% and the intension of fluorescence was 8.08 without PDGF. After stimulated by PDGF(1 ng/ml,50 ng/ml), the rates were 80% and 90% respectively, and the intension of fluorescence were 12.35 and 17.23. In 2%DMEM group, The rates of RPE of α-SMA expression were 85% without PDGF, and 95% ,100% respectively treated with PDGF (1 ng/ml,50 ng/ml). The intension of fluorescence was 14.79 without PDGF, and after stimulated by PDGF, they were 16.28 at 1 ng/ml and 21.36 at 50 ng/ml，which was 2 .7 times ber than that in DMEM group without PDGF. Conclusion PDGF could stimulate RPE cells to express α-SMA. (Chin J Ocul Fundus Dis，2003，19：201-268)
ObjectiveTo summarize the pathogenesis and epidemiology features of gastrointestinal stromal tumor(GIST), explore its diagnosis and therapy, and analyze its prognosis. MethodThe pertinent literatures about the pathogenesis, epidemiology features, diagnosis, therapy, and prognosis of GIST in recent years were reviewed. ResultsGIST was non-epithelial tumor which derived from interstitial cells of Cajal, was the most common mesenchymal tumor about accounting for 1%-3% in the digestive tract tumor. The median onset age of patients with GIST was 40-60 years. The gastric stromal tumor was about 60% in all the digestive tract tumor. The current consensus statement was that there was a relation between the pathogenesis of the GIST and proto-oncogene c-kit or platelet-derived growth factor receptor alpha(PDGFRα)gene mutation. But the mutations of PDGFRαand c-kit gene did not emerge at the same time in the same patient. The clinical manifestations of GIST were not specific, and the diagnosis mainly depended on endoscope and image technology, the correct diagnosis depended on pathological examination. The treatment of GIST was given priority of surgery and molecular targeted drug therapy, and the prognosis was closely related to risk assessment stratify of GIST. ConclusionsGISTs are mesenchymal tumors that has a potential of malignant transformation, the risk classification criteria for aggressive clinical course of primary GIST is an important indication for guiding the clinical therapy and prognostic evaluation. Further research would be needed in prevention, diagnosis, treatment, and relapse prevention of GIST.
Objective To study the effects of platelet-rich plasma(PRP) on the proliferation and osteogenetic activity of the marrow mesenchymal stem cells(MSCs) cultured in vitro to elucidate the cellular and molecularmechanism by which PRP accelerates bone repair.Methods The human MSCs were cultured in vitro and randomly divided into the experimental group(n=9) and control group(n=9). In the experimental group, the MSCs were interfused with PRP(10 μl/ml culture media). The proliferation ability of the cells was tested by flow cytometry and MTT, the osteogenetic activity by alkaline phosphatase(ALP) measuring and tetracycline fluorometry, and cbfal mRNA expression by reverse transcriptPCR.Results PRP could stimulate the MSCs proliferation. The flow cytometry assay showed that the MSCs proportion of S period of the experimental group significantly increased 14.5±0.4 in comparison with that of the control group 7.2±0.5 (P＜0.01) after 24 hours. MTT value showed that MSCs proliferatedto platform period earlier in the experimental group than in the control group. There was a significant increase in ALP activity of the experimental group 7.79±1.98,9.51±2.31and 14.03±3.02 when compared with that of the control group 2.06±0.77,2.84±0.82 and 2.58±0.84 after 3, 6 and 9 days(P＜0.05). The number of mineral nodes increased. Reverse transcript-PCR showed that the expression of cbfal mRNA were elevated gradually at 2,4 and 8 hours after interfused with PRP.Conclusion The effect of PRP on accelerating bone repair is related to its effects on stimulating the proliferation of MSCs, increasing the cbfal expression and promoting the osteogenetic activity.
Objective To investigate the effect of removing the implanted plate-rod system for scol iosis (PRSS) on maintaining scol iosis curve correction and preserving spinal mobil ity in patients with scol iosis. Methods From June 1998 to February 2002, 119 cases of scol iosis were treated with the implant of PRSS, which was removed 26-68 months later (average46.8 months). Complete follow-up data were obtained in 21 patients, including 6 males and 15 females aged 11-17 years old (average 13.8 years old). The disease course was 9-16 years (average 12.1 years). There were 2 cases of congenital scol iosis and 19 cases of idiopathic scol iosis, which included 5 cases of IA, 2 of IB, 1 of IIA, 2 of IIB, 2 of IIC, 2 of IIIA, 3 of IIIB, and 2 of IVA according to Lenke classification. There were 13 cases of thoracic scol iosis and 8 of thoracolumbar scol iosis. AP view and the lateral and anterior bending view of X-ray films before and at 3 to 6 months after removing PRSS were comparatively analyzed, the coronal and the sagittal Cobb angle were measured, and the height of vertebral body on the concave side and the convex side were measured, so as to know the effect of PRSS on the growth of the vertebral endplates. Results All the implants were removed successfully with an average operation time of 2.5 hours (range 2-4 hours) and a small amount of intraoperative blood loss. Twenty-one cases were followed up for 6-72 months (average 34.4 months). The coronal Cobb angle before and after the removal of PRSS was (20.25 ± 8.25)° and (23.63 ± 8.41)°, respectively, indicating there was no significant difference (P gt; 0.05); while the sagittal Cobb angle was (39.44 ± 12.38)° and (49.94 ± 10.42)°, respectively, indicating there was a significant difference (P lt; 0.05). The height of the top vertebral body on the concave side before and after the removal of PRSS was (1.78 ± 0.40) cm and (2.08 ± 0.35) cm, respectively, and there was a significant difference (P lt; 0.01); while the height on the convex side was (2.16 ± 0.47) cm and (2.18 ± 0.35) cm, respectively, indicating no significant difference was evident (P gt; 0.05). All the 21 patients had good prognosis and no major operative compl ication occurred. Conclusion PRSS is an effective instrumentation for the management of scol iosis. After the removal of the PRSS, the correction of scol iosis can be maintained, and the spinal mobil ity can be protected and restored.
【Abstract】 Objective To find out the best method to prepare platelet-rich plasma (PRP) and to evaluate the effect of PRP gel on skin flap survival and its mechanism. Methods Totally, 72 Wistar rats (aged 12 weeks, weighing 250-300 g) were used for the experiment. The arterial blood (8-10 mL) were collected from the hearts of 24 rats to prepare PRP with three kinds of centrifuge methods: in group A, 200 × g centrifuge for 15 minutes, and 500 × g centrifuge for 10 minutes；in group B, 312 × g centrifuge for 10 minutes, and 1 248 × g centrifuge for 10 minutes；and in group C, 200 × g centrifuge for 15 minutes, and 200 × g centrifuge for 10 minutes. The platelet was counted in the whole blood, PRP, and platelet-poor plasma (PPP) to determine an ideal centrifuge. PRP, PPP, and the serum after first centrifuge were collected. The concentrations of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor β1 (TGF-β1) were measured in the PRP, PPP, and serum using the enzyme-linked immunosorbent assay method, and PRP and PPP gels were prepared. The flaps of 11 cm × 3 cm in size were elevated on the back of 48 rats, which were divided into 3 groups: PRP gel (PRP group, n=16) and PPP gel (PPP group, n=16) were injected, no treatment was given in the control group (n=16). The flap survival rate was measured at 7 days. Histological and real-time PCR were used to count the inflammatory cells and blood vessel density, and to detect the expressions of vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), PDGF-AA, and PDGF-BB mRNA at 8 hours, 24 hours, 3 days, and 7 days. Results Platelet counting showed platelet in group A was the highest. ELISA evaluation showed that the concentrations of TGF-β1 and PDGF-BB were significantly higher in PRP than in PPP and serum (P lt; 0.05). The flap survival rate was 61.2% ± 9.1% in PRP group, showing significant differences (P lt; 0.05) when compared with that in PPP group (35.8% ± 11.3%) and control group (28.0% ± 5.4%). The inflammatory cells were significantly lower and the blood vessel density was significantly higher in PRP group than in PPP group and control group (P lt; 0.05). When compared with PPP group and control group, the expressions of VEGF and PDGF-BB increased at all time after operation in PRP group; the expression of EGF increased within 24 hours; and the expression of PDGF-AA increased after 3 days. There were significant differences in PDGF-AA mRNA at 3 days and 7 days, PDGF-BB mRNA at 8 hours, VEGF mRNA at 24 hours and 3 days, and EGF mRNA at 24 hours between PRP group and PPP and control groups (P lt; 0.05). Conclusion 200 × g centrifuge for 15 minutes and 500 × g centrifuge for 10 minutes is the best PRP preparation method. PRP can improve the skin flap survival by regulating the genes involved in angiogenesis.
Objective To investigate the interleukin-17 (IL-17) levels changes in both synovial fluid and venous plasma of patients with primary knee osteoarthritis (OA) after intra-articular injection of platelet-rich plasma (PRP). Methods Between January 2015 and January 2016, 30 patients with primary knee OA were treated by intra-articular injection of PRP once a week for 3 weeks (trial group). Thirty healthy individuals were recruited into the study as control. There was no significant difference in gender, age, and body mass index between 2 groups (P>0.05). Visual analogue scale (VAS) score and Knee Society Score (KSS) were used to evaluate pain level and function of the knee for patients with OA. The IL-17 levels in both venous plasma and synovial fluid were measured before injection and at 1, 3, 6, and 12 months after injection in trial group and the IL-17 levels in venous plasma were measured in control group. The levels were determined using ELISA method. Results There was no knee joint swelling, fever, local infection, or other uncomfortable symptoms for all patients in process of PRP injection. All patients were followed up 13.5 months on average (range, 12-15 months). In trial group, the VAS scores at different time points after injection were significantly lower than that before injection (P<0.05). And the KSS scores at different time points after injection were significantly higher than that before injection (P<0.05). There was no significant difference in VAS and KSS scores between different time points after injection (P>0.05). The IL-17 levels in venous plasma before and after injection in trial group were significantly higher than that in control group (P<0.05). The IL-17 levels in venous plasma at each time point after injection were significantly lower than that before injection (P<0.05). There was no significant difference in IL-17 levels in both venous plasma and synovial fluid between different time points after injection (P>0.05). Conclusion Intra-articular injection of PRP can significantly release the pain symptoms, improve joint function, and reduce IL-17 levels in both synovial fluid and venous plasma of the patients with knee OA, but IL-17 levels can not reduce to normal level.
Objective To investigate the effects of combined platelet-rich plasma (PRP) and arterial supercharging technique on the survival rate and functional restoration of cross-body region skin flaps in rabbits. MethodsTwelve healthy 6-month-old New Zealand White rabbits were randomly assigned to 4 groups (n=3): sham group, PRP group, anastomosis group, and combined treatment group. An axial skin flap with an area of 12 cm×6 cm on the inner side of the hind limbs of all animals were prepared, with the saphenous artery as the main blood supply. Following the ligation of both the proximal and distal ends of the saphenous artery across all groups, the sham group received no further intervention, the PRP group was subjected to PRP injection, the anastomosis group underwent in situ end-to-end anastomosis of the distal saphenous artery, and the combined treatment group received both in situ distal saphenous artery anastomosis and PRP administration. Flap survival was evaluated and recorded on postoperative days 1, 3, and 7, with survival rates calculated accordingly. On day 7, flap tissue samples were harvested for HE staining to assess basal tissue morphology. Additionally, immunohistochemical staining was conducted to detect the expression of α-smooth muscle actin (α-SMA), vascular endothelial growth factor (VEGF), and CD31 in the flap tissues. Results At postoperative day 1, no significant difference in flap survival rates were observed among the 4 groups (P>0.05). At day 3, the PRP group showed no significant difference compared to the sham group (P>0.05); however, both the anastomosis and combined treatment groups exhibited significantly higher survival rates than the sham group (P<0.05), the combined treatment group further demonstrated superior survival rates compared to both the PRP and anastomosis groups (P<0.05). At day 7, the combined treatment group maintained significantly higher survival rates than all other groups (P<0.05), while both the PRP and anastomosis groups exceeded the sham group (P<0.05). HE staining at day 7 revealed persistent inflammatory cell infiltration, sheet-like erythrocyte deposition, and disordered collagen fibers in the sham group. The PRP group showed nascent microvessel formation and early collagen reorganization, whereas the anastomosis group displayed mature microvasculature with resolved interstitial edema. The combined treatment group exhibited differentiated microvessels with densely packed collagen bundles. Immunohistochemical analysis at day 7 demonstrated significantly larger relative area percentages of α-SMA, VEGF, and CD31 positive cells in the combined treatment group compared to all other groups (P<0.05). Both the PRP and anastomosis groups also showed significantly higher values than the sham group (P<0.05). Conclusion The combination of PRP and arterial supercharging techniques significantly enhances flap healing, potentially through mechanisms involving augmented angiogenesis and improved blood supply.