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.
【摘要】 目的 探讨机采血小板在实验室内保存期间的生化改变及其意义。 方法 采用ROCHE cobas b 221和OLYMPUS AU400测定血小板样本的pH值、PO2、PCO2、葡萄糖（GLU）浓度、乳酸脱氢酶（LDH）释放量。 结果 保存期间pH值由7.311（第1天）降至7.116（第5天）；PO2波动于106.7～119.5 mm Hg（1 mm Hg=0.133 kPa）；PCO2波动于25.0～35.8 mm Hg；GLU浓度呈进行性下降，由18.8 mmol/L降至15.2 mmol/L；LDH浓度显著增加，由150 mmol/L（第1天）增至259 mmol/L（第5天）。 结论 血小板在保存期内，其生化参数会随着保存时间发生改变。而这些参数作为间接指标对机采血小板是否被细菌污染具有一定的指导意义。【Abstract】 Objective To observe the biochemistry changes of apheresis platelets (AP), including pH, PO2, PCO2, GLU, LDH, within the storage time and its significance. Methods The pH, PO2, PCO2, GLU and LDH of platelet samples were detected by ROCHE cobas b 221 and OLYMPUS AU400. Results During the storage time, the pH value of platelet decreased gradually from 7.311 (the first day) to 7.116 (the fifth day); the PO2 fluctuated from 106.7 mm Hg (1 mm Hg=0.133 kPa) to 119.5 mm Hg; the PCO2 fluctuated from 25.0 mm Hg to 35.8 mm Hg;the concentration of GLU decreased from 18.8 mmol/L to 15.2 mmol/L;the concentration of LDH increased observably from 150 mmol/L (the first day) to 259 mmol/L (the fifth day). Conclusion There were observably changes of biochemistry of the AP during storage time, and these changes may be the indicators of platelets contamination.
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.
Objective The tendon-bone heal ing is the key point to ensure the success of the anterior cruciate l igament (ACL) reconstruction. To observe the histological change in the tendon-bone heal ing after ACL reconstruction by different concentrations of platelet-rich plasma (PRP) combined with deproteinized bone (DPB) of calf as bone tunnel infill ing and to investigate the active effect of the complex on tendon-bone heal ing and to define the optimal concentration of PRP. Methods Eight mL blood was drawn from central artery of New Zealand rabbit ears; PRP was prepared by Landesbergmethod, and l iquid supernatant was used as thinner to prepare different concentrations of PRP (30%, 60%, and 100%). Fresh osteoepiphysis spongy bone was harvested from lower end of femur of newborn calf to prepare DPB by way of 30% H2O2 and ether alternating soaking for 24 hours continuous 6 times. DPB was soaked in different concentrations of PRP and mixed with activator to prepare the PRP/DPB complex. A total of 54 New Zealand white rabbits, aging 8-12 months, weighing (2.5 ± 0.4) kg, were divided randomly into 3 groups: group A (30%PRP/DPB complex, n=18), group B (60%PRP/DPB complex, n=18), and group C (100%PRP/DPB complex, n=18). The legs of the rabbits were randomly divided into experimental side and the control side; ACL was reconstructed by semitendinosus and PRP/DPB complex in bone tunnel in the experimental side, and only by semitendinosus in the control side. The general conditions of the rabbits were observed postoperatively and HE staining was used to observe the tendon-bone heal ing, then I-IV levels of semi-quantitative analysis of the tendon-bone heal ing were evaluated according to Demirag standard at 3, 6, and 12 weeks. Results General observation: Synovial fluid sl ightly increased in the specimens and no bony tissue was found in inner of femoral tunnel at 3 weeks; there was no synovial fluid in all the specimens and scar tissue was discovered in inner of femoral tunnel at 6 weeks; and there was no synovial fluid and the tendons became tighter with fibrous tissue at 12 weeks. Histological observation: New granulation tissue formed in the tendon-bone interface of group A experimental sides at 3 weeks; there was various widths of Sharpey type textile fiber in the tendon-bone interface at 6 weeks; Sharpey type textile fiber arranged regularly, which formed an irregular and blur “tidal l ine” at 12 weeks. Group B experimental sides were better than any other group at 3, 6, and 12 weeks; chondrocyte-l ike arranged regularly in the tendonboneinterface at 3 weeks; the number of chondrocyte-l ike per unit area was more than that of the other groups at 6 weeks;and chondrocyte-l ike prol iferated and matured in the tendon-bone interface, Sharpey type textile fiber became tighter andordered. Group C experimental sides were similar to both sides of group A at 3 weeks, however, the prol iferation of relatively mature dense connective tissue was worse than that of other groups at 6 and 12 weeks. According to Demirag grading, there were significant differences in tendon-bone heal ing between the experimental sides and the control sides of group B at 3 and 6 weeks, and between group B experimental sides and group C experimental sides at 12 weeks (P lt; 0.05). Conclusion The mixture of PRP/PRP has good biocompatibil ity and bone induction, so it can enhance tendon-bone heal ing after ACL reconstruction when the concentration of PRP is 60%.
Objective To investigate the clinical outcomes of autologous platelet rich plasma (PRP) for anterior cruciate ligament (ACL) reconstruction. Methods Between August 2014 and August 2016, 42 patients with ACL ruptures who underwent arthroscopic ACL reconstruction were randomly divided into 2 groups: 21 patients received graft soaked with PRP (trial group) and 21 patients received routine graft in ACL reconstruction (control group). Because 6 patients failed to be followed up, 17 patients of trial group and 19 of control group were enrolled in the study. There was no significant difference in gender, age, body mass index, side, injury reason, disease duration, Kellgren-Lawrence grade, and preoperative visual analogue scale (VAS), Lysholm score, and International Knee Documentation Committee (IKDC) activity scores between 2 groups (P>0.05). VAS score, Lysholm score, and IKDC activity scores were used to evaluate pain and function at 3 and 12 months postoperatively. Further, second arthroscopy and MRI examination were performed at 12 months postoperatively. Results The patients in both groups were followed up 3 to 12 months with an average of 9.83 months. The VAS score, Lysholm score, and IKDC activity scores were significantly improved at 3 and 12 months after operation in 2 groups (P<0.05), and the scores of trial group were significantly better than those of control group at 3 months (P<0.05), but no significant difference was found between 2 groups at 12 months (P>0.05). No complications of effusion, infection, and allergy were observed in 2 groups during follow-up. MRI showed good position of ACL grafts and good signal quality of the graft in the majority of the cases. However, mixed hyperintense and presence of synovial fluid at the femoral bone-tendon graft interface were found in 3 patients of trial group and 4 patients of control group, indicating poor remodeling ligamentation. MRI score was 3.53±1.13 in trial group and was 3.21±0.92 in control group, showing no significant difference (t=0.936,P=0.356). The second arthroscopy examination showed ligament remodeling score was higher in trial group than control group (t=3.248,P=0.014), but no significant difference was found in synovial coverage score and the incidence of cartilage repair (t=2.190,P=0.064;χ2=0.090,P=0.764). Conclusion PRP application in allograft ACL reconstruction can improve knee function and relieve pain after operation, which may also accelerate graft remodeling.
OBJECTIVE: To investigate the expression and distribution of platelet derived growth factor receptor-beta(PDGFR-beta) in normal skin and keloid and to discuss its biological function in keloid formation. METHODS: 1. To detect the expression and distribution of PDGFR-beta in normal skin and keloid tissue by immunohistochemistry; 2. To detect the receptor expression in vitro by Flow cytometry (FCM); 3. To detect the subcellular distribution of receptor by Laser confocal microscope. RESULTS: 1. Immunohistochemistry showed that normal skin and keloid tissue were almost the same in expression but different in distribution of PDGFR-beta; 2. There was more expression of PDGFR-beta in normal fibroblasts than that in keloid fibroblasts in vitro by FCM; 3. Laser confocal microscope revealed that the PDGFR-beta concentrated on the surface of cell membrane in keloid fibroblasts, but in normal skin fibroblasts, the receptors were coagulated on the nuclear membrane and intranucleus. CONCLUSION: Compared with the fibroblasts in vivo, there was a difference of the PDGFR-beta expression in fibroblasts in vitro, more expression of PDGFR-beta in normal fibroblast than that in keloid fibroblast in vitro; and the subcellular distribution of PDGFR-beta was different in normal skin and keloid fibroblasts. The characteristics of the expression and distribution of PDGFR-beta in keloid may contribute to the formation of keloid.
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 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.