ObjectiveTo evaluate the effect of leucocyte- and platelet-rich plasma (L-PRP) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in treating avascular necrosis of the femoral head (ANFH) in rabbits. MethodsTwenty-four New Zealand white rabbits (4-6 months old, both genders, weighing 2.0-3.0 kg) were used for the establishment of bilateral ANFH models and divided into 4 groups (n=6). BMSCs were isolated from the bone marrow of iliac crest, cultured and identified. L-PRP was prepared by Landesberg method. Core decompression only (group A), core decompression and L-PRP implantation (group B), core decompression and BMSCs implantation (group C), and core decompression and implantation of BMSCs and L-PRP were performed in 4 groups. To evaluate bone formation and remodeling of the defects, X-ray photography was taken at 2, 4, and 8 weeks postoperatively. The modified Lane-Sandhu scoring system was used to evaluate the bone formation. Two rabbits were sacrificed at 2, 4, 8 weeks after operation to harvest the specimens for histological observation, new blood vessel count and new bone area ratio. ResultsThe observations of radiology and histology displayed different degrees of bone regeneration at bone defect sites in each group. At 2, 4, and 8 weeks postoperatively, the results of Lane-Sandhu X-ray photography scoring, new blood vessel count, and new bone area ratio showed that groups C and D were significantly better than groups A and B, group D was significantly better than group C. and group B was significantly better than group A (P<0.05). ConclusionThese findings demonstrate that L-PRP can promote osteogenic differentiation of BMSCs in treating ANFH in rabbits, and core decompression associated with BMSCs and L-PRP is an effective and feasible method to treat ANFH.
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 [WTBZ]To assess the impact of dual antiplatelet therapy using aspirin and clopidogrel on postoperative bleeding and blood transfusion early after coronary artery bypass grafting (CABG). Methods [WTBZ]In this randomized controlled trial, 249 patients were randomly assigned to 2 groups after coronary artery bypass grafting from December 2007 to December 2008. Daily clopidogrel (75 mg) and aspirin (100 mg) were initiated in 124 patients (group AC) while aspirin (100 mg) alone was administered to 125 patients (group A). Antiplatelet therapy was initiated within 48h postoperatively. Demographic, operative, and postoperative data were compared between the two groups. Chest tube drainage and quantity of blood products used in both groups were recorded. The effects of the antiplatelet regimen on chest tube drainage were compared using a linear regression model. Results [WTBZ]No statistical difference of demographic, operative, and preoperative data was observed between the two groups （Pgt;0.05）. Chest tube drainage after patients received ntiplatelet agents was not significantly different between group A and group AC（495.00±270.89 ml vs. 489.25±316.68ml，t=0.146, P=0.884）. No statistical difference of cases of transfusion(81 cases vs. 91 cases，χ2=1.937, P=0.164) or quantity of red cells (2.51±2.88 U vs. 2.25±2.87 U, t=0.690, P=0.491) and plasma (195.45±300.88 ml vs. 223.01±238.68 ml,t=0.759, P=0.449) transfused was found between group A and group AC. No perioperative mortality, reexploration or extrathoracic bleeding occurred in either group. Early postoperative use of dual antiplatelet therapy was not associated with increased bleeding after coronary artery bypass grafting on multivariable analysis（r=2.297，95%CI:-64.526,69.121，P=0.946）. Conclusionpresent study suggests that according to a predefined administration protocol, dual antiplatelet therapy of aspirin and clopidogrel can safely be administered in the early postoperative period in CABG patients, without increasing the risk of bleeding complications.
Cardiopulmonary bypass(CPB) is associated with thrombocytopenia and platelet dysfunction. The primary cause of acquired platelet defect is thought to be activation and release of alpha granules during CPB. Before CPB, platelet-rich plasma (PRP) was prepared by obtaining the required amount of patient’s whole blood by autologous plateletpheresis. PRP could be reinfused after operation in order to protect the function and quantities of the platelets. On the other hand, PRP could be made into autologous platelet gel (APG). APG contains supraphysiologic amounts of growth factors, and has adequate tensile strength and adhesive ability. Therefore, it can be used for hemostasis in operation, sealing wound and enhancing incision or dehiscent sternal wounds healing.
Objective Platelet-rich plasma (PRP) can stimulate intervertebral disc cell proliferation, promote extracellular matrix synthesis, and inhibit annulus fibrosus cell apoptosis. To investigate the effects of autologous PRP on the treatment of the early intervertebral disc degeneration (IDD) so as to provide the experimental basis for its clinical application. Methods Forty-five healthy New Zealand white rabbits (male or female, weighing 2.5-3.0 kg) were randomly divided into the experimental group (n=15), the control group (n=15), and the sham group (n=15). PRP was prepared from the arterial blood of rabbit’s ears of the experimental group with Landesberg’s method. The platelet concentrations in both whole blood and PRP were detected. The rabbit model of early IDD was established by annulus fibrosus puncture (L4, 5, L5, 6) in both the experimental group and the control group; 100 ?L autologous PRP and 100 ?L PBS were injected into the degenerative intervertebral discs respectively after 2 weeks of models creation. In sham group, intervertebral discs were separated and exposed without treatment. The general conditions of the rabbits were observed after building models; at 2 weeks after degeneration, 1 and 2 weeks after intervention, 5 rabbits were selected randomly from each group respectively for MRI observation, histological observation by using HE staining and collagen type II immunohistochemical staining. The signal of lumbar MRI was assessed and the contents of collagen type II were detected. Results The platelet concentration of PRP was about 4.92 times as much as that of the whole blood. All the animals survived to the end of the experiment. At 2 weeks after degeneration, a lower T2 signal was observed in both the experimental group and the control group; the nucleus pulposus cells decreased and extracellular matrix degenerated; and the expression of collagen type II decreased in both the experimental group and control group. The degenerative grade of lumbar MRI in the experimental group and control group were significantly higher than that in the sham group (P lt; 0.05), and the content of collagen type II were significantly lower than that in the sham group (P lt; 0.05). At 1, 2 weeks after intervention, disc degeneration in the experimental group was significantly lower than that in control group (P lt; 0.05), and significant difference was found between experimental group and sham group (P lt; 0.05). The nucleus pulposus cells and chondroid matrix in the experimental group were more than those in the control group, showing slight stromal fibrosis; but the expression of collage type II was significantly higher than that in the control group (P lt; 0.05). Conclusion The disc injection of autologous PRP may terminate or even reverse the progress of rabbit early IDD, which may be associated with the role of multiple growth factors of PRP in regulating cell function, improving the tissue microenvironment, and promoting tissue regeneration.
Objective The use of autologous platelet-rich gel (APG) is a relatively new technology and a promising treatment method for infections, which is currently being used by a variety of surgical specialties. The mechanism of antibacterialeffect of APG is not yet fully discovered. Subsequent evidence suggests that platelets have multi ple functional attributes inantimicrobial host defense (including the capacity to generate antimicrobial oxygen metabol ites and the antimicrobial peptides) and interact directly with microorganisms, contribute to clearance of pathogens from the blood. To investigate the bacteriostasis of APG against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa in vitro. Methods Platelet-rich plasma (PRP) and platelet-poor plasma (PPP) were obtained from whole blood of 17 healthy donors. APG was prepared by mixing PRP with bovine thrombin in a 10% calcium gluconate solution or bovine thrombin in a 10% calcium gluconate solution and apocynin (APG-APO). Antibacterial effects of APG, PRP, and APG-APO on Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were evaluated by bacteriostasis assay. Results The culture results showed apparent decrease in the number of Staphylococcus aureus for both APG and APG-APO, which was maximal at first 4 hours and lasted to 24 hours and 8 hours, respectively; showing significant difference (P lt; 0.05) when compared APG with PRP and PPP, however no significant difference at first 8 hours (P gt; 0.05) and significant difference at 12 and 24 hours (P lt; 0.05) when compared APG with APG-APO; showing significant difference at first 4 hours (P lt; 0.05), no significant difference at 6, 8, 12, and 24 hours when compared APG-APO with PRP and PPP (P gt; 0.05). The bacteriostasis rates of APG and APG-APO were 27.36%-52.97% and 18.82%-51.52% against Escherichia coli, respectively; showing no significant difference (P gt; 0.05) when compared with PRP. The bacteriostasis rates of APG and APG-APO were less than 35% against Pseudomonas aeruginosa, showing no significant difference (P gt; 0.05) when compared with PRP; the bacteriostasis rates of PRP were less than 15% against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Conclusion APG may have potential bacteriostatic effect against Staphylococcus aureus by platelet mediating. Either APG or APG-APO has no obvious bacteriostatic effect against Escherichia coli or Pseudomonas aeruginosa. PRP has no antibacterial activity against Staphylococcus aureus, Escherichia coli or Pseudomonas aeruginosa.
To compare the platelet enrichment ratio of platelet-rich plasma (PRP) prepared by different centrifuge methods and to compare the concentration of growth factors released from autologous platelet-rich gel (APG) with the whole blood. Methods Thirteen diabetic patients with refractory skin lesions were enrolled in APG treatment. ① Three kinds of centrifuge methods were selected for PRP by 11 diabetic patients: A(n=6): 529 × g for 4 minutes in the first centrifugeand 854 × g for 6 minutes in the second centrifuge; B (n=5): 313 × g for 4 minutes in the first centrifuge and 1 252 × g for 6 minutes in the second centrifuge; C (n=5): 176 × g for 5 minutes in the first centrifuge and 1 252 × g for 5 minutes in the second centrifuge. Platelet counted on the whole blood and PRP was determined. The APG, produced by combining the PRPwith thrombin and calcium gluconate (10 ∶ 1) was used by patients. ② PDGF-BB, TGF-β1, VEGF, EGF, and IGF-1 were measured in the APG and the whole blood using the enzyme-l inked immunosorbent assay method. Results ① The average platelet concentration was higher in group B [(1 363.80 ± 919.74) × 109/ L] than in groups A[(779.67 ± 352.39) × 109/ L)] and C[(765.00 ± 278.78) × 109/ L] and the platelet recovery rate was 75.2% ± 21.0% in group B. ② The concentration of growth factors all increased with the increasing platelet number. On average, for the whole blood as compared with APG, the PDGF-BB concentration increased from (145.94 ± 133.24) pg/mL to (503.81 ± 197.86) pg/mL (P lt; 0.05); TGF-β1 concentration increased from (3.31 ± 2.27) ng/mL to (5.67 ± 4.80) ng/mL (P lt; 0.05); IGF-1concentration increased from (14.54 ± 35.34) ng/mL to (110.56 ± 84.36) ng/mL (P lt; 0.05); and EGF concentration increased from (160.73 ± 71.10) pg/mL to (265.95 ± 138.43) pg/mL (P lt; 0.05). No increase was found for VEGF(P gt; 0.05). ③ There was positive correlation between the platelet concentration and PDGF-BB and TGF-β1 (r = 0.627, r = 0.437, P lt; 0.05). ④ Thirteen diabetic repractory dermal ulcers received APG treatment for 18 times, 9 ulcers (69.2%) and 10 sinuses (88.3%) were cured at the end of 12-week treatment. Conclusion The method ofgroup B is the best centrifuge method. A variety of growth factors are detected and released from the platelets at significant levels in APG. There is positive correlation between the platelet concentration and PDGF-BB and TGF-β1 .
Objective To review and evaluate the clinical application of autologous plateletrich gel (APG) in refractory chronic diabetic cutaneous ulcers. Methods The latest literature was extensively reviewed, and the potential mechanism of APG for healing diabetic cutaneous ulcers was discussed. Results APG accelerated the ulcer healing, especially in healing the refractory diabetic cutaneous ulcers, and the high-level growth factors from the platelet releasate were thought to be the major potential mechanism of the APG action. Conclusion APG can promote the healing of refractory chronic diabetic cutaneous ulcers. Topical haemotherapy withAPG may be considered an adjuvant treatment of a multidisciplinary process and may be useful to the effective therapy of cutaneous ulcers.
To investigate the significance of activated platelet levels before and after surgical operation in patients with severed finger by severe injury, the P-selectin (CD62p) positive platelet percent in peripheral blood was studied in 32 patients before and after operation by flow cytometric immunology. At the second and third day after operation, seventeen patients, which in a high positive platelet percent, were measured again after anticoagulation treatment with a large dose of urokinase etc. The results were that the levels of CD62p positive platelet in peripheral blood before and after operation were significantly higher than that of the normal group (P lt; 0.01). The CD62p positive platelet percent was increased not much more in the anti-coagulation patients than in the normal ones (P gt; 0.05). There were a few vascular crisis observed. This study suggested that severed fingers by a crushing or wringing injury enhance the platelet activity and resulted in hypercoagulation, thus anti-coagulation measures should be taken to avoid possible thrombosis and improve the survival rate of replantation. CD62p was one of the sensitive indices of activated platelet, which could be used in calculating hypercoagulation accurately.
Objective To investigate the difference of curative effect of various surgical methods for the treatment of idiopathic senile macular hole. Methods A retrospective analysis was made for 86 eyes with stage Ⅱ-Ⅳ idiopathic full-thickness macular hole treated with various modes of operation,ie, single vitrectomy (7 eyes),vitrectomy combined with autologous platelet concentrate (APC) as an adjuvant (40 eyes), vitrectomy with internal limiting membrane (ILM) peeling (14 eyes), vitrectomy with both ILM peeling and APC treatment (25 eyes). The main outcome measures included anatomic reattachment rate，change of visual acuity，findings of optic coherence tomography (OCT), Amsler grid and intra or postoperative complication evaluations. Results (1) In visual acuity improvement, the APC group (80.0%) was significantly better than anyone of the other three groups (P＜0.05). (2) In anatomic success rate, the single vitrectomy group was significantly lower than the vitrectomy with APC treatment group（87.5%）or vitrectomy with both ILM peeling and APC as an adjuvant group(92.0%)(P＜0.05). (3) There was no significant difference in operative complication and improvement of distortion of vision. Conclusion Vitrectomy combined with APC as an adjuvant for the treatment of idiopathic macular hole is helpful to improve both the anatomic success rate and postoperative visual acuity. The usage of ILM peeling technique could improve the anatomic reattachment rate, but the vision prognosis of ILM peeling patients is not as good as the patients of APC as an adjuvant. (Chin J Ocul Fundus Dis, 2002, 18: 196-198)