scholarly journals Effectiveness of Two Methods for Preparation of Autologous Platelet-Rich Plasma: An Experimental Study in Rabbits

2010 ◽  
Vol 04 (04) ◽  
pp. 395-402 ◽  
Author(s):  
Maria J. H. Nagata ◽  
Michel R. Messora ◽  
Flávia A. C. Furlaneto ◽  
Stephen E. Fucini ◽  
Alvaro F. Bosco ◽  
...  

Objectives: The purpose of this study was to compare the quantity and quality of platelets in platelet- rich plasma (PRP) samples prepared using either the single- or the double-centrifugation protocol.Methods: Ten adult white New Zealand rabbits were used. Ten ml of blood were drawn from each animal via cardiac puncture. Each blood sample was divided into two equal parts for PRP preparation: 5 ml of blood were centrifuged according to a single-centrifugation protocol (Group I), and 5 ml were centrifuged according to a double-centrifugation protocol (Group II). Manual platelet counts were performed on the whole blood and PRP samples of each group. Smears were also done on all samples in order to see the morphology of the platelets. The data obtained in the manual platelet count were submitted to statistical analysis (repeated measures ANOVA, Tukey, P<.05).Results: The average whole blood platelet count was 446,389/μl. The PRP samples in Group II presented an average platelet amount significantly higher than that of Group I (1,986,875 ± 685,020/μl and 781,875 ± 217,693/μl, respectively). The PRP smears from Group II were the only one to present platelets with altered morphology (75% of the smears). A few lymphocytes with increased cytoplasm were observed in the PRP smears of both Groups I (25% of the smears) and II (62.5% of the smears).Conclusions: Within the limits of this study, it can be concluded that the double-centrifugation protocol resulted in higher platelet concentrations than did the single-centrifugation protocol. However, the double-centrifugation protocol caused alterations in platelet morphology and was more sensitive to small processing errors. (Eur J Dent 2010;4:395-402)

1994 ◽  
Vol 72 (04) ◽  
pp. 511-518 ◽  
Author(s):  
Valentine C Menys ◽  
Philip R Belcher ◽  
Mark I M Noble ◽  
Rhys D Evans ◽  
George E Drossos ◽  
...  

SummaryWe determined changes in platelet aggregability following cardiopulmonary bypass, using optical aggregometry to assess macroaggregation in platelet-rich plasma (PRP), and platelet counting to assess microaggregation both in whole blood and PRP. Hirudin was used as the anticoagulant to maintain normocalcaemia.Microaggregation (%, median and interquartile range) in blood stirred with collagen (0.6 µg/ml) was only marginally impaired following bypass (91 [88, 93] at 10 min postbypass v 95 (92, 96] prebypass; n = 22), whereas macroaggregation (amplitude of response; cm) in PRP stirred with collagen (1.0µg/ml) was markedly impaired (9.5 [8.0, 10.8], n = 41 v 13.4 [12.7,14.3], n = 10; p <0.0001). However, in PRP, despite impairment of macroaggregation (9.1 [8.5, 10.1], n = 12), microaggregation was near-maximal (93 [91, 94]), as in whole blood stirred with collagen. In contrast, in aspirin-treated patients (n = 14), both collagen-induced microaggregation in whole blood (49 [47, 52]) and macroaggregation in PRP (5.1 [3.8, 6.6]) were more markedly impaired, compared with control (both p <0.001).Similarly, in PRP, macroaggregation with ristocetin (1.5 mg/ml) was also impaired following bypass (9.4 [7.2, 10.7], n = 38 v 12.4 [10.0, 13.4]; p <0.0002, n = 20), but as found with collagen, despite impairment of macroaggregation (7.2 [3.5,10.9], n = 12), microaggregation was again near-maximal (96 [93,97]). The response to ristocetin was more markedly impared after bypass in succinylated gelatin (Gelo-fusine) treated patients (5.6 [2.8, 8.6], n = 17; p <0.005 v control), whereas the response to collagen was little different (9.3 v 9.5). In contrast to findings with collagen in aspirin-treated patients, the response to ristocetin was little different to that in controls (8.0 v 8.3). Impairment of macroaggregation with collagen or ristocetin did not correlate with the duration of bypass or the platelet count, indicating that haemodilution is not a contributory factor.In conclusion: (1) Macroaggregation in PRP, as determined using optical aggregometry, is specifically impaired following bypass, and this probably reflects impairment of the build-up of small aggregates into larger aggregates. (2) Impairment of aggregate growth and consolidation could contribute to the haemostatic defect following cardiac surgery.


2020 ◽  
Vol 47 (1) ◽  
Author(s):  
Samah M. Alian ◽  
Rabab S. Zaghlol ◽  
Sahar S. Khalil

Abstract Background To compare the clinical efficacy of platelet rich plasma (PRP) subacromial injection and extracorporeal shock wave therapy (ESWT) on refractory non-calcific partial thickness supraspinatous tendon tear. Results Significant improvement in all outcome measures achieved in both groups at 4 weeks follow-up period (the end of treatment course) (P< 0.05). At 12 weeks follow-up period, group I, showed significant improvement in all outcome measures compared to baseline and group II; however, in group II, there was no significant difference compared to baseline; moreover, deterioration in the previous improvement was noticed. Conclusion Both PRP (group I) and ESWT (group II) are effective therapeutic methods in the management of refractory non-calcific partial thickness supraspinatous tendon tear; however, PRP (group I) has better long-term effects on both pain and function.


2021 ◽  
Vol 26 (2) ◽  
pp. 150-157
Author(s):  
R. R. Farkhshatova ◽  
L. P. Gerasimova ◽  
I. T. Yunusov

Relevance. It is currently relevant to study and compare the effectiveness of the autologous connective tissue grafts and the combination of collagen-based and autologous platelet-rich plasma in the surgical treatment of Miller Class I gingival recessions.Materials and methods. We examined and treated 48 (20 male (41.67%) and 28 female (58.33%)) patients aged from 25 to 40 years with Miller Class I gingival recessions. All gingival recessions were treated surgically using a modified twolayer tunnel technique. The patients were divided into two groups according to the graft type. Group I (24 patients (50%) had a connective tissue graft from the hard palate. Group II (24 patients (50%) used the combination of the autologous platelet-rich plasma and 3D collagen matrix Fibromatrix for the regeneration of oral soft tissues. We removed the sutures on the 14th day. The patients were followed up on the 7th and 14th days and in 1.3 months.Results. 48 Miller Class I gingival recessions were treated between 2018 and 2020. The depth of gingival recessions averaged 3.5 ± 1.13 mm before treatment. The level of the attached keratinized gingiva regarding the cementoenamel junction significantly (p < 0.001) improved in both groups after the surgery. The width and thickness of the keratinized gingiva best increased in group II. The mean effectiveness of gingival recession treatment was 84% in study group I and 96% – in study group II. Pain syndrome, fibrinous plaque and soft tissue edema were insignificant in group II.Conclusion. The combination of the autologous platelet-rich plasma and Fibromatrix, collagen 3D matrix, for the regeneration of the oral soft tissues is a more effective technique for the treatment of Miller Class I gingival recessions. This technique has several advantages. It is minimally invasive, less painful, soft tissue postoperative swelling is less and the received volume of the attached keratinized gums is larger than with a connective tissue graft. 


1987 ◽  
Author(s):  
W L Nichols ◽  
S E Kaese ◽  
D A Gastineau ◽  
L A Otteman ◽  
E J W Bowie

Diagnosis of Bernard-Soulier syndrome (BSS) is complicated by the difficulty of separating the giant platelets from other blood cells to pursue analyses of platelet function and structure. We report on the utility of three whole blood assay techniques for diagnosis of a patient with BSS. To our knowledge, these three techniques have not been simultaneously applied or compared for efficacy in laboratory diagnosis of BSS. (1) Whole blood platelet aggregation responses, studied with an electrical impedence aggregometer, were equivalent to those more laboriously obtained using platelet-rich plasma prepared by unit gravity sedimentation, studied with an optical light transmittance aggregometer. Platelet aggregation responses were normal with ADP or collagen stimulation, and absent with Ristocetin or bovine plasma stimulation. (2) Whole blood radioimmunoassay of platelet glycoprotein (GP) expression was performed using iodinated murine monoclonal antibodies HP1-1D (anti-GP IIb/IIIa) and 6D1 (anti-GPlb, kindly supplied by Dr. Barry Coller, Stony Brook, NY). After incubation with citrated whole blood, centrifugation was used to separate cell-bound antibody which was quantitated with a gamma counter. The patient’s whole blood had a normal level of cell-bound GP Ilb/IIIa, but a markedly reduced level of cell-bound GP lb (5% of normal mean; n = 20). (3) Whole blood smear immunocytochemical staining with the monoclonals (indirect immuno-alkaline phosphatase technique), and qualitative analysis by light microscopy, revealed a marked reduction of GP lb expression by the patient’s giant platelets, whereas GP Ilb/IIIa expression was normal. This latter technique might be especially valuable as a screening technique when the patient is not directly available for laboratory study. Together with the patient’s life-long history of thrombocytopenia and moderate bleeding diathesis, and other laboratory observations including markedly prolonged bleeding times and reduced whole blood prothrombin consumption, these data established diagnosis of BSS. We conclude that these three relatively simple assays of platelets in whole blood should be of particular value in the laboratory differential diagnosis of patients with congenital thrombocytopenias and giant platelet syndromes.


Author(s):  
Barbara Nunn

The effect of aspirin on human platelet function is usually assessed using platelet-rich plasma (PRP). Some preliminary results in vitro suggested that the effect of aspirin appears to be greater in PRP than whole blood. To explore this possibility further, a comparison of the effect of aspirin in humans ex vivo has been made taking measurements simultaneously in whole blood and PRP at 2 platelet concentrations. Blood samples (36ml) were drawn from 7 male volunteers after a light breakfast. Each took 300mg soluble aspirin and blood samples were drawn again 2 hours later. Blood was mixed with 0.1 volumes 129nM trisodium citrate. Some (30ml) was then centrifuged to prepare PRP and platelet -poor plasma (PPP) by standard techniques. Platelet concentration of some PRP was adjusted with PPP to equal that of the corresponding blood sample; the rest was adjusted to 350,000 per μl. Aggregation in response to collagen (Horm, Munich) was measured photometrically at 37°. Aggregation in 0.5ml aliquots of whole blood was measured after 4 min stirring with 154mM NaCl (control) or collagen at 37° as the fall in single platelet count determined using an Ultraflo- 100 whole blood platelet counter (Clay Adams). The concentrations of collagen producing a 50% maximal response (EC50) in PRP and blood were determined. Dose-ratios for each volunteer were calculated by dividing the EC50 obtained after aspirin by the corresponding value obtained before aspirin.The effect of aspirin was significantly (p<0.001) less in blood than PRP. Whether or not the results in whole blood more closely reflect the effect of aspirin in vivo remains to be determined.


2011 ◽  
Vol 18 (03) ◽  
pp. 426-429
Author(s):  
SALMA TANWEER ◽  
TARIQ PERVEZ ◽  
IJAZ-UL-HAQUE TASEER ◽  
Abdul Qadir Khan ◽  
Muhammad Arshad

Objective: To see the association of platelet count, splenomegaly and development of oesophageal varices. Study Design: Observational study. Duration of study: One year from January 2010 to December 2010. Setting: Gastroenterology Department, Medical Unit III, Nishtar Hospital Multan. Methodology: One hundred ten cirrhotic patients were included. The record of these patients was scrutinized and data collected was entered and analyzed through SPSS 11. The patients were divided into three groups according to platelet count. Group I with platelet count less than 50000/mm3, group II with platelet count of 50000 to 100000/mm3 and group III with platelet count of 100000 to 150000/mm3. Similarly patients were also divided into three groups according to splenic size. Group I with splenic size 11 – 13 cm, group II with splenic size 13–16 cm and group III splenic size more than 16 cm. In each group presence of esophageal varices along with grading was noted. Results: The age of the patients varied from 15 to 80 years and mean age was 48.55 ± 13.88 years. Sixty five (59.1 %) were male and 45 (40.9%) were female. The hemoglobin level of these patients varied from 6.0 to 14.3 gm/100 ml with mean level of 9.23 ± 2.11gm/100ml. The platelet count varied from 22000 to 385000/mm3. The splenic size varied from 9 to 18 cm with mean of 12.53 ± 2.14 cm. Esophageal varices were detected in 102 cases. Seventeen cases were of grade I varices, 25 cases were of grade II varices, 40 cases were in grade III varices and 4 cases were in grade IV varices. Maximum number of grade-III (22 patients) and grade IV (3patients) esophageal varices occurred in patients having platelet count less than 50000/mm3. As the splenic size increases the grade of esophageal varices also increases. Maximum number of esophageal varices occur in grade II (25) followed by grade-III (37) in patients with splenic size in the range of 13 to 16 cm. Conclusions: A low platelet count and large splenic size are good non-invasive predictors of esophageal varices. These parameters can also accurately assess the presence of large varices. So a patient of cirrhosis with low platelet count and large spleen has an increased diagnostic yield of esophageal varices on upper GI endoscopy.


2021 ◽  
Vol 71 (4) ◽  
pp. 1377-81
Author(s):  
Humaira Sarwar ◽  
Irfan Shah ◽  
Ali Akhtar Khan ◽  
Muhammad Afzal ◽  
Adnan Babar ◽  
...  

Objective: To evaluate the role of combination therapy of platelet rich plasma (PRP) with arthrocentesis and to compare it with arthrocentesis alone in the temporomandibular dysfuction (TMD) patients.Study Design: Quasi experimental study.Place and Duration of Study: Oral and Maxillofacial Surgery department, Armed Forces Institute of Dentistry, Combined Military Hospital Rawalpindi, from Jan 2017 to Jun 2019.Methodology: Forty-two patients diagnosed with refractory unilateral temporomandibular dysfuction, were included in the study. Patients were divided in two groups with 21 temporomandibular joints in each. Arthrocentesis alone was the control group (group I) and arthrocentesis with intra-articular injection of platelet rich plasma (group II) was the study group. Treatment outcomes were assessed and compared for all patients based on clinical parameters of pre and post treatment; for pain, maximal mouth opening and temporomandibular joint clicking sounds. Results: Out of 42 patients, 33 (79%) were females and 9 (21%) were males with mean age of 34.3 ± 8.4 years. There was statistically significant difference in both groups for all variables. The p-value of maximum mouth opening of both groups before and after treatment was 0.746 and 0.01, joint clicking sounds were present in 69% of our patients before the treament and it reduced to 14% after the treatment. There was marked gradual decrease in pain of both groups, group I (6.48 ± 1.470 to 1.81 ± 0.602) and group II (7.29 ± 1.007 to 1.19 ± 0.402). Conclusion: Combination therapy of platelet rich plasma with arthrocentesis is more effective treatment method than.......


1990 ◽  
Vol 64 (02) ◽  
pp. 211-215 ◽  
Author(s):  
Dan S Sharp ◽  
Andrew D Beswick ◽  
John R O'Brien ◽  
Serge Renaud ◽  
John W G Yarnell ◽  
...  

SummaryThis epidemiological study was undertaken to explore possible relationships among various haematological indices, prevalent ischaemic heart disease and platelet “function” as measured by two rather different methods. ADP-induced platelet impedance changes in whole blood were strongly associated with prevalent ischaemic heart disease in a general population of 49-66 year men at increased risk. Adenosine diphosphate (ADP) induced platelet aggregation in platelet rich plasma (PRP) at a constant platelet count and also the whole blood platelet count and red cell (RBC) count were strongly and independently related to ADP-induced platelet impedance changes. Both platelet count and platelet aggregation in PRP assessed by changes in optical density were directly related to increasing platelet “sensitivity” as measured by impedance changes in whole blood but RBC count was inversely related. Positive independent relationships between platelet impedance changes and plasma viscosity and fibrinogen were markedly attenuated when platelet count was taken into account, but this finding does not discount a role for these factors in platelet aggregation. No relationship was noted between white blood cell (WBC) count and platelet impedance changes; however, a significant inverse relationship was noted with platelet aggregation in PRP. These findings indicate that laboratory-based experimental findings can be observed in population based studies, and that these haematological factors may be important indicators of ischaemic disease in the population.


1982 ◽  
Vol 48 (03) ◽  
pp. 327-329 ◽  
Author(s):  
S C Fox ◽  
M Burgess-Wilson ◽  
S Heptinstall ◽  
J R A Mitchell

SummaryThe Ultra-Flo 100 Whole Blood Platelet Counter has proved a useful tool for measuring platelet aggregation in whole blood, the extent of aggregation being deduced from the number of single platelets that remain. The technique has allowed us to show that platelets aggregate spontaneously in citrated blood and in heparinized blood but not in whole blood collected into EDTA. The aggregation occurs during storage but its rate is enhanced by stirring and it occurs more readily when the whole blood has been exposed to plastic rather than glass. It occurs much more readily in whole blood from some individuals than from others and the process may involve adenosine diphosphate (ADP). The rate of aggregation in whole blood is enhanced by several aggregating agents including collagen, ADP and sodium arachidonate which are more usually studied in platelet-rich plasma.


1987 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
C D Forbes

Spontaneous platelet aggregation (SPA) was studied in human whole blood at 3,5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter (Ultra Flo 100), SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of blood at 37°C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10-5M at each time interval; (b) 0.5-100 x 10-6M at 3 and 30 minutes, and (c) 15 x 10-6M in combination with 2 x 10-4M adenosine (Ad), 8 x 10-6M 2-chloradenosine (2ClAd, a specific ADP receptor blocker) and 5 x 10-5M aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy withAd, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when an effective concentration of Dipy and an ineffective concentration of Ad (10-4M) were addedtogether, the inhibitory effect of Dipy was not increased, suggesting that Dipy inhibits platelet aggregation independent of Ad.The increase in SPA with the time after venepuncture was abolished when bloodwas taken directly into the anticoagulant containing 2ClAd (5 x 10-6M). We conclude that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture, and makes a serious contribution to the artifacts ofin vitro platelet function studies. Furthermore, the decrease in the inhibitory action of Dipy with the time after venepuncture may explain why previously, it has not been possible to observe inhibition of platelet aggregation by Dipy in platelet rich plasma which requires time to prepare.


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