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 review the research progress of the preparation and components of the platelet rich plasma (PRP). MethodsThe recent literature concerning the biological mechanism, preparation, and components of PRP was analyzed and summarized. ResultsThe biological function of PRP depends on a series of intricate cascade of cellular and molecular events. PRP contains different concentrations of platelets, which would release a large number of the activated molecules, and also contains a small amount of white blood cells and red blood cells. The preparation of PRP is based on platelet concentration. Different preparation techniques would lead to different platelet concentrations, recovery ratios, and components. ConclusionThere is no uniform standard for the preparation of PRP. Different preparation methods and technical parameters of PRP will get different components and different concentrations of PRP, which also provide a reference for cl inicians to select the most appropriate PRP for individual patient.
ObjectiveTo investigate the effect of platelet rich plasma (PRP) in promoting wound healing of total hip arthroplasty (THA). MethodsBetween January 2011 and January 2012, 80 patients scheduled for THA and accorded with the inclusion criteria were divided into 2 groups:wounds were treated with PRP in 40 patients (PRP group) and with normal saline in 40 patients (control group). There was no significant difference in gender, age, disease duration, injury causes, sides, fracture type, and preoperative Harris hip scores between 2 groups (P>0.05). Routine drainage and functional exercise were performed after operation. ResultsThe postoperative drainage volume of PRP group[(137±26) mL] was significantly lower than that of control group[(424±39) mL] (t=38.726, P=0.000). At 4 days after operation, no inflammatory reaction was observed in 34 cases of PRP group and in 30 cases of control group, mild inflammatory reaction in 5 cases of PRP group and in 6 cases of control group, moderate inflammatory reaction in 1 case of PRP group and in 4 cases of control group; there was no significant difference between 2 groups (χ2=2.141, P=0.343). Wound healed by first intention in 40 patients of PRP group and in 39 patients of control group, showing no significant difference between 2 groups (P=1.000). The average follow-up period was 9 months (range, 6-12 months). The Harris hip scores of PRP group (90.2±2.5) and control group (89.3±3.1) at last follow-up were significantly better than those before operation (39.6±8.9 and 39.2±9.2 respectively) (t=34.618, P=0.000; t=32.638, P=0.000), but no significant difference was found between 2 groups (t=1.429, P=0.153). ConclusionUsing PRP in THA wound can reduce postoperative drainage volume, improve the healing of operation incision. It is a safe, effective, and promising procedure in treatment of THA wound.
Objective To explore the best centrifuge condition for preparing rabbit leukocyte-poor platelet-rich plasma (LP-PRP) by using single centrifugation method. Methods Sixteen healthy New Zealand rabbits, aged 3-4 months, were utilized in the investigation. A total of 15 mL anticoagulated blood was extracted from the central ear artery of each rabbit, with a repeat of the blood collection procedure after 1 and 2 months. The obtained blood specimens were individually subjected to centrifugation at a radius of 16.7 cm and speeds of 1 200, 1 300, 1 400, and 1 500 r/min (equivalent to centrifugal forces of 269×g, 315×g, 365×g, and 420×g) for durations of 2, 3, 4, and 5 minutes, resulting in a total of 16 groups. Following centrifugation, collect plasma from each group to a distance of 1.5 mL from the separation plane. The volumes, platelet enrichment coefficient, and platelet recovery rates of LP-PRP in each group, under varying centrifugation conditions, were methodically computed and subsequently compared. Results The volume of LP-PRP obtained under all centrifugation conditions ranged from 1.8 to 7.6 mL. At a consistent centrifugal speed, an extension of centrifugation time leaded to a significant increase in the volume of LP-PRP, accompanied by a declining trend in the platelet enrichment coefficient of LP-PRP. When centrifuged for 2 minutes, the volume of LP-PRP at speeds of 1 200 and 1 300 r/min was less than 2.0 mL, while the volume of LP-PRP obtained under other conditions was more than 2.0 mL. When centrifuged for 4 and 5 minutes, the volume of LP-PRP obtained at each speed was more than 4 mL. LP-PRP with a platelet enrichment coefficient more than 2.0 could be prepared by centrifuging at 1 200 r/min for each time group and 1 300 r/min for 2 and 3 minutes, and the highest LP-PRP platelet enrichment coefficient could be obtained by centrifugation for 2 minutes at a speed of 1 200 r/min. The platelet recovery rates of LP-PRP obtained by centrifugation at 1 200 r/min for 4 and 5 minutes, as well as centrifugation at 1 400 r/min for 5 minutes, were both greater than 60%. There was no significant difference between the groups when centrifuged at 1 200 r/min for 4 and 5 minutes (P>0.05). Conclusion In the process of preparing rabbit LP-PRP using a single centrifugation method, collecting 15 mL of blood and centrifuging at a radius of 16.7 cm and speed of 1 200 r/min for 4 minutes can prepare LP-PRP with a volume exceeding 2.0 mL, platelet enrichment coefficient exceeding 2.0, and platelet recovery rate exceeding 60%. This centrifugal condition can achieve the optimal LP-PRP action parameters in the shortest possible time.
ObjectiveTo investigate the effect of ultrasound-guided anterior injection of platelet-rich plasma (PRP) combined with routine physiotherapy on pain and functional improvement in patients with rotator cuff injury.MethodsThe patients with rotator cuff injury treated in the Department of Rehabilitation Medicine, the Affiliated Hospital of Southwest Medical University from August 2017 to June 2018 were randomly divided into control group and PRP group. The control group was treated with routine physiotherapy for 4 weeks, and the PRP group was treated with PRP injection guided by musculoskeletal ultrasound (once a week, two injections) combined with routine physiotherapy for 4 weeks. The Shoulder Pain and Disability Index (SPADI) and the European Shoulder Association Constant-Murley Score (CMS) were used to evaluate the changes of pain and function before intervention, 12 weeks after intervention and 24 weeks after intervention.ResultsFinally, 65 patients were included, with 33 in the control group and 32 in the PRP group. Compared with the control group, there were significant differences in the CMS and SAPDI scores between the PRP group and the control group at 12 and 24 weeks after intervention (P<0.05), except before intervention (P>0.05). Twelve and 24 weeks after intervention, the CMS scores of the two groups were higher than those before intervention, while the SPADI scores were lower than those before intervention (P<0.05). The results of repeated measures analysis of variances showed that there was no statistical significance in group effect of CMS or SAPDI scores (F=2.753, P=0.102; F=2.724, P=0.104), but the time effects of CMS and SAPDI scores were statistically significant (F=251.002, P<0.001; F= 1 846.753, P<0.001), and there was interaction between group and time (F=4.931, P=0.020; F=36.405, P<0.001).ConclusionAnterior injection of PRP guided by musculoskeletal ultrasound combined with conventional physiotherapy is more effective than conventional physiotherapy in relieving shoulder pain and improving shoulder joint function.
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.
Platelet-rich plasma (PRP) is a platelet-rich plasma protein concentrate extracted from autologous peripheral blood, which contains a variety of blood-derived growth factors and cytokines. As an autologous blood product, PRP is widely used in many fields such as tissue repair and regeneration because of its minimally invasive process, simple preparation process and good biological properties. The acquisition of PRP is mainly achieved by collecting peripheral blood through density gradient centrifugation. Various growth factors and cytokines in PRP can repair various tissues. With the deepening of PRP research, it is now gradually applied to rotator cuff injury, lateral epicondylitis of humerus, carpal tunnel syndrome, knee joint injury, gluteal muscle tendinopathy, achilles tendinopathy, plantar fasciitis, and other soft tissue injuries, and some progress has been made. This article reviews the progress on clinical applications of PRP in chronic soft tissue injuries to provide a theoretical basis.
Objective Platelet-rich plasma (PRP) can enhance the chondrocyte prol iferation and repair of cartilage defects. To explore the safety and efficacy of intra-knee-articular injection of PRP to treat knee articular cartilage degeneration by comparing with injecting sodium hyaluronate (SH). Methods Thirty consecutive patients (30 knees) with knee articular cartilage degeneration were selected between January 2010 and June 2010. According to different injections, 30 patients wererandomly divided into PRP group (test group, n=15) and SH group (control group, n=15). There was no significant difference in gender, age, body mass index, and Kellgren-Lawrence grade between 2 groups (P gt; 0.05). Test group received 3.5 mL of PRP intra-knee-articular injections while control group received 2 mL of SH during the same time period. Both treatments were administered in series of 3 intra-knee-articular injections at 3-week intervals. Then, adverse reactions were recorded. International Knee Documentation Committee (IKDC) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and Lequesne index were used for evaluation of treatment results. Results The patients of 2 groups were followed up 6 months. There were significant differences in IKDC score, WOMAC score, and Lequesne index between pre- and post-injection in 2 groups (P lt; 0.05); no significant difference was found between different time points (3, 4, and 6 months) in test group (P gt; 0.05), while significant differences were found between the postoperative 6th month and the postoperative 3rd and 4th months in control group (P lt; 0.05). There was no significant difference in IKDC score, WOMAC score, and Lequesne index between 2 groups within 4 months (P gt; 0.05), but the effectiveness of test group was significantly better than that of control group at 6 months after injection (P lt; 0.05). Adverse reactions occurred in 12 patients (31 injections) of test group and in 12 patients (30 injections) of control group. No significant difference in onset time, termination time, and duration of adverse reactions were found between 2 groups (P gt; 0.05). Conclusion Intra-knee-articular injection of PRP to treat knee articular cartilage degeneration is safe, which can alleviate symptoms of pain and swell ing and improve the qual ity of l ife of patients; however, further data of large samples and long-term follow-up are needed to confirm the safety and effectiveness.
Objective To explore the effectiveness and mechanism of pure platelet-rich plasma (P-PRP) on osteochondral injury of talus. Methods Thirty-six patients with osteochondral injury of talus selected between January 2014 and October 2017 according to criteria were randomly divided into control group (group A), leukocyte PRP (L-PRP) group (group B), and P-PRP group (group C), with 12 cases in each group. There was no significant difference in gender, age, disease duration, and Hepple classification among the three groups (P>0.05). Patients in the groups B and C were injected with 2.5 mL L-PRP or P-PRP at the bone graft site, respectively. Patients in the group A were not injected with any drugs. The American Orthopaedic Foot and Ankle Society (AOFAS) score and visual analogue scale (VAS) score were used to evaluate the effectiveness before operation and at 3, 6, and 12 months after operation. Study on the therapeutic mechanism of P-PRP: MC3T3-E1 cells were randomly divided into control group (group A), L-PRP group (group B), and P-PRP group (group C). Groups B and C were cultured with culture medium containing 5% L-PRP or P-PRP respectively. Group A was cultured with PBS of the same content. MTT assay was used to detect cell proliferation; ELISA was used to detect the content of matrix metalloprotein 9 (MMP-9) protein in supernatant; alkaline phosphatase (ALP) activity was measured; and real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of osteopontin (OPN), collagen type Ⅰ, and MMP-9 in cells. Western blot was used to detect the expression of MMP-9 in supernatant and phosphoinositide 3-kinase (PI3K), phosphorylated protein kinase B (pAKT), and phosphorylated c-Jun (p-c-Jun) in cells. ResultsAll patients were followed up 13-25 months, with an average of 18 months. No complication such as wound infection and internal fixation failure occurred. MRI showed that the degree of injury was similar between the three groups before operation, and patients in the three groups all recovered at 6 months after operation. Moreover, group C was superior to groups A and B. Compared with preoperation, AOFAS scores and VAS scores in the three groups were all significantly improved at each time point after operation (P<0.05). AOFAS score of group C was significantly higher than that of groups A and B at 3, 6, and 12 months after operation (P<0.05); there was no significant difference in VAS score between the three groups (P>0.05). Study on the therapeutic mechanism of P-PRP: The absorbance (A) value, ALP activity, the relative mRNA expression of OPN and collagen type Ⅰ in group C were significantly higher than those in groups A and B (P<0.05), and those in group B were significantly higher than those in group A (P<0.05). The relative expression of MMP-9 protein and mRNA and the content of MMP-9 protein detected by ELISA in group B were significantly higher than those in groups A and C, while those in group C were significantly lower than those in group A (P<0.05). Western blot detection showed that the relative expression of PI3K, pAKT, and p-c-Jun protein in group B was significantly higher than those in groups A and C (P<0.05), but there was no significant difference between groups A and C (P>0.05). Conclusion P-PRP is superior to L-PRP for osteochondral injury of talus, which may be related to the inhibition of PI3K/AKT/AP-1 signaling pathway in the osteoblast, thereby reducing the secretion of MMP-9.
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%.