ObjectiveTo summary the standardized management in research and applications of platelet derivatives for tissue regeneration.MethodsThe related literature about bottlenecks and standardized management of platelet derivatives in recent years was reviewed and analyzed.ResultsAlthough the platelet derivatives are increasingly used to accelerate the regenerative processes of injured joint, skin, nerve, ligament/tendon, and alveolar bone, etc., the large variation in preparation methods, diverse nomenclature, incomplete reporting system, and lack of quantitative and standardized management of the preparation process have caused uncertainty and incomparability of research and application results. In recent years, there has been a trend towards standardized research and management of platelet derivatives.ConclusionThe implementation of standardized research and quality management will contribute to promote the research and application of platelet derivatives in the field of tissue regeneration.
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.
Objective To compare effectiveness of injecting platelet-rich plasma (PRP) with different concentrations of leukocytes under ultrasound in treatment of supraspinatus tendon injury. Methods A clinical data of 30 patients with supraspinatus tendon injury, who met the selection criteria and were admitted between December 2022 and December 2023, was retrospectively analyzed. Thirty patients were treated with 4 injections of leukocyte-poor PRP (LP-PRP, n=10), leukocyte-rich PRP (LR-PRP, n=11), and triamcinolone (n=9), with an interval of 7-10 days between each injection. There was no significant difference between groups (P>0.05) in the age, gender, disease duration, affected shoulder side, Ellman classification, preoperative visual analogue scale (VAS) score, Constant score, Disabilities of the Arm, Shoulder, and Hand (DASH) score, and American Shoulder and Elbow Surgeons (ASES) score. At 1, 3, and 6 months after injection, the shoulder pain and function were evaluated by using the VAS score, Constant score, DASH score, and ASES score. MRI was conducted to observe supraspinatus tendon healing. Results No severe adverse reactions was observed in all groups. All patients were followed up 6-7 months, with an average of 6.5 months. After injection, the ASES score and Constant score gradually increased in the LR-PRP group and LP-PRP group, while the VAS score and DASH score decreased, with significant differences compared to before injection (P<0.05). Except for no significant difference between 3 and 6 months after injection in LR-PRP group (P>0.05), the above scores showed significant differences between different time points (P<0.05). At 1 month after injection, the Constant score in triamcinolone group significantly increased compared to before injection, while the VAS score significantly decreased (P<0.05). There was no significant difference in all scores between other time points in the triamcinolone group (P>0.05). Except for 1 month after injection, there was no significant difference in Constant score and VAS score between groups (P>0.05). At all other time points, the LR-PRP group and LP-PRP group had better scores than the triamcinolone group (P<0.05). There was no significant difference between the LR-PRP group and the LP-PRP group (P>0.05). MRI showed that only 4 patients in the LP-PRP group had signs of repair at the supraspinatus tendon injury site at 6 months after injection, while no significant tendon repair sign was observed in the other patients. Conclusion Compared with triamcinolone treatment, multiple injections of LP-PRP and LR-PRP under ultrasound can promote the recovery of shoulder joint function and significantly relieve pain in patients with supraspinatus tendon injury, and imaging improvement can be seen after LP-PRP treatment.
ObjectiveTo investigate the effectiveness of the treatment under the guidance of “diamond concept” for femoral shaft fractures nonunion after intramedullary fixation.MethodsBetween January 2014 and December 2016, 21 cases of femoral shaft fractures nonunion after intramedullary fixation were treated with auxiliary plate fixation combined with autogenous iliac graft, and autologous bone marrow concentrate and platelet-rich plasma (PRP) gel under the guidance of the “diamond concept”. There were 13 males and 8 females, with an average age of 32.5 years (range, 17-48 years). All fractures were closed femoral shaft fractures. Four patients underwent internal fixation with plate and resulted in nonunion, then they were fixed with intramedullary nails, but did not heal either. The rest 17 patients were fixed with intramedullary nailing. Fracture nonunion classification: 4 cases of hypertrophic nonunion, 17 cases of atrophic nonunion; the length of bone defect was 1-3 mm; the duration from the last treatment to the current treatment was 10-23 months (mean, 14.3 months). The operation time, intraoperative blood loss, the time between operation and full loading, fracture healing time, and complications were recorded. The visual analogue scale (VAS) score and the imaging system of fracture healing of the extremities (RUST) of patients before operation and at last follow-up were recorded to evaluate the fracture healing; the function of the affected limb was evaluated according to the Schatzker-Lambert efficacy score standard at last follow-up.ResultsThe operation time was 105-160 minutes, with an average of 125.6 minutes; the intraoperative blood loss was 160-580 mL, with an average of 370.5 mL. All incisions healed by first intention, without vascular or nerve injury. All patients were followed up 22-46 months (mean, 26.5 months). All the fractures healed, with a fracture healing time of 3-7 months (mean, 4.8 months). During the follow-up, there was no infection, loosening, implant breakage, re-fracture, and other complications. The VAS score at last follow-up was 0.8±0.3, showing significant difference (t=7.235, P=0.000) when compared with preoperative score (5.2±3.7); the RUST score was 3.4±0.3, which was significantly higher than the preoperative score (1.5±0.7) (t=8.336, P=0.000). According to the Schatzker-Lambert effectiveness evaluation standard, the limb function was excellent in 16 cases, good in 4 cases, fair in 1 case, and the excellent and good rate was 95.42%.ConclusionNonunion after intramedullary fixation of femoral fracture treated with auxiliary plate combined with autogenous iliac graft, autogenous bone marrow concentration and PRP gel in accordance with the “diamond concept” can not only restore the stability of the fracture ends, but also improves the biological environment of the fracture site, and can improve the rate of fracture healing.
ObjectiveTo investigate the effect of human adipose-derived stem cells (hADSCs) on pressure ulcers in mouse.MethodsThe subcutaneous adipose tissue from voluntary donation was harvested. Then the hADSCs were isolated and cultured by mechanical isolation combined with typeⅠcollagenase digestion. The 3rd generation cells were identified by osteogenic, adipogenic, chondrogenic differentiations and flow cytometry. The platelet rich plasma (PRP) from peripheral blood donated by healthy volunteers was prepared by centrifugation. The pressure ulcer model was established in 45 C57BL/6 mice by two magnets pressurized the back skin, and randomly divided into 3 groups (n=15). The wounds were injected with 100 μL of hADSCs (1×106 cells) transfected with a green fluorescent protein (GFP)-carrying virus, 100 μL human PRP, and 100 μL PBS in hADSCs group, PRP group, and control group, respectively. The wound healing was observed after injection. The wound healing rate was calculated on the 5th, 9th, and 13th days. On the 5th, 11th, and 21st day, the specimens were stained with HE staing, Masson staining, and CD31 and S100 immunohistochemical staining to observe the vascular and nerve regeneration of the wound. In hADSCs group, fluorescence tracer method was used to observe the colonization and survival of the cells on the 11th day.ResultsThe cultured cells were identified as hADSCs by induced differentiation and flow cytometry. The platelet counting was significantly higher in PRP group than in normal peripheral blood group (t=5.781, P=0.029). General observation showed that the wound healing in hADSCs group was superior to those in PRP group and control group after injection. On the 5th, 9th, and 13th days, the wound healing rate in hADSCs group was significantly higher than those in PRP group and control group (P<0.05). Histological observation showed that compared with PRP group and control group, inflammatory cell infiltration and inflammatory reaction were significantly reduced in hADSCs group, collagen deposition was significantly increased, and skin appendage regeneration was seen on the 21st day; at each time point, the expression of collagen was significantly higher in hADSCs group than in PRP group and control group (P<0.05). Immunohistochemical staining showed that the number of neovascularization and the percentage of S100-positive cells in hADSCs group were significantly better than those in PRP group and control group on the 5th, 9th, and 13th days (P<0.05). Fluorescent tracer method showed that the hADSCs could colonize the wound and survive during 11 days after injection.ConclusionLocal transplantation of hADSCs can accelerate healing of pressure ulcer wounds in mice and improve healing quality by promoting revascularization and nerve regeneration.
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 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.
ObjectiveTo investigate the effectiveness of extracorporeal shock wave therapy combined with platelet-rich plasma (PRP) injection in treatment of knee osteoarthritis (KOA) by prospective clinical study.MethodsBetween June 2015 and June 2018, 180 patients with KOA met the inclusion criteria were included in study and randomly allocated to group A (n=60), group B (n=60), and group C (n=60). The patients were treated with autologous PRP intra-articular injection in group A, extracorporeal shock wave therapy in group B, and extracorporeal shock wave therapy combined with autologous PRP intra-articular injection in group C, once a week and 5 times a duration of treatment. There was no significant difference in age, gender, disease duration, side of KOA, and Kellgren-Lawrence grading between groups (P>0.05). The pain and function of knee joint were assessed by visual analogue scale (VAS) score, Lequesne Index score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and knee joint activity before treatment and at 1, 3, and 5 weeks after the first treatment.ResultsThere were significant differences in VAS score, Lequesne Index score, WOMAC score, and knee joint activity between pre- and post-treatment in all groups (P<0.05). VAS score, Lequesne Index score, and WOMAC score gradually decreased with the prolongation of treatment time (P<0.05); but there was no significant difference in knee joint activity between different time points (P>0.05). There was no significant difference in VAS score, Lequesne Index score, WOMAC score, and knee joint activity between groups before treatment (P>0.05); the scores of group C were superior to groups A and B (P< 0.05) at different time points after treatment; while the knee joint activities of 3 groups were similar (P>0.05).ConclusionThe extracorporeal shock wave therapy combined with PRP injection can relieve the pain synergistically for KOA.
In recent years, regenerative medical technology and modern rehabilitation technology complement each other and develop rapidly. Regenerative rehabilitation with tissue regeneration and functional recovery as the core concept arises at the historic moment. Regenerative rehabilitation can quickly repair damaged or diseased tissues and organs, and restore or improve the function of patients to the greatest extent. This paper introduces the origin and development of regenerative rehabilitation, discusses the research progress and significance of related strategies from three aspects of neurological, motor and circulatory diseases, and stress the importance of regenerative rehabilitation in helping patients to obtain the best curative effect.
ObjectiveTo analyze the effectiveness of percutaneous injection of autologous concentrated bone marrow aspirate (cBMA) combined with platelet-rich plasma (PRP) in the treatment of delayed fracture healing.MethodsA prospective, randomized, controlled, single-blind case study was conducted. Between March 2016 and July 2018, 66 patients who met the inclusion and exclusion criteria for delayed fracture healing but had solid internal fixation of the fracture end were randomly divided into control group (31 cases, treated with percutaneous autogenous bone marrow blood injection) and study group (35 cases, treated with percutaneous autogenous cBMA+PRP injection). General data such as gender, age, body mass index, site of delayed fracture healing, length of bone defect at fracture end, and preoperative radiographic union score for tibia (RUST) showed no significant difference between the two groups (P>0.05). Before injection, Kirschner wire was used in both groups to stimulate the fracture end and cause minor injury. The fracture healing time, treatment cost, and adverse reactions were recorded and compared between the two groups. Visual analogue scale (VAS) score was used to evaluate pain improvement. The tibial RUST score was extended to the tubular bone healing evaluation.ResultsNo infection of bone marrow puncture needle eyes occurred in both groups. In the control group, local swelling was obvious in 5 cases and pain was aggravated at 1 day after operation in 11 cases. In the study group, postoperative swelling and pain were not obvious, but 2 cases presented local swelling and pain. All of them relieved after symptomatic treatment. Patients in both groups were followed up, the follow-up time of the control group was 16-36 months (mean, 21.8 months), and the study group lasted 14-33 months (mean, 23.2 months). The amount of bone marrow blood was significantly lower in the study group than in the control group (t=4.610, P=0.000). The degree of postoperative pain in the study group was less than that in the control group, and the treatment cost was higher than that in the control group. But the differences between the two groups in VAS score at 1 day after operation and treatment cost were not significant (P>0.05). Fracture healing was achieved in 19 cases (61.3%) in the control group and 30 cases (85.7%) in the study group. The difference in fracture healing rate between the two groups was significant (χ2=5.128, P=0.024). Fracture healing time and RUST score at last follow-up were significantly better in the study group than in the control group (P<0.05). At last follow-up, RUST scores in both groups were significantly improved when compared with those before operation (P<0.05).ConclusionAutogenous cBMA combined with PRP percutaneous injection can provide high concentration of BMSCs and growth factors, and can improve the fracture healing rate and shorten the fracture healing time better than autogenous bone marrow blood injection.