Intra-Articular Leukocyte-Poor Platelet-Rich Plasma Injections for Japanese Patients With Osteoarthritis of the Knee: A Three-Year Observational Retrospective Study After Phase 1 and Phase 2a Trials.

BACKGROUND: We have previously confirmed the safety and feasibility of intra-articular (IA) platelet-rich plasma (PRP) injections in Japanese patients with osteoarthritis (OA) of the knee. This study aimed to investigate the clinical and radiological outcomes in patients who were followed up for three years. METHODS: Nine patients were evaluated in this observational study. All the patients were women with a mean age of 60.6 years. PRP was prepared by single centrifugation and classified as leukocyte-poor (LP)-PRP, and was administered via three IA injections at weekly intervals. Patients were evaluated at baseline and final follow-up using the Kellgren-Lawrence (KL) grade and the Japanese Orthopedics Association (JOA) score. The need for additional treatment was also investigated at the final follow-up. RESULTS: At a mean follow-up of three years, the JOA score improved from 75 points (p) to 83.8 p. The sub-categorical scores changed as follows: gait, 23.1-26.9 p; stairs, 12.5-17.5 p; range of motion, 30-30 p; and swelling, 9.4-10 p. While the KL grade was maintained in six patients, it progressed in two patients from I to II and II to III, respectively. Two patients received additional treatment at the final follow-up. CONCLUSIONS: At the final follow-up, the functional knee score, especially gait and the ability to go up and down the stairs, improved in six out of eight patients without additional treatment. These results suggest that LP-PRP injections produced safe outcomes without OA-worsening in most patients at three-year follow-up.

Comparison Between Concentrated Autologous Bone Marrow Aspirate Transplantation as a Hip Preserving Surgery and Natural Course in Idiopathic Osteonecrosis of the Femoral Head.

Purpose The purpose is to compare the therapeutic efficacy of concentrated autologous bone marrow aspirate transplantation (CABMAT) with that of observation alone for osteonecrosis of the femoral head (ONFH). Methods This single-center study included patients with idiopathic ONFH that were either treated with CABMAT (CABMAT group) or managed through observation alone (observation group) over a >2-year follow-up period. The Japanese Investigation Committee classification was used to diagnose and classify ONFH. The collapse rates for stages 1 and 2 ONFH (i.e., pre-collapse stages) and the THA conversion rates were compared between the CABMAT and observation groups. Results The CABMAT and observation groups comprised 232 (mean follow-up: 8.2 years) and 106 (mean follow-up: 6.0 years) patients, respectively. No significant intergroup differences were noted in the stages, types, and associated factors of ONFH. The collapse rates for pre-collapse stages in the CABMAT and observation groups were 67.1% and 65.3%, respectively. For stage 1, the collapse rates were significantly lower in the observation group than in the CABMAT group (p<0.05). The overall THA conversion rates in the CABMAT and observation groups were 24.3% and 41.5%, respectively (p<0.0001). For ONFH of stages 3A and 3B (collapse stages), the THA conversion rates were significantly lower in the CABMAT group (p<0.05). Conclusion Collapse rates were significantly higher for stage 1 ONFH; for collapse stages, the THA conversion rates were significantly lower in the CABMAT group than in the observation group. Therefore, observation and CABMAT are recommended for ONFH of stage 1 and for ONFH of higher stages, respectively.

Ten-year results of concentrated autologous bone marrow aspirate transplantation for osteonecrosis of the femoral head: a retrospective study.

BACKGROUND: Idiopathic osteonecrosis of the femoral head (ONFH) occurs at a relatively younger age. It is therefore important to prevent the resultant femoral head collapse and requirement of total hip arthroplasty in these patients. In 2003, we initiated concentrated autologous bone marrow aspirate transplantation (CABMAT), a joint-preserving treatment for ONFH, at our institution. Here, we report the long-term results of CABMAT treatment. METHODS: We retrospectively collated and analyzed the demographic and treatment data of 69 patients (109 hips) with idiopathic ONFH treated with CABMAT between April 2003 and April 2008. RESULTS: Totally, 44 patients (21 men, 23 women, 80 hips) completed the 10-year follow-up. The follow-up rate was 73.4%, and the mean follow-up period was 12.0 (range, 10.0-15.4) years. The mean age of the patients was 42.2 (range, 16.3-70.5) years. Using the Association Research Circulation Osseous (ARCO) classification system for preoperative analysis, 12, 31, 32, and 5 hips were classified as stages 1, 2, 3, and 4, respectively. The overall rate of conversion to total hip arthroplasty (THA) was 34% (27/80 hips). In a multivariate regression analysis, the preoperative stage of ONFH and the body mass index were found to correlate significantly with conversion to THA. Totally, 43 hips (of 80) were classified as belonging to the pre-collapse stage (i.e., stages 1 or 2). The overall collapse rate and the THA-conversion rate of these hips were estimated to be 49% (21/43) and 14% (6/43), respectively. CONCLUSIONS: On the basis of our long-term findings, the minimally invasive and feasible CABMAT therapy can be utilized as one of a joint-preserving treatment for ONFH.

Growth factor levels in leukocyte-poor platelet-rich plasma and correlations with donor age, gender, and platelets in the Japanese population.

BACKGROUND: Clinical application of platelet-rich-plasma (PRP) has been accelerated to investigate early recovery from various musculoskeletal conditions. It involves the promotion of tissue damage repair through the action of multiple growth factors at physiological concentrations. The composition of PRP differs based on many factors, which may include age and gender. Therefore, we analyzed correlations between age, gender, and platelet counts in PRP with growth factors in Japanese subjects. METHOD: Peripheral blood was drawn from 39 healthy volunteers between 20 and 49 years of age (age, mean ± standard deviation = 33 ± 8.7 years; gender ratio, male:female = 19:20; BMI, mean ± standard deviation = 22 ± 4.0) and prepared through centrifugation (volume, 6 mL per sample). After being activated with CaCl2, the supernatant was stored. The mean platelet count in PRP was 41.4 ± 12.2 × 104/µL. PRP concentration rate (i.e., PRP/peripheral platelet counts) was 1.8 ± 0.4 times. Growth factor levels (platelet-derived growth factor-BB, transforming growth factor-ß1, vascular endothelial growth factor, epidermal growth factor, fibroblast growth factor, insulin-like growth factor-1, and hepatocyte growth factor) were measured using enzyme-linked immunosorbent assay (ELISA), and correlations with age, gender, and PRP platelet counts were statistically analyzed by calculating Spearman's rank correlation coefficients (r). RESULTS: Age was negatively correlated with platelet-derived growth factor-BB and insulin-like growth factor-1 (r = - 0.32, - 0.39), and gender had no influence on growth factors. Platelet counts in PRP positively correlated with platelet-derived growth factor-BB, transforming growth factor-ß1, epidermal growth factor, and hepatocyte growth factor (r = 0.39, 0.75, 0.71, and 0.48, respectively). CONCLUSIONS: This clinical study shows a significant variation of PRP among individual patients and that this variation is influenced by the age and the platelet counts of the subjects. Our data demonstrate that patient characteristics account for the differences in PRP physiological activity.

Mid-term results of concentrated autologous bone marrow aspirate transplantation for corticosteroid-associated osteonecrosis of the femoral head in systemic lupus erythematosus.

PURPOSE: We had previously established concentrated autologous bone marrow aspirate transplantation (CABMAT), a one-step, low-invasive, joint-preserving surgical technique for treating osteonecrosis of the femoral head (ONFH). This study aimed to evaluate the effects of CABMAT as a hip-preserving surgical approach, preventing conversion to total hip arthroplasty (THA) and femoral head collapse in patients with systemic lupus erythematosus (SLE). METHODS: Since 2003, 52 SLE patients (8 male, 44 female, 92 hips, mean age 35.3 (16-77) (years) were treated with CABMAT. The mean follow-up period was 5.5 (0.7-14) years. Conversion rate to THA and its predicting factors were analyzed. RESULTS: The overall conversion rate to THA was 29% (27/92). Conversion rate to THA was 0% (0/3), 0% (0/4), 22% (9/41), and 41% (18/44) in types A, B, C1, and C2, respectively. Conversion rate to THA was 26% (5/19), 26% (6/23), 28% (11/39), 44% (4/9), and 50% (1/2) in stages 1, 2, 3A, 3B, and 4, respectively. In multivariate logistic regression analysis, sex, body mass index (BMI), pre-operative type, and pre-operative stage were significantly correlated with conversion to THA. CONCLUSION: The conversion rate to THA was lower than that in the natural course and core decompression, but was higher than that seen in other bone marrow transplantation and osteotomy. Since sex, pre-operative type, and pre-operative stage were significantly correlated with conversion to THA, it is suggested that the higher proportion of women, advanced stage (stage 3A or above), and advanced type (type C or above) in this study affected the THA conversion rate.

Intra-articular platelet-rich plasma (PRP) injections for treating knee pain associated with osteoarthritis of the knee in the Japanese population: a phase I and IIa clinical trial.

Intra-articular platelet-rich plasma (PRP) injection has been found to be effective for treating osteoarthritis in patients from Western countries; however, the safety and efficacy of PRP have not been sufficiently investigated in Japanese patients. The present study aimed to evaluate the safety and feasibility of intra-articular PRP injection in Japanese patients with knee osteoarthritis. PRP without white blood cells was prepared using a single-spin centrifuge (PRGF-Endoret; BTI Biotechnology Institute, Vitoria, Spain). A 6-mL PRP volume was injected in the knee joint three times at 1 week intervals. All patients were prospectively evaluated before intervention and at 1, 3, and 6 months after the treatment. Adverse events, the Visual Analog Scale (VAS) pain score, Japanese Knee Osteoarthritis Measure (JKOM) score and Japanese Orthopedic Association score were evaluated. Ten patients (all women; average age, 60.6 years) were treated. Only minor adverse events after injection were noted, and symptoms resolved within 48 hours after the injection. The average VAS pain scores were 71.6 mm and 18.4 mm at baseline and the 6-month follow-up, respectively (P < 0.05). At the 6-month follow-up, 80% of patients had a decrease in the VAS pain score of 50% or more. The average JKOM scores were 35.2 and 14.3 at baseline and at the 1-month follow-up, respectively (P < 0.05). Intra-articular PRP injection likely represents a safe treatment option for Japanese patients with mild-to-moderate knee osteoarthritis, and has the potential to relieve pain for up to 6 months, but further study is needed to verify the efficacy.

Hip preserving surgery with concentrated autologous bone marrow aspirate transplantation for the treatment of asymptomatic osteonecrosis of the femoral head: retrospective review of clinical and radiological outcomes at 6 years postoperatively.

BACKGROUND: We had previously established concentrated autologous bone marrow aspirate transplantation (CABMAT), a one-step, low-invasive, joint-preserving surgical technique for treating osteonecrosis of the femoral head (ONFH). The present study aimed to evaluate the effects of CABMAT as a hip preserving surgical approach, preventing femoral head collapse in asymptomatic ONFH. METHODS: In total, 222 patients (341 hips) with ONFH were treated with CABMAT between April 2003 and March 2013. Based on magnetic resonance imaging, we determined that 119 of these patients had bilateral asymptomatic ONFH (238 hips), and 38 further patients had unilateral asymptomatic ONFH (38 hips). In this series, we retrospectively examined 31 hips in 31 patients with unilateral asymptomatic ONFH treated surgically between 2003 and 2012 and followed up for more than 2 years. Clinical and radiological evaluation were performed immediately before the procedure and at the final follow-up. The two-year follow-up rate among patients with unilateral ONFH was 82% (31/38). Therefore, the present study included 31 patients (19 males and 12 females), with a mean age and follow-up period of 40 and 5.8 years, respectively. Of the 31 asymptomatic hips, 5, 6, 10, and 10 had osteonecrosis of types A, B, C1, and C2, respectively. The diagnosis, classification, and staging of ONFH were based on the 2001 Japanese Orthopaedic Association (JOA) classification. RESULTS: Secondary collapse of the femoral head was observed in 6/10 hips and 5/10 hips with osteonecrosis of types C1 and C2, respectively. Total hip arthroplasty was performed in 9.6% of patients (3/31 hips), at an average of 33 months after surgery. Clinical symptoms improved after surgery, and the secondary collapse rate at a mean of 5.8 years after CABMAT was lower than that reported in several previous studies on the natural course of asymptomatic ONFH. CONCLUSIONS: Early diagnosis of ONFH (i.e., before femoral head collapse) and early intervention with CABMAT could improve the clinical outcome of corticosteroid and alcohol-induced ONFH.

Feasibility and Efficacy of Autologous Bone Marrow Aspirate Transplantation Combined with Human Parathyroid Hormone 1-34 Administration to Treat Osteonecrosis in a Rabbit Model.

No studies have examined the transplantation of a bone marrow aspirate (BMA) containing mesenchymal stem cells (MSCs) combined with human parathyroid hormone 1-34 (hPTH1-34) administration. Therefore, we evaluated the feasibility and efficacy of autologous BMA transplantation combined with hPHT1-34 administration in a bone necrosis model. The metatarsal bones of rabbits were necrotized using liquid nitrogen, and the rabbits received a BMA transplantation or saline injection followed by hPTH1-34 (30 µg/kg) or saline administration three times per week (n = 3-4 per group). The rabbits were euthanized at 12 weeks after the initiation of treatment. No systemic adverse effects or local neoplastic lesions were observed. Importantly, the rabbits in the BMA transplantation plus hPTH1-34 group showed the highest bone volumes and histological scores of new bone. These data confirmed the feasibility of BMA transplantation combined with hPTH1-34, at least during the experimental period. The observed efficacy may be explained by a synergistic effect from the stimulation of MSC differentiation to osteoblasts with hPTH1-34-mediated suppression of apoptosis in osteoblasts. These results indicate the promising potential for BMA transplantation combined with hPTH1-34 administration in bone necrosis treatment. Longer term experiments are needed to confirm the safety of this therapeutic strategy.

2. The Effect of Combined Therapy, Percutaneous Autologous Concentrated Bone Marrow Grafting and Low-Intensity Pulsed Ultrasound (LIPUS), on the Treatment of Non-Unions.

We discuss the effect of combined therapy of percutaneous autologous concentrated bone graft and LIPUS on complex non-union treatment. Seventeen of 27 treated patients who had received the therapy at least 1 year before were discussed (10 femurs, 5 tibiae, 1 humerus, and 1 ulna). The average age of the patients was 40.7, and atrophic degeneration was observed in all cases. After 12 months of treatment, bone union was recognized in 76% in all cases, and in 87% of lower long bones. It was reported that LIPUS was effective at improving blood flow, accelerating cytokines which induce angiogenesis, promoting the transport of nutrition and enzymes to living cells, developing the differentiation of osteoblast from mesenchymal stem cells (MSC), inhibiting the differentiation and development of osteoclast, and promoting endochondral ossification. In this study, all patients had been treated with LIPUS for more than 3 months before the grafting was conducted, but the bone union seemed to stop. It was thought that this combined therapy provided a bone marrow cell growth factor sufficient to enable new bone formation to re-start bone union, and then LIPUS worked effectively to promote the initial differentiation, contributing to new bone formation. This combination therapy-less invasive, safe, and low cost-was considered one useful treatment option for non-union.

The safety and efficacy of combined autologous concentrated bone marrow grafting and low-intensity pulsed ultrasound in the treatment of osteonecrosis of the femoral head.

Osteonecrosis of the femoral head (ONFH) is commonly treated with total hip arthroplasty; however, the disadvantages of this form of treatment, especially in young patients, include the need for revision arthroplasty. Here we describe a novel, combined approach to the treatment of ONFH based on autologous concentrated bone marrow grafting and low-intensity pulsed ultrasound (LIPUS). The 7 male and 7 female patients (mean age: 40 years; 22 hips) underwent autologous concentrated bone marrow grafting followed by 6 months of continuous LIPUS. The mean follow-up period was 26 months. We evaluated site-specific bacterial infection of the grafted bone marrow concentrate microbiologically and site-specific cancer by magnetic resonance imaging 24 months after grafting. All patients were assessed using the visual analogue scale (VAS) for pain and the Japanese Orthopedic Association (JOA) hip score. Clinical and plain radiographic evaluations were performed before grafting and at the most recent follow-up. Computed tomography (CT) scans were obtained before and 12 months after grafting. None of the grafted bone marrow concentrates were infected, and none of the patients developed a tumor at the treatment site. The VAS and JOA scores improved in all patients. Collapse progressed in 8 of the 22 hips, but none required total hip arthroplasty. The mean volume of new bone formation 12 months post-grafting as seen on CT was 1256 mm(3). New bone formation was observed in all patients. Our study demonstrates the safety and efficacy of autologous concentrated bone marrow grafting and LIPUS as a joint-preserving procedure for patients with ONFH.

Fate of bone marrow mesenchymal stromal cells following autologous transplantation in a rabbit model of osteonecrosis.

BACKGROUND AIMS: Internalizing quantum dots (i-QDs) are a useful tool for tracking cells in vivo in models of tissue regeneration. We previously synthesized i-QDs by conjugating QDs with a unique internalizing antibody against a heat shock protein 70 family stress chaperone. In the present study, i-QDs were used to label rabbit mesenchymal stromal cells (MSCs) that were then transplanted into rabbits to assess differentiation potential in an osteonecrosis model. METHODS: The i-QDs were taken up by bone marrow-derived MSCs collected from the iliac of 12-week-old Japanese white rabbits that were positive for cluster of differentiation (CD)81 and negative for CD34 and human leukocyte antigen DR. The average rate of i-QD internalization was 93.3%. At 4, 8, 12, and 24 weeks after transplantation, tissue repair was evaluated histologically and by epifluorescence and electron microscopy. RESULTS: The i-QDs were detected at the margins of the drill holes and in the necrotized bone trabecular. There was significant colocalization of the i-QD signal in transplanted cells and markers of osteoblast and mineralization at 4, 8, and 12 weeks post-transplantation, while i-QDs were detected in areas of mineralization at 12 and 24 weeks post-transplantation. Moreover, i-QDs were observed in osteoblasts in regenerated tissue by electron microscopy, demonstrating that the tissue was derived from transplanted cells. CONCLUSION: These results indicate that transplanted MSCs can differentiate into osteoblasts and induce tissue repair in an osteonecrosis model and can be tracked over the long term by i-QD labeling.

Novel unidirectional porous hydroxyapatite used as a bone substitute for open wedge high tibial osteotomy.

PURPOSE: The purpose of this study was to clinically and radiologically evaluate the availability, osteoconductivity, and resorption of a novel unidirectional porous hydroxyapatite (UDPHAp) used as an artificial substitute for open wedge high tibial osteotomy (OWHTO). Our hypothesis was that UDPHAp is a safe and useful bone substitute for OWHTO. MATERIALS AND METHODS: Seven patients (2 men and 5 women aged 34-72years) who underwent OWHTO and were followed up for more than 12months were retrospectively studied. After the osteotomy, the gap created was filled with UDPHAp(REGENOS® Kuraray Co.Ltd). Radiography and computed tomography(CT) were performed, and gap healing was assessed postoperatively. The Japanese Orthopaedic Association (JOA) knee score was determined pre- and post-operatively for clinical evaluation. RESULTS: Neither gross displacement nor collapse of the UDPHAp block graft was observed within 12 months after surgery. Both radiographs and CT showed attenuation of lucency and increasing sclerosis over time. JOA score improved from 71.2 (65-80) to 95.8 (85-100). CONCLUSIONS: Short term results for OWHTO using UDPHAp was satisfactory. Clinical improvement of JOA scores were seen, besides osteogenesis was progressing in and around the artificial bone grafts.

Circadian variation of growth factor levels in platelet-rich plasma.

OBJECTIVE: The influence of circadian rhythms has recently been considered in advanced studies of chronopharmacology and chronotherapeutics. Although emerging studies have reported the usefulness of platelet-rich plasma (PRP), no reports have described the diurnal variations in PRP growth factor levels. In the present study, we hypothesized that growth factor levels in PRP show a significant diurnal variation. DESIGN: Controlled laboratory study. SETTING: Institutional. PARTICIPANTS: Ten adult male volunteers (mean age, 30.0 ± 2.6 years; range, 26-34 years). INTERVENTIONS: Each subject donated a 20-mL sample of fasting blood, 4 times per day (7 AM, 11 AM, 3 PM, and 7 PM). MAIN OUTCOME MEASURES: Each blood sample was processed to produce PRP. Complete blood counts of peripheral blood (PB) and PRP, as well as quantitative determination of PRP growth factor levels (platelet-derived growth factor and transforming growth factor) were performed. Statistical analyses were conducted using a one-way repeated measures analysis of variance. RESULTS: No significant differences were observed in platelet, white blood cell, and red blood cell counts in the PB or in the PRP samples throughout the course of the day. In addition, there were no significant differences in PRP growth factor levels at the various time points. CONCLUSIONS: Significant diurnal variations in platelet counts and growth factor levels were not observed in PRP. Therefore, adjustments for diurnal variation in chronotherapy may not be important when PRP is used in clinical practice.

Percutaneous autologous concentrated bone marrow grafting in the treatment for nonunion.

The purpose of this study was to evaluate the clinical and radiographic treatment effects of percutaneous autologous concentrated bone marrow grafting in nonunion cases and to evaluate the effectiveness of this grafting procedure. We enrolled 17 cases those had atrophic changes due to continuous nonunion for over 9 months after injury and had undergone low-intensity pulsed ultrasound treatment for more than 3 months. The site of nonunion was the femur in 10 cases, the tibia in 5 cases, the humerus in 1 case, and the ulna in 1 case. They underwent percutaneous autologous concentrated bone marrow grafting and continued low-intensity pulsed ultrasound stimulation treatment after grafting. Patients were evaluated using the visual analogue scale for pain at immediately before the procedure, 3, 6, and 12 months after grafting. Plain radiographs of the affected site were taken and evaluated about the healing of the nonunion site at each clinical evaluation. As quantitative assessment, CT scans were undertaken before the procedure and 6 months after grafting. The visual analogue scale pain score was reduced consistently after grafting in all patients. About the healing at the nonunion site, 11 and 13 cases of bone union were observed at 6 and 12 months after grafting. The mean volume of callus formation based on CT images was 4,147 (262-27,392) mm3 total between grafting and 6 months. Percutaneous autologous concentrated bone marrow grafting is an effective procedure for the treatment of patients with nonunion.

The effects of plasma rich in growth factors (PRGF-Endoret) on healing of medial collateral ligament of the knee.

PURPOSE: Platelet-rich plasma (PRP) has been increasingly used in sports-related injuries for therapeutic applications. However, there are numerous manufacturing procedures and treatment protocols of PRP use, which make difficult to assess its real efficacy for tissue healing. This study addressed to evaluate the therapeutic effects of locally delivered plasma rich in growth factors (PRGF-Endoret) on the early healing of medial collateral ligament (MCL) in rabbit knees. METHODS: Thirty-one Japanese white rabbits were subjected to a mop-end tear in the MCL of the left knee. PRGF-Endoret was prepared using Anitua's technique. Two groups were set up. In 17 knees, prepared 1.0 ml of PRGF-Endoret after clotting was applied on the tear site, while in 14 knees the tear site was untreated serving as a control. Quantitative aspects of PRGF-Endoret, the concentration of platelets, leukocytes and erythrocytes and therapeutic growth factors such as PDGF-BB and TGF-ß1 were measured. Rabbits were sacrificed at 3 and 6 weeks after the operation and histological and biomechanical evaluation were performed. RESULTS: No leukocytes were measured and certain amount of growth factors such as PDGF-BB and TGF-ß1 were confirmed in the PRGF-Endoret. PRGF-Endoret stimulated proliferation of fibroblasts and neovascularization, and induced statistically better structural properties in repaired MCL. CONCLUSIONS: Our findings provide evidence that local administration of PRGF-Endoret promotes early steps in ligament healing and the repair of structural properties in a rabbit model. PRGF-Endoret would be a useful product in clinical treatment of ligament injuries.