Introduction: Before proceeding to long-term studies, we studied early clinical results of combined Autologous Chondrocyte Implantation (ACI) and Allogenic Meniscus Transplantation (AMT). Meniscus deficient knees develop early osteo-arthritis (OA) of the knee joint. Autologous Cartilage Implantation (ACI) is contraindicated in case of meniscus deficient knees. And on contrary the Allogenic Meniscus Transplantation (AMT) is contraindicated in cartilage defects in the knee joint. But a combination of the two procedures for bone on bone OA might be a solution for this problem. This was the main purpose of our study. Methods: We studied a consecutive series of eight patients (7 males and 1 female), with an average age= 43 years (29–58), presenting with painful secondary arthritis, due to premature loss of meniscus and chondral defect/s. Median size of the femoral defects was 8.16 cm2 and of the tibial side 2.69 cm2 All patients were treated with a combination of Autologous Chondrocyte implantation (ACI) and Allogenic Meniscus Transplantation (AMT). Chondral defects were covered with periosteum/ Chondroguide membrane, secured in place with in-vitro cultured autologous chondrocytes injected underneath the path. Meniscus placed as load-bearing washer on the surface of ACI of tibia. ACI rehabilitation protocol followed post-operatively. Assessment at the end of one year was done with self-assessed Lysholm score, histology and the MRI scan. Results: Mean pre-operaive Lysholm score was 49 (17–75). This increased to a mean of 66 (26–87) at 1 year, an average increase of 16.4 points. Average one-year satisfaction score was 3 and they were back to all active life style. Five out of eight patients showed significant functional improvement at last post-operative follow-up (2 to 6 years; mean of 3.2 years). Complications were aseptic synovitis in 3 cases. Three failures were noted showig persistant pain and swelling in one, rupture of meniscus in second and third patient had a knee replacement. Arthroscopy at 1 year showed a stable meniscus with all healed peripheral margins in all except in one case with some thinning with no evidence of rejection. Histology of meniscus showed a fibrocartilage well populated with viable cells and the peripheral zone was well vascularised and integrated with capsule. Biopsy of ACI site was predominantly of fibrocartilage with good basal integration with subchondral bone. On MRI scan, allogenic meniscus was well integrated with capsule along the line of repair, showing foci of variable signal intensities within the meniscus. There was no evidence of meniscal subluxation in all but one case showing mild extrusion. ACI graft site showed a varied appearance, with 3 grafts showing focal grade 3to 4 changes. Conclusion: Seven out of eight patients improved post-operatively at one year, in terms of pain relief and increased activity. It’s possible to combine these two techniques together. Short-term outcomes are satisfactory. We could not find any deleterious effects of combining these two techniques together. So we conclude that, this might act as a one step towards a biological knee replacement

1 . The results of transplanting the greater trochanter in 225 "low-friction" arthroplasties of the hip have been examined. 2. Non-union occurred in an average of 7 per cent of cases. 3. When non-union occurred the results still showed improvement. 4. Four different methods of fixation were used, of which that using two wires, crossed in the horizontal and coronal planes, never failed to secure union. 5. Transplantation of the greater trochanter to the best position is only possible if the neck of the femur is shortened or if the centre of motion of the arthroplasty is displaced medially by deepening the acetabulum, or by a combination of both. 6. In the best position the transplanted trochanter considerably improved active abduction against gravity

1 . A ten-year study of fifty-four operations for transplantation of the hamstring muscles in thirty-one spastic patients has been made. 2. Twenty-one patients were improved after operation, six were not improved, and in four the duration of follow-up was too short for proper assessment. One patient died from other causes. 3. Greatest benefit was not obtained until one year after operation. 4. The results varied considerably in different grades of spastic patient. Important factors affecting the results were age, sex, personality, balance and function of arm and hip. Hand and major hip operations should be carried out before hamstring transplantation. 5. The objectives of operation are discussed. These were most consistently achieved in older, more ambitious or more responsible males with good balance and with good arm and hip function. 6. Factors which were of less direct importance were mental capacity, minor degrees of limb length inequality, and foot deformities. 7. Operative correction of foot deformities should not be done before hamstring transplantation. 8. Serial plaster correction of the knee flexion deformity before operation is preferred to division of the patellar retinacula. 9. Transplantation of the hamstrings to the patella in spastics is under trial

MACI Cartilage Transplantation has been performed in New Zealand for the last five years for patients with isolated articular cartilage defects who have failed a microfracture technique. Thirteen patients have undergone this procedure. Patients were evaluated by an independent research assistant preoperatively and at 6 months, 1, 2, 3 and 5 years Visual analogue pain score, ICRS score, WOMAC score, KOOS score, IKDC score and SF 36 were collected at all time points. An MRI scan was performed in all patients at all time periods. Biopsies are taken at three years. There is an improvement in visual analogue pain score, ICRS score, WOMAC score, KOOS score, IKDC score and SF 36 at all time points. Results improve with time. No procedures have failed. All MRI scans showed restoration of articular cartilage. Histology shows hyaline like cartilage. Medium term results are encouraging. The rehabilitation is long thus 12 month data does not give an accurate outcome measure. Ongoing evaluation is occurring

The technique of Matrix Induced Autologous Chondrocyte Transplantation (MACI) is well established with satisfactory outcomes up to five years in the knee. Fewer series describe the outcomes of this technique in the ankle. We present the functional outcomes of the technique for a single surgeon series in a general hospital setting. Twenty-seven patients, mean age 41, were reviewed at 3.7 (range, 1 to 5) years. Patients were assessed using the American Orthopaedic Foot and Ankle Society (AOFAS) hind-foot scale, Tegener activity score and University of California lower extremity activity scale. MRI findings were also reviewed. While most patients report a significant improvement in symptoms with full return to activities of daily living, 36% of those under 40 and 78% of those over 40 reported restricted recreational activity. Of the patients under 40 years of age, 86% were able to run compared with 23% of those over 40. Of patients over 40, 64% continued to have moderate or severe pain. Careful preoperative counselling is required for patients of all ages regarding likely outcomes. In patients over 40 the procedure is unlikely to give good pain relief and alternative options should be considered

Background. Autologous Chondrocyte Implantation (ACI) is frequently used to treat chondral defects in the knee with a good long-term outcome. This is contraindicatd in meniscal deficient knees. Allogenic Menicsal Transplantation (AMT) has been shown to give good symptomatic relief in meniscus deficient knees. However this is contraindicated in advanced cartilage degeneration. We hypothesized that combination of these two might be a solution for bone-on-bone arthritis in young individuals. Methods. We studied a consecutive series of 12 patients who underwent combined ACI and AMT between 1998 and 2005. Pre operative and post operative comparisons of lysholm scores were recorded. Magnetic Resonance Imaging was performed to assess the integration ACI & AMT. Arthroscopy was performed at one year for assessment and obtain biopsy for histological examination. Results. Out of the twelve patients only eleven were included as one had died at three months after surgery. The median pre-operative lysholm score was 45 which rose to 64 at one year. Magnetic Resonance Imaging showed good integration of both ACI and menisci. Most of the patients were able to lead an active lifestyle. Conclusion. The combination of both ACI & AMT could give a good result and defer a total knee replacement in young indi

The Royal National Orthopaedic Hospital has completed an extensive trial of ACI versus MACI in the treatment of symptomatic osteochondral defects of the knee. A new technique has now been proposed which is quicker and easier to perform. This is the Gel-Type Autologous Chondrocyte Transplantation, CHONDRONTM. At Stanmore CHONDRON has been used for the past 17 months. Our aim was to assess the short term functional outcome of patients who have undergone CHONDRONTM using validated outcome scoring questionnaires. We retrospectively reviewed the notes of 43 patients that had undergone CHONDRONTM over one year ago and scored them using the Modified Cincinnati Score, the Visual Analogue Score and the Benltey Stanmore Functional Rating Score. RESULTS. The mean pre-operative Modified Cincinnati Score was 39.9, which improved to a mean of 59.8 post-operatively. The mean Visual Analogue Score improved from 6.7 to 5.1 post-operatively. The median Bentley Functional Rating Score was 3 pre-operatively and 2 post-operatively. CONCLUSIONS. These early results show that 76% of the patients who were treated with CHONDRONTM experienced a reduction in pain and improvement in post-operative function. In the patients in whom the symptoms were worse, the deterioration in score could be partly explained by numerous previous procedures on the same site, presence of early osteoarthritis or the presence of multiple osteochondral lesions. This highlights the importance of careful patient selection in order to gain maximum benefit from the procedure

It has been reported that the total steroid dose and acute rejection episodes after organ transplantation is one of the risk factors for the development of osteonecrosis of the femoral head (ONFH), and ONFH in steroid-iduced subgroup may progress more aggressively to femoral head collapse requiring total hip arthroplasty. Despite inherent medical co-morbidities of solid organ transplantation patients, most authors recently have reported successful outcomes of THAs in those patients. But there are few comparative studies on the outcome of THAs for ONFH after different organ transplantations. The purpose of this study was to evaluate and compare a single tertiary referral institution's experience of performing primary THAs in kidney transplantation (KT) and liver transplantation (LT) patients with specific focus on the total steroid dose, clinical outcomes, and relationship between ONFH and absence or presence of acute rejection (AR).

Between 1999 and 2010, 4,713 patients underwent organ transplantations (1,957 KT and 2,756 LT) and AR was occurred in 969 patients (20.6%) after transplantation. Among these patients, 131 patients (191 hips) underwent THA for ONFH, and they were retrospectively reviewed. In KT groups, there were 57 men and 36 women with a mean age of 43.7 years. In LT groups, there were 26 men and 13 women with a mean age of 50.4 years. We investigated the dose of steroid administration on both groups, the time period from transplantation to THA, Harris hips score (HHS), visual analogue scale (VAS) and complications. The mean follow up period was 8.1 years (range, 5 to 14 years).

One-hundred and thirty-one (2.8%) patients [93 KT and 38 LT] underwent THA after transplantation. The total steroid dose after transplantations was significantly higher in KT group (10,420 mg) than that in LT group (4,567 mg), but the total steroid dose in the first 2 weeks after transplantation was significantly higher in LT group (3,478 mg) than that in KT group (2,564 mg). Twenty-three (2.4%) patients (19 KT and 4 LT) who underwent THA had an episode of AR. In LT group, the total steroid dose in AR groups was significantly higher than that in non-AR groups, whereas in KT group, there was no significant difference of the total steroid dose between AR group and non-AR group. The rate of THAs for ONFH was similar in both groups (2.4% in AR group, 2.9% in non-AR group). The mean time period from transplantation to THA was 986 days for KT and 1,649 days for LT patients. Both groups showed satisfactory HHS and VAS at final follow up, revealed no differences between the groups.

The rate of THAs for ONFH was three times higher in KT patients than that in LT patients, but it was similar in both AR group and non-AR group. The total steroid dose was also higher in KT patients compared to LT patients. The clinical outcomes of THA were satisfactory with few complications in both KT and LT patients. Therefore, THAs seems to be a good option for the patients with symptomatic steroid-induced ONFH after KT and LT.

We investigated the clinical, arthroscopic and biomechanical outcome of transplanting autologous chondrocytes, cultured in atelocollagen gel, for the treatment of full-thickness defects of cartilage in 28 knees (26 patients) over a minimum period of 25 months. Transplantation eliminated locking of the knee and reduced pain and swelling in all patients. The mean Lysholm score improved significantly. Arthroscopic assessment indicated that 26 knees (93%) had a good or excellent outcome. There were few adverse features, except for marked hypertrophy of the graft in three knees, partial detachment of the periosteum in three and partial ossification of the graft in one. Biomechanical tests revealed that the transplants had acquired a hardness similar to that of the surrounding cartilage. We conclude that transplanting chondrocytes in a newly-formed matrix of atelocollagen gel can promote restoration of the articular cartilage of the knee

Introduction. Tunnelwidening in failed anterior cruciate ligament reconstruction (ACLR) can result in the staged revision procedures with a need for bone transplantation prior to revision reconstruction. Limited knowledge exist regarding to quality of different transplantation methods. The present study used CT-scanning to evaluate tunnel bone density after allogenic bone chips and bone cylinder transplantation. We hypothesized that bone chips transplantation resulted in higher bone density than bone cylinder transplantation due to possible voids between individual cylinders in the tunnels. Methods. The records of 24 patients operated for 1st stage revision ACLR from April 2003 to march 2010 were included in the study. twelve patients had their tunnels transplanted with bone chips and twelve patients with bone cylinders from allogenic femoral heads. Bone chips were created by fine bone milling and cylinders were extracted by 7–8 mm core drilling. Bone density 3–4 months after transplantation were evaluated by CT scanning reconstruction slides with 5 mm intervals throughout the tunnel length using histomorphometry. Results. There were 15 females and 9 males with an average age of 32 yrs. Using bone chips the bone density in the tibial tunnels was 55% and the femoral tunnels the bone density was 68% Using bone cylinders bone density was 60% in the tibial tunnels and 53% in the femoral tunnels. The femoral bone density in the bone chip group was significantly higher than the bone cylinders (p < 0.05). Conclusion. Transplantation with bone chips results in superior bone quality in the femoral tunnels where as no difference where demonstrated in the tibial tunnels

Abstract

Many factors have been reported to affect the functional survival of OCA transplants, including chondrocyte viability at time of transplantation, rate and extent of allograft bone integration, transplantation techniques, and postoperative rehabilitation protocols and adherence. The objective of this study was to determine the optimal subchondral bone drilling technique by evaluating the effects of hole diameter on the material properties of OCAs while also considering total surface area for potential biologic benefits for cell and vascular ingrowth.

Using allograft tissues that would be otherwise discarded in combination with deidentified diagnostic imaging (MRI and CT), a model of a large shell osteochondral allograft was recreated using LS-PrePost and FEBio based on clinically relevant elastic material properties for cortical bone, trabecular bone, cartilage, and hole ingrowth tissue. The 0.8 mesh size model consisted of 4 mm trabecular bone, 4 mm cortical bone, and 3 mm cartilage sections that summed to a cross-sectional area of 1600 mm2 (40 mm x 40 mm). Holes were modeled to be 4mm deep in relation to clinical practice where holes are drilled from the deep margin of subchondral trabecular bone to the cortical subchondral bone plate. To test the biomechanic variations between drill hole sizes, models with hole sizes pertinent to standard-of-care commercially available orthopaedic drill sizes of 1.1mm, 2.4 mm, or 4.0 mm holes were loaded across the top surface over a one second duration and evaluated for effective stress, effective strain, 1st principal strain, and 3rd principal strain in compressive conditions.

Results measured effective stress and strain and 1st and 3rd principal strain increased with hole depth.

The results of the present FEA modeling study indicate that the larger 4.0 mm diameter holes were associated with greater stresses and strains within OCA shell graft, which may render the allograft at higher risk for mechanical failure. Based on these initial results, the smaller diameter 2.4 mm and 1.1 mm holes will be further investigated to determine optimal number, configuration, and depth of subchondral drilling for OCA preparation for transplantation

Abstract

1. Experiments to examine the antigenicity of homologous bone tissues in rats are reported. The tissues studied included fresh marrow-free cortical bone blocks and chips, fresh, boiled, frozen and freeze-dried marrow-containing iliac bone, fresh iliac bone devoid of marrow, and fresh red marrow.

2. The various tissues were transplanted from hooded to Wistar rats. Three weeks later a skin graft from each donor was transplanted to its respective host to detect the presence of transplantation immunity, which was indicated by the early rejection of the skin graft.

3. Homografts of fresh cortical bone evoked transplantation immunity indicating that it contained transplantation antigens which were also in the skin.

4. Homografts of fresh marrow-containing iliac bone also evoked transplantation immunity, which was shown to be caused by the red marrow.

5. Fresh iliac homografts devoid of marrow did not elicit transplantation immunity. This suggests that iliac bone tissue may not contain transplantation antigens or that the small amount of iliac bone inserted was insufficient.

6. Microscopy of the grafts, removed after three weeks, showed that the inflammatory infiltrations around the bone homografts and autografts were not very different, but that the amount of new bone formed was different. The autografts produced a lot of new bone, the homografts only a little.

7. It is suggested that the immune response evoked in the host by the foreign graft impairs the formation of new bone by fresh homografts of cortical blocks, cortical chips and marrow-containing iliac bone.

8. The impairment of new bone formation by homografts of marrow-free iliac bone is discussed. Such bone grafts fail to evoke detectable transplantation immunity. Why these grafts do not form more new homologous bone than the other homografts studied, is not clear.

9. Homografts of boiled and frozen iliac bone do not evoke any detectable change in the sensitivity of the host to donor tissue.

10. Homografts of freeze-dried marrow-containing iliac bone elicit a slight but significant prolongation of the survival of skin homografts. The implication, in terms of modern immunological theory, is that in such grafts certain tissue antigens still persist.

Skeletal muscle injuries often lead to severe functional deficits. Mesenchymal stem cell (MSC) therapy is a promising but still experimental tool in the regeneration of muscle function after severe trauma. One of the most important questions, which has to be answered prior to a possible future clinical application is the ideal time of transplantation. Due to the initial inflammatory environment we hypothesized that a local injection of the cells immediately after injury would result in an inferior functional outcome compared to a delayed transplantation.

Twenty-seven female Sprague Dawley rats were used for this study. Bone marrow was aspirated from both tibiae of each animal and autologous MSC cultures obtained from the material. The animals were separated into three groups (each n=9) and the left soleus muscles were bluntly crushed in a standardized manner. In group 1 2×106 MSCs were transplanted into the injured muscle immediately after trauma, whereas group 2 and 3 received an injection of saline. Another week later the left soleus muscles of the animals of group 2 were transplanted with the same number of MSCs. Group 1 and 3 received a sham treatment with the application of saline solution in an identical manner. In vivo functional muscle testing was performed four weeks after trauma to quantify muscle regeneration.

Maximum contraction forces after twitch stimulation decreased to 39 ± 18 % of the non injured right control side after crush trauma of the soleus muscles as measured in group 3. Tetanic stimulation showed a reduction of the maximum contraction capacity of 72 ± 12 % of the value obtained from intact internal control muscles. The transplantation of 2 x 106 MSCs one week after trauma improved the functional regeneration of the injured muscles as displayed by significantly higher contraction forces in group 2 (twitch: p = 0.014, tetany: p = 0.018). Local transplantation of the same number of MSCs immediately after crush injury was able to enhance the regeneration process to a similar extent with an increase of maximum twitch contraction forces by 73.3 % (p = 0.006) and of maximum tetanic contraction forces by 49.6 % (p = 0.037) compared to the control group.

The presented results underline the effectivity of MSC transplantation in the treatment of severe skeletal muscle injuries. The most surprising finding was that despite of the fundamental differences of the local environment into which MSCs had been transplanted, similar results could be obtained in respect to functional skeletal muscle regeneration. We assume that the effect of the MSC after immediate injection can partly be explained by their known immunomodulatory competences. The data of our study provide evidence for a large time window of MSC transplantation after muscle trauma.

In this study, we aimed to investigate tibiofemoral and allograft loading parameters after OCA transplantation using tibial plateau shell grafts to characterize the clinically relevant biomechanics that may influence joint kinematics and OCA osseointegration after transplantation. The study was designed to test the hypothesis that there are significant changes in joint loading after tibial plateau OCA transplantation that may require unique post-operative rehabilitation regimens in patients to restore balance in the knee joint.

Fresh-frozen cadaveric knees (n=6) were thawed and mounted onto a 6 DOF KUKA robot. Specimens were size matched to +2 mm for the medial-to-lateral width of the medial tibial hemiplateaus. Three specimens served as allograft recipient knees and three served as donor knees. Recipient knees were first tested in their native state and then tested with size-matched medial tibial hemiplateau shell grafts (n=3) prepared from the donor knees using custom-cut tab-in-slot and subchondral drilling techniques. Tekscan sensors were placed in the joint spaces to evaluate the loading conditions under 90N biaxial loading at full extension of the knee before and after graft placement. The I-Scan system used in conjunction analyzed the total force, pressure distribution, peak pressure, and center of force within the joint space.

Data demonstrated significant difference (p<0.05) in joint space loading after graft implantation compared to controls in both lateral and medial tibial plateaus. The I-Scan pressure mapping system displayed changes in femoral condylar contact points as well.

The results demonstrated that joint space loading was significantly different (p<0.05) between all preoperative and postoperative cadaveric specimens. Despite the best efforts to size match grafts, slight differences in the host's joint geometry resulted in shifts of contact areas between the tibial plateau and femoral condyle therefore causing either an increase or decrease in pressure measured by the sensor. This concludes that accuracy in graft size matching is extremely important to restoring close to normal loading across the joint and this can be further ensured through postoperative care customized to the patient after OCA surgery.

Autologous osteochondral transplantation (AOT) is an effective treatment for large Osteochondral Lesions of the Talus (OLT), however little is reported on an athletic population, who are likely to place higher demands on the reconstruction. The aim is to report the outcomes of large OLT (>150mm²) within an athletic population. The study population was limited to professional or amateur athletes (Tegner score >6) with an OLT of size 150mm² or greater. The surgical intervention was AOT with a donor site from the lateral femoral condyle. Clinical outcomes at a minimum of 24 months included Return to Sport, VAS and FAOS Scores. In addition, graft incorporation was evaluated by MRI using MOCART scores at 12 months post-surgery. 38 athletes including 11 professional athletes were assessed. Mean follow-up was 46 months. Mean lesion size was 249mm². 33 patients returned to sport at their previous level and one did not return to sport (mean return to play 8.2 months). Visual analogue scores improved from 4.53 pre-operatively to 0.63 post-operatively (p=0.002). FAOS Scores improved significantly in all domains (p< 0.001). Two patients developed knee donor site pain, and both had three osteochondral plugs harvested. Univariant analysis demonstrated no association between pre-operative patient or lesion characteristics and ability to return to sport. However, there was a strong correlation between MOCART scores and ability to return to sport (AUC=0.89). Our study suggests that AOT is a viable option in the management of large osteochondral talar defects in an athletic population, with favourable return to sport levels, patient satisfaction, and FAOS/VAS scores. The ability to return to sport is predicated upon good graft incorporation and further research is required to optimise this technique. Our data also suggests that patients should be aware of the increased risk of developing knee donor site pain when three osteochondral plugs are harvested.

Introduction

The purpose of this study was to examine trends in patient characteristics and clinical outcomes that occur with age as a statistical variable when performing autologous osteochondral transplantation (AOT) for the treatment of osteochondral lesions of the talus (OLT).

Introduction

Osteochondral lesions (OCLs) of the talus are a challenging and increasingly recognized problem in chronic ankle pain. Many novel techniques exist to attempt to treat this challenging entity. Difficulties associated with treating OCLs include lesion location, size, chronicity, and problems associated with potential graft harvest sites. Matrix associated stem cell transplantation (MAST) is one such treatment described for larger lesions >15mm2 or failed alternative therapies. This cohort study describes a medium-term review of the outcomes of talar lesions treated with MAST.

Aim of study: Comparison of clinical and radiological results of the applied methods.

Material and method: 20 patients with osteochondritis dissecans of the talus were treated. Autologous chondrocyte transplantation was applied to a group of 10 patients (Group A) and autologous osteochondral transplantation to the rest 10 patients (Group B).Group A: Average-age=30,8 years, -follow up time= 33,6 (12–48) months, -lesion size= 20×16,2 (35–15 × 25–15) mm and depth =7 (20–5) mmGroup B: Average –age =33, 7 years, follow up time =32,4 (12–48) months, – lesion size=16,5×15 (25–10 × 20–10) mm and depth=4 (5–3) mm.All patients underwent clinical und radiological investigation and the symptoms were classified in accordance with the Finsen classification. Group A was treated with autologous chondrocyte and Group B with autologous osteochondral transplantation. Osteotomy of the medial or lateral malleolus was necessary by 6 Group A – patients and 4 Group B – patients. Postoperative treatment : Non-weight bearing for 6 weeks, continuous passive motion of the joint, clinical and radiological follow up 3rd, 6th, 12th and yearly basis

Results: Postoperatively the average Finsen score showed for Group A an improvement from 3,5 (very bad) to 1,1 (excellent) and for Group B from 3,6 (very bad) to 1 (excellent). MRI follow up showed defect coverage by 7 patients of Group A and 9 patients of Group B after 12 months. Second look arthroscopy was performed by 4 patient of Group A and 3 patients of Group B showing full coverage of the defect site. Complications: persisting swelling for an average time of 2,7 months in Group A and 4,3 months in Group B. Additionally 10 degrees loss of joint extension in 4 patients of Group A and 3 patients of Group B.

Conclusion: Autologous chondrocyte transplantation as well as autologous osteochondral transplantation present very good clinical results. As methods of articular surface restoration they have their advantages and disadvantages playing an important role in chosing one of them, in combination always with the surgeons philosophy. The existence of these methods is important in influencing the course of the ankle joint towards a good prognosis, in cases of osteochondral lesion. Further investigation is necessary.