Multiple intra-articular injections of allogeneic bone marrow-derived stem cells potentially improve knee lesions resulting from surgically induced osteoarthritis: an animal study.

AIM: Mesenchymal stem cells (MSCs) have several properties that may support their use as an early treatment option for osteoarthritis (OA). This study investigated the role of multiple injections of allogeneic bone marrow-derived stem cells (BMSCs) to alleviate the progression of osteoarthritic changes in the various structures of the mature rabbit knee in an anterior cruciate ligament (ACL)-deficient OA model. MATERIALS AND METHODS: Two months after bilateral section of the ACL of Japanese white rabbits aged nine months or more, either phosphate buffered saline (PBS) or 1 x 106 MSCs were injected into the knee joint in single or three consecutive doses. After two months, the articular cartilage and meniscus were assessed macroscopically, histologically, and immunohistochemically using collagen I and II. RESULTS: Within the PBS injection (control group), typical progressive degenerative changes were revealed in the various knee structures. In the single MSC injection (single group), osteoarthritic changes were attenuated, but still appeared, especially in the medial compartments involving fibrillation of the articular cartilage, osteophyte formation in the medial plateau, and longitudinal tear of the meniscus. In the multiple-injections group, the smoothness and texture of the articular cartilage and meniscus were improved. Histologically, absence or reduction in matrix staining and cellularity were noticeable in the control and single-injection groups, respectively, in contrast to the multiple-injections group, which showed good intensity of matrix staining and chondrocyte distribution in the various cartilage zones. Osteoarthritis Research Society International (OARSI) scoring showed significantly better results in the multiple-injections group than in the other groups. Immunohistochemically, collagen I existed superficially in the medial femoral condyle in the single group, while collagen II was more evident in the multiple-injections group than the single-injection group. CONCLUSION: A single injection of MSCs was not enough to restore the condition of osteoarthritic joints. This is in contrast to multiple injections of MSCs, which had the ability to replace lost cells, as well as reducing inflammation. Cite this article: Bone Joint J 2019;101-B:824-831.

Risk factors for progression of articular cartilage damage after anatomical anterior cruciate ligament reconstruction: a second-look arthroscopic evaluation.

Aims: To investigate the risk factors for progression of articular cartilage damage after anatomical anterior cruciate ligament (ACL) reconstruction. Patients and Methods: A total of 174 patients who underwent second-look arthroscopic evaluation after anatomical ACL reconstruction were enrolled in this study. The graded condition of the articular cartilage at the time of ACL reconstruction was compared with that at second-look arthroscopy. Age, gender, body mass index (BMI), ACL reconstruction technique, meniscal conditions, and other variables were assessed by regression analysis as risk factors for progression of damage to the articular cartilage. Results: In the medial compartment, multivariable logistic regression analysis indicated that partial medial meniscectomy (odds ratio (OR) 6.82, 95% confidence interval (CI) 2.11 to 22.04, p = 0.001), pivot-shift test grade at the final follow-up (OR 3.53, CI 1.39 to 8.96, p = 0.008), BMI (OR 1.15, CI 1.03 to 1.28, p = 0.015) and medial meniscal repair (OR 3.19, CI 1.24 to 8.21, p = 0.016) were significant risk factors for progression of cartilage damage. In the lateral compartment, partial lateral meniscectomy (OR 10.94, CI 4.14 to 28.92, p < 0.001) and side-to-side differences in anterior knee laxity at follow-up (OR 0.63, p = 0.001) were significant risk factors. Conclusion: Partial meniscectomy was found to be strongly associated with the progression of articular cartilage damage despite r anatomical ACL reconstruction. Cite this article: Bone Joint J 2018;100-B:285-93.

Revision single-bundle anterior cruciate ligament reconstruction with over-the-top route procedure.

PURPOSE: In revision anterior cruciate ligament reconstruction (ACLR), the single-stage technique and the over-the-top route (OTTR) procedure were usually selected for cases where the bone tunnel cannot be created at an anatomical position due to tunnel enlargement and overlap with the mal-positioned tunnel of primary reconstruction. The purpose of this study was to evaluate the clinical results of revision single-bundle ACL reconstruction using OTTR procedure and to compare the clinical results of OTTR procedure with those of anatomical single-bundle revision reconstruction (SBR). HYPOTHESIS: The results of OTTR procedure are equivalent to that of SBR. METHODS: Seventy-six revision ACL reconstruction knees from April 2002 to December 2012 were involved in our study. We focused on 21 knees which underwent surgery with SBR and 22 knees with OTTR using hamstring tendon. The clinical results were evaluated by means of the Lysholm score and the knee stability was assessed by the Lachman test, pivot-shift test and side-to-side difference by KT-2000 pre-operatively and after 1 year post-operatively. AP translation and rotational laxity using a navigation system were evaluated before and after revision ACL reconstruction under anesthesia in 8 cases of OTTR and in 6 cases of SBR. RESULTS: There was no statistically significant difference between the OTTR and SBR regarding Lysholm score, Lachman test, pivot-shift test, ATT by KT-2000, and AP translation and rotational laxity with a navigation system. CONCLUSIONS: The clinical results of OTTR are almost equivalent to those of SBR. For the cases in which it is impossible to create the femoral tunnel in an anatomical position, OTTR is a valuable revision ACL reconstruction method. LEVEL OF EVIDENCE: Case-control study. Level III.

Clinical outcomes of second-look arthroscopic evaluation after anterior cruciate ligament augmentation: comparison with single- and double-bundle reconstruction.

We report the clinical outcome and findings at second-look arthroscopy of 216 patients (mean age 25 years (11 to 58)) who underwent anterior cruciate ligament (ACL) reconstruction or augmentation. There were 73 single-bundle ACL augmentations (44 female, 29 male), 82 double-bundle ACL reconstructions (35 female, 47 male), and 61 single-bundle ACL reconstructions (34 female, 27 male). In 94 of the 216 patients, proprioceptive function of the knee was evaluated before and 12 months after surgery using the threshold to detect passive motion test. Second-look arthroscopy showed significantly better synovial coverage of the graft in the augmentation group (good: 60 (82%), fair: 10 (14%), poor: 3 (4%)) than in the other groups (p = 0.039). The mean side-to-side difference measured with a KT-2000 arthrometer was 0.4 mm (-3.3 to 2.9) in the augmentation group, 0.9 mm (-3.2 to 3.5) in the double-bundle group, and 1.3 mm (-2.7 to 3.9) in the single-bundle group: the result differed significantly between the augmentation and single-bundle groups (p = 0 .013). No significant difference in the Lysholm score or pivot-shift test was seen between the three groups (p = 0.09 and 0.65, respectively). In patients with good synovial coverage, three of the four measurements used revealed significant improvement in proprioceptive function (p = 0.177, 0.020, 0.034, and 0.026). We conclude that ACL augmentation is a reasonable treatment option for patients with favourable ACL remnants.

Proprioceptive function after isolated single-bundle posterior cruciate ligament reconstruction with remnant preservation for chronic posterior cruciate ligament injuries.

INTRODUCTION: Posterior cruciate ligament (PCL) reconstruction using the remnant preserving technique may contribute to improved postoperative posterior stability, graft healing, and proprioception recovery. Although there have been several reports on remnant preserving PCL reconstruction, no study has yet evaluated the proprioceptive functions before and after PCL reconstruction with remnant preservation. The purpose of this study is to retrospectively evaluate the clinical outcomes and proprioceptive function after isolated single-bundle PCL reconstruction with remnant preservation for chronic PCL injuries. HYPOTHESIS: Isolated single-bundle PCL reconstruction with remnant preservation surgery for chronic PCL injuries provides satisfactory clinical outcomes and good recovery of the proprioceptive function. METHODS: Nineteen patients who had undergone isolated single-bundle PCL reconstruction with remnant preservation for chronic PCL injuries were followed up for more than 2 years. The posterior laxity was measured by the gravity sag view, stress radiography and the KT-2000 knee arthrometer. The proprioceptive function was defined as the threshold to detect passive motion (TTDPM). RESULTS: The average Lysholm score significantly improved from 63.7±13.2 preoperatively to 94.4±4.6 at final follow-up. The postoperative posterior laxity significantly improved. Regarding TTDPM, there were no significant differences between the preoperative score and the score at every given time point, regardless of the starting angles and the moving directions of the knees. CONCLUSIONS: The proprioceptive function, defined as TTDPM, is maintained after single-bundle PCL reconstruction with remnant preservation, and the postoperative clinical scores and posterior laxity significantly improve. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

Unsuccessful regeneration of the semitendinosus tendon harvested for anterior cruciate ligament reconstruction: report of two cases.

Recent magnetic resonance imaging (MRI) and three-dimensional (3D) computed tomography (CT) analyses have demonstrated that semitendinosus tendon can regenerate at a high rate following harvesting the tendon for anterior cruciate ligament (ACL) reconstruction. Although it is known that the regeneration of the semitendinosus tendon does not occur in all the patients, the reason for this unsuccessful regeneration of the tendon in certain patients remains unknown. We recently encountered two cases in which regeneration of the semitendinosus tendon was unsuccessful because of apparent reasons. These patients experienced a sudden sharp pain in the posterior aspect of their thighs when their hamstring muscles were subjected to aggressive load at 3 and 4 weeks after surgery. At the follow-up examination conducted after 12 months, 3D CT imaging revealed unsuccessful regeneration of the semitendinosus tendons in both cases. Severe proximal retraction of the semitendinosus muscle belly was also confirmed.

Acceleration of tendon-bone healing in anterior cruciate ligament reconstruction using an enamel matrix derivative in a rat model.

We examined whether enamel matrix derivative (EMD) could improve healing of the tendon-bone interface following reconstruction of the anterior cruciate ligament (ACL) using a hamstring tendon in a rat model. ACL reconstruction was performed in both knees of 30 Sprague-Dawley rats using the flexor digitorum tendon. The effect of commercially available EMD (EMDOGAIN), a preparation of matrix proteins from developing porcine teeth, was evaluated. In the left knee joint the space around the tendon-bone interface was filled with 40 µl of EMD mixed with propylene glycol alginate (PGA). In the right knee joint PGA alone was used. The ligament reconstructions were evaluated histologically and biomechanically at four, eight and 12 weeks (n = 5 at each time point). At eight weeks, EMD had induced a significant increase in collagen fibres connecting to bone at the tendon-bone interface (p = 0.047), whereas the control group had few fibres and the tendon-bone interface was composed of cellular and vascular fibrous tissues. At both eight and 12 weeks, the mean load to failure in the treated specimens was higher than in the controls (p = 0.009). EMD improved histological tendon-bone healing at eight weeks and biomechanical healing at both eight and 12 weeks. EMD might therefore have a human application to enhance tendon-bone repair in ACL reconstruction.

Prospective study of kinesthesia after ACL reconstruction.

Previous studies used a variety of methods to assess kinesthesia, thus no consensus exists regarding kinesthetic adaptation after anterior cruciate ligament (ACL) reconstruction. This study prospectively examined whether kinesthesia is adapted after ACL reconstruction, and then discussed the actual angular velocity required to properly assess kinesthesia in ACL-reconstructed patients. 31 patients were evaluated using the threshold to detect passive motion (TTDPM) test, which was applied preoperatively, and at 3, 6, and 12 months following surgery. TTDPMs were measured at 15° or 45° of knee flexion toward both extension and flexion with angular velocities of 0.1°/s or 0.2°/s. ACL-reconstructed knees showed significantly impaired TTDPMs compared to healthy knees before the operation at 15° of knee flexion toward extension and at 45° of knee flexion toward both extension and flexion at 0.2°/s (15° of knee flexion toward extension, P=0.036; 45° of knee flexion toward extension, P=0.015; 45° of knee flexion toward flexion, P=0.030). However, there were no significant differences after 3 months of follow-up. On the basis of these results, applying 0.2°/s seems appropriate to assess TTDPM for patients with an ACL reconstruction, and kinesthesia is adapted within 12 months after the operation. Sensory function and biomechanical stability are also adapted following ACL reconstruction.

Atelocollagen-associated autologous chondrocyte implantation for the repair of chondral defects of the knee: a prospective multicenter clinical trial in Japan.

BACKGROUND: New tissue-engineering technology was developed to create a cartilage-like tissue in a three-dimensional culture using atelocollagen gel. The minimum 2-year followup outcome of transplanting autologous chondrocytes cultured in atelocollagen gel for the treatment of full-thickness defects of cartilage in knees was reported from the single institution. The present multicenter study was conducted to determine clinical and arthroscopic outcomes in patients who underwent atelocollagen-associated autologous chondrocyte implantation for the repair of chondral defects of the knees. METHODS: At six medical institutes in Japan, we prospectively evaluated the clinical and arthroscopic outcomes of transplanting autologous chondrocytes cultured in atelocollagen gel for the treatment of full-thickness defects of cartilage in 27 patients (27 knees) with cartilage lesions on a femoral condyle or on a patellar facet over 24 months. RESULTS: The Lysholm score significantly increased from 60.0 +/- 13.7 points to 89.8 +/- 9.5 points (P = 0.001). Concerning the ICRS grade for arthroscopic appearance, 6 knees (24%) were assessed as grade I (normal) and 17 knees (68%) as grade II (nearly normal). There were few adverse features, except for detachment of the graft in two cases. CONCLUSIONS: We concluded that transplanting chondrocytes in a newly formed matrix of atelocollagen gel can promote restoration of the articular cartilage of the knee.

Histological evaluation of internally-fixed osteochondral lesions of the knee.

We evaluated the histological changes before and after fixation in ten knees of ten patients with osteochondritis dissecans who had undergone fixation of the unstable lesions. There were seven males and three females with a mean age of 15 years (11 to 22). The procedure was performed either using bio-absorbable pins only or in combination with an autologous osteochondral plug. A needle biopsy was done at the time of fixation and at the time of a second-look arthroscopy at a mean of 7.8 months (6 to 9) after surgery. The biopsy specimens at the second-look arthroscopy showed significant improvement in the histological grading score compared with the pre-fixation scores (p < 0.01). In the specimens at the second-look arthroscopy, the extracellular matrix was stained more densely than at the time of fixation, especially in the middle to deep layers of the articular cartilage. Our findings show that articular cartilage regenerates after fixation of an unstable lesion in osteochondritis dissecans.

Histological and biomechanical study of impacted cancellous allografts with cement in the femur: a canine model.

INTRODUCTION: An experimental canine model was used for quantitative histological study using bone morphometry and biomechanical study on impacted cancellous allografting with cement in revision total hip arthroplasty. MATERIALS AND METHODS: The prosthesis had a collarless polished titanium alloy tapered stem with a head diameter of 15 mm. Twelve Beagle dogs were used. Cancellous bone grafts were obtained aseptically from the femoral condyles of other beagles. Histological examinations were performed up to 1 year after surgery. Biomechanical examinations were performed at 8 weeks after surgery. RESULTS: The new bone formation proceeded from the proximal part to the distal site of the femur in the long axis as well as from the host cortical bone to the cement layer. Bone morphometry showed that the mineral apposition rate was higher in the proximal site of the femur than in the distal site at 8 weeks after surgery, but no significant differences were observed at other times. The bone formation rate was higher in the proximal site of the femur at 8 weeks after surgery, while it increased in the distal site at 16 weeks postoperatively but had decreased at 1 year after surgery significantly. The load test at 8 weeks after surgery showed that there was no significant difference at all sites attached strain gauge between impacted cancellous allograft group and control group. CONCLUSION: We concluded that biological replacement of the grafted bone by new bone settled at 1 year after surgery in this experimental model. However, the process was not completed.

Repair of chronic osteochondral defects in the rat. A bone marrow-stimulating procedure enhanced by cultured allogenic bone marrow mesenchymal stromal cells.

Bone marrow mesenchymal stromal cells were aspirated from immature male green fluorescent protein transgenic rats and cultured in a monolayer. Four weeks after the creation of the osteochondral defect, the rats were divided into three groups of 18: the control group, treated with an intra-articular injection of phosphate-buffered saline only; the drilling group, treated with an intra-articular injection of phosphate-buffered saline with a bone marrow-stimulating procedure; and the bone marrow mesenchymal stromal cells group, treated with an intra-articular injection of bone marrow mesenchymal stromal cells plus a bone marrow-stimulating procedure. The rats were then killed at 4, 8 and 12 weeks after treatment and examined. The histological scores were significantly better in the bone marrow mesenchymal stromal cells group than in the control and drilling groups at all time points (p < 0.05). The fluorescence of the green fluorescent protein-positive cells could be observed in specimens four weeks after treatment.

Tophaceous gout in the bipartite patella with intra-osseous and intra-articular lesions: a case report.

We report a case of a 34-year-old man who had an acute gouty attack with tophi around the accessory bone of the bipartite patella with intra-osseous lesions. As the symptoms did not respond to conservative treatments, the patient was treated surgically. Arthroscopy revealed urate crystals surrounded by inflammatory synovitis on the surface of the bipartite patella. Arthrotomy exposed the abundant crystals around the accessory bone and in the intra-osseous lesion. These findings made us speculate that some pre-existing inflammatory conditions around the accessory bone induced deposition of urate crystals, and the destruction of the barrier between the deposits and the joint led to an acute arthritis. Curettage of the tophi resulted in immediate improvement of knee function. There had been no recurrence of symptoms at the 2-year follow-up.

Reconstruction of the medial patellofemoral ligament for the treatment of habitual or recurrent dislocation of the patella in children.

We investigated the clinical outcome of a reconstructive procedure of the medial patellofemoral ligament for the treatment of habitual or recurrent dislocation of the patella in four children (6 knees), with a minimum follow-up of four years. The technique involves transfer of the tendon of semitendinosus to the patella using the posterior one-third of the femoral insertion of the medial collateral ligament as a pulley. There was no recurrence of dislocation after surgery. The mean Kujala score at follow-up was 96.3 points. Radiological assessment showed that the congruence angle, the tilt angle and the lateral shift radio were restored to normal. The lateral and medial stress shift ratios and the Insall-Salvati ratio remained abnormal. We conclude that this technique can be recommended for the treatment of habitual or recurrent patellar dislocation in children, although hypermobility and patella alta are not fully corrected.

Pretibial cyst formation after anterior cruciate ligament reconstruction using auto hamstring grafts: two case reports in a prospective study of 89 cases.

Eighty-nine cases after anterior cruciate ligaments (ACL) reconstruction were followed prospectively with magnetic resonance imaging (MRI). The patients were examined using axial and sagittal MRI at least twice during the postoperative evaluation of reconstructed ACL. Two cases of pretibial cyst formation were observed. At the time of cyst formation, neither patient had any subjective or objective evidence of knee instability. The cyst of one case communicated with the intra-articular. The minimum follow-up period after the surgical excision was 9 months, with no evidence of recurrence. We might speculate that the critical period for cyst formation in both patients occurred at less than 12 months after their ACL reconstruction. We concluded that the cyst formation was most likely due to incomplete graft tendon incorporation within the osseous tunnel.

Appearance of anterior cruciate ligament autografts in their tibial bone tunnels on oblique axial MRI.

The objective of this study was to observe the changing appearance of human anterior cruciate ligament (ACL) grafts in their tibial bone tunnels by MRI using oblique axial images. One-hundred and eight knees in 75 patients were studied by MRI at 1-33 months after arthroscopic ACL reconstructions using double-looped, autogenous semitendinosus and/or gracilis tendons. Knees with poor stability were excluded from this study. The examinations were performed at 0.2T with spin echo proton density and T2-weighted oblique axial images. Appearances of grafts were mainly described on spin echo proton density images based upon time after surgery. The grafts appeared as homogeneous, low signal intensity areas in the bone tunnels at 1 month after the surgery. Ring-shaped low signal intensity areas were observed along the wall of the bone tunnels in the 2- to 3-month group. In many grafts from this group, each tendinous bundle appeared as a low signal area separated by a high signal intensity area. In all cases in the 4- to 6-month group, the thickness of the ring-shaped low signal intensity area had increased, whereas the thickness of the high signal intensity area had decreased. In almost all of the cases, the interior of the bone tunnel gradually became a homologous low signal intensity region by 7 to 12 months after the surgery. According to these results, it is suggested that the maturation of the tendon-bone interface was completed from 6 to 12 months after the ACL reconstruction.

Ganglion cysts of the cruciate ligaments detected by MRI.

Eight patients with ganglion cysts arising from the cruciate ligaments of the knee joint underwent arthroscopic excision after the MR examination. The MR findings, clinical features and arthroscopic findings were evaluated comparatively.

Early expression of marker genes in the rabbit medial collateral and anterior cruciate ligaments: the use of different viral vectors and the effects of injury.

Gene therapy is a technique that may offer advantages over current methods of cytokine delivery to ligaments. To determine if implanted genes could be expressed in normal and injured knee ligaments, the medial collateral ligament and anterior cruciate ligament were studied in 18 rabbits. A retroviral ex vivo technique using allograft medial collateral ligament and anterior cruciate ligament fibroblasts and an adenoviral in vivo technique were compared as methods for delivering the LacZ marker gene to knee ligaments. Bilateral knee surgeries were performed, and the rabbits were equally divided into three groups. Group 1 received the retrovirus and the medial collateral ligament was ruptured, Group 2 received the adenovirus and the medial collateral ligament was ruptured, and Group 3 received the adenovirus and the medial collateral ligament was not injured. The anterior cruciate ligament was not injured in any group. The medial collateral and anterior cruciate ligaments of the right knees received 10(6) allografted, transduced ligament fibroblasts or 10(9) adenovirus particles, whereas the ligaments of the left knee received a similar volume of saline solution only. Equal numbers of rabbits were killed at 10 days, 3 weeks, and 6 weeks following the procedure. Ligament samples were stained with X-gal to detect the expression of the LacZ gene product, beta-galactosidase. LacZ gene expression was evident in ruptured and uninjured medial collateral ligaments as well as in the anterior cruciate ligament. The expression lasted between 10 days and 3 weeks in the medial collateral and anterior cruciate ligaments with use of the retrovirus and between 3 and 6 weeks in the medial collateral ligament and at least 6 weeks in the anterior cruciate ligament with the adenovirus. The length of gene expression in the ruptured and uninjured medial collateral ligaments did not differ. These preliminary studies indicate that gene transfer to normal and injured knee ligaments is possible.

The effects of platelet-derived growth factor-BB on healing of the rabbit medial collateral ligament. An in vivo study.

We report a biologic approach to improve medial collateral ligament healing using growth factors normally expressed in healing tissue. Our previous in vitro work demonstrated that platelet-derived growth factor-BB and transforming growth factor-beta 1 promoted fibroblast proliferation and matrix synthesis, respectively. There-fore, these growth factors were used in vivo to determine whether they could improve medial collateral ligament healing, whether this effect was dose-dependent, and if combinations of growth factors could improve healing more than individual growth factors. Thirty-seven rabbits had various doses of growth factors applied to the ruptured right medial collateral ligaments using a fibrin sealant delivery vehicle. The five groups consisted of 1) two groups receiving two doses of platelet-derived growth factor-BB, 2) two groups receiving two doses of this growth factor plus transforming growth factor-beta 1, and 3) one group receiving fibrin sealant only. After sacrifice at 6 weeks, biomechanical and histologic evaluations of the healing ligament were performed. Femur-medial collateral ligament-tibia complexes of the knees given the higher dose of platelet-derived growth factor-BB had ultimate load, energy absorbed to failure, and ultimate elongation values that were 1.6, 2.4, and 1.6 times greater than the same complexes of the control group. Adding transforming growth factor-beta 1 did not lead to any further increase in the structural properties of the complex compared with treatment with platelet-derived growth factor-BB. These encouraging results suggest that use of platelet-derived growth factor-BB may improve the quality of the healing medial collateral ligament, and that it may also have a similar potential for promoting healing of other ligaments.

The effects of age on rabbit MCL fibroblast matrix synthesis in response to TGF-beta 1 or EGF.

In this study, we examined the effects of age on collagen and total protein synthesis by ligament fibroblasts in response to growth factors. Three different doses of transforming growth factor-beta 1 (TGF-beta 1) or epidermal growth factor (EGF) were individually added to in vitro fibroblast cultures from the medial collateral ligament (MCL) of skeletally immature (age 3 months), mature (age 12 months) and senescent (age 48-51 months) rabbits. Analysis of the effects of age revealed that fibroblasts from senescent rabbits produced significantly less collagen in response to TGF-beta 1 or EGF stimulation when compared to fibroblasts from immature rabbits. Furthermore, increased age was found to result in significant reductions in the baseline levels of collagen synthesis but not total protein synthesis. Additionally, collagen and total protein synthesis by MCL fibroblasts were significantly affected by the TFG-beta 1 dose, but not by the EGF dose. When fibroblasts were normalized to their own controls, the increase in collagen and total protein synthesis due to TGF-beta 1 and EGF for the senescent group were found to be greater than those for the skeletally immature rabbits at all doses. This demonstrates that MCL fibroblasts from senescent rabbits are responsive to growth factors.