Increased migratory activity and cartilage regeneration by superficial-zone chondrocytes in enzymatically treated cartilage explants.

BACKGROUND: Limited chondrocyte migration and impaired cartilage-to-cartilage healing is a barrier in cartilage regenerative therapy. Collagenase treatment and delivery of a chemotactic agent may play a positive role in chondrocyte repopulation at the site of cartilage damage. This study evaluated chondrocyte migratory activity after enzymatic treatment in cultured cartilage explant. Differential effects of platelet-derived growth factor (PDGF) dimeric isoforms on the migratory activity were investigated to define major chemotactic factors for cartilage. METHODS: Full-thickness cartilage (4-mm3 blocks) were harvested from porcine femoral condyles and subjected to explant culture. After 15 min or 60 min of actinase and collagenase treatments, chondrocyte migration and infiltration into a 0.5-mm cartilage gap was investigated. Cell morphology and lubricin, keratan sulfate, and chondroitin 4 sulfate expression in superficial- and deep-zone chondrocytes were assessed. The chemotactic activities of PDGF-AA, -AB, and -BB were measured in each zone of chondrocytes, using a modified Boyden chamber assay. The protein and mRNA expression and histological localization of PDGF-ß were analyzed by western blot analysis, real-time reverse transcription polymerase chain reaction (RT-PCR), and immunohistochemistry, and results in each cartilage zone were compared. RESULTS: Superficial-zone chondrocytes had higher migratory activity than deep-zone chondrocytes and actively bridged the cartilage gap, while metachromatic staining by toluidine blue and immunoreactivities of keratan sulfate and chondroitin 4 sulfate were detected around the cells migrating from the superficial zone. These superficial-zone cells with weak immunoreactivity for lubricin tended to enter the cartilage gap and possessed higher migratory activity, while the deep-zone chondrocytes remained in the lacuna and exhibited less migratory activity. Among PDGF isoforms, PDGF-AB maximized the degree of chemotactic activity of superficial zone chondrocytes. Increased expression of PDGF receptor-ß was associated with higher migratory activity of the superficial-zone chondrocytes. CONCLUSIONS: In enzymatically treated cartilage explant culture, chondrocyte migration and infiltration into the cartilage gap was higher in the superficial zone than in the deep zone. Preferential expression of PDGF receptor-ß combined with the PDGF-AB dimeric isoform may explain the increased migratory activity of the superficial-zone chondrocytes. Cells migrating from superficial zone may contribute to cartilage regeneration.

Morphometric measurement of cortical bone trajectory for lumbar pedicle screw insertion using computed tomography.

STUDY DESIGN: A morphometric measurement of cortical bone trajectory (CBT) for the lumbar pedicle screw insertion using computed tomography (CT). OBJECTIVE: The aim of this study was to conduct a detailed morphometric measurement of the CBT. SUMMARY OF BACKGROUND DATA: The CBT is a novel lumbar pedicle screw trajectory, which follows a caudocephalad path sagittally and a laterally directed path in the transverse plane. The advantage associated with this modified technique is increased cortical bone contact, providing an enhanced screw purchase. However, little is known about the possible screw size or detailed direction of the trajectory. METHODS: The CT scans of 100 adults who underwent examination for spinal problems were studied. A total of 470 lumbar vertebrae excluding spondylosis, malformation, and tumor were observed. In this trajectory, the starting point was supposed to be the junction of the center of the superior articular process and 1 mm inferior to the inferior border of the transverse process. The CT images were analyzed using 3-dimensional reconstruction software. The diameter, length, lateral angle to the vertebral sagittal plane, and cephalad angle to the vertebral horizontal plane of the trajectory were measured. RESULTS: The mean diameter gradually increased from L1 to L5 (from 6.2 mm at L1 to 8.4 mm at L5). The mean length from L1 to L5 were 36.8, 38.2, 39.3, 39.8, and 38.3 mm, respectively. The lateral angle from L1 to L5 were 8.6, 8.5, 9.1, 9.1, and 8.8 degrees, respectively. The cephalad angle from L1 to L5 were 26.2, 25.5, 26.2, 26.0, and 25.8 degrees, respectively. CONCLUSIONS: The morphology of the pedicle, such as shape and pedicle axis angle, differed over the lumbar levels, our measurements demonstrated similar data excluding the diameter of the trajectory. There were no significant differences between each level of the lateral and cephalad angles.

Acquired diaphragmatic hernia following a peritoneal biopsy for gastric cancer dissemination in the diaphragm: a case report.

BACKGROUND: Acute diaphragmatic hernia is a life-threatening condition caused by prolapse of an abdominal organ into the thoracic cavity through a defect in the diaphragm. We present herein a case of acquired diaphragmatic hernia following a peritoneal biopsy for gastric cancer dissemination in the diaphragm. CASE PRESENTATION: A 72-year-old, female patient presented with a complaint of acute abdomen 10 months after receiving a diagnosis of stage IV gastric cancer with peritoneal dissemination based on peritoneal biopsy findings during staging laparoscopy. Computed tomography demonstrated herniation of the small intestine into the thoracic cavity. Emergency surgery was performed, and a full-thickness diaphragmatic defect was found intraoperatively at the same location as the previous, peritoneal biopsy. The incarcerated small intestine was atraumatically repositioned into the abdominal cavity, and the defect was closed laparoscopically using an absorbable barbed suture. CONCLUSIONS: Although complications of staging laparoscopy are extremely rare, excising disseminated nodules from the diaphragm carries the risk of diaphragmatic hernia. For this reason, avoiding excision is desirable unless a diaphragmatic biopsy is needed.

Photodynamic therapy using intra-articular Photofrin for murine MRSA arthritis: biphasic light dose response for neutrophil-mediated antibacterial effect.

BACKGROUND AND OBJECTIVE: Bacterial arthritis does not respond well to antibiotics and moreover multidrug resistance is spreading. We previously tested photodynamic therapy (PDT) mediated by systemic Photofrin® in a mouse model of methicillin-resistant Staphylococcus aureus (MRSA) arthritis, but found that neutrophils were killed by PDT and therefore the infection was potentiated. STUDY DESIGN/MATERIALS AND METHODS: The present study used an intra-articular injection of Photofrin® and optimized the light dosimetry in order to maximize bacterial killing and minimize killing of host neutrophils. MRSA (5 × 10(7) CFU) was injected into the mouse knee followed 3 days later by 1 µg of Photofrin® and 635-nm diode laser illumination with a range of fluences within 5 minutes. Synovial fluid was sampled 6 hours or 1-3, 5, and 7 days after PDT to determine MRSA colony-forming units (CFU), neutrophil numbers, and levels of cytokines. RESULTS: A biphasic light dose response was observed with the greatest reduction of MRSA CFU seen with a fluence of 20 J cm(-2), whereas lower antibacterial efficacy was observed with fluences that were either lower or higher. Consistent with these results, a significantly higher concentration of macrophage inflammatory protein-2, a CXC chemokine, and greater accumulation of neutrophils were seen in the infected knee joint after PDT with a fluence of 20 J cm(-2) compared to fluences of 5 or 70 J cm(-2). CONCLUSION: PDT for murine MRSA arthritis requires appropriate light dosimetry to simultaneously maximize bacterial killing and neutrophil accumulation into the infected site, while too little light does not kill sufficient bacteria and too much light kills neutrophils and damages host tissue as well as bacteria and allows bacteria to grow unimpeded by host defense.

Nonviral retrograde gene transfer of human hepatocyte growth factor improves neuropathic pain-related phenomena in rats.

Peripheral nerve injury occasionally causes chronic neuropathic pain with hyperalgesia and allodynia. However, its treatment is difficult. Here, we used a chronic constriction injury (CCI) model in rats to investigate the effects on experimental neuropathic pain of the human hepatocyte growth factor (HGF) gene delivered into the nervous system by retrograde axonal transport following its repeated intramuscular transfer, using liposomes containing the hemagglutinating virus of Japan (HVJ). CCI (control) rats exhibited marked mechanical allodynia and thermal hyperalgesia, and decreased blood flow in sciatic nerve and hind paw. All these changes were significantly reversed by HGF gene transfer. In the sciatic nerve in HGF-treated rats, the size-frequency distributions for myelinated and unmyelinated axons each showed a rightward shift, the number of myelinated axons >5 microm in diameter was significantly increased, and the mean diameter of unmyelinated axons was significantly increased (versus CCI rats). Levels of P2X3, P2X4, and P2Y1 receptor mRNAs, and of interleukin-6 (IL-6) and activating transcription factor 3 (ATF3) mRNAs, were elevated in the ipsilateral dorsal root ganglia and/or sciatic nerve by CCI, and these levels were decreased by HGF gene transfer. These results may point toward a potential new treatment strategy for chronic neuropathic pain in this model.

Efficacy of nonviral gene transfer of human hepatocyte growth factor (HGF) against ischemic-reperfusion nerve injury in rats.

Ischemic neuropathy is common in subjects with critical limb ischemia, frequently causing chronic neuropathic pain. However, neuropathic pain caused by ischemia is hard to control despite the restoration of an adequate blood flow. Here, we used a rat model of ischemic-reperfusion nerve injury (IRI) to investigate possible effects of hepatocyte growth factor (HGF) against ischemic neuropathy. Hemagglutinating virus of Japan (HVJ) liposomes containing plasmids encoded with HGF was delivered into the peripheral nervous system by retrograde axonal transport following its repeated injections into the tibialis anterior muscle in the right hindlimb. First HGF gene transfer was done immediately after IRI, and repeated at 1, 2 and 3 weeks later. Rats with IRI exhibited pronounced mechanical allodynia and thermal hyperalgesia, decreased blood flow and skin temperature, and lowered thresholds of plantar stimuli in the hind paw. These were all significantly improved by HGF gene transfer, as also were sciatic nerve conduction velocity and muscle action potential amplitudes. Histologically, HGF gene transfer resulted in a significant increase of endoneurial microvessels in sciatic and tibial nerves and promoted nerve regeneration which were confirmed by morphometric analysis. Neovascularization was observed in the contralateral side of peripheral nerves as well. In addition, IRI elevated mRNA levels of P2X3 and P2Y1 receptors, and transient receptor potential vanilloid receptor subtype 1 (TRPV1) in sciatic nerves, dorsal root ganglia and spinal cord, and these elevated levels were inhibited by HGF gene transfer. In conclusion, HGF gene transfer is a potent candidate for treatment of acute ischemic neuropathy caused by reperfusion injury, because of robust angiogenesis and enhanced nerve regeneration.

Photocrosslinkable chitosan hydrogel can prevent bone formation in both rat skull and fibula bone defects.

UV light irradiation to a photocrosslinkable chitosan (Az-CH-LA) resulted in an insoluble and flexible hydrogel within 30 s. The purpose of this study was to evaluate the ability of the photocrosslinkable chitosan to inhibit bone formation in the bone defects. A 5-mm-diameter defect was made in the rat calvarium, and then photocrosslinkable chitosan was implanted and irradiated with UV for 30 s. Furthermore, a 2-mm defect was made in the fibula of a rat hind leg, and then photocrosslinkable chitosan was implanted and irradiated with UV. Bone formations in the rat skull and fibula defects with photocrosslinkable chitosan hydrogel were significantly prevented for 8 weeks. Thus, the chitosan hydrogel has an inhibitory effect on bone formation.

Bone mass assessment in naval crew members by quantitative ultrasound technique.

OBJECTIVE: Of necessity, naval crews live in confined spaces when on board warships, which may lead to decreased bone mass and to subsequent bone fractures. Therefore, we investigated the bone mass of crew members and the relationship between bone mass and lifestyle factors. METHODS: We selected 1510 crew members of the Japan Maritime Self-Defense Force. All were men between 18 and 58 years of age. We measured their bone mass by applying quantitative ultrasound (QUS) to the calcaneus. In addition, we reviewed daily milk consumption, levels of physical exercise, type of on-board job, fracture history, nutritional supplementation habits, and body mass index (BMI). RESULTS: Bone mass values were lower than the published mean values for Japanese men across the twenties to forties age groups. CONCLUSIONS: The factors found to be related to bone mass in this study were age, type of on-board job, exercise level, and milk consumption. Lifestyle factors are usually within our control. Our data suggest that moderate levels of regular exercise and milk consumption may maintain bone mass.

The growth rate of the humerus: long-term follow-up of treatment of solitary bone cyst of the proximal humerus using cannulated screws: a case report.

Although the growth of the proximal epiphysis of the humerus is thought to contribute 80% of the total increase in humeral length, few articles have provided evidence for this. A 9-year-old boy with a pathological fracture at the humeral neck, owing to a solitary bone cyst, was treated by the decompression method using cannulated screws for 7.5 years. During this period, we measured the longitudinal humeral bone growth. The value obtained for the longitudinal growth contribution of the proximal humerus was 88% in this particular case.

Epidemiological patterns of traumatic musculoskeletal injuries and non-traumatic disorders in Japan Self-Defense Forces.

BACKGROUND: The epidemiological patterns of musculoskeletal injuries or disorders in military personnel have not been well documented and a better understanding is required for proper preventative measures and treatment. Here, we investigated musculoskeletal injuries or disorders among members of the Japan Self-Defense Forces. METHODS: All orthopedic patients (n = 22,340) who consulted to Japan Self-Defense Forces Hospitals were investigated for their type of injury or disorder, the injured body part, the mechanism, and the cause of injuries. RESULTS: Thirty-nine percent of the cases were classified as traumatic injuries, and 61% were classified as non-traumatic disorders. Of the traumatic injury patients, the injured body part was the upper extremity in 32%, the trunk in 23%, and the lower extremities in 45% of the cases. The most common injured body location was the knee followed by the hand/finger and ankle. Exercise was the most common cause of injury, followed by traffic accident and military training. Contusions were the most common traumatic injuries, followed by sprains and fractures. Of non-traumatic disorders, the lower extremities were reported as the injured part in 43% of the disorders. Lumbar spine disorders were the most common non-traumatic disorders, followed by tendon and joint disorders. CONCLUSIONS: Over one-third of orthopedic cases among members of the Japan Self-Defense Forces are traumatic injuries, with the knee being the body part most commonly injured and exercise being the leading cause of injury.

Retrospective cohort study between selective and standard C3-7 laminoplasty. Minimum 2-year follow-up study.

A total of 64 patients with cervical spondylotic myelopathy (CSM) were assessed in this study. Forty-two patients underwent selective expansive open-door laminoplasty (ELAP). Twenty-two patients who underwent conventional C3-7 ELAP served as controls. There were no significant differences in recovery rate of JOA scores, C2-C7 angle or cervical range of motion between two groups. Incidence of axial symptoms and segmental motor paralysis in selective ELAP was significantly lower than those in the C3-7 ELAP. Size of anterior compression mass, postoperative spinal cord positions and decompression conditions were evaluated using preoperative or postoperative MRI in 50 of 64 patients. There was a positive correlation between number of expanded laminae and maximum anterior spaces of spinal cord. Incomplete decompression was developed in three of 37 patients in selective ELAP and in two of 13 patients in C3-7ELAP. Mean size of anterior compression mass at incomplete decompression levels was significantly greater than that at complete decompression levels. Since, there was less posterior movement of the spinal cord in selective ELAP than that in C3-7ELAP, minute concerns about size of anterior compression mass is necessary to decide the number of expanded laminae. Overall, selective ELAP was less invasive and useful in reducing axial symptoms and segmental motor paralysis. This new surgical strategy was effective in improving the surgical outcomes of CSM, and short-term results were satisfactory.

Spontaneous resorption in recurrent intradural lumbar disc herniation. Case report.

The authors report a case of spontaneous resorption of intradural disc material in a patient with recurrent intradural lumbar disc herniation and review magnetic resonance (MR) imaging and histopathological findings. Intradural lumbar disc herniation is rare, and most patients with this condition require surgical intervention due to severe leg pain and vesicorectal disturbance. In the present case, however, the recurrent intradural herniated mass had completely disappeared by 9 months after onset. Histological examination of intradural herniated disc tissue demonstrated infiltrated macrophages and angiogenesis within the herniated tissue, and Gd-enhanced MR images showed rim enhancement not only at the initial presentation, but also at recurrence. The authors conclude that when rim enhancement is present on Gd-enhanced MR images, there is a possibility of spontaneous resorption even though the herniated mass may be located within the intradural space. Moreover, when radiculopathy is controllable and cauda equina syndrome is absent, conservative therapy can be selected.

A new operative procedure using a Küntcher nail for a periprosthetic supracondylar femoral fracture after revision total knee arthroplasty: a case report.

Periprosthetic supracondylar femoral fractures after total knee arthroplasty (TKA) are difficult surgical problems. We report a case of an 84-year-old female, in which an original method was applied to treat a periprosthetic supracondylar femoral nonunion just proximal a femoral component. The new method features extending the stem of the femoral component with a Küntcher nail. Eventually bony union was obtained and the patient is now able to walk without any support.

Recovery of Shoulder Rotational Muscle Strength After Arthroscopic Bankart Repair.

BACKGROUND: Shoulder rotational muscles act as dynamic stabilizers of the glenohumeral joint, and the recovery of muscle strength plays an important role in stabilizing the joint during postoperative rehabilitation. However, temporal changes in muscle strength after arthroscopic Bankart repair have not been clarified. PURPOSE: To better understand the temporal recovery of shoulder rotational muscle strength after arthroscopic Bankart repair. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Isokinetic concentric shoulder rotational muscle strength was evaluated in 50 patients who were diagnosed with recurrent dislocations of the glenohumeral joint and treated with arthroscopic Bankart repair. RESULTS: The mean peak torque/weight and total work were reduced significantly at 1.5 months after surgery (P < .0001) and returned to preoperative levels by 6 months for external rotation and 4.5 months for internal rotation. The contralateral peak torque ratios reached preoperative levels by 6 months after surgery. The ipsilateral peak torque ratios were reduced between 1.5 and 3 months after surgery and returned to preoperative levels at 6 months for external rotation and 4.5 months for internal rotation. CONCLUSION: Isokinetic shoulder rotational muscle strength after arthroscopic Bankart repair recovered to preoperative levels by 6 months for external rotation and 4.5 months for internal rotation.

Comparison of Pedicle Screw Fixation Strength Among Different Transpedicular Trajectories: A Finite Element Study.

STUDY DESIGN: Comparative biomechanical study by finite element (FE) method. OBJECTIVE: To investigate the pullout strength of pedicle screws using different insertional trajectories. SUMMARY OF BACKGROUND DATA: Pedicle screw fixation has become the gold standard for spinal fusion, however, not much has been done to clarify how the fixation strength of pedicle screws are affected by insertional trajectories and bone properties. MATERIALS AND METHODS: Three-dimensional FE models of 20 L4 vertebrae were constructed from the computed tomographic data. Five different transpedicular trajectories were compared: the traditional trajectory, the vertical trajectory, and the 3 lateral trajectories with different sagittal directions (caudal, parallel, cranial). For a valid comparison, screws of the same shape and size were inserted into the same pedicle in each subject, and the pullout strength were compared with nonlinear FE analyses. In addition, the pullout strength was correlated with bone mineral density (BMD). RESULTS: The mean pullout strength showed a 3.9% increase for the vertical trajectory relative to the traditional trajectory, 6.1% for the lateral-caudal trajectory, 21.1% for the lateral-parallel trajectory, and 34.7% for the lateral-cranial trajectory. The lateral-cranial trajectory demonstrated the highest value among all trajectories (P<0.001). In each trajectory, the correlation coefficient between the pullout strength and BMD of the femoral neck (r=0.74-0.83, P<0.01) was higher than the mean BMD of all the lumbar vertebrae (r=0.49-0.75, P<0.01), BMD of the L4 vertebra (r=0.39-0.64, P<0.01), and regional BMD of the L4 pedicle (r=0.53-0.76, P<0.01). CONCLUSIONS: Regional variation in the vertebral bone density and the amount of denser bone-screw interface contribute to the differences of stiffness among different screw trajectories. BMD of the femoral neck is considered to be a better objective predictor of pedicle screw stability than that of the lumbar vertebra.

Cortical Bone Trajectory for Thoracic Pedicle Screws: A Technical Note.

STUDY DESIGN: A morphometric measurement of new thoracic pedicle screw trajectory using computed tomography and a biomechanical study on cadaveric thoracic vertebrae using insertional torque. OBJECTIVE: To introduce a new thoracic pedicle screw trajectory which maximizes engagement with denser bone. SUMMARY OF BACKGROUND DATA: Cortical bone trajectory (CBT) which maximizes the thread contact with cortical bone provides enhanced screw purchase. Despite the increased use of CBT screws in the lumbar spine, no study has yet reported the insertional technique for thoracic CBT. METHODS: First, the computed tomography scans of 50 adults were studied for morphometric measurement of lower thoracic CBT. The starting point was determined to be the intersection of the lateral two thirds of the superior articular process and the inferior border of the transverse process. The trajectory was straight forward in the axial plane angulated cranially targeting the posterior third of the superior endplate. The maximum diameter, length, and the cephalad angle were investigated. Next, the insertional torque of pedicle screws using this new technique was measured and compared with that of the traditional technique on 24 cadaveric thoracic vertebrae. RESULTS: All morphometric parameters of thoracic CBT increased from T9 to T12 (the mean diameter: from 5.8 mm at T9 to 8.5 mm at T12; the length: from 29.7 mm at T9 to 32.0 mm at T12; and the cephalad angle: from 21.4 degrees at T9 to 27.6 degrees at T12). The mean maximum insertional torque of CBT screws and traditional screws were 1.02±0.25 and 0.66±0.15 Nm, respectively. The new technique demonstrated average 53.8% higher torque than the traditional technique (P<0.01). CONCLUSIONS: The detailed morphometric measurement and favorable screw fixation stability of thoracic CBT are reported. The insertional torque using thoracic CBT technique was 53.8% higher than that of the traditional technique.

Nonviral gene transfer of human hepatocyte growth factor improves streptozotocin-induced diabetic neuropathy in rats.

Peripheral neuropathy is common and ultimately accounts for significant morbidity in diabetes. Recently, several neurotrophic factors have been used to prevent progression of diabetic neuropathy. In this study, we gave repeated intramuscular injections of the human hepatocyte growth factor (HGF) gene percutaneously, using liposomes containing the hemagglutinating virus of Japan (HVJ), to examine therapeutic efficacy of nonviral gene transfer of HGF for experimental diabetic sensorimotor neuropathy in rats. Experimental diabetes induced by intraperitoneal injection of streptozotocin resulted in a marked tactile allodynia (but not in a thermal hyperalgesia), in a reduction of both the conduction velocity and the amplitude, and in a decreased laser Doppler flux of the nerve and the muscle at 6 weeks after the induction. All these changes were significantly reversed by repeated gene transfer of HGF. Furthermore, we analyzed the density of endoneurial capillaries and morphometrical changes of the nerve. The density of endoneurial capillaries, disclosing marked reduction in diabetic rats, was also reversed significantly by repeated gene transfer of HGF; however, no considerable differences were observed morphometrically in either myelinated or unmyelinated axons. These results suggest that nonviral HVJ liposome-mediated gene transfer of human HGF has potential for the safe effective treatment of diabetic sensorimotor neuropathy.

Upregulated expression of inducible nitric oxide synthase plays a key role in early apoptosis after anterior cruciate ligament injury.

The reason that the anterior cruciate ligament (ACL) has a very poor healing potential after injury is not well understood. In this study, we investigated the role of nitric oxide (NO) in the apoptotic cell death of ACL cells using a rabbit model and in vitro cell culture. The apoptosis of ACL cells in vivo was analyzed by TUNEL assay and electron microscopy. NO synthase (NOS) expression was observed by immunohistochemical analysis. ACL cells were cultured and the susceptibility to NO-induced apoptosis was tested. Inducible NOS (iNOS) expression after treatment with cytokines was examined by immunohistochemical and RT-PCR analyses. Mitogen-activated protein kinase (MAPK) inhibitors were used for the analysis of downstream signals. A significant number of apoptotic cells were observed on days 1 to 3 after injury; the apoptotic rate returned to the control level by day 7. Upregulation of iNOS in the ACL remnant was observed at day 1. Intraarticular injection of NOS inhibitor suppressed the apoptotic rate. Isolated ACL cells showed much higher susceptibility to NO-induced apoptosis than did medial collateral ligament cells. IL-1beta stimulated ACL cells to upregulate iNOS mRNA and increase NO production. p38 MAPK inhibitor decreased NO-induced apoptosis. Rapid iNOS induction after injury contributes to the high apoptotic rate of ACL cells, and this may partly account for the poor healing capacity of this ligament. iNOS and NO production is suggested to be stimulated by IL-1beta, and NO activates the p38 MAPK pathway and triggers an apoptotic signal in ACL cells.

Tissue engineering of articular cartilage with autologous cultured adipose tissue-derived stromal cells using atelocollagen honeycomb-shaped scaffold with a membrane sealing in rabbits.

Adipose tissue derived stromal cells (ATSCs), which were isolated from adipose tissue of rabbit, have shown to possess multipotential, that is, they differentiate into osteoblasts and adipocytes in plate-culturing and into chondrocytes in an established aggregate culture using defined differentiation-inductive medium. The aim of this study was to evaluate the utility of ATSCs in tissue engineering procedures for repair of articular cartilage-defects using the atelocollagen honeycomb-shaped scaffold with a membrane sealing (ACHMS-scaffold). We intended to repair full-thickness articular cartilage defects in rabbit knees using autologously cultured ATSCs embedded in the ACHMS-scaffold. ATSCs were incubated within the ACHMS-scaffold to allow a high density and three-dimensional culture with control medium. An articular cartilage defect was created on the patellar groove of the femur, and the defect was filled with the ATSCs-containing ACHMS-scaffold, ACHMS-scaffold alone, or empty (control). Twelve weeks after the operation, the histological analyses showed that only the defects treated with the ATSCs-containing ACHMS-scaffold were filled with reparative hyaline cartilage, highly expressed Type II collagen. These results indicate that transplantation of autologous ATSCs-containing ACHMS-scaffold is effective in repairing articular cartilage defects.