Bone Repair with Differentiated Osteoblasts from Adipose-derived Stem Cells in Hydroxyapatite/Tricalcium Phosphate In vivo.

BACKGROUND: Recently, tissue engineering has developed approaches for repair and restoration of damaged skeletal system based on different scaffolds and cells. This study evaluated the ability of differentiated osteoblasts from adipose-derived stem cells (ADSCs) seeded into hydroxyapatite/tricalcium phosphate (HA-TCP) to repair bone. METHODS: In this study, ADSCs of 6 canines were seeded in HA-TCP and differentiated into osteoblasts in osteogenic medium in vitro and bone markers evaluated by reverse transcription polymerase chain reaction (RT-PCR). Scanning electron microscopy (SEM) was applied for detection of cells in the pores of scaffold. HA-TCP with differentiated cells as the test group and without cells as the cell-free group were implanted in separate defected sites of canine's tibia. After 8 weeks, specimens were evaluated by histological, immunohistochemical methods, and densitometry test. The data were analyzed using the SPSS 18 version software. RESULTS: The expression of Type I collagen and osteocalcin genes in differentiated cells were indicated by RT-PCR. SEM results revealed the adhesion of cells in scaffold pores. Formation of trabecular bone confirmed by histological sections that revealed the thickness of bone trabecular was more in the test group. Production of osteopontin in extracellular matrix was indicated in both groups. Densitometry method indicated that strength in the test group was similar to cell-free group and natural bone (P > 0.05). CONCLUSIONS: This research suggests that ADSCs-derived osteoblasts in HA-TCP could be used for bone tissue engineering and repairing.

An animal model study for bone repair with encapsulated differentiated osteoblasts from adipose-derived stem cells in alginate.

OBJECTIVES: Adipose derived stem cells (ADSCs) can be engineered to express bone specific markers. The aim of this study is to evaluate repairing tibia in animal model with differentiated osteoblasts from autologous ADSCs in alginate scaffold. MATERIALS AND METHODS: In this study, 6 canine's ADSCs were encapsulated in alginate and differentiated into osteoblasts. Alkaline phosphatase assay (ALP) and RT-PCR method were applied to confirm the osteogenic induction. Then, encapsulated differentiated cells (group 1) and cell-free alginate (group 2) implanted in defected part of dog's tibia for 4 and 8 weeks. Regenerated tissues and compressive strength of samples were evaluated by histological and Immunohistochemical (IHC) methods and Tensometer Universal Machine. RESULTS: Our results showed that ADSCs were differentiated into osteoblasts in vitro, and type I collagen and osteocalcin genes expression in differentiated osteoblasts was proved by RT-PCR. In group 2, ossification and thickness of trabecula were low compared to group 1, and in both groups woven bone was observed instead of control group's compact bone. Considering time, we found bone trabeculae regression and ossification reduction after 8 weeks compared with 4 weeks in group 2, but in group 1 bone formation was increased in 8 weeks. Presence of differentiated cells caused significantly more compressive strength in comparison with group 2 (P-value ≤0.05). CONCLUSION: This research showed that engineering bone from differentiated adipose-derived stem cells, encapsulated in alginate can repair tibia defects.

Comparison between Chondrogenic Markers of Differentiated Chondrocytes from Adipose Derived Stem Cells and Articular Chondrocytes In Vitro.

OBJECTIVE(S): Osteoarthritis is one of the most common diseases in middle-aged population in the world. Cartilage tissue engineering (TE) has been presented as an effort to introduce the best combination of cells, biomaterial scaffolds and stimulating growth factors to produce a cartilage tissue similar to the natural articular cartilage. In this study, the chondrogenic potential of adipose derived stem cells (ADSCs) was compared with natural articular chondrocytes cultured in alginate scaffold. MATERIALS AND METHODS: Human ADSCs were obtained from subcutaneous adipose tissue and human articular chondrocytes from non-weight bearing areas of knee joints. Cells were seeded in 1.5% alginate and cultured in chondrogenic media for three weeks with and without TGFß3. The genes expression of types II and X collagens was assessed by Real Time PCR and the amount of aggrecan (AGC) and type I collagen measured by ELISA and the content of glycosaminoglycan evaluated by GAG assay. RESULTS: Our findings showed that type II collagen, GAG and AGC were expressed, in differentiated ADSCs. Meanwhile, they produced a lesser amount of types II and X collagens but more AGC, GAG and type I collagen in comparison with natural chondrocytes (NCs). CONCLUSION: Further attempt should be carried out to optimize achieving type II collagen in DCs, as much as, natural articular chondrocytes and decline of the production of type I collagen in order to provide efficient hyaline cartilage after chondrogenic induction, prior to the usage of harvested tissues in clinical trials.

Comparison of Phenotypic Characterization between Differentiated Osteoblasts from Stem Cells and Calvaria Osteoblasts In vitro.

BACKGROUND: Characteristics of differentiated osteoblasts from adipose derived stem cells (ADSCs) in compared with isolated osteoblasts from normal bone such as calvaria are unknown. The aim of this study was determination and comparison of phenotypic characterization between differentiated osteoblasts from stem cells and calvaria osteoblasts in vitro. METHODS: In this study, mesenchymal stem cells were isolated from adipose tissue of human by enzymatic digestion and were differentiated into osteoblasts using osteogenic medium. Characteristics of these cells at first, second, third and fourth weeks were comprised with calvaria osteoblasts that were isolated from human calvaria by explanation culture method. To screen the characteristics of both calvaria and the differentiated osteoblasts, we used western blot to identify protein levels, von Kossa staining for mineral matrix detection and alkaline phosphatase (ALP) assay kit (Sigma) for ALP activity measurement. Difference between calvaria and differentiated osteoblast cells were analyzed by one-way ANOVA and P < 0.05 was considered as statistically significant. RESULTS: Alkaline phosphatase activity, collagen and mineral material production in differentiated osteoblasts at third week were more significantly than calvaria cells (P < 0.05). Our results indicated that there was no significant different in osteocalcin (OC) production between differentiated osteoblast at first, second and third weeks and calvaria cells but declined at fourth week (P < 0.05). CONCLUSIONS: Our survey showed that cellular traits of differentiated osteoblasts presented better than calvaria osteoblasts in vitro conditions. Therefore, we suggest that ADSCs could be used in next studies for bone tissue engineering.

Large retroperitoneal paraganglioma concurrent with periampullary adenocarcinoma.

Paragangliomas are tumors that originate from extra-adrenal medullary neural crest derivatives. They are rarely located in retroperitoneal space. These tumors are often discovered incidentally during imaging studies performed for other reasons. Periampullary cancers include adenocarcinomas arising from the pancreas, ampulla of Vater, duodenum or distal common bile duct. The exact site of origin of periampullary tumors is often difficult to ascertain pre-operatively. We report the case of a patient who had a retroperitoneal non-functional paraganglioma, concurrent with periampullary adenocarcinoma. An 81-year-old woman was admitted with progressive abdominal fullness. There was an upper paramedian, left sided, large, palpable mass on the physical examination. Laboratory investigations showed an increase in liver enzyme levels. On abdominal computed tomography the patient found to have a large retroperitoneal mass and dilation in biliary tract, which was confirmed by magnetic resonance cholangiopancreatography. She had a tumoral papi in Endoscopic Retrograde cholangiopancreatography. Which biopsy revealed adenocarcinoma. She underwent surgery for excision of abdominal mass and pancreaticoduodenectomy. And pathologic study showed paraganglioma. This is the first ever reported case of concurrent paraganglioma and periampullary adenocarcinoma.

An animal model study for repair of tracheal defects with autologous stem cells and differentiated chondrocytes from adipose-derived stem cells.

BACKGROUND: Stenosis of trachea with mucosal and cartilage lesions is a challenging problem in tracheal surgery. Owing to ease of harvest and abundance, adipose-derived stem cells (ADSCs) are attractive and increasingly used in tissue engineering. The aim of this study was to evaluate the repair of trachea with autologous stem cells and differentiated chondrocytes from adipose-derived stem cells in an animal model. METHODS AND MATERIAL: Six canine ADSCs were isolated and proliferated in monolayer culture and CD44; CD90 markers were investigated by flow cytometry. ADSCs were seeded in alginate beads and were differentiated into chondrocytes by TGF-ß3. Cartilage-specific markers with reverse transcriptase polymerase chain reaction were demonstrated in differentiated cells. These differentiated cells and stem cells in alginate scaffold were separately transferred to a defect created in canine's trachea. After 8 weeks, the healing and cartilage formation in the trachea was evaluated by histological methods. RESULTS: We identified formed cartilage pieces and chondrocytes with lacuna and extracellular matrix in defects implanted with differentiated cells, but in other groups, staining of the sections did not show the presence of cartilage in the engineered tracheal wall. CONCLUSION: We showed that cartilage- engineered from differentiated adipose-derived stem cells in alginate biodegradable scaffold could repair tracheal cartilage defects.

A comparison between subpleural patient-controlled analgesia by bupivacaine and intermittent analgesia in post-operative thoracotomy: A double-blind randomized clinical trial.

BACKGROUND: The efficacy of subpleural analgesia to reduce postoperative pain intensity in patients after lateral thoracotomy is controversial. In this study, we demonstrated the efficacy of two types of subpleural analgesia. METHODS: This prospective, controlled, randomized, double-blind trial was performed in Department of Thoracic Surgery of Alzahra Hospital associated with Isfahan University of Medical Sciences from June 2009 until August 2010. After posterolateral thoracotomy and admission to the ICU, patients were randomly assigned into two groups of subpleural patient-controlled analgesia (SPCA) (0.02 cc/kg/h of 0.5% bupivacaine) and subpleural intermittent analgesia (SIA) (0.1cc/kg/6h of 0.5% bupivacaine). The data regarding age, sex, visual analog scale (VAS) (at 8, 16 and 24 hours after initiation of analgesia), morphine consumption, systemic adverse effects, length of ICU and hospital stay, complications, public health service (PHS) criteria, and cost was recorded. Data was analyzed by Mann-Whitney U-test, repeated measured test, chi-square test and the Fisher's exact test. A p < 0.05 was considered significant. RESULTS: The study population consisted of 90 patients. There were no significant differences in sex, age, weight, intraoperative analgesics, duration of one-lung ventilation, and adverse effects between the SPCA and SIA groups. Although pain scores were significantly reduced at 16 hours after the first subpleural instillation of bupivacaine 0.5% with patient-controlled analgesia, comparison between mean pain scores in the two groups at 8 and 24 hours after the first subpleural instillation of bupivacaine 0.5% revealed no significant difference. In addition, no significant difference was found in VAS scores at the three evaluated times (p < 0.05). CONCLUSIONS: Optimal use of SPCA bupivacaine for postoperative pain treatment is more effective in pain reduction than SIA bupivacaine. The consumption rate of opioid and bupivacaine was also decreased in SPCA group.

Pneumatic balloon dilation therapy is as effective as esophagomyotomy for achalasia.

For many years different treatments have been used for achalasia. However, esophagomyotomy (ESM) and pneumatic balloon dilation (PBD) have been considered the treatments of choice. Despite new research, some controversies still exist. We compared patients who underwent open ESM (n=19) with those who underwent PBD (n=45). Data on age, gender, pre- and postprocedure symptoms, clinical manifestations at the time of research, clinical relief, type of surgery, and costs were collected via questionnaire. Open ESMs were performed by two expert surgeons, and PBDs were performed by one gastroenterologist. There was no significant difference in clinical symptoms and in patient satisfaction between the groups before and after the procedures except for chest pain. Clinical relief status (excellent, good, moderate, or poor) was comparable (26%, 42%, 15%, 15% for open ESM group and 40%, 20%, 24%, 15% for PBD group). Postprocedure complications were not significantly different between the two groups. Clinical rates of relapse for open ESM and PBD groups were 38.6% and 25%, respectively. There were no serious complications. There was no significant difference between the clinical outcomes of the two methods of achalasia treatment. Considering other important factors such as a shorter period of hospitalization, fewer sick days off, risk of general anesthesia, and cosmetic sequels, PBD is preferable for the majority of patients.