A comparative study of aggrecan synthesis between natural articular chondrocytes and differentiated chondrocytes from adipose derived stem cells in 3D culture

Introduction: The main obstacle for tissue engineering is to find the most appropriate cell which is able to produce extracellular matrix (ECM) similar or better than natural chondrocytes in vitro. This study compared aggrecan synthesis′s potential between differentiated chondrocytes (DCs) from adipose-derived stem cells (ADSCs) and natural articular chondrocytes (NCs) in 3D culture in vitro. Materials and Methods: Human ADSCs were isolated from sub-cutaneous adipose tissue and then the surface markers including CD 14, 45 CD105, CD90, CD44 were analyzed by flow cytometry. Also human articular chondrocytes were yielded of non-weight bearing area of Knee cartilage. Both types of the cells were encapsulated in alginate scaffolds and cultured in chondrogenic medium with and without TGFβ3 for 3 weeks. Then the extent of aggercan (AGC) production was evaluated by ELISA on days 14 and 21. Results: Our findings indicated that differentiated chondrocytes (DCs) with and without TGFβ3 synthesized more AGC than natural chondrocytes (NCs) on day 14. But DCs without TGFβ3 had higher production than other groups on day 21. Application of TGFβ3 resulted in an increase of amount of AGC in DCs on day 14 but a decrease on day 21 than same group. Conclusion: Since, aggrecan is an important chondrogenic marker, it was concluded that ADSCs can be possible reliable alternative cell source for cartilage tissue engineering in future

شیوع آرکوئت فورامن در بیماران مراجعه کننده به بخش اورژانس؛ یک مطالعه اپیدمیولوژیک

Introduction: Arcuate foramen anomaly is an anatomic variation in the first thoracic vertebrae that forms a space for passing of vertebral artery. Considering the role of this bony bridge in migraine development and its effect on prevalence of various types of pain in head, shoulder, and neck areas, as well as its importance in screw placement on the lateral mass of atlas, the present study evaluated the prevalence of this anomaly among those presented to the emergency department (ED) of Poursina Hospital, Rasht, Iran. Methods: The present study is a cross-sectional one, carried out on patients over 18 years of age that visited ED of Poursina Hospital, Rasht, Iran, in 2015 and underwent lateral neck radiography using census method. Data of the patients were gathered using their medical profile or by asking them on the phone, and recorded in a pre-designed checklist. Data were analyzed with the aid of SPSS version 24 and chi-square and independent t-test statistical tests. P-values less than 0.05 were considered as significant. Results: 1146 patients with the mean age of 37.95 ± 15.88 years were studied (69.2% male). Mean age was 36.05 ± 15.26 years among men and 14.21 ± 16.44 years among women (p < 0.001). The overall prevalence of arcuate foramen was calculated to be 9.6% [95% confidence interval 7.89 – 11.30; 110 patients (74.5% male)]. This anomaly had a prevalence of 10.34% in men and 7.93% in women (p > 0.05). Conclusion: Based on the results of the present study, the prevalence of arcuate foramen anomaly was estimated to be 9.6% in Guilan province using lateral neck radiography. Its prevalence rate was 10.34% in men and 7.93% in women. Considering this prevalence rate, presence of this anomaly should be considered as a possibility in patients with complaints of head, shoulder, and neck pain. مقدمه: آنومالی آرکوئت فورامن یک واریاسیون آناتومیکی در مهره اول گردنی می باشد که محلی را برای عبور شریان ورتبرال ایجاد می کند. با توجه به نقشی که این پل استخوانی در پیشرفت میگرن و تاثیر آن در بروز انواع دردهای مربوط به نواحی سر، شانه و گردن و همچنین اهمیتی که در زمان تعبیه پیچ در توده جانبی اطلس دارد، پژوهش حاضر به بررسی میزان شیوع این آنومالی در مراجعه کنندگان به بخش اورژانس بیمارستان پورسینا رشت پرداخته است. روش کار: پژوهش حاضر یک مطالعه مقطعی آینده نگر می باشد که بر روی بیماران بالای 18 سال که در سال 1394 به بخش اورژانس بیمارستان پورسینا رشت مراجعه کرده بودند و به هر دلیلی برای آنها گرافی لترال گردن درخواست شده بود به روش در دسترس صورت پذیرفت. داده های بیماران از طریق بررسی پرونده بالینی، پرسش بصورت تلفنی و یا با استفاده از چک لیستی که از پیش طراحی شده بود جمع آوری و ثبت گردیدند. با کمک نرم افزار آماری SPSS نسخه 24 و آزمون های آماری کای دو و تی تست مستقل داده ها مورد تجزیه آماری قرار گرفتند. مقدار p کمتر از 05/0 معنی دار در نظر گرفته شد. يافته ها: در مجموع 1146 بیمار با میانگین سنی 88/15±95/37 سال مورد مطالعه قرار گرفتند (2/69 درصد مرد). میانگین سنی مردان 26/15±05/36 سال و زنان 44/16±21/14 سال بود (001/0>p ). میزان شیوع کلی آرکوئت فورامن، 6/9 درصد (110 بیمار) با فاصله اطمینان 30/11- 89/7 بدست آمد (5/74 درصد مرد). این آنومالی در مردان شیوعی معادل 34/10 درصد و در زنان 93/7 درصد داشت (05/0<p ). نتيجه گيری: بر اساس نتایج مطالعه حاضر شیوع آنومالی آرکوئت فورامن با کمک گرافی لترال گردن در استان گیلان 6/9 درصد بدست آمد. این میزان شیوع در مردان 34/10 و در زنان 93/7 درصد بود. با توجه به این میزان شیوع، وجود این آنومالی باید به عنوان یک گزینه در بیماران با شکایت درد در نواحی سر،  شانه و گردن مد نظر قرار گیرد

Prevalence of Arcuate Foramen among Emergency Department Visitors; an Epidemiologic Study

Introduction: Arcuate foramen anomaly is an anatomic variation in the first thoracic vertebrae that forms a space for passing of vertebral artery. Considering the role of this bony bridge in migraine development and its effect on prevalence of various types of pain in head, shoulder, and neck areas, as well as its importance in screw placement on the lateral mass of atlas, the present study evaluated the prevalence of this anomaly among those presented to the emergency department (ED) of Poursina Hospital, Rasht, Iran. Methods: The present study is a cross-sectional one, carried out on patients over 18 years of age that visited ED of Poursina Hospital, Rasht, Iran, in 2015 and underwent lateral neck radiography using census method. Data of the patients were gathered using their medical profile or by asking them on the phone, and recorded in a pre-designed checklist. Data were analyzed with the aid of SPSS version 24 and chi-square and independent t-test statistical tests. P-values less than 0.05 were considered as significant. Results: 1146 patients with the mean age of 37.95 ± 15.88 years were studied (69.2% male). Mean age was 36.05 ± 15.26 years among men and 14.21 ± 16.44 years among women (p < 0.001). The overall prevalence of arcuate foramen was calculated to be 9.6% [95% confidence interval 7.89 – 11.30; 110 patients (74.5% male)]. This anomaly had a prevalence of 10.34% in men and 7.93% in women (p > 0.05). Conclusion: Based on the results of the present study, the prevalence of arcuate foramen anomaly was estimated to be 9.6% in Guilan province using lateral neck radiography. Its prevalence rate was 10.34% in men and 7.93% in women. Considering this prevalence rate, presence of this anomaly should be considered as a possibility in patients with complaints of head, shoulder, and neck pain

Cryopreservation of Rat Bone Marrow Derived Mesenchymal Stem Cells by Two Conventional and Open-pulled Straw Vitrification Methods

Objective: Mesenchymal stem cells (MSCs) are obtained from a variety of sources, mainlythe bone marrow. These cells have a great potential for clinical research, however they cannotstay alive for long periods in culture. The aim of this study is to determine whether vitrificationcan be a useful freezing method for the storage of MSCs.Materials and Methods: Mesenchymal stem cells were isolated from rat bone marrow basedon their capacity to adhere to plastic culture surfaces. MSCs were cryopreserved using boththe vitrification method and open-pulled straw (OPS) vitrification and stored in liquid nitrogenwith ethylene glycol ficoll (EFS) as a cryoprotectant for two months. The morphology andviability of thawed MSCs were evaluated by trypan blue staining. Furthermore, pre and postcryopreserved MSCs were induced to osteocyte and adipocyte with corresponding osteogenicand adipogenic medium.Results: After thawing, the viability rates were 81.33% ± 6.83 for the vitrification method and80.83% ± 6.4 for OPS vitrification, while the values in the pre-vitrification control group were88.16% ± 6.3 (Mean ± SD, n = 6). Post-cryopreserved cells from both the vitrification methodand OPS vitrification also had a similar cellular morphology and colony-formation that wasindistinguishable from non-vitrified fresh MSCs. In addition, the resuscitated cells cultured ininduction medium showed osteogenesis. Mineral production and deposition was detectableby alizarine red S staining. Moreover, by applying an adipogenic differentiation condition,both pre and post cryopreserved cells differentiated into adipocyte and lipid vacuole accumulationthat was stained by oil red O.Conclusion: Vitrification is a reliable and effective method for the cryopreservation of MSCs

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 TGFβ3. 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