Novel Autologous Therapy for Long-Gap Peripheral Nerve Injury Using Human Sk-SCs

Losses in vital functions of the somatic motor and sensory nervous system are induced by severe long-gap peripheral nerve transection injury. In such cases, autologous nerve grafts are the gold standard treatment, despite the unavoidable sacrifice of other healthy functions, whereas the prognosis is not always favorable. Here, we use human skeletal muscle-derived stem cells (Sk-SCs) to reconstitute the function after long nerve-gap injury. Muscles samples were obtained from the amputated legs from 9 patients following unforeseen accidents. The Sk-SCs were isolated using conditioned collagenase solution, and sorted as CD34+/45- (Sk-34) and CD34-/45-/29+ (Sk-DN/29+) cells. Cells were separately cultured/expanded under optimal conditions for 2 weeks, then injected into the athymic nude mice sciatic nerve long-gap model (7-mm) bridging an acellular conduit. After 8-12 weeks, active cell engraftment was observed only in the Sk-34 cell transplanted group, showing preferential differentiation into Schwann cells and perineurial/endoneurial cells, as well as formation of the myelin sheath and perineurium/endoneurium surrounding regenerated axons, resulted in 87% of numerical recovery. Differentiation into vascular cell lineage (pericyte and endothelial cells) were also observed. A significant tetanic tension recovery (over 90%) of downstream muscles following electrical stimulation of the sciatic nerve (at upper portion of the gap) was also achieved. In contrast, Sk-DN/29+ cells were completely eliminated during the first 4 weeks, but relatively higher numerical (83% vs. 41% in axon) and functional (80% vs. 60% in tetanus) recovery than control were observed. Noteworthy, significant increase in the formation of vascular networks in the conduit during the early stage (first 2 weeks) of recovery was observed in both groups with the expression of key factors (mRNA and protein levels), suggesting the paracrine effects to angiogenesis. These results suggested that the human Sk-SCs may be a practical source for autologous stem cell therapy following severe peripheral nerve injury

Implementasi manajemen sarana dan prasarana dalam meningkatkan mutu pendidikan pada madrasah tsanawiyah negeri (MTSN) Rantauprapat Kabupaten Labuhanbatu

Penelitian ini bertujuan untuk mengetahui perencanaan, pengorganisasian, pelaksanaan, pengawasan manajemen sarana dan prasarana dalam meningkatkan mutu pendidikan pada Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu. Penelitian ini dilakukan dengan menggunakan pendekatan kualitatif, Teknik pengumpulan data dalam penelitian ini menggunakan observasi, wawancara dan dokumen. Data yang didapat kemudian dianalisis dengan menggunakan analisis data kualitatif yang terdiri dari: (a).reduksi data, (b).penyajian data, dan (c) penarikan kesimpulan. Temuan penelitian: (1) perencanaan manajemen sarana dan prasarana di Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu terlebih dahulu dilakukan analisis kebutuhan riil baik yang menyangkut kebutuhan administrasi maupun pendukung kegiatan proses pembelajaran, seperti ruang kelas, moubilair, dan lain sebagainya. Yang melibatkan: Kepala Madrasah, KTU, bendahara, PKM, dan bahkan utusan dari komite sekolah. (2).Pengorganisasian manajemen sarana dan prasarana pada Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu dilakukan berdasarkan rumpun (kelompok) dari setiap jenis sarana itu sendiri, misalnya: bangunan fisik, moubilair, ATK, lingkungan, dan lain sebagainya yang kesemuanya itu di arsiparis berdasarkan ketentuan yang berlaku. (3) Pelaksanaan manajemen sarana dan prasarana di Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu berjalan baik dan lancar. Pelaksanaannya masing-masing pihak bekerja sesuai job/pekerjaan masing-masing dan sesuai kepentingannya, sehingga sistem kerja tidak ada tumpang tindih antara satu sama lain. Dan pertanggung jawabannya langsung kepada Kepala madrasah MTsN Rantauprapat walaupun tetap di bawah koordinasi PKM sarana dan prasarana. (4).Pengawasan manajemen sarana dan prasarana pada Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu dilakukan dengan cara: a) Pengawasan rutin setiap harinya yang dilakukan oleh PKM sarana jika menyangkut persoalan sarana pendukung pembelajaran, sedangkan yang menyangkut administrasi dilakukan oleh KTU. b) Secara berkala yakni setiap 6 (enam) bulan sekali diadakan rapat evaluasi tentang keadaan sarana dan prasarana. (5) Terkait dengan evaluasi diketahui bahwa sarana dan prasarana di Madrasah Tsanawiyah Negeri (MTsN) Rantauprapat Kabupaten Labuhanbatu sudah terpenuhi dan sesuai dengan standar pendidikan nasional.

Cardiomyocyte Formation by Skeletal Muscle-Derived Multi-Myogenic Stem Cells after Transplantation into Infarcted Myocardium

BACKGROUND: Cellular cardiomyoplasty for myocardial infarction has been developed using various cell types. However, complete differentiation and/or trans-differentiation into cardiomyocytes have never occurred in these transplant studies, whereas functional contributions were reported. METHODS AND RESULTS: Skeletal muscle interstitium-derived CD34(+)/CD45(-) (Sk-34) cells were purified from green fluorescent protein transgenic mice by flowcytometory. Cardiac differentiation of Sk-34 cells was examined by in vitro clonal culture and co-culture with embryonic cardiomyocytes, and in vivo transplantation into a nude rat myocardial infarction (MI) model (left ventricle). Lower relative expression of cardiomyogenic transcription factors, such as GATA-4, Nkx2-5, Isl-1, Mef2 and Hand2, was seen in clonal cell culture. However, vigorous expression of these factors was seen on co-culture with embryonic cardiomyocytes, together with formation of gap-junctions and synchronous contraction following sphere-like colony formation. At 4 weeks after transplantation of freshly isolated Sk-34 cells, donor cells exhibited typical cardiomyocyte structure with formation of gap-junctions, as well as intercalated discs and desmosomes, between donor and recipient and/or donor and donor cells. Fluorescence in situ hybridization (FISH) analysis detecting the rat and mouse genomic DNA and immunoelectron microscopy using anti-GFP revealed donor-derived cells. Transplanted Sk-34 cells were incorporated into infarcted portions of recipient muscles and contributed to cardiac reconstitution. Significant improvement in left ventricular function, as evaluated by transthoracic echocardiography and micro-tip conductance catheter, was also observed. CONCLUSIONS AND SIGNIFICANCE: Skeletal muscle-derived multipotent Sk-34 cells that can give rise to skeletal and smooth muscle cells as reported previously, also give rise to cardiac muscle cells as multi-myogenic stem cells, and thus are a potential source for practical cellular cardiomyoplasty

Qualitative alteration of peripheral motor system begins prior to appearance of typical sarcopenia syndrome in middle-aged rats. Front Aging Neurosci

Qualitative changes in the peripheral motor system were examined using young, adult, middle-aged, and old-aged rats in order to assess before and after the appearance of sarcopenia symptoms. Significant loss of muscle mass and strength, and slow-type fiber grouping with a loss of innervated nerve fibers were used as typical markers of sarcopenia. Dynamic twitch and tetanus tension and evoked electromyogram (EEMG) were measured via electrical stimulation through the sciatic nerve under anesthesia using our force-distance transducer system before and after sciatectomy. Digital and analog data sampling was performed and shortening and relaxing velocity of serial twitches was calculated with tension force. Muscle tenderness in passive stretching was also measured as stretch absorption ability, associated with histological quantitation of muscle connective tissues. The results indicated the validity of the present model, in which old-aged rats clearly showed the typical signs of sarcopenia, specifically in the fast-type plantaris muscles, while the slow-type soleus showed relatively mild syndromes. These observations suggest the following qualitative alterations as the pathophysiological mechanism of sarcopenia: (1) reduction of shortening and relaxing velocity of twitch; (2) decline of muscle tenderness following an increase in the connective tissue component; (3) impaired recruitment of motor units (MUs) (sudden depression of tetanic force and EEMG) in higher stimulation frequencies over 50-60 Hz; and (4) easy fatigability in the neuromuscular junctions. These findings are likely to be closely related to significant losses in fast-type MUs, muscle strength and contraction velocity, which could be a causative factor in falls in the elderly. Importantly, some of these symptoms began in middle-aged rats that showed no other signs of sarcopenia. Thus, prevention should be started in middle age that could be retained relatively higher movement ability

Clinical Features and Outcomes of Bilateral Atypical Femoral Fractures

Bilateral atypical femoral fractures (AFFs) are relatively rare. In this report, we retrospectively researched clinical features and outcomes of bilateral AFFs treated at our institution. We previously treated 4 patients (8 limbs) with intramedullary nailing for complete AFFs (6 limbs) and incomplete AFFs (2 limbs). The mean age at the first operation was 53.3 years, and all patients were female. Of the 4 patients, two had breast cancer, and another two had systemic lupus erythematosus. Three of them received bisphosphonates, and 2 received denosumab, proton pump inhibitor, or glucocorticoid therapy. Only 2 of 6 cases of incomplete AFFs had prodromal pain before progressing to complete fracture. The mean interval from the first surgery to contralateral fracture or prophylactic surgery was 16 months. Radiographically, complete bone union was achieved in 6 limbs. However, a small gap at the lateral cortex of fracture site remained in 2 limbs. Finally, all of the patients were pain-free and able to walk without a cane. It is absolutely necessary to confirm contralateral femoral conditions; however, prediction of progression to complete fracture based solely on prodromal pain was difficult. Therefore, we should advise patients about the danger of progression to complete AFFs even if they are asymptomatic, and a prophylactic surgery should be performed after obtaining informed consent

Reconstitution of the complete rupture in musculotendinous junction using skeletal muscle-derived multipotent stem cell sheet-pellets as a “bio-bond”

Background. Significant and/or complete rupture in the musculotendinous junction (MTJ) is a challenging lesion to treat because of the lack of reliable suture methods. Skeletal muscle-derived multipotent stem cell (Sk-MSC) sheet-pellets, which are able to reconstitute peripheral nerve and muscular/vascular tissues with robust connective tissue networks, have been applied as a “bio-bond”. Methods. Sk-MSC sheet-pellets, derived from GFP transgenic-mice after 7 days of expansion culture, were detached with EDTA to maintain cell–cell connections. A completely ruptured MTJ model was prepared in the right tibialis anterior (TA) of the recipient mice, and was covered with sheet-pellets. The left side was preserved as a contralateral control. The control group received the same amount of the cell-free medium. The sheet-pellet transplantation (SP) group was further divided into two groups; as the short term (4–8 weeks) and long term (14–18 weeks) recovery group. At each time point after transplantation, tetanic tension output was measured through the electrical stimulation of the sciatic nerve. The behavior of engrafted GFP+ tissues and cells was analyzed by fluorescence immunohistochemistry. Results. The SP short term recovery group showed average 64% recovery of muscle mass, and 36% recovery of tetanic tension output relative to the contralateral side. Then, the SP long term recovery group showed increased recovery of average muscle mass (77%) and tetanic tension output (49%). However, the control group showed no recovery of continuity between muscle and tendon, and demonstrated increased muscle atrophy, with coalescence to the tibia during 4–8 weeks after operation. Histological evidence also supported the above functional recovery of SP group. Engrafted Sk-MSCs primarily formed the connective tissues and muscle fibers, including nerve-vascular networks, and bridged the ruptured tendon–muscle fiber units, with differentiation into skeletal muscle cells, Schwann cells, vascular smooth muscle, and endothelial cells. Discussion. This bridging capacity between tendon and muscle fibers of the Sk-MSC sheet-pellet, as a “bio-bond,” represents a possible treatment for various MTJ ruptures following surgery

RESEARCH Open Access

Clinical results of a surgical technique using endobuttons for complete tendon tear of pectoralis major muscle: report of five case