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"Pendahuluan: Cedera saraf tepi dapat menyebabkan cacat fungsional yang parah. Denervasi yang berkepanjangan menyebabkan perubahan permanen pada organ target sekalipun dilakukan operasi dengan teknik rekonstruksi saraf bedah mikro. Oleh karena itu, banyak penelitian regeneratif telah dilakukan untuk meningkatkan luaran fungsional setelah tindakan rekonstruksi saraf pada cedera saraf tepi.Metode: Dilakukan transeksi komplet saraf iskiadikus tungkai belakang kanan pada 20 tikus Sprague-Dawley jantan. Setiap ujung saraf dijahit ke otot terdekat untuk mencegah pertumbuhan saraf spontan. Setelah 3 minggu denervasi, dilakukan penyambungan saraf dan implantasi sel punca mesenkimal (SPM) ke neuromuscular junction (NMJ) otot gastroknemius pada kelompok perlakuan (n=10). Sepuluh tikus lainnya (kelompok kontrol) menerima plasebo (NaCL). Delapan minggu setelah penyambungan saraf, seluruh sampel dinilai luaran fungsional dengan analisis walking track dan studi neurofisiologi yang dilakukan sebelum terminasi. Berat otot basah dievaluasi kemudian dilakukan pemeriksaan histomorfometri.Hasil dan Diskusi: Denervasi saraf iskiadikus selama tiga minggu menghasilkan model cedera saraf perifer kronis yang secara klinis mengakibatkan gangguan berjalan dan secara histologis menyebabkan degenerasi otot gastroknemius. Tidak ada perbedaan analisis walking track, compound muscle action potential (CMAP) dan berat otot basah antara kedua kelompok. Namun, penyambungan saraf yang dikombinasikan dengan implantasi SPM memberikan gambaran preservasi NMJ dan regenerasi otot yang lebih baik sebagai organ target saraf yang terbukti secara histologis dengan fragmentasi reseptor asetilkolin (AChR) yang jauh lebih rendah, jumlah dan kepadatan AChR yang lebih besar, serta diameter serat otot yang lebih besar.Kesimpulan: Implantasi SPM di NMJ berpotensi menunda degenerasi organ target dan meningkatkan regenerasi

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Introduction: Peripheral nerve injury is a devastating condition that can lead to severe functional disabilities. Despite the use of advanced microsurgical nerve reconstruction techniques, prolonged denervation causes irreversible changes in target organs. Thus, many regenerative studies have been conducted to increase functional outcomes in animal models.

Methods: Complete sciatic nerve transection was performed on the right hind limb of 20 male Sprague-Dawley rats. Each nerve end was sutured to the approximate muscle at a distance to prevent spontaneous nerve regrowth. After 3 weeks of denervation, the nerve was repaired and mesenchymal stem cells was injected directly to the gastrocnemius neuromuscular junction (NMJ) in MSCs group (n=10) and the rest of the rats (control group) received placebo (normal saline). Clinically, all sample were observed by walking track analysis and a neurophysiology study which was done before termination (8 weeks after nerve repair). Postmortem wet muscle weight was evaluated, and histological examination was performed

Result and Discussion: Three weeks sciatic nerve denervation produce the chronic peripheral nerve injury model which resulted clinically as gait disturbance and histologically as gastrocnemius muscle degeneration. There was no difference in walking track analysis, compound muscle action potential (CMAP) and muscle weight between two groups. However, the delayed nerve repair combined with stem cell implantation gives a better NMJ and muscle regeneration as the nerve target organ that proven histologically with a significantly lesser acetylcoline receptor (AChR) fragmentation, greater AChR amount and density, as well as larger muscle fiber diameter.

Conclusion: Mesenchymal stem cell direct implantation in neuromuscular junction possibly delayed the end organ degeneration and enhanced regeneration proven by a better morphometry profile in MSCs group."

"Pendahuluan: Cedera saraf tepi dapat menyebabkan cacat fungsional yang parah. Denervasi yang berkepanjangan menyebabkan perubahan permanen pada organ target sekalipun dilakukan operasi dengan teknik rekonstruksi saraf bedah mikro. Oleh karena itu, banyak penelitian regeneratif telah dilakukan untuk meningkatkan luaran fungsional setelah tindakan rekonstruksi saraf pada cedera saraf tepi.Metode: Dilakukan transeksi komplet saraf iskiadikus tungkai belakang kanan pada 20 tikus Sprague-Dawley jantan. Setiap ujung saraf dijahit ke otot terdekat untuk mencegah pertumbuhan saraf spontan. Setelah 3 minggu denervasi, dilakukan penyambungan saraf dan implantasi sel punca mesenkimal (SPM) ke neuromuscular junction (NMJ) otot gastroknemius pada kelompok perlakuan (n=10). Sepuluh tikus lainnya (kelompok kontrol) menerima plasebo (NaCL). Delapan minggu setelah penyambungan saraf, seluruh sampel dinilai luaran fungsional dengan analisis walking track dan studi neurofisiologi yang dilakukan sebelum terminasi. Berat otot basah dievaluasi kemudian dilakukan pemeriksaan histomorfometri.Hasil dan Diskusi: Denervasi saraf iskiadikus selama tiga minggu menghasilkan model cedera saraf perifer kronis yang secara klinis mengakibatkan gangguan berjalan dan secara histologis menyebabkan degenerasi otot gastroknemius. Tidak ada perbedaan analisis walking track, compound muscle action potential (CMAP) dan berat otot basah antara kedua kelompok. Namun, penyambungan saraf yang dikombinasikan dengan implantasi SPM memberikan gambaran preservasi NMJ dan regenerasi otot yang lebih baik sebagai organ target saraf yang terbukti secara histologis dengan fragmentasi reseptor asetilkolin (AChR) yang jauh lebih rendah, jumlah dan kepadatan AChR yang lebih besar, serta diameter serat otot yang lebih besar.Kesimpulan: Implantasi SPM di NMJ berpotensi menunda degenerasi organ target dan meningkatkan regenerasi

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Introduction: Peripheral nerve injury is a devastating condition that can lead to severe functional disabilities. Despite the use of advanced microsurgical nerve reconstruction techniques, prolonged denervation causes irreversible changes in target organs. Thus, many regenerative studies have been conducted to increase functional outcomes in animal models.

Methods: Complete sciatic nerve transection was performed on the right hind limb of 20 male Sprague-Dawley rats. Each nerve end was sutured to the approximate muscle at a distance to prevent spontaneous nerve regrowth. After 3 weeks of denervation, the nerve was repaired and mesenchymal stem cells was injected directly to the gastrocnemius neuromuscular junction (NMJ) in MSCs group (n=10) and the rest of the rats (control group) received placebo (normal saline). Clinically, all sample were observed by walking track analysis and a neurophysiology study which was done before termination (8 weeks after nerve repair). Postmortem wet muscle weight was evaluated, and histological examination was performed

Result and Discussion: Three weeks sciatic nerve denervation produce the chronic peripheral nerve injury model which resulted clinically as gait disturbance and histologically as gastrocnemius muscle degeneration. There was no difference in walking track analysis, compound muscle action potential (CMAP) and muscle weight between two groups. However, the delayed nerve repair combined with stem cell implantation gives a better NMJ and muscle regeneration as the nerve target organ that proven histologically with a significantly lesser acetylcoline receptor (AChR) fragmentation, greater AChR amount and density, as well as larger muscle fiber diameter.

Conclusion: Mesenchymal stem cell direct implantation in neuromuscular junction possibly delayed the end organ degeneration and enhanced regeneration proven by a better morphometry profile in MSCs group."

"Cedera saraf perifer, seperti cedera saraf skiatik dapat mengakibatkan disabilitas. Metode pengobatan dalam memicu regenerasi saraf yang mengalami cedera dapat dilakukan melalui terapi berbasis sel punca mesenkimal (SPM) dengan potensi regenerasi dan memicu efek parakrin. Regenerasi saraf skiatik dianalisis secara histologi melalui analisis deskriptif otot gastroknemius tikus galur Sprague– Dawley dengan pewarnaan hematoksilin–eosin dan jumlah kluster neuromuscular junction (NMJ) pada serat otot dengan pewarnaan Palmgren silver. Analisis secara molekuler juga dilakukan untuk mengamati tingkat ekspresi gen esensial yang mampu meregulasi regenerasi dari saraf skiatik dengan qRT-PCR. Kelompok yang digunakan dalam penelitian ini adalah kelompok kontrol (sehat) dan kelompok perlakuan cedera saraf skiatik yang diinjeksikan dengan SPM adiposa manusia. Penelitian dilakukan untuk mengamati pengaruh injeksi SPM adiposa manusia terhadap regenerasi saraf skiatik dari aspek histologi dan mengamati pengaruh injeksi SPM adiposa manusia melalui ekspresi gen Agrin dan PROM1. Hasil penelitian menunjukkan bahwa injeksi SPM adiposa manusia menunjukkan aspek regenerasi histologi pada otot gastroknemius dengan mengurangi tingkat atrofi otot, tetapi belum adanya regenerasi jumlah kluster NMJ yang terbentuk pada serat otot. Ekspresi gen Agrin menurun sebesar 1,1 kali lipat, sedangkan gen PROM1 menurun sebesar 2,1 kali lipat. Uji statistik pada jumlah kluster NMJ dan ekspresi gen Agrin tidak menunjukkan perbedaan nyata antarkelompok, sedangkan ekspresi gen PROM1 menunjukkan adanya perbedaan nyata antarkelompok. Berdasarkan hasil penelitian, injeksi SPM adiposa manusia belum menunjukkan aspek regenerasi pada saraf skiatik.

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Peripheral nerve injury, such as sciatic nerve injury, can result in disability. Treatment to induce nerve regeneration can be conducted with mesenchymal stem cells (MSC) therapy, which has regenerative potential and triggers paracrine effects. Sciatic nerve regeneration is analyzed histologically through descriptive analysis of the gastrocnemius muscle of Sprague–Dawley strain rat with hematoxylin-eosin staining and the number of neuromuscular junction (NMJ) clusters in muscle fibers with Palmgren silver staining. Molecular analysis is conducted to observe the expression levels of essential genes that can regulate the regeneration of the sciatic nerve using qRT-PCR. The groups used are the control group (healthy) and the treatment group of injection with human adipose tissue derived MSC. This study aims to observe the effect of human adipose tissue derived MSC injection on sciatic nerve regeneration with histological perspective and to observe its effect through the expression of Agrin and PROM1 genes. Results showed that the injection of human adipose tissue derived MSC demonstrates histological regeneration in the gastrocnemius muscle by reducing muscle atrophy. However, there is no regeneration in the number of NMJ clusters formed on muscle fibers. The expression of the Agrin gene decreased by 1.1-fold, while the PROM1 gene decreased by 2.1-fold. Statistical analysis revealed no significant differences in NMJ clusters and Agrin gene expression between groups, but a significant difference was found in PROM1 gene expression. Based on the results, the injection of human adipose tissue derived MSC has not yet shown aspects of regeneration in the sciatic nerve."